Heterocyclic Compounds
20250353846 ยท 2025-11-20
Assignee
Inventors
- Guoliang ZHANG (Beijing, CN)
- Zhikun NI (Beijing, CN)
- Jianzhuang MIAO (Beijing, CN)
- Zhi Zhang (Beijing, CN)
- Ce Wang (Beijing, CN)
Cpc classification
A61K31/522
HUMAN NECESSITIES
A61K31/436
HUMAN NECESSITIES
A61K31/4355
HUMAN NECESSITIES
C07D491/052
CHEMISTRY; METALLURGY
C07D491/056
CHEMISTRY; METALLURGY
A61K31/4188
HUMAN NECESSITIES
C07D491/048
CHEMISTRY; METALLURGY
C07D519/00
CHEMISTRY; METALLURGY
International classification
A61K31/4188
HUMAN NECESSITIES
A61K31/4355
HUMAN NECESSITIES
A61K31/436
HUMAN NECESSITIES
A61K31/522
HUMAN NECESSITIES
C07D491/048
CHEMISTRY; METALLURGY
C07D491/052
CHEMISTRY; METALLURGY
C07D491/056
CHEMISTRY; METALLURGY
Abstract
Disclosed herein is a compound of Formula (I) having the following structure for activating T cells, promoting T cell proliferation, and/or exhibiting antitumor activity, a method of using the compounds disclosed herein for treating cancer, and a pharmaceutical composition comprising the same.
##STR00001##
Claims
1. A compound of formula (I), ##STR00715## or a pharmaceutically acceptable salt, a tautomer, a stereoisomer, an enantiomer, an isotopologue, or a prodrug thereof, wherein X.sup.1 is C or N, each of X.sup.2 and X.sup.3 is independently selected from N or CH; X.sub.4 is N, O or S; R.sup.1 is hydrogen, deuterium, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl; R.sup.2 is hydrogen, halogen, cyano, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl, provided that R.sup.2 is absent when X.sup.1 is N; R.sup.4 is hydrogen, halogen, cyano, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl, provided that R.sup.4 is absent when X.sup.4 is O or S; R.sup.5 is hydrogen, halogen, cyano, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl, R.sup.5aC(O), R.sup.5aC(O)O, R.sup.5aOC(O), R.sup.5aC(O)NR.sup.5b, R.sup.5aNR.sup.5bC(O), or R.sup.5aSO.sub.2, wherein R.sup.5a and R.sup.5b are each independently hydrogen, alkyl, or cycloalkyl; each of R.sup.7, R.sup.9, R.sup.8, and R.sup.10 is independently hydrogen, cyano, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxyl, substituted or unsubstituted cycloalkyl, C(O)R.sup.7a, or -alkyl-C(O)R.sup.7a, and wherein R.sup.7a is hydrogen, alkyl, or alkoxy, provided that at least one of R.sup.7 and R.sup.9 is not hydrogen; or R.sup.7 and R.sup.9 are each hydrogen and R.sup.8 and R.sup.10 together form a bridge containing at least one CH.sub.2-moiety in addition to the two bridgehead atoms; or R.sup.8 and R.sup.10 are each hydrogen and R.sup.7 and R.sup.9 together form a bridge containing at least one CH.sub.2 moiety in addition to the two bridgehead atoms; L.sup.1 is a direct bond, O, N(R.sup.L), substituted or unsubstituted alkyl, -alkylene or C(O), wherein R.sup.L is hydrogen or alkyl; Cy.sup.1 is unsubstituted or substituted cycloalkyl, unsubstituted or substituted aryl, unsubstituted or substituted heterocyclyl, or unsubstituted or substituted heteroaryl, wherein Cy.sup.1 is optionally substituted with 1 to 5 substituents R.sup.3a, wherein R.sup.3a is selected from deuterium, alkoxy, alkyl, halogen, oxo, R.sup.3bSO.sub.2, cycloalkyl, cyano, R.sup.3bC(O)N(R.sup.3c), N(R.sup.3bR.sup.3c)C(O), N(R.sup.3bR.sup.3c), R.sup.3bOC(O), cycloalkyl or heterocyclyl; optionally wherein two R.sup.3a connect to the same carbon and together form a spirocyclic ring; optionally wherein two R.sup.3a form a fused ring with Cy.sup.1, wherein R.sup.3a is unsubstituted or substituted with cyano, alkoxy, alkyl, halogen, or hydroxy; and wherein R.sup.3b and R.sup.3c are each independently hydrogen or alkyl.
2. The compound according to claim 1, which is a compound of any one of formulas (IA1), (IB1) or (IC1): ##STR00716## wherein the variables are defined as herein.
3. The compound according to claim 1, which is a compound of any one of formula (IA2), (IB2) or (IC2): ##STR00717## wherein the variables are defined as herein.
4. The compound according to claim 1, which is a compound of any one of formula (IA3), (IB3) or (IC3): ##STR00718## wherein the variables are defined as herein.
5. The compound according to claim 1, which is a compound of any one of formula (IA4), (IB4) or (IC4): ##STR00719## wherein the variables are defined as herein.
6. The compound according to claim 1, which is a compound of formula (II): ##STR00720## or a pharmaceutically acceptable salt, a tautomer, a stereoisomer, an enantiomer, an isotopologue, or a prodrug thereof, wherein X.sup.1 is C or N, X.sup.4 is N, O or S; R.sup.1 is hydrogen, deuterium, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl; R.sup.2 is hydrogen, halogen, cyano, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl, provided that R.sup.2 is absent when X.sup.1 is N; R.sup.4 is hydrogen, halogen, cyano, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl, provided that R.sup.4 is absent when X.sup.4 is O or S; R is hydrogen, halogen, cyano, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl, R.sup.5aC(O), R.sup.5aC(O)O, R.sup.5aOC(O), R.sup.5aC(O)NR.sup.5b, R.sup.5aNR.sup.5bC(O), or R.sup.5aSO.sub.2, wherein R.sup.5a and R.sup.5b are each independently hydrogen, alkyl, or cycloalkyl; each of R.sup.7, and R.sup.9 is independently hydrogen, cyano, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxyl, substituted or unsubstituted cycloalkyl, C(O)R.sup.7a, or -alkyl-C(O)R.sup.7a, and wherein R.sup.7a is hydrogen, alkyl, or alkoxy, provided that at least one of R.sup.7 and R.sup.9 is not hydrogen; or R.sup.7 and R.sup.9 together form a bridge containing at least one CH.sub.2 moiety in addition to the two bridgehead atoms; L.sup.1 is a direct bond, O, N(R.sup.L), substituted or unsubstituted alkyl, -alkylene or C(O), wherein R.sup.L is hydrogen or alkyl; m is 0, 1, 2 or 3, n is 0 or 1 wherein R.sup.3a is selected from deuterium, alkoxy, alkyl, halogen, oxo, R.sup.3bSO.sub.2, cycloalkyl, cyano, R.sup.3bC(O)N(R.sup.3c), N(R.sup.3bR.sup.3c)C(O), N(R.sup.3bR.sup.3c), R.sup.3bOC(O), cycloalkyl or heterocyclyl; optionally wherein two R.sup.3a connect to the same carbon and together form a spirocyclic ring; optionally wherein two R.sup.3a form a fused ring with Cy.sup.1, wherein R.sup.3a is unsubstituted or substituted with cyano, alkoxy, alkyl, halogen, or hydroxy; and wherein R.sup.3b and R.sup.3c are each independently hydrogen or alkyl.
7. The compound according to any one of claims 1 to 6, wherein R.sup.1 is hydrogen, or substituted or unsubstituted alkyl.
8. The compound according to any one of claims 1 to 7, wherein R.sup.1 is hydrogen, or C.sub.1-4alkyl optionally substituted with deuterium, halogen, hydroxy, alkoxy or cycloalkyl; preferably R.sup.1 is hydrogen, or C.sub.1-4alkyl optionally substituted with deuterium, halogen, hydroxy, alkoxy or cycloalkyl; more preferably R.sup.1 is hydrogen, or C.sub.1-3alkyl optionally substituted with deuterium, or halogen; even more preferably R.sup.1 is hydrogen, methyl, methyl-d3, ethyl, isopropyl, 2-hydroxyethyl, 2-methoxyethyl, 2,2,2-trifluoroethyl, 2,2-difluoroethyl, or cyclopropylmethyl.
9. The compound according to claim 8, wherein R.sup.2 is hydrogen, halogen, C.sub.1-4alkyl, C.sub.1-4 alkoxyl or cyano; preferably R.sup.2 is hydrogen, F, Br, Cl or CN.
10. The compound according to any one of claims 1 to 9, wherein R.sup.4 is hydrogen, halogen or alkyl, wherein the alkyl is optionally substituted with deuterium, halogen or OR.sup.4a, wherein R.sup.4a is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl or heterocycloalkyl, wherein alkyl, alkenyl, alkynyl, cycloalkyl or heterocycloalkyl is optionally substituted with C.sub.1-6alkyl, C.sub.1-6alkoxy or C.sub.3-8cycloalkyl; preferably R.sup.4 is hydrogen, or C.sub.1-3alkyl optionally substituted with deuterium, or halogen; more preferably R.sup.4 is hydrogen, fluoro, chloro, bromo, methyl, methyl-d3, trifluoromethyl, ethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl, propyl, cyclopropyl.
11. The compound according to any one of claims 1 to 10, wherein R.sup.5 is hydrogen, alkyl, alkenyl or alkynyl, wherein said alkyl is unsubstituted or substituted with cyano; preferably R.sup.5 is C.sub.1-4alkyl, C.sub.2-4 alkenyl or C.sub.2-4alkynyl, wherein said alkyl is substituted with cyano; more preferably R.sup.5 is C.sub.1-4alkyl, wherein said alkyl is substituted with cyano.
12. The compound according to any one of claims 1 to 11, wherein each of R.sup.7 and R.sup.9 is independently hydrogen, alkyl, or C(O)R.sup.7a, wherein said alkyl is unsubstituted or substituted with halogen, cyano, hydroxy, or alkoxy, and wherein R.sup.7a is hydrogen, alkyl, or alkoxy; preferably each of R.sup.7 and R.sup.9 is independently C.sub.1-2alkyl.
13. The compound according to claim 12, wherein R.sup.7 and R.sup.9 are each independently hydrogen, methyl, ethyl, methoxymethyl, 1-hydroxyethyl, 2-methoxyethyl, cyanomethyl, hydroxyethyl, hydroxymethyl, methoxycarbonyl, difluoromethyl, provided that at least one of R.sup.7 and R.sup.9 is not hydrogen.
14. The compound according to any one of claims 1 to 13, wherein R.sup.8 and R.sup.10 are each hydrogen; R.sup.7 is methyl; and R.sup.9 is methyl; R.sup.8 and R.sup.10 are each hydrogen; R.sup.7 is ethyl; and R.sup.9 is ethyl; R.sup.8 and R.sup.10 are each hydrogen; R.sup.7 is methyl; and R.sup.9 is ethyl; or R.sup.8 and R.sup.10 are each hydrogen; R.sup.7 is ethyl, and R.sup.9 is methyl; R.sup.8 and R.sup.10 are each hydrogen; R.sup.7 is methoxymethyl, and R.sup.9 is methyl, ethyl or methoxymethyl.
15. The compound according to any one of claims 1 to 14, wherein L.sup.1 is a direct bond, O, N(R.sup.L), -alkylene- or C(O), wherein R.sup.L is hydrogen or alkyl and wherein said -alkylene- is unsubstituted or substituted with deuterium, halogen, alkoxy, alkynyl or heterocyclyl; preferably L.sup.1 is a direct bond, O, N(R.sup.L), -alkylene- or C(O), wherein R.sup.L is hydrogen or alkyl; preferably L.sup.1 is C.sub.1-4alkylene, preferably C.sub.1-2alkylene; more preferably L.sup.1 is a direct bond, CH.sub.2, CH(CH.sub.3), CH(CD.sub.3), CH(CH.sub.2CH.sub.3), CH(CHF.sub.2), CH(prop-1-ynyl)-, N(H), N(CH.sub.3), O, CH(C(O)NHCH.sub.2CH.sub.2OCH.sub.3)or C(CH.sub.3).sub.2.
16. The compound according to any one of claims 1 to 5 and 7 to 15, wherein Cy.sup.1 is aryl, heterocyclyl, heterocyclyl, heteroaryl, or cycloalkyl, each of which is unsubstituted or substituted with one, two, three, or four substituents R.sup.3a, wherein R.sup.3a is selected from deuterium, alkoxy, alkyl, halogen, R.sup.3bSO.sub.2, cycloalkyl, cyano, R.sup.3b-C(O)N(R.sup.3c), N(R.sup.3bR.sup.3c)C(O), N(R.sup.3bR.sup.3c), R.sup.3bOC(O), or heterocyclyl, wherein said alkyl moiety in the group alkyl or alkoxy is unsubstituted or substituted with halogen, alkoxy, hydroxy, or cyano; said cycloalkyl or heterocyclyl is unsubstituted or substituted with alkoxy, alkyl, halogen, or hydroxy, wherein R.sup.3b and R.sup.3c are each independently hydrogen or alkyl.
17. The compound according to claim 16, wherein Cy.sup.1 is unsubstituted or substituted cycloalkyl, unsubstituted or substituted aryl, unsubstituted or substituted heterocyclyl, or unsubstituted or substituted heteroaryl; preferably Cy.sup.1 is optionally substituted with one, two or three substituents R.sup.3a, wherein R.sup.3a is selected from deuterium, fluoro, bromo, chloro, methyl, difluoromethyl, trifluoromethoxy, methoxy, methoxymethyl, trifluoromethyl, methylsulfonyl, difluoro, ethoxy, isopropyl, isopropoxy, difluoromethoxy, cyclopropyl, 2,2-difluorocyclopropyl, 1-fluorocyclopropyl, 2-fluorocyclopropyl, cyclobutyl, 1-hydroxyethyl, 2-hydroxyethyl, ethyl, 1,1-difluoroethyl, cyano, dimethoxy, dichloro, acetamido, 1-methoxyethyl, cyanomethyl, carbamoyl, methoxycarbonyl, dimethylcarbamoyl, (difluoromethoxy)methyl, amino, 1-(difluoromethoxy)ethyl, azetidin-1-yl, 2-methoxypropan-2-yl, 1-methoxycyclopropyl, oxetan-3-yl, 1-methylazetidin-3-yl, 1-hydroxyazetidin-3-yl, 2-cyanoethyl, 2-cyanopropan-2-yl, hydroxymethyl, 3-hydroxypropyl, 2-hydroxypropan-2-yl, or 2-fluoropropan-2-yl.
18. The compound according to claim 16, wherein Cy.sup.1 is aryl, which is unsubstituted or substituted with one, two or three substituents R.sup.3a, wherein R.sup.3a is selected from alkoxy, halogen-substituted alkoxy, alkoxyalkyl-, alkyl, alkenyl, halogen-substituted alkyl, halogen, R.sup.3bSO.sub.2, cycloalkyl, hydroxyalkyl-, cyano, R.sup.3bC(O)N(R.sup.3c), cyano-substituted alkyl, N(R.sup.3bR.sup.3c)C(O), R.sup.3bOC(O), heterocyclylalkyl- or heterocyclyl, said cycloalkyl or heterocyclyl is unsubstituted or substituted with alkoxy, alkyl, halogen, or hydroxy, wherein R.sup.3b and R.sup.3c are each independently hydrogen or alkyl.
19. The compound according to claim 18, wherein Cy.sup.1 is phenyl, which is substituted with one or two R.sup.3a, wherein R.sup.3a is fluoro, chloro, bromo; methyl, difluoromethyl, trifluoromethyl, 1,1-difluoroethyl, methoxymethyl, 1-methoxyethyl, ethyl, propyl, isopropyl, tert-butyl, 2-hydroxypropan-2-yl, 1-hydroxy-2-methylpropan-2-yl, 2-cyanopropan-2-yl, morpholinomethyl; prop-1-en-2-yl; cyclopropyl, 1-methylcyclopropyl; methoxy, difluoromethoxy, trifluoromethoxy, 1,1-difluoroethoxy, isopropoxy; oxetan-3-yl, morpholino, 2-oxa-6-azaspiro[3.3]heptan-6-yl; or methylcarbamoyl.
20. The compound according to claim 16, wherein Cy.sup.1 is a monocyclic 5- to 9-membered heterocyclyl or a bicyclic 7- to 10-membered heterocyclyl which is unsubstituted or substituted with one, two, three or four R.sup.3a (provided that the valency theory has been met), wherein R.sup.3a is selected from deuterium, alkoxy, alkyl, halogen, R.sup.3bSO.sub.2, cycloalkyl, cyano, R.sup.3bC(O)N(R.sup.3c), N(R.sup.3bR.sup.3c)C(O), N(R.sup.3bR.sup.3c), R.sup.3bOC(O), oxo or heterocyclyl, wherein said alkyl moiety in the group alkyl or alkoxy is unsubstituted or substituted with halogen, alkoxy, hydroxy, or cyano; said cycloalkyl or heterocyclyl is unsubstituted or substituted with alkoxy, alkyl, halogen, or hydroxy, wherein R.sup.3b and R.sup.3c are each independently hydrogen or alkyl; preferably, R.sup.3a is selected from alkoxy, halogen-substituted alkoxy, alkoxyalkyl-, alkyl, halogen-substituted alkyl, halogen, R.sup.3bSO.sub.2, cycloalkyl, hydroxyalkyl-, cyano, R.sup.3bC(O)N(R.sup.3c), cyano-substituted alkyl, N(R.sup.3bR.sup.3c)C(O), R.sup.3bOC(O), or heterocyclyl, said cycloalkyl or heterocyclyl is unsubstituted or substituted with alkoxy, alkyl, halogen, or hydroxy, wherein R.sup.3b and R.sup.3c are each independently hydrogen or alkyl; preferably, said monocyclic 5- to 9-membered heterocyclyl is tetrahydrofuranyl, tetrahydropyranyl, 1,4-dioxanyl, piperidinyl, piperazinyl, or dihydropyridinyl, each of which is unsubstituted or substituted with one, two or three R.sup.3a as disclosed herein; preferably Cy.sup.1 is tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydro-2H-pyran-2-yl, tetrahydro-2H-pyran-3-yl, tetrahydro-2H-pyran-4-yl, 1,4-dioxan-2-yl, 1,4-dioxan-3-yl, piperidin-1-yl, piperidin-2-yl, piperidin-3-yl, piperidin-4-yl, piperazin-1-yl, piperazin-2-yl, piperazin-3-yl, piperazin-4-yl, 1,2-dihydropyridin-3-yl, 1,2-dihydropyridin-4-yl, 1,2-dihydropyridin-5-yl, or 1,2-dihydropyridin-6-yl.
21. The compound according to claim 16, wherein Cy.sup.1 is dihydrobenzodioxinyl, benzodioxolyl, chromanyl, or chromenyl, which is unsubstituted or substituted with one, two, three or three R.sup.3a, wherein R.sup.3a is selected from alkoxy, alkyl, halogen, and oxo; preferably, Cy.sup.1 is 2,3-dihydrobenzo[b][1,4]dioxin-5-yl, 2,3-dihydrobenzo[b][1,4]dioxin-6-yl, benzo[d][1,3]dioxol-4-yl, benzo[d][1,3]dioxol-5-yl, chroman-2-yl, chroman-3-yl, chroman-4-yl, chroman-5-yl, chroman-6-yl, chroman-7-yl, chromen-2-yl, chromen-3-yl, chromen-4-yl, chromen-5-yl, chromen-6-yl, chromen-7-yl, each of which is unsubstituted or substituted with one, two, three or three R.sup.3a, wherein R.sup.3a is selected from alkoxy, alkyl, halogen, and oxo; preferably R.sup.3a is methyl, ethyl, propyl, isopropyl, fluoro, chloro, bromo or oxo.
22. The compound according to claim 16, wherein Cy.sup.1 is a monocyclic 5- to 9-membered heteroaryl or a bicyclic 7- to 10-membered heteroaryl which is unsubstituted or substituted with one, two or three R.sup.3a, wherein R.sup.3a is selected from deuterium, alkoxy, alkyl, halogen, R.sup.3bSO.sub.2, cycloalkyl, cyano, R.sup.3bC(O)N(R.sup.3c), N(R.sup.3bR.sup.3c)C(O), N(R.sup.3bR.sup.3c), R.sup.3bOC(O), oxo, phenyl or heterocyclyl, wherein said alkyl moiety in the group alkyl or alkoxy is unsubstituted or substituted with halogen, alkoxy, hydroxy, or cyano; said cycloalkyl or heterocyclyl is unsubstituted or substituted with alkoxy, alkyl, halogen, or hydroxy, wherein R.sup.3b and R.sup.3c are each independently hydrogen or alkyl; preferably, R.sup.3a is selected from deuterium, alkoxy, halogen-substituted alkoxy, alkoxyalkyl-, alkyl, halogen-substituted alkyl, halogen, oxo, hydroxyalkyl-, cyano, cyano-substituted alkyl, N(R.sup.3bR.sup.3c)C(O), N(R.sup.3bR.sup.3c), R.sup.3bOC(O), oxo, phenyl or heterocyclyl, said cycloalkyl or heterocyclyl is unsubstituted or substituted with alkoxy, alkyl, halogen, or hydroxy, wherein R.sup.3b and R.sup.3c are each independently hydrogen or alkyl; more preferably R.sup.3a is selected from deuterium, fluoro, bromo, chloro, methyl, difluoromethyl, trifluoromethyl, trifluoromethoxy, methoxy, methoxymethyl, methylsulfonyl, ethoxy, isopropoxy, isopropyl, tert-butyl, cyclopropyl, 1-hydroxyethyl, ethyl, 1,1-difluoroethyl, cyano, cyclopropyl-, acetamido, 1-methoxyethyl, cyanomethyl, carbamoyl, methoxycarbonyl, methylcarbamoyl, dimethylcarbamoyl, (difluoromethoxy)methyl, amino, isopropylamino, (difluoromethoxy)ethyl, azetidin-1-yl, 2-methoxypropan-2-yl, 1-methoxycyclopropyl, oxetan-3-yl, 1-methylazetidin-3-yl, 1-hydroxyazetidin-3-yl, phenyl or oxo.
23. The compound according to claim 16, wherein Cy.sup.1 is pyrazolyl, imidazolyl, thiazolyl, pyridinyl, pyrimidinyl, pyrazinyl or pyridazinyl, each of which is unsubstituted or substituted with one, two or three R.sup.3a as disclosed herein; preferably Cy.sup.1 is 1H-pyrazol-1-yl, 1H-pyrazol-3-yl, 1H-pyrazol-4-yl, thiazol-4-yl, thiazol-5-yl, 1H-pyrazol-5-yl, 1H-imidazol-2-yl, 1H-imidazol-4-yl, 1H-imidazol-5-yl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, 1H-indol-2-yl, 1H-indol-3-yl, 1H-indol-4-yl, 1H-indol-5-yl, 1H-indol-6-yl, 1H-indol-7-yl, pyrazin-2-yl, or pyridazin-4-yl, each of which is unsubstituted or substituted with one, two or three R.sup.3a as disclosed herein; preferably R.sup.3a is selected from deuterium, fluoro, bromo, chloro, methyl, difluoromethyl, trifluoromethyl, trifluoromethoxy, methoxy, ethoxy, isopropoxy, isopropyl, tert-butyl, carbamoyl, methylcarbamoyl, dimethylcarbamoyl, isopropylamino, azetidin-1-yl, phenyl or oxo.
24. The compound according to claim 16, wherein Cy.sup.1 is indolyl, triazolopyridinyl, imidazopyridinyl, benzooxazolyl, benzo[d]thiazolyl, quinolinyl, isoquinolinyl, naphthyridinyl, dioxinopyridineyl, quinoxalinyl, benzo[d]imidazolyl, benzofuranyl, benzoxazinyl, imidazo[4,5-b]pyridinyl, thiazolo[5,4-b]pyridinyl, thiazolo[4,5-b]pyridinyl, thieno[2,3-b]pyridinyl, dihydrodioxinopyridinyl, dioxinopyridinyl, dihydro-2H-pyranopyridinyl, 2H-pyranopyridinyl, dihydrofuropyridinyl, furopyridinyl, benzo[d]thiazolyl, isoindolinyl or thieno[3,2-b]pyridinyl, each of which is unsubstituted or substituted with one, two or three R.sup.3a as disclosed herein; preferably Cy.sup.1 is quinoxalin-6-yl, quinoxalin-2-yl, isoquinolin-3-yl, isoquinolin-7-yl, quinolin-6-yl, 1H-benzo[d]imidazol-5-yl, benzofuran-6-yl, benzofuran-5-yl, benzo[e][1,3]oxazin-6-yl, 2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl, 2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-7-yl, 3,4-dihydro-2H-pyrano[3,2-b]pyridin-6-yl, 3,4-dihydro-2H-pyrano[3,2-b]pyridin-8-yl, 2,3-dihydrofuro[2,3-b]pyridin-6-yl, 2,3-dihydrofuro[3,2-b]pyridin-5-yl, benzo[d]thiazol-5-yl, benzo[d]thiazol-6-yl, isoindolin-5-yl, 1H-benzo[d]imidazol-2-yl, 1H-benzo[d]imidazol-4-yl, 1H-benzo[d]imidazol-6-yl, 1H-benzo[d]imidazol-7-yl, [1,2,4]triazolo[1,5-a]pyridin-2-yl, [1,2,4]triazolo[1,5-a]pyridin-5-yl, [1,2,4]triazolo[1,5-a]pyridin-6-yl, [1,2,4]triazolo[1,5-a]pyridin-7-yl, [1,2,4]triazolo[1,5-a]pyridin-8-yl, 3H-imidazo[4,5-b]pyridine-2-yl, 3H-imidazo[4,5-b]pyridine-5-yl, 3H-imidazo[4,5-b]pyridine-6-yl, 3H-imidazo[4,5-b]pyridine-7-yl, 1H-imidazo[4,5-b]pyridin-2-yl, 1H-imidazo[4,5-b]pyridin-5-yl, 1H-imidazo[4,5-b]pyridin-6-yl, 1H-imidazo[4,5-b]pyridin-7-yl, benzo[d]oxazol-2-yl, benzo[d]oxazol-4-yl, benzo[d]oxazol-5-yl, benzo[d]oxazol-6-yl, benzo[d]oxazol-7-yl, benzo[d]thiazol-2-yl, benzo[d]thiazol-4-yl, benzo[d]thiazol-7-yl, quinolin-2-yl, quinolin-3-yl, quinolin-4-yl, quinolin-5-yl, quinolin-7-yl, isoquinolin-1-yl, isoquinolin-4-yl, isoquinolin-5-yl, isoquinolin-6-yl, isoquinolin-8-yl, quinoxalin-3-yl, quinoxalin-4-yl, quinoxalin-5-yl, quinoxalin-7-yl, quinoxalin-8-yl, 1,8-naphthyridin-2-yl, 1,8-naphthyridin-3-yl, 1,8-naphthyridin-4-yl, 2,3-dihydro-[1,4]dioxino[2,3-b]pyridine-6-yl, 2,3-dihydro-[1,4]dioxino[2,3-b]pyridine-7-yl, 2,3-dihydro-[1,4]dioxino[2,3-b]pyridine-8-yl, quinoxalin-6-yl-2,3-d2, 1H-indol-2-yl, 1-methyl-1H-benzo[d]imidazol-6-yl, 3H-imidazo[4,5-b]pyridin-2-yl, 4,5,6,7-tetrahydro-1H-benzo[d]imidazol-2-yl, thiazolo[5,4-b]pyridin-2-yl, thiazolo[5,4-b]pyridin-5-yl, thiazolo[5,4-b]pyridin-6-yl, thiazolo[5,4-b]pyridin-7-yl, thiazolo[4,5-b]pyridin-2-yl, thiazolo[4,5-b]pyridin-5-yl, thiazolo[4,5-b]pyridin-6-yl, thiazolo[4,5-b]pyridin-7-yl, thieno[2,3-b]pyridin-2-yl, thieno[2,3-b]pyridin-3-yl, thieno[2,3-b]pyridin-4-yl, thieno[2,3-b]pyridin-5-yl, thieno[2,3-b]pyridin-6-yl, thieno[3,2-b]pyridin-2-yl, thieno[3,2-b]pyridin-3-yl, thieno[3,2-b]pyridin-5-yl, thieno[3,2-b]pyridin-6-yl, thieno[3,2-b]pyridin-7-yl, each of which is unsubstituted or substituted with one, two or three R.sup.3a as disclosed herein.
25. The compound according to claim 16, wherein Cy.sup.1 is quinoxalin-6-yl, quinoxalin-2-yl, benzo[d]thiazol-5-yl, benzo[d]thiazol-6-yl, isoindolin-4-yl, indolin-5-yl, 1H-benzo[d]imidazol-5-yl, benzofuran-6-yl, benzofuran-5-yl, or benzo[e][1,3]oxazin-6-yl, isoquinolin-3-yl, isoquinolin-7-yl, quinolin-6-yl, 2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl, 2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-7-yl, 3-dihydro-[1,4]dioxino[2,3-c]pyridin-7-yl, 3,4-dihydro-2H-pyrano[3,2-b]pyridin-6-yl, 3,4-dihydro-2H-pyrano[3,2-b]pyridin-8-yl, 2,3-dihydrofuro[2,3-b]pyridin-6-yl, 2,3-dihydrofuro[3,2-b]pyridin-5-yl, each of which is unsubstituted or substituted with one, two or three R.sup.3a, wherein R.sup.3a is deuterium, methyl, ethyl, isopropyl, cyanomethyl, 2-cyanoethyl, 2-cyanopropan-2-yl, hydroxymethyl, 2-hydroxyethyl, 1-hydroxyethyl, 3-hydroxypropyl, 2-hydroxypropan-2-yl, difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, 2-fluoropropan-2-yl, methoxy, isopropoxy, difluoromethoxy, fluoro, chloro, bromo, oxo, cyano, amino, cyclopropyl, 2,2-difluorocyclopropyl, 1-fluorocyclopropyl, 2-fluorocyclopropyl, or cyclobutyl; preferably deuterium, methyl, ethyl, trifluoromethyl, methoxy, isopropoxy, difluoromethoxy, fluoro, chloro, cyano, amino, or cyclopropyl.
26. The compound according to claim 16, wherein Cy.sup.1 is 4-fluoro-2-methoxyphenyl, p-tolyl, 4-fluoro-2-(trifluoromethyl)phenyl, 2-(difluoromethoxy)-4-fluorophenyl, 4-fluoro-2-(trifluoromethoxy)phenyl, 4-fluoro-2-(methoxymethyl)phenyl, 4-fluoro-2-(1-methoxyethyl)phenyl, 1-(difluoromethoxy-4-fluorophenyl, 2-(difluoromethyl)-4-fluorophenyl, 2-(1,1-difluoroethoxy)-4-fluorophenyl, 3-(trifluoromethyl)phenyl, 3-isopropoxyphenyl, 3-(trifluoromethoxy)phenyl, 3-isopropylphenyl, 3-(tert-butyl)phenyl, 3-(2-hydroxypropan-2-yl)phenyl, 3-(difluoromethyl)-4-fluorophenyl, 4-fluoro-3-(trifluoromethyl)phenyl, 2-fluoro-5-(trifluoromethyl)phenyl, 4-methyl-3-(trifluoromethyl)phenyl, 4-methyl-2-(trifluoromethyl)phenyl, 3-cyclopropylphenyl, 3-(1-methylcyclopropyl)phenyl, 3-(oxetan-3-yl)phenyl, 4-fluoro-3-(methylcarbamoyl)phenyl, 3-(1-hydroxy-2-methylpropan-2-yl)phenyl, 3-(2-cyanopropan-2-yl)phenyl, 4-fluoro-3-(prop-1-en-2-yl)phenyl, 4-fluoro-3-(1-methylcyclopropyl)phenyl, 3-methyl-4-(trifluoromethyl)phenyl, 4-(morpholinomethyl)phenyl, 3-(2-oxa-6-azaspiro[3.3]heptan-6-yl)phenyl, 3-morpholinophenyl, 3-(morpholinomethyl)phenyl, 4-morpholinophenyl, 4-(trifluoromethyl)phenyl, 4-fluorophenyl, 3-methyl-2-(trifluoromethyl)phenyl, 3-(difluoromethyl)-5-fluorophenyl, 3-(1,1-difluoroethyl)-4-methoxyphenyl, m-tolyl, 3-methoxyphenyl, or 3-(difluoromethyl)phenyl; or 3,3-dimethyl-2,3-dihydro-1H-inden-5-yl; or 4,4-difluorochroman-6-yl, 2,2-dimethyl-4-oxochroman-7-yl, 3-methyl-4-oxo-4H-chromen-6-yl, 2,2-dimethylchroman-7-yl, or 2-methyl-4-oxo-4H-chromen-7-yl; or 2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 8-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 7-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 5-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 8-chloro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 2,2,3,3-tetrafluoro-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 3,3-difluoro-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 7-fluoro-2,2-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, or 7-chloro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl; or 2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 2,2-difluorobenzo[d][1,3]dioxol-5-yl, 2-ethyl-2-methylbenzo[d][1,3]dioxol-5-yl, spiro[benzo[d][1,3]dioxole-2,1-cyclobutan]-5-yl, 6-chloro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 7-chloro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 4-chloro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 7-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 4-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 4,6-difluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 2,2,7-trimethylbenzo[d][1,3]dioxol-5-yl, 7-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-4-yl, 2,2-dimethylbenzo[d][1,3]dioxol-4-yl, 2,2-difluorobenzo[d][1,3]dioxol-4-yl, 4-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, or 2,2-dimethylbenzo[d][1,3]dioxol-5-yl; or 2-(trifluoromethyl)pyridin-3-yl, 6-isopropoxypyridin-2-yl, 6-(trifluoromethyl)pyridin-2-yl, 2-(trifluoromethyl)pyridin-4-yl, 4-(trifluoromethyl)pyridin-2-yl, 2-(azetidin-1-yl)pyridin-4-yl, 2-(isopropylamino)pyridin-4-yl, 6-isopropylpyridin-2-yl, 2-isopropoxypyridin-4-yl, 6-(trifluoromethoxy)pyridin-2-yl, 2-(tert-butyl)pyridin-4-yl, 2-(methylcarbamoyl)pyridin-4-yl, 2-(2,2-dimethylcarbamoyl)pyridin-4-yl, 5,6-dimethylpyridin-2-yl, 2-phenylpyridin-4-yl, 5-(difluoromethyl)pyridin-2-yl, 5-methylpyridin-2-yl, 5-(trifluoromethyl)pyridin-2-yl, 2-methoxypyridin-4-yl, 1-methyl-6-oxo-1,6-dihydropyridin-3-yl, 1-methyl-2-oxo-1,2-dihydropyridin-4-yl, 2-(trifluoromethyl)thiazol-4-yl, or 1-methyl-5-(trifluoromethyl)-1H-pyrazol-4-yl; or quinoxalin-6-yl, 3-methylquinoxalin-6-yl, 3-(trifluoromethyl)quinoxalin-6-yl, or quinoxalin-2-yl; or isoquinolin-3-yl, 2-methyl-1-oxo-1,2-dihydroisoquinolin-7-yl, 2-methyl-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl, 1,2-dimethyl-4-oxo-1,4-dihydroquinolin-7-yl, 1,3-dimethyl-4-oxo-1,4-dihydroquinolin-6-yl, or 2-methylquinolin-7-yl; or 2-methyl-1H-benzo[d]imidazol-5-yl, 2,2-dimethyl-2,3-dihydrobenzofuran-6-yl, 2,2-dimethyl-3-oxo-2,3-dihydrobenzofuran-6-yl, 3,3-dimethyl-2,3-dihydrobenzofuran-5-yl, or 3-methyl-4-oxo-3,4-dihydro-2H-benzo[e][1,3]oxazin-6-yl; 2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl, 3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl, 4,4-difluoro-3,4-dihydro-2H-pyrano[3,2-b]pyridin-6-yl, 4,4-difluoro-3,4-dihydro-2H-pyrano[3,2-b]pyridin-8-yl, 7-chloro-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl, 7-fluoro-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl, 8-fluoro-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl, 2,2-dimethyl-2,3-dihydrofuro[2,3-b]pyridin-6-yl, 3,3-dimethyl-2,3-dihydrofuro[3,2-b]pyridin-5-yl, 3,3-dimethyl-2,3-dihydrofuro[3,2-b]pyridine-5-yl, 2,2-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-c]pyridin-7-yl, 2,2-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-7-yl, 3-methyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl, or 3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl; or 2-methylbenzo[d]thiazol-5-yl, 2-methylbenzo[d]thiazol-6-yl, 2-ethylbenzo[d]thiazol-5-yl, 2-isopropylbenzo[d]thiazol-5-yl, or 2-(trifluoromethyl)thiazol-5-yl; or 2-methyl-1,3-dioxoisoindolin-5-yl, 2-isopropyl-1,3-dioxoisoindolin-5-yl, 1-oxo-2-(2,2,2-trifluoroethyl)isoindolin-5-yl, 2,3,3-trimethyl-1-oxoisoindolin-5-yl, 2-methyl-3-oxoisoindolin-5-yl, or 2-methyl-1-oxoisoindolin-5-yl.
27. The compound of claim 6, wherein the moiety ##STR00721## is ##STR00722## wherein m is 0, 1, 2 or 3 and R.sup.3a is alkoxy, alkyl, halogen, and oxo.
28. The compound of claim 27, wherein the moiety ##STR00723## is 2,2-dimethylbenzo[d][1,3]dioxol-5-yl or 3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, wherein m is 0, 1, 2 or 3 and R.sup.3a is alkoxy, alkyl, halogen, and oxo.
29. The compound of claim 27, wherein the moiety ##STR00724## is 2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 2,2-difluorobenzo[d][1,3]dioxol-5-yl, 2-ethyl-2-methylbenzo[d][1,3]dioxol-5-yl, spiro[benzo[d][1,3]dioxole-2,1-cyclobutan]-5-yl, 6-chloro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 7-chloro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 4-chloro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 7-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 4-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 4,6-difluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 2,2,7-trimethylbenzo[d][1,3]dioxol-5-yl, 2,2-difluorobenzo[d][1,3]dioxol-4-yl, 4-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 2,2-dimethylbenzo[d][1,3]dioxol-5-yl; 2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 8-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 7-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 5-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 8-chloro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 2,2,3,3-tetrafluoro-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 3,3-difluoro-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 7-fluoro-2,2-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, or 7-chloro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl.
30. The compound of claim 1, wherein the compound is selected from: 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(quinoxalin-6-yl)ethyl)piperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (A1); 2-(7-((2S,5R)-4-(1-(4-fluoro-2-methoxyphenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (A4); 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(p-tolyl)ethyl)piperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (A5); 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(1-methyl-5-(trifluoromethyl)-1H-pyrazol-4-yl)ethyl)piperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (A6); 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(2-(trifluoromethyl)pyridin-3-yl)ethyl)piperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (A7); 2-(7-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (A9); 2-(6-((2R,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2-(methoxymethyl)-5-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A10); 2-(6-((2R,5R)-4-(1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2-(methoxymethyl)-5-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A11); 2-(6-((2R,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2-(methoxymethyl)-5-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A12); 2-(6-((2R,5R)-2-(methoxymethyl)-5-methyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A13); 2-(6-fluoro-7-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (A14); 2-(6-fluoro-7-((2S,5R)-4-(1-(4-fluoro-2-methoxyphenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (A16); 2-(7-((2S,5R)-4-(1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-6-fluoro-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (A18); 2-(7-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-6-fluoro-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (A19); 2-(7-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-ethyl-6-fluoro-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (A20); 2-(7-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-ethyl-6-fluoro-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (A21); 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(quinoxalin-6-yl)ethyl)piperazin-1-yl)-6-fluoro-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (A22); 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-6-fluoro-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (A23); 2-(6-((2S,5R)-4-(1-(4-fluoro-2-methoxyphenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A24); 2-(6-((2S,5R)-4-(1-(2-(difluoromethoxy)-4-fluorophenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A25); 2-(6-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethoxy)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A26); 2-(6-((2S,5R)-4-(1-(4-fluoro-2-(methoxymethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A27); 2-(6-((2S,5R)-4-(1-(4-fluoro-2-(1-methoxyethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A28); 2-(6-((2S,5R)-4-(1-(2-(1-(difluoromethoxy)ethyl)-4-fluorophenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A29); 2-(6-((2S,5R)-4-(1-(2-(difluoromethyl)-4-fluorophenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A30); 2-(6-((2S,5R)-4-(1-(2-(1,1-difluoroethoxy)-4-fluorophenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A31); 2-(6-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A33); 2-(6-((2S,5R)-2,5-diethyl-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A34); 2-(9-ethyl-6-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A35); 2-(6-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-bis(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A36); 2-(6-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-9-(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A37); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A41); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A42); 2-(9-ethyl-6-((2S,5R)-4-(1-(3-isopropoxyphenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A43); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-(trifluoromethoxy)phenyl)ethyl)piperazin-1-yl)9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A44); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(quinoxalin-6-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A48); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A49); 2-(6-((2S,5R)-5-ethyl-2-methyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A50); 2-(6-((2S,5R)-2,5-diethyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A51); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A52); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-3-ethyl-9-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A53); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-(trifluoromethyl)quinoxalin-6-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A54); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-methylbenzo[d]thiazol-5-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A55); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-methylbenzo[d]thiazol-6-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A56); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-methyl-1H-benzo[d]imidazol-5-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A57); 2-(6-((2S,5R)-4-(1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A58); 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A59); 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A60); 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-ethyl-9-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A61); 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-diethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A62); 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A63); 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-diethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A64); 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A65); 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-3-ethyl-9-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A66); 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-diethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A67); 2-(6-((2S,5R)-4-(1-(8-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A68); 2-(6-((2S,5R)-4-(1-(7-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,4,5,9-tetrahydro-2H-purin-8-yl)acetonitrile (A69); 2-(6-((2S,5R)-4-(1-(5-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A70); 2-(9-ethyl-6-((2S,5R)-4-(1-(5-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A71); 2-(6-((2S,5R)-4-(1-(8-chloro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A72); 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A74); 2-(6-((2S,5R)-4-(1-(4,4-difluorochroman-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A77); 2-(6-((2S,5R)-4-(1-(4,4-difluorochroman-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A78); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2,2,3,3-tetrafluoro-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A79); 2-(6-((2S,5R)-4-(1-(3,3-difluoro-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A80); 2-(6-((2S,5R)-4-(1-(2,2-dimethyl-4-oxochroman-7-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A81); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-methyl-1,3-dioxoisoindolin-5-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A83); 2-(6-((2S,5R)-4-(1-(2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A85); 2-(9-cyclopropyl-6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A86); 2-(9-(2,2-difluoroethyl)-6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A87); 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-9-propyl-3,9-dihydro-2H-purin-8-yl)acetonitrile (A88); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-methyl-4-oxo-4H-chromen-6-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A91); 2-(6-((2S,5R)-4-(1-(2,2-dimethylchroman-7-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A94); 2-(9-ethyl-6-((2S,5R)-4-(1-(2-ethylbenzo[d]thiazol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A95); 2-(9-ethyl-6-((2S,5R)-4-(1-(2-isopropylbenzo[d]thiazol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A96); 2-(6-((2S,5R)-4-(1-(2,2-dimethyl-2,3-dihydrobenzofuran-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A97); 2-(6-((2S,5R)-4-(1-(2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A98); 2-(6-((2S,5R)-4-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A99); 2-(6-((2S,5R)-4-(1-(4,4-difluoro-3,4-dihydro-2H-pyrano[3,2-b]pyridin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A102); 2-(6-((2S,5R)-4-(1-(4,4-difluoro-3,4-dihydro-2H-pyrano[3,2-b]pyridin-8-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A103); 2-(9-ethyl-6-((2S,5R)-4-(1-(3-isopropylphenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A108); 2-(6-((2S,5R)-4-(1-(3-(tert-butyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A110); 2-(9-ethyl-6-((2S,5R)-4-(1-(3-(2-hydroxypropan-2-yl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A111); 2-(6-((2S,5R)-4-(1-(3-(difluoromethyl)-4-fluorophenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A112); 2-(9-ethyl-6-((2S,5R)-4-(1-(4-fluoro-3-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A113); 2-(9-ethyl-6-((2S,5R)-4-(1-(2-fluoro-5-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A114); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(4-methyl-3-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A115); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(4-methyl-2-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A116); 2-(9-ethyl-6-((2S,5R)-4-(1-(6-isopropoxypyridin-2-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A117); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(6-(trifluoromethyl)pyridin-2-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A118); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-(trifluoromethyl)pyridin-4-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A119); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(4-(trifluoromethyl)pyridin-2-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A120); 2-(6-((2S,5R)-4-(1-(2-(azetidin-1-yl)pyridin-4-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A121); 2-(9-ethyl-6-((2S,5R)-4-(1-(2-(isopropylamino)pyridin-4-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A122); 2-(9-ethyl-6-((2S,5R)-4-(1-(6-isopropylpyridin-2-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A123); 2-(9-ethyl-6-((2S,5R)-4-(1-(2-isopropoxypyridin-4-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A124); 2-(6-((2S,5R)-4-(1-(3-cyclopropylphenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A125); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-(1-methylcyclopropyl)phenyl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A126); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-(oxetan-3-yl)phenyl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A127); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(6-(trifluoromethoxy)pyridin-2-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A128); 2-(6-((2S,5R)-4-(1-(2-(tert-butyl)pyridin-4-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A129); 4-(1-((2R,5S)-4-(8-(cyanomethyl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazin-1-yl)ethyl)-N-methylpicolinamide (A130); 4-(1-((2R,5S)-4-(8-(cyanomethyl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazin-1-yl)ethyl)-N,N-dimethylpicolinamide (A131); 5-(1-((2R,5S)-4-(8-(cyanomethyl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazin-1-yl)ethyl)-2-fluoro-N-methylbenzamide (A132); 2-(9-ethyl-6-((2S,5R)-4-(1-(3-(1-hydroxy-2-methylpropan-2-yl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A133); 2-(3-(1-((2R,5S)-4-(8-(cyanomethyl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazin-1-yl)ethyl)phenyl)-2-methylpropanenitrile (A134); 2-(9-ethyl-6-((2S,5R)-4-(1-(4-fluoro-3-(prop-1-en-2-yl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A135); 2-(9-ethyl-6-((2S,5R)-4-(1-(4-fluoro-3-(1-methylcyclopropyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A136); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-methyl-4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A137); 2-(6-((2S,5R)-4-(1-(5,6-dimethylpyridin-2-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A138); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(4-(morpholinomethyl)phenyl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A139); 2-(6-((2S,5R)-4-(1-(3-(2-oxa-6-azaspiro[3.3]heptan-6-yl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A140); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-morpholinophenyl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A141); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-(morpholinomethyl)phenyl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A142); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(4-morpholinophenyl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A143); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-phenylpyridin-4-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A144); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(p-tolyl)but-2-yn-1-yl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A145); 2-(6-((2S,5R)-4-(1-(5-(difluoromethyl)pyridin-2-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A146); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(5-methylpyridin-2-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A147); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(5-(trifluoromethyl)pyridin-2-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A148); 2-(6-((2S,5R)-2,5-diethyl-4-(1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A149); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A150); 2-(6-((2S,5R)-2,5-diethyl-4-(1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A151); 8-(but-2-yn-1-yl)-6-((2S,5R)-2,5-diethyl-4-(1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-3,9-dimethyl-3,9-dihydro-2H-purin-2-one (A152); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(quinoxalin-2-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A153); 2-(6-((2S,5R)-4-(1-(isoquinolin-3-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A154); 2-(6-((2S,5R)-4-(1-(1,2-dimethyl-4-oxo-1,4-dihydroquinolin-7-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A155); 2-(9-ethyl-6-((2S,5R)-4-(1-(7-fluoro-2,2-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A156); 2-(9-ethyl-6-((2S,5R)-4-(1-(7-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A157); 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A158); 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A159); 2-(6-((2S,5R)-4-(1-(7-chloro-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A160); 2-(9-ethyl-6-((2S,5R)-4-(1-(7-fluoro-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A161); 2-(6-((2S,5R)-4-(1-(7-fluoro-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A162); 2-(9-ethyl-6-((2S,5R)-4-(1-(8-fluoro-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A163); 2-(6-((2S,5R)-4-(1-(8-fluoro-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A164); 2-(9-(2,2-difluoroethyl)-6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A165); 2-(9-(2,2-difluoroethyl)-6-((2S,5R)-4-(1-(2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A166); 2-(6-((2S,5R)-4-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A167); 2-(9-ethyl-6-((2S,5R)-4-(1-(2-ethyl-2-methylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A168); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(spiro[benzo[d][1,3]dioxole-2,1-cyclobutan]-5-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A169); 2-(6-((2S,5R)-4-(1-(6-chloro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A170); 2-(6-((2S,5R)-4-(1-(7-chloro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A171); 2-(6-((2S,5R)-4-1-(4-chloro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A172); 2-(9-ethyl-6-((2S,5R)-4-(1-(7-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A173); 2-(9-ethyl-6-((2S,5R)-4-(1-(6-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A174); 2-(9-ethyl-6-((2S,5R)-4-(1-(4-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A175); 2-(6-((2S,5R)-4-(1-(4,6-difluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A176); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2,2,7-trimethylbenzo[d][1,3]dioxol-5-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A177); 2-(9-ethyl-6-((2S,5R)-4-(1-(2-isopropyl-1,3-dioxoisoindolin-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A178); 2-(6-((2S,5R)-4-(1-(2,2-dimethyl-3-oxo-2,3-dihydrobenzofuran-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A179); 2-(6-((2S,5R)-4-(1-(2,2-dimethyl-2,3-dihydrofuro[2,3-b]pyridin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A180); 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrofuro[3,2-b]pyridin-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A181); 2-(6-((2S,5R)-4-(3,3-dimethyl-2,3-dihydrofuro[3,2-b]pyridine-5-carbonyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A182); 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzofuran-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A183); 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydro-1H-inden-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A184); 2-(9-ethyl-6-((2S,5R)-4-(1-(7-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-4-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A185); 2-(6-((2S,5R)-4-(1-(2,2-dimethylbenzo[d][1,3]dioxol-4-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A186); 2-(6-((2S,5R)-4-(1-(2,2-difluorobenzo[d][1,3]dioxol-4-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A187); 2-(3-ethyl-6-fluoro-7-((2S,5R)-4-(1-(4-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (A190); 2-(7-((2S,5R)-4-(1-(2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-ethyl-6-fluoro-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (A188); 2-(3-ethyl-6-fluoro-7-((2S,5R)-4-(1-(7-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (A189); 2-(3-ethyl-6-fluoro-7-((2S,5R)-4-(1-(4-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (A190); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-methyl-4-oxo-4H-chromen-7-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A191); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-methyl-4-oxo-3,4-dihydro-2H-benzo[e][1,3]oxazin-6-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A192); 2-(6-((2S,5R)-4-(1-(1,3-dimethyl-4-oxo-1,4-dihydroquinolin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,4,5,9-tetrahydro-2H-purin-8-yl)acetonitrile (A193); 2-(7-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (A2); 2-(7-((2S,5R)-4-(1-(4-fluorophenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (A3); 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(4-methyl-2-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (A8); 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(4-methyl-2-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-6-fluoro-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (A15); 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(3-methyl-2-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-6-fluoro-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (A17); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(4-methyl-2-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A32); 2-(6-((2S,5R)-4-(1-(3-(difluoromethyl)-5-fluorophenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A38); 2-(6-((2S,5R)-4-(1-(3-(1,1-difluoroethyl)-4-methoxyphenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A39); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A40); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(m-tolyl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A45); 2-(9-ethyl-6-((2S,5R)-4-(1-(3-methoxyphenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A46); 2-(9-ethyl-6-((2S,5R)-4-(1-(2-methoxypyridin-4-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A47); 2-(6-((2S,5R)-4-(1-(7-chloro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A73); 2-(6-((2S,5R)-4-(1-(2,2-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-c]pyridin-7-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A75); 2-(6-((2S,5R)-4-(1-(2,2-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-7-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A76); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(1-oxo-2-(2,2,2-trifluoroethyl)isoindolin-5-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A82); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2,3,3-trimethyl-1-oxoisoindolin-5-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A84); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-methyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A89); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-methylquinolin-7-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A90); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-methyl-1-oxo-1,2-dihydroisoquinolin-7-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A92); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-methyl-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A93); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-methyl-3-oxoisoindolin-5-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A100); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-methyl-1-oxoisoindolin-5-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A101); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-(trifluoromethyl)thiazol-4-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A104); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-(trifluoromethyl)thiazol-5-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A105); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A106); 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(1-methyl-2-oxo-1,2-dihydropyridin-4-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A107); 2-(6-((2S,5R)-4-(1-(3-(difluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (A109); 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(quinoxalin-6-yl)ethyl)piperazin-1-yl)-4-methyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridin-2-yl)acetonitrile (A194); 2-(7-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-4-methyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridin-2-yl)acetonitrile (A195); or 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(4-methyl-2-(trifluoromethyl)phenyl) ethyl)piperazin-1-yl)-4-methyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridin-2-yl)acetonitrile (A196).
31. A pharmaceutical composition comprising one or more compounds of any one of claims 1-30, or a stereoisomer or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.
32. A method of treating a disease, comprising administering to a patient in need of such treatment a therapeutically effective amount of a compound of any one of claims 1-30, wherein the disease is cancer.
Description
DETAILED DESCRIPTION OF THE DISCLOSURE
Definitions
[0016] The following terms have the indicated meanings throughout the specification:
[0017] As used herein, including the appended claims, the singular forms of words such as a, an, and the, include their corresponding plural references unless the context clearly dictates otherwise.
[0018] The term or is used to mean, and is used interchangeably with, the term and/or unless the context clearly dictates otherwise.
[0019] The term alkyl refers to a hydrocarbon group selected from linear and branched saturated hydrocarbon groups derived from an alkane by removal of one hydrogen atom from the same carbon atom, which comprises from 1 to 18, such as from 1 to 12, further such as from 1 to 10, more further such as from 1 to 8, or from 1 to 6, or from 1 to 4, carbon atoms. Examples of alkyl groups comprising from 1 to 6 carbon atoms (i.e., C.sub.1-6 alkyl) include, but not limited to, methyl, ethyl, 1-propyl or n-propyl (n-Pr), 2-propyl or isopropyl (i-Pr), 1-butyl or n-butyl (n-Bu), 2-methyl-1-propyl or isobutyl (i-Bu), 1-methylpropyl or s-butyl (s-Bu), 1,1-dimethylethyl ort-butyl (t-Bu), 1-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl and 3,3-dimethyl-2-butyl groups. The alkyl group can be optionally enriched in deuterium, e.g., CD.sub.3, CD.sub.2CD.sub.3 and the like. The term alkylene refers to a hydrocarbon group selected from linear and branched saturated hydrocarbon groups derived from an alkane by removal of two hydrogen atoms from the same carbon atom, which comprises from 1 to 6, such as from 1 to 4, carbon atoms, further such as from 1 to 3, more further such as 1, 2 or 3 carbon atoms, include, but not limited to, methylene (CH.sub.2), ethylene (CH.sub.2CH.sub.2), 1-methymethylene (CH(CH.sub.3)), or trimethylene (CH.sub.2CH.sub.2CH.sub.2). When the alkyl groups described herein are said to be substituted, they may be substituted with any substituent or substituents as those found in the exemplary compounds and embodiments disclosed herein, as well as halogen (chloro, iodo, bromo, or fluoro); alkyl; hydroxyl; alkoxy; alkoxyalkyl; amino; alkylamino; carboxy; nitro; cyano; thiol; thioether; imine; imide; amidine; guanidine; enamine; aminocarbonyl; acylamino; phosphonato; phosphine; thiocarbonyl; sulfonyl; sulfone; sulfonamide; ketone; aldehyde; ester; urea; urethane; oxime; hydroxyl amine; alkoxyamine; aralkoxyamine; N-oxide; hydrazine; hydrazide; hydrazone; azide; isocyanate; isothiocyanate; cyanate; thiocyanate; B(OH).sub.2, O(alkyl)aminocarbonyl, aryl, heterocyclyl, or heteroaryl.
[0020] The term halogen refers to fluoro (F), chloro (Cl), bromo (Br) and iodo (I).
[0021] The term haloalkyl refers to an alkyl group in which one or more hydrogen is/are replaced by one or more halogen atoms such as fluoro, chloro, bromo, and iodo. Examples of the haloalkyl include haloC.sub.1-6alkyl, haloC.sub.1-6alkyl or halo C.sub.1-4alkyl, but not limited to CF.sub.3, CH.sub.2Cl, CH.sub.2CF.sub.3, CCl.sub.2, CF.sub.3, and the like.
[0022] The term alkyloxy or alkoxy refers to an alkyl group as defined above attached to the parent molecular moiety through an oxygen atom. Examples of an alkyloxy, e.g., C.sub.1-6alkyloxy or C.sub.1-4 alkyloxy include, but not limited to, methoxy, ethoxy, isopropoxy, propoxy, n-butoxy, tert-butoxy, pentoxy and hexoxy and the like.
[0023] The term amino refers to NH.sub.2.
[0024] The term alkenyl herein refers to a hydrocarbon group selected from linear and branched hydrocarbon groups comprising at least one CC double bond and from 2 to 18, such as from 2 to 8, further such as from 2 to 6, carbon atoms. Examples of the alkenyl group, e.g., C2-6 alkenyl, include, but not limited to ethenyl or vinyl, prop-1-enyl, prop-2-enyl, 2-methylprop-1-enyl, but-1-enyl, but-2-enyl, but-3-enyl, buta-1,3-dienyl, 2-methylbuta-1,3-dienyl, hex-1-enyl, hex-2-enyl, hex-3-enyl, hex-4-enyl, and hexa-1,3-dienyl groups.
[0025] The term alkynyl herein refers to a hydrocarbon group selected from linear and branched hydrocarbon group, comprising at least one CC triple bond and from 2 to 18, such as 2 to 8, further such as from 2 to 6, carbon atoms. Examples of the alkynyl group, e.g., C2-6 alkynyl, include, but not limited to ethynyl, 1-propynyl, 2-propynyl (propargyl), 1-butynyl, 2-butynyl, and 3-butynyl groups.
[0026] The term cycloalkyl refers to a hydrocarbon group selected from saturated cyclic hydrocarbon groups, comprising monocyclic and polycyclic (e.g., bicyclic and tricyclic) groups including fused, bridged or spiro cycloalkyl. For example, the cycloalkyl group may comprise from 3 to 12, such as from 3 to 10, further such as 3 to 8, further such as 3 to 6, 3 to 5, or 3 to 4 carbon atoms. Even further for example, the cycloalkyl group may be selected from monocyclic group comprising from 3 to 12, such as from 3 to 10, further such as 3 to 8, 3 to 6 carbon atoms. Examples of the monocyclic cycloalkyl group include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl, 1-cyclohex-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, and cyclododecyl groups. In particular, Examples of the saturated monocyclic cycloalkyl group, e.g., C.sub.3-8cycloalkyl, include, but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl groups. In a preferred embedment, the cycloalkyl is a monocyclic ring comprising 3 to 6 carbon atoms (abbreviated as C.sub.3-6 cycloalkyl), including but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Examples of the bicyclic cycloalkyl groups include those having from 7 to 12 ring atoms arranged as a fused bicyclic ring selected from [4,4], [4,5], [5,5], [5,6] and [6,6] ring systems, or as a bridged bicyclic ring selected from bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, and bicyclo[3.2.2]nonane. Further Examples of the bicyclic cycloalkyl groups include those arranged as a bicyclic ring selected from [5,6] and [6,6] ring systems.
[0027] The term deuterated compound refers to a compound wherein one or more carbon-bound hydrogen(s) are replaced by one or more deuterium(s). The term deuterated is used herein to modify a chemical structure or an organic group or radical, wherein one or more carbon-bound hydrogen(s) are replaced by one or more deuterium(s), e.g., deuterated-alkyl, deuterated-cycloalkyl, deuterated-heterocycloalkyl, deuterated-aryl, deuterated-morpholinyl, and the like. For example, the term deuterated-alkyl defined above refers to an alkyl group as defined herein, wherein at least one hydrogen atom bound to carbon is replaced by a deuterium. In a deuterated alkyl group, at least one carbon atom is bound to a deuterium; and it is possible for a carbon atom to be bound to more than one deuterium; it is also possible that more than one carbon atom in the alkyl group is bound to a deuterium.
[0028] The term aryl used alone or in combination with other terms refers to an aromatic carbocyclic group of from 6 to 14 carbon atoms having a single ring (e.g., phenyl) or multiple condensed rings (e.g., naphthyl or anthryl). In some embodiments, aryl groups contain 6-14 carbons, and in others from 6 to 12 or even 6 to 10 carbon atoms in the ring portions of the groups. Particular aryls include phenyl, biphenyl, naphthyl and the like. An aryl group can be substituted or unsubstituted. The phrase aryl groups also includes groups containing fused rings, such as fused aromatic-aliphatic ring systems (e.g., indanyl, tetrahydronaphthyl, and the like). In some embodiments, an aryl group refers to a group selected from: 5- and 6-membered carbocyclic aromatic rings, e.g., phenyl; bicyclic ring systems such as 7 to 12-membered bicyclic ring systems, wherein at least one ring is carbocyclic and aromatic, e.g., naphthyl and indanyl; and, tricyclic ring systems such as 10 to 15 membered tricyclic ring systems wherein at least one ring is carbocyclic and aromatic, e.g., fluorenyl.
[0029] The terms aromatic hydrocarbon ring and aryl are used interchangeably throughout the disclosure herein. In some embodiments, a monocyclic or bicyclic aromatic hydrocarbon ring has 5 to 10 ring-forming carbon atoms (i.e., C.sub.5-10 aryl). Examples of a monocyclic or bicyclic aromatic hydrocarbon ring include, but not limited to, phenyl, naphth-1-yl, naphth-2-yl, anthracenyl, phenanthrenyl, and the like. In some embodiments, the aromatic hydrocarbon ring is a naphthalene ring (naphth-1-yl or naphth-2-yl) or phenyl ring. In some embodiments, the aromatic hydrocarbon ring is a phenyl ring.
[0030] The term heteroaryl herein refers to a group selected from: [0031] 5-, 6- or 7-membered aromatic, monocyclic rings comprising at least one heteroatom, for example, from 1 to 4, or, in some embodiments, from 1 to 3, in some embodiments, from 1 to 2, heteroatoms, selected from nitrogen (N), sulfur (S) and oxygen (O), with the remaining ring atoms being carbon; [0032] 7- to 12-membered bicyclic rings comprising at least one heteroatom, for example, from 1 to 4, or, in some embodiments, from 1 to 3, or, in other embodiments, 1 or 2, heteroatoms, selected from nitrogen, oxygen or optionally oxidized sulfur as ring member(s), with the remaining ring atoms being carbon and wherein at least one ring is aromatic and at least one heteroatom is present in the aromatic ring; and [0033] 11- to 14-membered tricyclic rings comprising at least one heteroatom, for example, from 1 to 4, or in some embodiments, from 1 to 3, or, in other embodiments, 1 or 2, heteroatoms, selected from nitrogen, oxygen or optionally oxidized sulfur as ring member(s), with the remaining ring atoms being carbon and wherein at least one ring is aromatic and at least one heteroatom is present in an aromatic ring.
[0034] When the total number of S and O atoms in the heteroaryl group exceeds 1, those heteroatoms are not adjacent to one another. In some embodiments, the total number of S and O atoms in the heteroaryl group is not more than 2. In some embodiments, the total number of S and O atoms in the aromatic heterocycle is not more than 1. When the heteroaryl group contains more than one heteroatom ring member, the heteroatoms may be the same or different. The nitrogen atoms in the ring(s) of the heteroaryl group can be oxidized to form N-oxides.
[0035] The term optionally oxidized sulfur used herein refers to S, SO or SO.sub.2.
[0036] The terms aromatic heterocyclic ring and heteroaryl are used interchangeably throughout the disclosure herein. In some embodiments, a monocyclic or bicyclic aromatic heterocyclic ring has 5-, 6-, 7-, 8-, 9- or 10-ring forming members with 1, 2, 3, or 4 heteroatom ring members independently selected from nitrogen (N), sulfur (S) and oxygen (O) and the remaining ring members being carbon. In some embodiments, the monocyclic or bicyclic aromatic heterocyclic ring is a monocyclic or bicyclic ring comprising 1 or 2 heteroatom ring members independently selected from nitrogen (N), sulfur (S) and oxygen (O). In some embodiments, the monocyclic or bicyclic aromatic heterocyclic ring is a 5- to 6-membered heteroaryl ring, which is monocyclic and which has 1 or 2 heteroatom ring members independently selected from nitrogen (N), sulfur (S) and oxygen (O). In some embodiments, the monocyclic or bicyclic aromatic heterocyclic ring is a 8- to 10-membered heteroaryl ring, which is bicyclic and which has 1 or 2 heteroatom ring members independently selected from nitrogen, sulfur and oxygen.
[0037] Examples of the heteroaryl group or the monocyclic or bicyclic aromatic heterocyclic ring include, but are not limited to, (as numbered from the linkage position assigned priority 1) 1H-pyrazolyl (such as 1H-pyrazol-3-yl, 1H-pyrazol-4-yl or 1H-pyrazol-5-yl), pyridyl or pyridinyl (such as 2-pyridyl, 3-pyridyl, or 4-pyridyl), cinnolinyl, pyrazinyl, pyrimidinyl (such as pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl or 2,4-pyrimidinyl, 3,5-pyrimidinyl), imidazolyl (such as 1H-imidazol-2-yl, 1H-imidazol-4-yl, 1H-imidazol-5-yl, or 2,4-imidazolyl), imidazopyridinyl, isoxazolyl, oxazolyl, thiazolyl, isothiazolyl, thiadiazolyl (such as 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, or 1,3,4-thiadiazolyl), tetrazolyl, thienyl (such as thien-2-yl, thien-3-yl), triazinyl, benzothienyl, furyl or furanyl, benzofuryl, benzoimidazolyl, indolyl (such as 1H-indol-2-yl, 1H-indol-3-yl, 1H-indol-4-yl, 1H-indol-5-yl, 1H-indol-6-yl or 1H-indol-7-yl), isoindolyl, indolinyl, oxadiazolyl (such as 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, or 1,3,4-oxadiazolyl), phthalazinyl, pyrazinyl (such as pyrazin-2-yl), pyridazinyl, pyrrolyl, triazolyl (such as 1,2,3-triazolyl, 1,2,4-triazolyl, or 1,3,4-triazolyl), quinolinyl (such as quinolin-2-yl, quinolin-3-yl, quinolin-4-yl, quinolin-5-yl, quinolin-6-yl, or quinolin-7-yl), isoquinolinyl (such as isoquinolin-1-yl, isoquinolin-3-yl, isoquinolin-4-yl, isoquinolin-5-yl, isoquinolin-6-yl, isoquinolin-7-yl, or isoquinolin-8-yl), pyrazolyl, pyrrolopyridinyl (such as 1H-pyrrolo[2,3-b]pyridin-5-yl), pyrazolopyridinyl (such as 1H-pyrazolo[3,4-b]pyridin-5-yl), pteridinyl, purinyl, 1-oxa-2,3-diazolyl, 1-oxa-2,4-diazolyl, 1-oxa-2,5-diazolyl, 1-oxa-3,4-diazolyl, 1-thia-2,3-diazolyl, 1-thia-2,4-diazolyl, 1-thia-2,5-diazolyl, 1-thia-3,4-diazolyl, furazanyl (such as furazan-2-yl, furazan-3-yl), benzofurazanyl, benzothiophenyl, benzothiazolyl, benzoxazolyl (such as benzo[d]oxazol-2-yl, benzo[d]oxazol-4-yl, benzo[d]oxazol-5-yl, benzo[d]oxazol-6-yl or benzo[d]oxazol-7-yl), quinazolinyl, quinoxalinyl (such as quinoxalin-2-yl, quinoxalin-3-yl, quinoxalin-4-yl, quinoxalin-5-yl, quinoxalin-6-yl, quinoxalin-7-yl or quinoxalin-8-yl), naphthyridinyl (such as 1,8-naphthyridin-2-yl, 1,8-naphthyridin-3-yl, or 1,8-naphthyridin-4-yl), 2,3-dihydro-[1,4]dioxino[2,3-b]pyridinyl (such as 2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl, 2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-7-yl, or 2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-8-yl), furopyridinyl, benzothiazolyl (such as benzo[d]thiazol-2-yl, benzo[d]thiazol-4-yl, benzo[d]thiazol-5-yl, benzo[d]thiazol-6-yl or benzo[d]thiazol-7-yl), benzo[d]imidazolyl (such as 1H-benzo[d]imidazol-2-yl, 1H-benzo[d]imidazol-4-yl, 1H-benzo[d]imidazol-5-yl, 1H-benzo[d]imidazol-6-yl or 1H-benzo[d]imidazol-7-yl), [1,2,4]triazolo[1,5-a]pyridinyl (such as [1,2,4]triazolo[1,5-a]pyridin-2-yl, [1,2,4]triazolo[1,5-a]pyridin-5-yl, [1,2,4]triazolo[1,5-a]pyridin-6-yl, [1,2,4]triazolo[1,5-a]pyridin-7-yl, or [1,2,4]triazolo[1,5-a]pyridin-8-yl), 3H-imidazo[4,5-b]pyridinyl (such as 3H-imidazo[4,5-b]pyridin-2-yl, 3H-imidazo[4,5-b]pyridin-5-yl, 3H-imidazo[4,5-b]pyridin-6-yl or 3H-imidazo[4,5-b]pyridin-7-yl), 1H-imidazo[4,5-b]pyridinyl (such as 1H-imidazo[4,5-b]pyridin-2-yl, 1H-imidazo[4,5-b]pyridin-5-yl, 1H-imidazo[4,5-b]pyridin-6-yl, 1H-imidazo[4,5-b]pyridin-7-yl), [1,2,4]triazolo[1,5-a]pyridinyl (such as [1,2,4]triazolo[1,5-a]pyridin-2-yl, 1,2,4]triazolo[1,5-a]pyridin-5-yl, [1,2,4]triazolo[1,5-a]pyridin-6-yl, [1,2,4]triazolo[1,5-a]pyridin-7-yl or [1,2,4]triazolo[1,5-a]pyridin-8-yl), indazolyl (such as 1H-indazol-5-yl), 5,6,7,8-tetrahydroisoquinoline, thiazolo[5,4-b]pyridinyl (such as thiazolo[5,4-b]pyridin-2-yl, thiazolo[5,4-b]pyridin-5-yl, thiazolo[5,4-b]pyridin-6-yl or thiazolo[5,4-b]pyridin-7-yl), thiazolo[4,5-b]pyridinyl (such as thiazolo[4,5-b]pyridin-2-yl, thiazolo[4,5-b]pyridin-5-yl, thiazolo[4,5-b]pyridin-6-yl or thiazolo[4,5-b]pyridin-7-yl), thieno[2,3-b]pyridinyl (such as thieno[2,3-b]pyridin-2-yl, thieno[2,3-b]pyridin-3-yl, thieno[2,3-b]pyridin-4-yl, thieno[2,3-b]pyridin-5-yl or thieno[2,3-b]pyridin-6-yl), thieno[3,2-b]pyridinyl (such as thieno[3,2-b]pyridin-2-yl, thieno[3,2-b]pyridin-3-yl, thieno[3,2-b]pyridin-5-yl, thieno[3,2-b]pyridin-6-yl, or thieno[3,2-b]pyridin-7-yl).
[0038] Also, a heteroaryl fused with a Heterocyclyl is defined as heteroaryl.
[0039] Heterocyclyl, heterocycle or heterocyclic are interchangeable and refer to a non-aromatic heterocyclyl group comprising one or more heteroatoms selected from nitrogen, oxygen or optionally oxidized sulfur as ring members, with the remaining ring members being carbon, including monocyclic, fused, bridged, and spiro ring, i.e., containing monocyclic heterocyclyl, bridged heterocyclyl, spiro heterocyclyl, and fused heterocyclic groups.
[0040] The term monocyclic heterocyclyl refers to monocyclic groups in which at least one ring member is a heteroatom selected from nitrogen, oxygen or optionally oxidized sulfur. A heterocycle may be saturated or partially saturated.
[0041] Exemplary monocyclic 4 to 9-membered heterocyclyl groups include, but not limited to, (as numbered from the linkage position assigned priority 1) pyrrolidin-1-yl, pyrrolidin-2-yl, pyrrolidin-3-yl, imidazolidin-2-yl, imidazolidin-4-yl, pyrazolidin-2-yl, pyrazolidin-3-yl, piperidin-1-yl, piperidin-2-yl, piperidin-3-yl, piperidin-4-yl, 2,5-piperazinyl, pyranyl, morpholinyl, morpholino, morpholin-2-yl, morpholin-3-yl, oxiranyl, aziridin-1-yl, aziridin-2-yl, azocan-1-yl, azocan-2-yl, azocan-3-yl, azocan-4-yl, azocan-5-yl, thiiranyl, azetidin-1-yl, azetidin-2-yl, azetidin-3-yl, oxetanyl, oxetan-3-yl, thietanyl, 1,2-dithietanyl, 1,3-dithietanyl, dihydropyridinyl, tetrahydropyridinyl, thiomorpholinyl, thioxanyl, piperazinyl, homopiperazinyl, homopiperidinyl, azepan-1-yl, azepan-2-yl, azepan-3-yl, azepan-4-yl, oxepanyl, thiepanyl, 1,4-oxathianyl, 1,4-dioxepanyl, 1,4-oxathiepanyl, 1,4-oxaazepanyl, 1,4-dithiepanyl, 1,4-thiazepanyl and 1,4-diazepanyl, 1,4-dithianyl, 1,4-azathianyl, oxazepinyl, diazepinyl, thiazepinyl, dihydrothienyl, dihydropyranyl, dihydrofuranyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, tetrahydrothiopyranyl, 1-pyrrolinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, 1,4-dioxanyl, 1,3-dioxolanyl, pyrazolinyl, pyrazolidinyl, dithianyl, dithiolanyl, pyrazolidinyl, imidazolinyl, pyrimidinonyl, or 1,1-dioxo-thiomorpholinyl.
[0042] The term spiro heterocyclyl refers to a 5 to 20-membered polycyclic heterocyclyl with rings connected through one common carbon atom (called a spiro atom), comprising one or more heteroatoms selected from nitrogen, oxygen or optionally oxidized sulfur as ring members, with the remaining ring members being carbon. One or more rings of a spiro heterocyclyl group may contain one or more double bonds, but none of the rings has a completely conjugated pi-electron system. Preferably a spiro heterocyclyl is 6 to 14-membered, and more preferably 7 to 12-membered. According to the number of common spiro atoms, a spiro heterocyclyl is divided into mono-spiro heterocyclyl, di-spiro heterocyclyl, or poly-spiro heterocyclyl, and preferably refers to mono-spiro heterocyclyl or di-spiro heterocyclyl, and more preferably 4-membered/4-membered, 3-membered/5-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered, or 5-membered/6-membered mono-spiro heterocyclyl. Examples of spiro heterocyclyl groups include, but not limited to, spiro[2.2]pentanyl, spiro[2.3]hexanyl, spiro[2.4]heptanyl, spiro[3.3]heptanyl, spiro[2.5]octanyl, spiro[3.4]octanyl, spiro[2.6]nonanyl, spiro[3.5]nonanyl, spiro[4.4]nonanyl, spiro[2.7]decanyl, spiro[3.6]decanyl, spiro[4.5]decanyl, spiro[3.7]undecanyl, spiro[4.6]undecanyl, spiro[5.5]undecanyl, spiro[4.7]dodecanyl, and spiro[5.6]dodecanyl, wherein one or two carbon atoms are replaced with oxygen, or nitrogen, for example, 2-oxa-6-azaspiro[3.3]heptanyl (e.g., 2-oxa-6-azaspiro[3.3]heptan-6-yl), 1,7-dioxaspiro[4.5]decyl, 2-oxa-7-aza-spiro[4.4]nonyl, 7-oxa spiro[3.5]nonyl and 5-oxa-spiro[2.4]heptyl, spiro[benzo[d][1,3]dioxole-2,1-cyclobutan]-yl (e.g., spiro[benzo[d][1,3]dioxole-2,1-cyclobutan]-5-yl).
[0043] The term fused heterocyclic group refers to a 5 to 20-membered polycyclic heterocyclyl group, wherein each ring in the system shares an adjacent pair of atoms (carbon and carbon atoms or carbon and nitrogen atoms) with another ring, comprising one or more heteroatoms selected from nitrogen, oxygen or optionally oxidized sulfur as ring members, with the remaining ring members being carbon. One or more rings of a fused heterocyclic group may contain one or more double bonds, but none of the rings has a completely conjugated pi-electron system. Preferably, a fused heterocyclyl is 6 to 14-membered, and more preferably 7 to 10-membered. According to the number of membered rings, a fused heterocyclyl is divided into bicyclic, tricyclic, tetracyclic, or polycyclic fused heterocyclyl, preferably refers to bicyclic or tricyclic fused heterocyclyl, and more preferably 5-membered/5-membered, or 5-membered/6-membered bicyclic fused heterocyclyl. Representative examples of fused heterocycles include, but not limited to, the following groups octahydrocyclopenta[c]pyrrole (e.g., octahydrocyclopenta[c]pyrrol-2-yl), octahydropyrrolo[3,4-c]pyrrolyl, octahydroisoindolyl, isoindolinyl (e.g., isoindoline-2-yl), octahydro-benzo[b][1,4]dioxin.
[0044] The term bridged heterocyclyl refers to a 5- to 14-membered polycyclic heterocyclic alkyl group, wherein every two rings in the system share two disconnected atoms, comprising one or more heteroatoms selected from nitrogen, oxygen or optionally oxidized sulfur as ring members, with the remaining ring members being carbon. One or more rings of a bridged heterocyclyl group may contain one or more double bonds, but none of the rings has a completely conjugated pi-electron system. Preferably, a bridged heterocyclyl is 6 to 14-membered, and more preferably 7 to 10-membered. According to the number of membered rings, a bridged heterocyclyl is divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclyl, and preferably refers to bicyclic, tricyclic or tetracyclic bridged heterocyclyl, and more preferably bicyclic or tricyclic bridged heterocyclyl. Representative examples of bridged heterocyclyls include, but not limited to, the following groups: 2-azabicyclo[2.2.1]heptyl, azabicyclo[3.1.0]hexyl, 2-azabicyclo[2.2.2]octyl and 2-azabicyclo[3.3.2]decyl.
[0045] A cycloalkylalkyl group is a radical of the formula: -alkyl-cycloalkyl, wherein alkyl and cycloalkyl are as defined above. Substituted cycloalkylalkyl groups may be substituted at the alkyl, the cycloalkyl, or both the alkyl and the cycloalkyl portions of the group. Representative cycloalkylalkyl groups include but are not limited to methylcyclopropyl, methylcyclobutyl, methylcyclopentyl, methylcyclohexyl, ethylcyclopropyl, ethylcyclobutyl, ethylcyclopentyl, ethylcyclohexyl, propylcyclopentyl, propylcyclohexyl and the like.
[0046] An aralkyl group is a radical of the formula: -alkyl-aryl, wherein alkyl and aryl are defined above. Substituted aralkyl groups may be substituted at the alkyl, the aryl, or both the alkyl and the aryl portions of the group. Representative aralkyl groups include but are not limited to benzyl and phenethyl groups and fused (cycloalkylaryl)alkyl groups such as 4-ethyl-indanyl.
[0047] An heterocyclylalkyl group is a radical of the formula: -alkyl-heterocyclyl, wherein alkyl and heterocyclyl are defined above. Substituted heterocyclylalkyl groups may be substituted at the alkyl, the heterocyclyl, or both the alkyl and the heterocyclyl portions of the group. Representative heterocylylalkyl groups include but are not limited to 4-ethyl-morpholinyl, 4-propylmorpholinyl, furan-2-yl methyl, furan-3-yl methyl, pyridin-3-yl methyl, tetrahydrofuran-2-yl ethyl, and indol-2-yl propyl.
[0048] A halogen is fluorine, chlorine, bromine or iodine.
[0049] A hydroxyalkyl group is an alkyl group as described above substituted with one or more hydroxy groups.
[0050] An alkoxy group is O-(alkyl), wherein alkyl is defined above.
[0051] An alkoxyalkyl group is -(alkyl)-O-(alkyl), wherein alkyl is defined above.
[0052] An amino group is a radical of the formula: NH.sub.2.
[0053] An alkylamino group is a radical of the formula: NH-alkyl or N(alkyl).sub.2, wherein each alkyl is independently as defined above.
[0054] A carboxy group is a radical of the formula: C(O)OH.
[0055] An aminocarbonyl group is a radical of the formula: C(O)N(R.sup.#).sub.2, C(O)NH(R.sup.#) or C(O)NH.sub.2, wherein each R.sup.# is independently a substituted or unsubstituted alkyl, cycloalkyl, aryl, aralkyl, heterocyclyl or heterocyclyl group as defined herein.
[0056] An acylamino group is a radical of the formula: NHC(O)(R.sup.#) or N(alkyl)C(O)(R.sup.#), wherein each alkyl and Rare independently as defined above.
[0057] A sulfonylamino group is a radical of the formula: NHSO.sub.2(R.sup.#) or N(alkyl)SO.sub.2(R.sup.#), wherein each alkyl and Rare defined above.
[0058] A urea group is a radical of the formula: N(alkyl)C(O)N(R.sup.#).sub.2, N(alkyl)C(O)NH(R.sup.#), N(alkyl)C(O)NH.sub.2, NHC(O)N(R.sup.#).sub.2, NHC(O)NH(R.sup.#), or NH(CO)NHR.sup.#, wherein each alkyl and R.sup.# are independently as defined above.
[0059] When the groups described herein, with the exception of alkyl group, are said to be substituted, they may be substituted with any appropriate substituent or substituents. Illustrative examples of substituents are those found in the exemplary compounds and embodiments disclosed herein, as well as halogen (chloro, iodo, bromo, or fluoro); alkyl; hydroxyl; alkoxy; alkoxyalkyl; amino; alkylamino; carboxy; nitro; cyano; thiol; thioether; imine; imide; amidine; guanidine; enamine; aminocarbonyl; acylamino; phosphonato; phosphine; thiocarbonyl; sulfonyl; sulfone; sulfonamide; ketone; aldehyde; ester; urea; urethane; oxime; hydroxyl amine; alkoxyamine; aralkoxyamine; N-oxide; hydrazine; hydrazide; hydrazone; azide; isocyanate; isothiocyanate; cyanate; thiocyanate; oxygen (O); B(OH).sub.2, O(alkyl)aminocarbonyl; cycloalkyl, which may be monocyclic or fused or non-fused polycyclic (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), or a heterocyclyl, which may be monocyclic or fused or non-fused polycyclic (e.g., pyrrolidyl, piperidyl, piperazinyl, morpholinyl, orthiazinyl); monocyclic or fused or non-fused polycyclic aryl or heteroaryl (e.g., phenyl, naphthyl, pyrrolyl, indolyl, furanyl, thiophenyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, pyrazolyl, pyridyl, quinolinyl, isoquinolinyl, acridinyl, pyrazinyl, pyridazinyl, pyrimidyl, benzimidazolyl, benzothiophenyl, or benzofuranyl) aryloxy; aralkyloxy; heterocyclyloxy; and heterocyclyl alkoxy.
[0060] Compounds disclosed herein may contain an asymmetric center and may thus exist as enantiomers. Enantiomers refer to two stereoisomers of a compound which are non-superimposable mirror images of one another. Where the compounds disclosed herein possess two or more asymmetric centers, they may additionally exist as diastereomers. Enantiomers and diastereomers fall within the broader class of stereoisomers. All such possible stereoisomers as substantially pure resolved enantiomers, racemic mixtures thereof, as well as mixtures of diastereomers are intended to be included. All stereoisomers of the compounds disclosed herein and/or pharmaceutically acceptable salts thereof are intended to be included. Unless specifically mentioned otherwise, reference to one isomer applies to any of the possible isomers. Whenever the isomeric composition is unspecified, all possible isomers are included.
[0061] The term substantially pure as used herein means that the titled stereoisomer contains no more than 35%, such as no more than 30%, further such as no more than 25%, even further such as no more than 20%, by weight of any other stereoisomer(s). In some embodiments, the term substantially pure means that the titled stereoisomer contains no more than 10%, for example, no more than 5%, such as no more than 1%, by weight of any other stereoisomer(s).
[0062] When compounds disclosed herein contain olefinic double bonds, unless specified otherwise, such double bonds are meant to include both E and Z geometric isomers.
[0063] When compounds disclosed herein contain a di-substituted cyclohexyl or cyclobutyl group, substituents found on cyclohexyl or cyclobutyl ring may adopt cis and trans formations. Cis formation means that both substituents are found on the upper side of the 2 substituent placements on the carbon, while trans would mean that they were on opposing sides.
[0064] Diastereomers refers to stereoisomers of a compound with two or more chiral centers but which are not mirror images of one another. Diastereomeric mixtures can be separated into their individual diastereomers on the basis of their physical chemical differences by methods well known to those skilled in the art, such as by flash column chromatography and/or fractional crystallization. Enantiomers can be separated by converting the enantiomeric mixture into a diastereomeric mixture by reaction with an appropriate optically active compound (e.g., chiral auxiliary such as a chiral alcohol or Mosher's acid chloride), separating the diastereomers and converting (e.g., hydrolyzing) the individual diastereoisomers to the corresponding pure enantiomers. Enantiomers can also be separated by use of a chiral HPLC column.
[0065] Pharmaceutically acceptable salts refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio. A pharmaceutically acceptable salt may be prepared in situ during the final isolation and purification of the compounds disclosed herein, or separately by reacting the free base function with a suitable organic acid or by reacting the acidic group with a suitable base.
[0066] Selective inhibitory activity or selectivity refers to the difference in the degree of inhibition against DGK and DGK; In some embodiments, a compound showing selective inhibitory activity of DGK over DGK refers a compound which shows a ratio of IC50 against DGK and IC50 against DGK larger than or equal to about 20. In some embodiments, a compound showing selective inhibitory activity of DGK over DGK refers a compound which shows a ratio of IC50 against DGK and IC50 against DGK larger than or equal to about 20. In some embodiments, a compound showing selective inhibitory activity of DGK over DGK refers a compound which shows an IC50 against DGK is not larger than about 2000 nM with the ratio of IC50 against DGK and IC50 against DGK larger than or equal to about 20; a compound showing selective inhibitory activity of DGK over DGK refers a compound which shows an IC50 against DGK is not larger than about 2000 nM with the ratio of IC50 against DGK and IC50 against DGK larger than or equal to about 20; and a compound showing dual inhibitory activity refers to a compound which shows inhibitory activities against both DGK and DGK with IC50 no larger than 500 nM and the ratio of the two IC50 values no more than 20. In some embodiments, a compound showing dual inhibitory activity refers to a compound which shows inhibitory activities against both DGK and DGK with IC50 no larger than 1000 nM and the ratio of the two IC50 values no more than 20. In some embodiments, a compound showing dual inhibitory activity refers to a compound which shows inhibitory activities against both DGK and DGK with IC50 no larger than 2000 nM and the ratio of the two IC50 values no more than 20.
[0067] As defined herein, a pharmaceutically acceptable salt thereof include salts of at least one compound of Formulas (I), (IA1), (IA2), (IA3), (IA4), (IB1), (IB2), (IB3), (IB4), (IC1), (IC2), (IC3), (IC4) and (II) (collectively referred to as formulas (I) to (II)), and salts of the stereoisomers of the compound of Formulas (I) to (II), such as salts of enantiomers, and/or salts of diastereomers.
[0068] The terms administration, administering, treating and treatment herein, when applied to an animal, human, experimental subject, cell, tissue, organ, or biological fluid, mean contact of an exogenous pharmaceutical, therapeutic, diagnostic agent, or composition to the animal, human, subject, cell, tissue, organ, or biological fluid. Treatment of a cell encompasses contact of a reagent to the cell, as well as contact of a reagent to a fluid, where the fluid is in contact with the cell. The term administration and treatment also means in vitro and ex vivo treatments, e.g., of a cell, by a reagent, diagnostic, binding compound, or by another cell. The term subject herein includes any organism, preferably an animal, more preferably a mammal (e.g., rat, mouse, dog, cat, rabbit) and most preferably a human.
[0069] The term effective amount or therapeutically effective amount refers to an amount of the active ingredient, such as compound that, when administered to a subject for treating a disease, or at least one of the clinical symptoms of a disease or disorder, is sufficient to affect such treatment for the disease, disorder, or symptom. The therapeutically effective amount can vary with the compound, the disease, disorder, and/or symptoms of the disease or disorder, severity of the disease, disorder, and/or symptoms of the disease or disorder, the age of the subject to be treated, and/or the weight of the subject to be treated. An appropriate amount in any given instance can be apparent to those skilled in the art or can be determined by routine experiments. In some embodiments, therapeutically effective amount is an amount of at least one compound and/or at least one stereoisomer thereof, and/or at least one pharmaceutically acceptable salt thereof disclosed herein effective to treat as defined above, a disease or disorder in a subject. In the case of combination therapy, the therapeutically effective amount refers to the total amount of the combination objects for the effective treatment of a disease, a disorder or a condition.
[0070] All Formulations of the pharmaceutical composition disclosed herein can be produced by the conventional methods in the pharmaceutical field. For example, the active ingredient can be mixed with one or more excipients, then to make the desired Formulation. The pharmaceutically acceptable excipient refers to conventional pharmaceutical carriers suitable for the desired pharmaceutical Formulation, for example: a diluent, a vehicle such as water, various organic solvents, etc., a filler such as starch, sucrose, etc. a binder such as cellulose derivatives, alginates, gelatin and polyvinylpyrrolidone (PVP); a wetting agent such as glycerol; a disintegrating agent such as agar, calcium carbonate and sodium bicarbonate; an absorption enhancer such as quaternary ammonium compound; a surfactant such as hexadecanol; an absorption carrier such as Kaolin and soap clay; a lubricant such as talc, calcium stearate, magnesium stearate, polyethylene glycol, etc. In addition, the pharmaceutical composition further comprises other pharmaceutically acceptable excipients such as a decentralized agent, a stabilizer, a thickener, a complexing agent, a buffering agent, a permeation enhancer, a polymer, aromatics, a sweetener, and a dye.
[0071] The term disease refers to any disease, discomfort, illness, symptoms or indications, and can be interchangeable with the term disorder or condition.
[0072] Throughout this specification and the claims which follow, unless the context requires otherwise, the term comprise, and variations such as comprises and comprising are intended to specify the presence of the features thereafter, but do not exclude the presence or addition of one or more other features. When used herein the term comprising can be substituted with the term containing, including or sometimes having.
[0073] Throughout this specification and the claims which follow, the term Cn-m indicates a range which includes the endpoints, wherein n and m are integers and indicate the number of carbons. Examples include C.sub.1-8, C.sub.1-6, and the like.
[0074] The term Tautomers refers to isomeric forms of a compound that are in equilibrium with each other. The concentrations of the isomeric forms will depend on the environment the compound is found in and may be different depending upon, for example, whether the compound is a solid or is in an organic or aqueous solution. For example, in aqueous solution, pyrazoles may exhibit the following isomeric forms, which are referred to as tautomers of each other:
##STR00007##
[0075] As readily understood by one skilled in the art, a wide variety of functional groups and other structures may exhibit tautomerism and all tautomers of compounds of any one of formulas (I) to (II) are within the scope of the present invention.
[0076] Unless specifically defined elsewhere in this document, all other technical and scientific terms used herein have the meaning commonly understood by one of ordinary skill in the art to which this invention belongs.
[0077] It should also be noted the compounds provided herein can contain unnatural proportions of atomic isotopes at one or more of the atoms. For example, the compounds may be radiolabeled with radioactive isotopes, such as for example tritium (H), iodine-125 (.sup.125I), sulfur-35 (.sup.35S), or carbon-14 (.sup.4C), or may be isotopically enriched, such as with deuterium (H), carbon-13 (.sup.13C), or nitrogen-15 (.sup.15N). As used herein, an isotopologue is an isotopically enriched compound. The term isotopically enriched refers to an atom having an isotopic composition other than the natural isotopic composition of that atom. Isotopically enriched may also refer to a compound containing at least one atom having an isotopic composition other than the natural isotopic composition of that atom. The term isotopic composition refers to the amount of each isotope present for a given atom. Radiolabeled and isotopically enriched compounds are useful as therapeutic agents, e.g., cancer and inflammation therapeutic agents, research reagents, e.g., binding assay reagents, and diagnostic agents, e.g., in vivo imaging agents. All isotopic variations of the compounds as described herein, whether radioactive or not, are intended to be encompassed within the scope of the embodiments provided herein. In some embodiments, there are provided isotopologues of the compounds, for example, the isotopologues are deuterium, carbon-13, or nitrogen-15 enriched compounds.
[0078] The term DGK or DAGK (Diacylglycerol kinase) refers to a family of enzymes that catalyzes the conversion of diacylglycerol (DAG) to phosphatidic acid (PA), utilizing ATP as a source of the phosphate. In non-stimulated cells, DGK activity is low, allowing DAG to be used for glycerophospholipid biosynthesis, but on receptor activation of the phosphoinositide pathway, DGK activity increases, driving the conversion of DAG to PA. As both lipids are thought to function as bioactive lipid signaling molecules with distinct cellular targets, DGK therefore occupies an important position, effectively serving as a switch by terminating the signalling of one lipid while simultaneously activating signalling by another. See Mdrida I, Avila-Flores A, Merino E (January 2008). Diacylglycerol kinases: at the hub of cell signalling, The Biochemical Journal, 409 (1): 1-18. Currently, ten members of the DGK family have been cloned and identified. Although all family members have conserved catalytic domains and two cysteine rich domains, they are further classified into five groups according to the presence of additional functional domains and substrate specificity. See van Blitterswijk W J, Houssa B (October 2000), Properties and functions of diacylglycerol kinases, Cellular Signalling, 12 (9-10): 595-605. These are as follows: [0079] a) Type 1DGK-, DGK-, DGK-contain EF-hand motifs and a recoverin homology domain [0080] b) Type 2DGK-, DGK-, DGK-contain a pleckstrin homology domain [0081] c) Type 3DGK-has specificity for arachidonate-containing DAG [0082] d) Type 4DGK-, DGK-tcontain a MARCKS homology domain, ankyrin repeats, a C-terminal nuclear localisation signal, and a PDZ-binding motif. [0083] e) Type 5DGK-contains a third cysteine-rich domain, a pleckstrin homology domain and a proline rich region.
[0084] The term treating as used herein, means an alleviation, in whole or in part, of a disorder, disease or condition, or one or more of the symptoms associated with a disorder, disease, or condition, or slowing or halting of further progression or worsening of those symptoms, or alleviating or eradicating the cause(s) of the disorder, disease, or condition itself. In one embodiment, treating means and alleviation, in whole or in part, of a disorder, disease or condition, or symptoms associated with a condition, treatable or preventable by inhibition of a DGK pathway. In another embodiment, treating means and alleviation, in whole or in part, of a disorder, disease or condition, or symptoms associated with a condition, treatable or preventable by inhibition of a DGK- pathway. In another embodiment, treating means and alleviation, in whole or in part, of a disorder, disease or condition, or symptoms associated with a condition, treatable or preventable by inhibition of a DGK- pathway. In one embodiment, the disorder is cancer.
[0085] Preventing as used herein, means a method of delaying and/or precluding the onset, recurrence or spread, in whole or in part, of a disorder, disease or condition; barring a subject from acquiring a disorder, disease, or condition; or reducing a subject's risk of acquiring a disorder, disease, or condition. In another, the disorder is a condition, treatable or preventable by inhibition of a DGK pathway. In another, the disorder is a condition, treatable or preventable by inhibition of a DGK- pathway. In another, the disorder is a condition, treatable or preventable by inhibition of a DGK- pathway. In one embodiment, the disorder is cancer.
[0086] The term subject includes an animal, including, but not limited to, an animal such a cow, monkey, horse, sheep, pig, chicken, turkey, quail, cat, dog, mouse, rat, rabbit or guinea pig, in one embodiment a mammal, in another embodiment a human. In one embodiment, a subject is a human having or at risk for having liver fibrotic disorders or diabetes or metabolic syndrome leading to liver fibrotic disorders, or a condition, treatable or preventable by inhibition of a JNK pathway, or a symptom thereof.
[0087] As used herein, the term Compound refers to compounds of formulas (I), (IA1), (IA2), (IA3), (IA4), (IB1), (IB2), (IB3), (IB4), (IC1), (IC2), (IC3), (IC4), (II) and salts of the stereoisomers of the compound of Formulas (I), (IA1), (IA2), (IA3), (IA4), (IB1), (IB2), (IB3), (IB4), (IC1), (IC2), (IC3), (IC4) and (II) (collectively referred to as formulas (I) to (II)) as well as to further embodiments provided herein. In one embodiment, a Compound is a compound exemplified herein. The term Compound includes pharmaceutically acceptable salts, tautomers, isotopologues, stereoisomers, deuterated compounds and prodrugs of the compounds provided herein.
[0088] As used herein and unless otherwise indicated, the term prodrug means a compound that can hydrolyze, oxidize, or otherwise react under biological conditions (in vitro or in vivo) to provide an active compound, particularly a compound of any one of formulas (I) to (II). Examples of prodrugs include, but are not limited to, derivatives and metabolites of a compound that include biohydrolyzable moieties such as biohydrolyzable amides, biohydrolyzable esters, biohydrolyzable carbamates, biohydrolyzable carbonates, biohydrolyzable ureides, and biohydrolyzable phosphate analogues. In certain embodiments, prodrugs of compounds with carboxyl functional groups are the lower alkyl esters of the carboxylic acid. The carboxylate esters are conveniently formed by esterifying any of the carboxylic acid moieties present on the molecule. Prodrugs can typically be prepared using well-known methods, such as those described by Burger's Medicinal Chemistry and Drug Discovery 6.sup.th ed. (Donald J. Abraham ed., 2001, Wiley) and Design and Application of Prodrugs (H. Bundgaard ed., 1985, Harwood Academic Publishers Gmfh).
[0089] As used herein and unless otherwise indicated, the term stereoisomer or stereomerically pure means one stereoisomer of a Compound that is substantially free of other stereoisomers of that compound. For example, a stereomerically pure compound having one chiral center will be substantially free of the opposite enantiomer of the compound. A stereomerically pure compound having two chiral centers will be substantially free of other diastereomers of the compound. A typical stereomerically pure compound comprises greater than about 80% by weight of one stereoisomer of the compound and less than about 20% by weight of other stereoisomers of the compound, greater than about 90% by weight of one stereoisomer of the compound and less than about 10% by weight of the other stereoisomers of the compound, greater than about 95% by weight of one stereoisomer of the compound and less than about 5% by weight of the other stereoisomers of the compound, or greater than about 97% by weight of one stereoisomer of the compound and less than about 3% by weight of the other stereoisomers of the compound. The compounds can have chiral centers and can occur as racemates, individual enantiomers or diastereomers, and mixtures thereof. All such isomeric forms are included within the embodiments disclosed herein, including mixtures thereof.
Compounds
[0090] The compounds provided herein have a novel core structure and show the desired inhibition of DGK and DGK. In some embodiments, the compounds disclosed herein show the dual inhibitory activity of both DGK and DGK. In some embodiments, the compounds disclosed herein show the selective inhibitory activity of DGK over DGK. In some embodiments, the compounds disclosed herein show the selective inhibitory activity of DGK over DGK.
[0091] Disclosed herein provides a compound of formula (I),
##STR00008##
or a pharmaceutically acceptable salt, a tautomer, a stereoisomer, an enantiomer, an isotopologue, or a prodrug thereof,
wherein [0092] X.sup.1 is C or N, [0093] each of X.sup.2 and X.sup.3 is independently selected from N or CH; [0094] X.sup.4 is N, O or S; [0095] R.sup.1 is hydrogen, deuterium, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl; R.sup.2 is hydrogen, halogen, cyano, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl, provided that R.sup.2 is absent when X.sup.1 is N; [0096] R.sup.4 is hydrogen, halogen, cyano, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl, provided that R.sup.4 is absent when X.sup.4 is O or S; [0097] R.sup.5 is hydrogen, halogen, cyano, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl, R.sup.5aC(O), R.sup.5aC(O)O, R.sup.5aOC(O), R.sup.5aC(O)NR.sup.5b, R.sup.5aNR.sup.5bC(O), or R.sup.5aSO.sub.2, wherein R.sup.5a and R.sup.5b are each independently hydrogen, alkyl, or cycloalkyl; [0098] each of R.sup.7, R.sup.9, R.sup.8, and R.sup.10 is independently hydrogen, cyano, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxyl, substituted or unsubstituted cycloalkyl, C(O)R.sup.7a, or -alkyl-C(O)R.sup.7a, and wherein R.sup.a is hydrogen, alkyl, or alkoxy, provided that at least one of R.sup.7 and R.sup.9 is not hydrogen; or R.sup.7 and R.sup.9 are each hydrogen and R.sup.9 and R.sup.10 together form a bridge containing at least one CH.sub.2-moiety in addition to the two bridgehead atoms; or R.sup.8 and R.sup.10 are each hydrogen and R.sup.7 and R.sup.9 together form a bridge containing at least one CH.sub.2 moiety in addition to the two bridgehead atoms; [0099] L.sup.1 is a direct bond, O, N(R.sup.L), substituted or unsubstituted alkyl, -alkylene or C(O), wherein R.sup.L is hydrogen or alkyl; [0100] Cy.sup.1 is unsubstituted or substituted cycloalkyl, unsubstituted or substituted aryl, unsubstituted or substituted heterocyclyl, or unsubstituted or substituted heteroaryl, wherein Cy.sup.1 is optionally substituted with 1 to 5 substituents R.sup.3a; [0101] wherein R.sup.3a is selected from deuterium, alkoxy, alkyl, halogen, oxo, R.sup.3bSO.sub.2, cycloalkyl, cyano, R.sup.3bC(O)N(R.sup.3c), N(R.sup.3bR.sup.3c)C(O), N(R.sup.3bR.sup.3c), R.sup.3bOC(O), cycloalkyl or heterocyclyl; optionally wherein two R.sup.3a connect to the same carbon and together form a spirocyclic ring; optionally wherein two R.sup.3a form a fused ring with Cy.sup.1, wherein R.sup.3a is unsubstituted or substituted with cyano, alkoxy, alkyl, halogen, or hydroxy; and [0102] wherein R.sup.3b and R.sup.3c are each independently hydrogen or alkyl.
The definitions of X.sup.1 and X.sup.4
[0103] In some embodiments, X.sup.1 is C. In some embodiments, X.sup.1 is N. In some embodiments, X.sup.4 is N. In some embodiments, X.sup.4 is S. In some embodiments, X.sup.4 is O. In some embodiments, X.sup.1 is C and X.sup.4 is N. In some embodiments, X.sup.1 is N and X.sup.4 is N. In some embodiments, X.sup.1 is C and X.sup.4 is S.
[0104] In some embodiments, the compound of formula (I) is a compound of any one of formulas (IA1), (IB1) or (IC1):
##STR00009##
wherein the variables are defined as herein.
[0105] In some embodiments, the compound of formula (I) is a compound of any one of formula (IA2), (IB2) or (IC2):
##STR00010##
wherein the variables are defined as herein.
[0106] In some embodiments, the compound of formula (I) is a compound of any one of formula (IA3), (IB3) or (IC3):
##STR00011##
wherein the variables are defined as herein.
[0107] In some embodiments, the compound of formula (I) is a compound of any one of formula (IA4), (IB4) or (IC4):
##STR00012##
wherein the variables are defined as herein.
[0108] In some embodiments, the compound of formula (I) is a compound of formula (II):
##STR00013##
or a pharmaceutically acceptable salt, a tautomer, a stereoisomer, an enantiomer, an isotopologue, or a prodrug thereof, [0109] wherein [0110] X.sup.1 is C or N, [0111] X.sup.4 is N, O or S; [0112] R.sup.1 is hydrogen, deuterium, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl; [0113] R.sup.2 is hydrogen, halogen, cyano, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl, provided that R.sup.2 is absent when X.sup.1 is N; [0114] R.sup.4 is hydrogen, halogen, cyano, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl, provided that R.sup.4 is absent when X.sup.4 is O or S; [0115] R.sup.5 is hydrogen, halogen, cyano, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl, R.sup.5aC(O), R.sup.5aC(O)O, R.sup.5aOC(O), R.sup.5aC(O)NR.sup.5b, R.sup.5aNR.sup.5bC(O), or R.sup.5aSO.sub.2, wherein R.sup.5a and R.sup.5b are each independently hydrogen, alkyl, or cycloalkyl; [0116] each of R.sup.7, and R.sup.9 is independently hydrogen, cyano, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxyl, substituted or unsubstituted cycloalkyl, C(O)R.sup.7a, or -alkyl-C(O)R.sup.7a, and wherein R.sup.7a is hydrogen, alkyl, or alkoxy, provided that at least one of R.sup.7 and R.sup.9 is not hydrogen; [0117] or R.sup.7 and R.sup.9 together form a bridge containing at least one CH.sub.2 moiety in addition to the two bridgehead atoms; [0118] L.sup.1 is a direct bond, O, N(R.sup.L), substituted or unsubstituted alkyl, -alkylene or C(O), wherein R.sup.L is hydrogen or alkyl; [0119] m is 0, 1, 2 or 3, [0120] n is 0 or 1 [0121] wherein R.sup.3a is selected from deuterium, alkoxy, alkyl, halogen, oxo, R.sup.3bSO.sub.2, cycloalkyl, cyano, R.sup.3bC(O)N(R.sup.3c), N(R.sup.3bR.sup.3c)C(O), N(R.sup.3bR.sup.3c), R.sup.3bOC(O), cycloalkyl or heterocyclyl; [0122] optionally wherein two R.sup.3a connect to the same carbon and together form a spirocyclic ring; [0123] optionally wherein two R.sup.3a form a fused ring with Cy.sup.1, wherein R.sup.3a is unsubstituted or substituted with cyano, alkoxy, alkyl, halogen, or hydroxy; and [0124] wherein R.sup.3b and R.sup.3c are each independently hydrogen or alkyl.
[0125] Unless specifically indicated otherwise, embodiment or embodiments hereinafter refer to embodiments concerning a compound of any one of formulas (I) to (II).
The Definitions of R.SUP.1
[0126] In some embodiments, R.sup.1 is hydrogen, or substituted or unsubstituted alkyl. In some embodiments, R.sup.1 is hydrogen, or substituted or unsubstituted C.sub.1-4alkyl.
[0127] In some embodiments, R.sup.1 is hydrogen, or C.sub.1-4alkyl optionally substituted with deuterium, halogen, hydroxy, alkoxy or cycloalkyl. In some embodiments, R.sup.1 is hydrogen, or C.sub.1-4alkyl optionally substituted with deuterium, halogen, hydroxy, alkoxy or cycloalkyl. In some embodiments, R.sup.1 is hydrogen, or C.sub.1-3alkyl optionally substituted with deuterium, or halogen. In some embodiments, R.sup.1 is hydrogen, or C.sub.1-3alkyl optionally substituted with deuterium.
[0128] In some embodiments, R.sup.1 is hydrogen, methyl, methyl-d3, ethyl, isopropyl, 2-hydroxyethyl, 2-methoxyethyl, 2,2,2-trifluoroethyl, 2,2-difluoroethyl, or cyclopropylmethyl. In some embodiments, R.sup.1 is hydrogen, methyl, ethyl or methyl-d3. In some embodiments, R.sup.1 is methyl or methyl-d3. In some embodiments, R.sup.1 is methyl.
The Definitions of R.SUP.2
[0129] In some embodiments, R.sup.2 is hydrogen, halogen, alkyl, alkoxyl, or cyano, provided that R.sup.2 is absent when X.sup.1 is N. In some embodiments, R.sup.2 is hydrogen, halogen, C.sub.1-4alkyl, C.sub.1-4 alkoxyl or cyano. In some embodiments, R.sup.2 is hydrogen, F, Br, Cl or CN. In some embodiments, R.sup.2 is hydrogen. In some embodiments, R.sup.2 is F, Br, or Cl.
[0130] In some embodiments, R.sup.2 is hydrogen, halogen, C.sub.1-4alkyl, C.sub.1-4 alkoxyl or cyano; preferably R.sup.2 is hydrogen, F, Br, Cl or CN; more preferably R.sup.2 is hydrogen, F, or CN; even more preferably R.sup.2 is hydrogen.
The Definitions of R.SUP.4
[0131] In some embodiments, R.sup.4 is hydrogen, halogen or alkyl, wherein the alkyl is optionally substituted with deuterium, halogen or OR.sup.4a, wherein R.sup.4a is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl or heterocycloalkyl, wherein alkyl, alkenyl, alkynyl, cycloalkyl or heterocycloalkyl is optionally substituted with C.sub.1-6alkyl, C.sub.1-6alkoxy or C.sub.3-8cycloalkyl. In some embodiments, R.sup.4 is hydrogen, halogen or C.sub.1-4alkyl, wherein the alkyl is optionally substituted with halogen or OR.sup.4a. In some embodiments, R.sup.4 is hydrogen, halogen or C.sub.1-4alkyl, wherein the alkyl is optionally substituted with deuterium or halogen. In some embodiments, R.sup.4 is hydrogen, or C.sub.1-3alkyl optionally substituted with deuterium, or halogen. In some embodiments, R.sup.4 is hydrogen, or C.sub.1-3alkyl optionally substituted with deuterium.
[0132] In some embodiments, R.sup.4 is hydrogen, fluoro, chloro, bromo, methyl, methyl-d3, trifluoromethyl, ethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl, propyl, cyclopropyl. In some embodiments, R.sup.4 is hydrogen, methyl, methyl-d3, ethyl, 2,2-difluoroethyl, propyl, or cyclopropyl.
The Definitions of R.SUP.5
[0133] In some embodiments, R.sup.5 is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, R.sup.5aC(O), R.sup.5aC(O)O, R.sup.5aOC(O), R.sup.5aC(O)NR.sup.5b, R.sup.5aNR.sup.5bC(O), R.sup.5aSO.sub.2or heterocyclyl, wherein said alkyl is unsubstituted or substituted with cyano, C(O)OR.sup.5c, C(O)R.sup.5c, C(O)NR.sup.5cR.sup.5d, heterocyclyl, alkoxy, hydroxy, cycloalkyl, or NR.sup.5cR.sup.5d; and wherein each of said cycloalkyl and heterocyclyl is unsubstituted or substituted with alkyl, cyano or halogen substituted alkyl, cyano, C(O)OR.sup.5c, C(O)R.sup.5c, C(O)NR.sup.5cR.sup.5d heterocyclyl, alkoxy, hydroxy, cycloalkyl, NR.sup.5cR.sup.5d, or R.sup.5cSO.sub.2, wherein R.sup.5a and R.sup.5b are each independently hydrogen, alkyl, or cycloalkyl; and wherein R.sup.5c and R.sup.5d are hydrogen or alkyl.
[0134] In some embodiments, R.sup.5 is hydrogen, alkyl, alkenyl or alkynyl, wherein said alkyl is unsubstituted or substituted with cyano. In some embodiments, R.sup.5 is C.sub.1-4alkyl, C.sub.2-4alkenyl or C.sub.2-4alkynyl, wherein said alkyl is substituted with cyano. In some embodiments, R.sup.5 is C.sub.1-4alkyl, wherein said alkyl is substituted with cyano.
[0135] In some embodiments, R.sup.5 is C.sub.1-4alkyl, wherein said alkyl is substituted with cyano, alkoxy, hydroxy, NR.sup.5cR.sup.5d, NOCH.sub.3, S(O)CH.sub.3, S(O).sub.2CH.sub.3, or S(O)(=NH)CH.sub.3.
[0136] In some embodiments, R.sup.5 is hydrogen, CH.sub.2CN, CH.sub.2C(O)OMe, CH(CH.sub.3)CN, oxiran-2-ylmethyl-, prop-2-yn-1-yl, but-2-yn-1-yl, prop-1-en-2-yl, methyl, isopropyl, CH.sub.2CH.sub.2OCH.sub.3, CH.sub.2OCH.sub.3, CH(CH.sub.3)OCH.sub.3, CH.sub.2C(O)NH.sub.2, CH.sub.2CH.sub.2OH, CH.sub.2OH, cyclopropyl-CH.sub.2, CH.sub.2CH.sub.2N(CH.sub.3).sub.2, CH.sub.3SO.sub.2, cyclopropyl, cyclobutyl, cyclopropyl-C(O), 1-cyanocyclopropyl, 2-cyanocyclopropyl, 2-cyanocyclobutyl, 3-(cyanomethyl)-1-(ethylsulfonyl)azetidine-3-yl, 1-cyano-2-cyclopentyleth-2-yl, CH.sub.2N(CH.sub.3).sub.2, CHNOCH.sub.3, CH.sub.2S(O)CH.sub.3, CH.sub.2S(O).sub.2CH.sub.3, or CH.sub.2S(O)(=NH)CH.sub.3; preferably, R is hydrogen, CNCH.sub.2, CH.sub.2C(O)OMe, CH(CH.sub.3)CN, oxiran-2-ylmethyl-, prop-2-yn-1-yl, but-2-yn-1-yl, prop-1-en-2-yl, CH.sub.2OH, CH(CH.sub.3)CN, CH.sub.2OCH.sub.3, CH(CH.sub.3)OCH.sub.3, CH.sub.2N(CH.sub.3).sub.2, CHNOCH.sub.3, CH.sub.2S(O)CH.sub.3, CH.sub.2S(O).sub.2CH.sub.3, or CH.sub.2S(O)(=NH)CH.sub.3; more preferably, R.sup.5 is CNCH.sub.2, prop-2-yn-1-yl, but-2-yn-1-yl, or prop-1-en-2-yl, CH.sub.2OH, CH(CH.sub.3)CN, CH.sub.2OCH.sub.3, CH(CH.sub.3)OCH.sub.3, CH.sub.2N(CH.sub.3).sub.2, CHNOCH.sub.3, CH.sub.2S(O)CH.sub.3, CH.sub.2S(O).sub.2CH.sub.3, or CH.sub.2S(O)(=NH)CH.sub.3; more preferably R.sup.5 is but-2-yn-1-yl, CNCH.sub.2, CH.sub.2OH, CH(CH.sub.3)CN, CH.sub.2OCH.sub.3, CH(CH.sub.3)OCH.sub.3, CH.sub.2N(CH.sub.3).sub.2, CHNOCH.sub.3, CH.sub.2S(O)CH.sub.3, CH.sub.2S(O).sub.2CH.sub.3, or CH.sub.2S(O)(=NH)CH.sub.3; more preferably R.sup.5 is CNCH.sub.2or but-2-yn-1-yl.
The definitions of R.sup.7/R.sup.9, R.sup.8/R.sup.10
[0137] In some embodiments, each of R.sup.7, R.sup.9, R, and R.sup.10 is independently hydrogen, alkyl, or C(O)R.sup.7a, wherein said alkyl is unsubstituted or substituted with halogen, cyano, hydroxy, or alkoxy, and wherein R.sup.7a is hydrogen, alkyl, or alkoxy, provided that at least one of R.sup.7 and R.sup.9 is not hydrogen.
[0138] In some embodiments, each of R.sup.7 and R.sup.9 is independently hydrogen, alkyl, or C(O)R.sup.7a, wherein said alkyl is unsubstituted or substituted with halogen, cyano, hydroxy, or alkoxy, and wherein R.sup.7a is hydrogen, alkyl, or alkoxy. In some embodiments, each of R.sup.7 and R.sup.9 is independently C.sub.1-4alkyl. In some embodiments, each of R.sup.7 and R.sup.9 is independently C.sub.1-2alkyl.
[0139] In some embodiments, R.sup.7 and R.sup.9 are each independently hydrogen, methyl, ethyl, methoxymethyl, 1-hydroxyethyl, 2-methoxyethyl, cyanomethyl, hydroxyethyl, hydroxymethyl, methoxycarbonyl, difluoromethyl, provided that at least one of R.sup.7 and R.sup.9 is not hydrogen.
[0140] In some embodiments, R.sup.8 and R.sup.10 are each hydrogen.
[0141] In some embodiments, R.sup.7 is methyl, and R.sup.9 is methyl; or R.sup.7 is ethyl, and R.sup.9 is ethyl; or R.sup.7 is methyl, and R.sup.9 is ethyl; or R.sup.7 is methyl, and R.sup.9 is methoxycarbonyl; or R.sup.7 is hydrogen, and R.sup.9 is methyl; or R.sup.7 is hydrogen, and R.sup.9 is ethyl.
[0142] In some embodiments, R.sup.7 and R.sup.9 are each hydrogen and R.sup.8 and R.sup.10 together form a bridge containing at least one CH.sub.2 moiety in addition to the two bridgehead atoms. In some embodiments, R.sup.7 and R.sup.9 are each hydrogen and R.sup.8 and R.sup.10 together form a bridge containing one CH.sub.2moiety in addition to the two bridgehead atoms. In some embodiments, R.sup.7 and R.sup.9 are each hydrogen and R.sup.8 and R.sup.10 together form a bridge containing two CH.sub.2moieties in addition to the two bridgehead atoms.
[0143] In some embodiments, R.sup.8 and R.sup.10 are each hydrogen and R.sup.7 and R.sup.9 together form a bridge containing at least one CH.sub.2 moiety in addition to the two bridgehead atoms. In some embodiments, R and R.sup.10 are each hydrogen and R.sup.7 and R.sup.9 together form a bridge containing one CH.sub.2moiety in addition to the two bridgehead atoms. In some embodiments, R.sup.8 and R.sup.10 are each hydrogen and R.sup.7 and R.sup.9 together form a bridge containing two CH.sub.2moieties in addition to the two bridgehead atoms.
[0144] In some embodiments, R.sup.8 and R.sup.10 are each hydrogen.
[0145] In some embodiments, R.sup.8 and R.sup.10 are each hydrogen; R.sup.7 is methyl; and R.sup.9 is methyl. In some embodiments, R.sup.8 and R.sup.10 are each hydrogen; R.sup.7 is ethyl; and R.sup.9 is ethyl. In some embodiments, R.sup.8 and R.sup.10 are each hydrogen; R.sup.7 is methyl; and R.sup.9 is ethyl. In some embodiments, R.sup.8 and R.sup.10 are each hydrogen; R.sup.7 is ethyl, and R.sup.9 is methyl. In some embodiments, R.sup.8 and R.sup.10 are each hydrogen; R.sup.7 is methoxymethyl, and R.sup.9 is methyl, ethyl or methoxymethyl.
The Definitions of L.SUP.1
[0146] In some embodiments, L.sup.1 is a direct bond, O, N(R.sup.L), -alkylene- or C(O), wherein R.sup.L is hydrogen or alkyl and wherein said -alkylene- is unsubstituted or substituted with deuterium, halogen, alkoxy, alkynyl or heterocyclyl. In some embodiments, L.sup.1 is a direct bond, O, N(R.sup.L), -alkylene- or C(O), wherein R.sup.L is hydrogen or alkyl. In some embodiments, L.sup.1 is C.sub.1-4alkylene, preferably C.sub.1-2alkylene. In some embodiments, L.sup.1 is a direct bond, CH.sub.2, CH(CH.sub.3), CH(CD.sub.3)-, CH(CH.sub.2CH.sub.3), CH(CHF.sub.2), CH(prop-1-ynyl)-, N(H), N(CH.sub.3), O, CH(C(O)NHCH.sub.2CH.sub.2OCH.sub.3)or C(CH.sub.3).sub.2. In some embodiments, L.sup.1 is CH.sub.2, CH(CH.sub.3)or CH(CD.sub.3)-. In some embodiments, L.sup.1 is CH(CH.sub.3)or CH(CD.sub.3)-.
The Definition of X.sup.2 and X.sup.3
[0147] In some embodiments, X.sup.2 and X.sup.3 are independently N or CH. In some embodiments, X.sup.2 is N, and X.sup.3 is N. In some embodiments, X.sup.2 is N, and X.sup.3 is CH. In some embodiments, X.sup.2 is CH, and X.sup.3 is N. In some embodiments, X.sup.2 is CH, and X.sup.3 is CH.
The Definitions of Cy.SUP.1
[0148] In some embodiments, Cy.sup.1 is aryl, heterocyclyl, heterocyclyl, heteroaryl, or cycloalkyl, each of which is unsubstituted or substituted with one, two, three, or four substituents R.sup.3a, wherein R.sup.3a is selected from deuterium, alkoxy, alkyl, halogen, R.sup.3bSO.sub.2, cycloalkyl, cyano, R.sup.3bC(O)N(R.sup.3c), N(R.sup.3bR.sup.3c)C(O), N(R.sup.3bR.sup.3c), R.sup.3bOC(O), or heterocyclyl, wherein said alkyl moiety in the group alkyl or alkoxy is unsubstituted or substituted with halogen, alkoxy, hydroxy, or cyano; said cycloalkyl or heterocyclyl is unsubstituted or substituted with alkoxy, alkyl, halogen, or hydroxy, wherein R.sup.3b and R.sup.3c are each independently hydrogen or alkyl.
[0149] In some embodiments, Cy.sup.1 is unsubstituted or substituted cycloalkyl, unsubstituted or substituted aryl, unsubstituted or substituted heterocyclyl, or unsubstituted or substituted heteroaryl. In some embodiments, Cy.sup.1 is optionally substituted with one, two or three substituents R.sup.3a, wherein R.sup.3a is selected from deuterium, fluoro, bromo, chloro, methyl, difluoromethyl, trifluoromethoxy, methoxy, methoxymethyl, trifluoromethyl, methylsulfonyl, difluoro, ethoxy, isopropyl, isopropoxy, difluoromethoxy, cyclopropyl, 2,2-difluorocyclopropyl, 1-fluorocyclopropyl, 2-fluorocyclopropyl, cyclobutyl, 1-hydroxyethyl, 2-hydroxyethyl, ethyl, 1,1-difluoroethyl, cyano, dimethoxy, dichloro, acetamido, 1-methoxyethyl, cyanomethyl, carbamoyl, methoxycarbonyl, dimethylcarbamoyl, (difluoromethoxy)methyl, amino, 1-(difluoromethoxy)ethyl, azetidin-1-yl, 2-methoxypropan-2-yl, 1-methoxycyclopropyl, oxetan-3-yl, 1-methylazetidin-3-yl, 1-hydroxyazetidin-3-yl, 2-cyanoethyl, 2-cyanopropan-2-yl, hydroxymethyl, 3-hydroxypropyl, 2-hydroxypropan-2-yl, or 2-fluoropropan-2-yl.
[0150] In some embodiments, Cy.sup.1 is aryl, which is unsubstituted or substituted with one, two or three substituents R.sup.3a, wherein R.sup.3a is selected from deuterium, alkoxy, alkyl, halogen, R.sup.3bSO.sub.2, cycloalkyl, cyano, R.sup.3bC(O)N(R.sup.3c), N(R.sup.3bR.sup.3c)C(O), N(R.sup.3bR.sup.3c), R.sup.3bOC(O), or heterocyclyl, wherein said alkyl moiety in the group alkyl or alkoxy is unsubstituted or substituted with halogen, alkoxy, hydroxy, or cyano; said cycloalkyl or heterocyclyl is unsubstituted or substituted with alkoxy, alkyl, halogen, or hydroxy, wherein R.sup.3b and R.sup.3c are each independently hydrogen or alkyl.
[0151] In some embodiments, Cy.sup.1 is aryl, which is unsubstituted or substituted with one, two or three substituents R.sup.3a, wherein R.sup.3a is selected from alkoxy, halogen-substituted alkoxy, alkoxyalkyl-, alkyl, alkenyl, halogen-substituted alkyl, halogen, R.sup.3bSO.sub.2, cycloalkyl, hydroxyalkyl-, cyano, R.sup.3bC(O)N(R.sup.3c), cyano-substituted alkyl, N(R.sup.3bR.sup.3c)C(O), R.sup.3bOC(O), heterocyclylalkyl- or heterocyclyl, said cycloalkyl or heterocyclyl is unsubstituted or substituted with alkoxy, alkyl, halogen, or hydroxy, wherein R.sup.3b and R.sup.3c are each independently hydrogen or alkyl.
[0152] In some embodiments, Cy.sup.1 is phenyl, which is substituted with one or two R.sup.3a, wherein R.sup.3a is fluoro, chloro, bromo; methyl, difluoromethyl, trifluoromethyl, 1,1-difluoroethyl, methoxymethyl, 1-methoxyethyl, ethyl, propyl, isopropyl, tert-butyl, 2-hydroxypropan-2-yl, 1-hydroxy-2-methylpropan-2-yl, 2-cyanopropan-2-yl, morpholinomethyl; prop-1-en-2-yl; cyclopropyl, 1-methylcyclopropyl; methoxy, difluoromethoxy, trifluoromethoxy, 1,1-difluoroethoxy, isopropoxy; oxetan-3-yl, morpholino, 2-oxa-6-azaspiro[3.3]heptan-6-yl; or methylcarbamoyl.
[0153] In some embodiments, Cy.sup.1 is an aryl of from 6 to 14 carbon atoms having a single ring or multiple condensed rings which is unsubstituted or substituted with one, two or three R.sup.3a. In some embodiments, said aromatic carbocyclic group is phenyl, naphthyl or anthryl, indanyl, indenyl, dihydroindenyl or tetrahydronaphthyl, which is unsubstituted or substituted with one, two or three R.sup.3a.
[0154] In some embodiments, Cy.sup.1 is phenyl. In some embodiments, Cy.sup.1 is phenyl, which is substituted with one R.sup.3a as disclosed herein at position 4 and optionally substituted with R.sup.3a on the other position.
[0155] In some embodiments, Cy.sup.1 is naphthalenyl. In some embodiments, Cy.sup.1 is naphthalen-1-yl, naphthalen-2-yl, naphthalen-3-yl, naphthalen-4-yl.
[0156] In some embodiments, Cy.sup.1 is a monocyclic 5- to 9-membered heterocyclyl or a bicyclic 7- to 10-membered heterocyclyl which is unsubstituted or substituted with one, two, three or four R.sup.3a (provided that the valency theory has been met), wherein R.sup.3a is selected from deuterium, alkoxy, alkyl, halogen, R.sup.3bSO.sub.2, cycloalkyl, cyano, R.sup.3bC(O)N(R.sup.3c), N(R.sup.3bR.sup.3c)C(O), N(R.sup.3bR.sup.3c), R.sup.3bOC(O), oxo or heterocyclyl, wherein said alkyl moiety in the group alkyl or alkoxy is unsubstituted or substituted with halogen, alkoxy, hydroxy, or cyano; said cycloalkyl or heterocyclyl is unsubstituted or substituted with alkoxy, alkyl, halogen, or hydroxy, wherein R.sup.3b and R.sup.3c are each independently hydrogen or alkyl; preferably, R.sup.3a is selected from alkoxy, halogen-substituted alkoxy, alkoxyalkyl-, alkyl, halogen-substituted alkyl, halogen, R.sup.3bSO.sub.2, cycloalkyl, hydroxyalkyl-, cyano, R.sup.3bC(O)N(R.sup.3c), cyano-substituted alkyl, N(R.sup.3bR.sup.3c)C(O), R.sup.3bOC(O), or heterocyclyl, said cycloalkyl or heterocyclyl is unsubstituted or substituted with alkoxy, alkyl, halogen, or hydroxy, wherein R.sup.3b and R.sup.3c are each independently hydrogen or alkyl. In some embodiments, said monocyclic 5- to 9-membered heterocyclyl is tetrahydrofuranyl, tetrahydropyranyl, 1,4-dioxanyl, piperidinyl, piperazinyl, or dihydropyridinyl, each of which is unsubstituted or substituted with one, two or three R.sup.3a as disclosed herein. In some embodiments, Cy.sup.1 is tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydro-2H-pyran-2-yl, tetrahydro-2H-pyran-3-yl, tetrahydro-2H-pyran-4-yl, 1,4-dioxan-2-yl, 1,4-dioxan-3-yl, piperidin-1-yl, piperidin-2-yl, piperidin-3-yl, piperidin-4-yl, piperazin-1-yl, piperazin-2-yl, piperazin-3-yl, piperazin-4-yl, 1,2-dihydropyridin-3-yl, 1,2-dihydropyridin-4-yl, 1,2-dihydropyridin-5-yl, or 1,2-dihydropyridin-6-yl.
[0157] In some embodiments, Cy.sup.1 is piperidinyl (e.g., piperidin-1-yl) or piperazinyl (e.g., piperazin-4-yl), which is substituted with one R.sup.3a as disclosed herein at position 4 and optionally substituted with R.sup.3a on the other position.
[0158] In some embodiments, Cy.sup.1 is a bicyclic 7- to 10-membered heterocyclyl which is unsubstituted or substituted with one, two, three or four R.sup.3a (provided that the valency theory has been met). In some embodiments, Cy.sup.1 is dihydrobenzodioxinyl, benzodioxolyl, chromanyl, or chromenyl, which is unsubstituted or substituted with one, two, three or three R.sup.3a, wherein R.sup.3a is selected from alkoxy, alkyl, halogen, and oxo. In some embodiments, Cy.sup.1 is 2,3-dihydrobenzo[b][1,4]dioxin-5-yl, 2,3-dihydrobenzo[b][1,4]dioxin-6-yl, benzo[d][1,3]dioxol-4-yl, benzo[d][1,3]dioxol-5-yl, chroman-2-yl, chroman-3-yl, chroman-4-yl, chroman-5-yl, chroman-6-yl, chroman-7-yl, chromen-2-yl, chromen-3-yl, chromen-4-yl, chromen-5-yl, chromen-6-yl, chromen-7-yl, each of which is unsubstituted or substituted with one, two, three or three R.sup.3a, wherein R.sup.3a is selected from alkoxy, alkyl, halogen, and oxo. In some embodiments, R.sup.3a is methyl, ethyl, propyl, isopropyl, fluoro, chloro, bromo or oxo. In some embodiments, two R.sup.3a connect to the same carbon and together form a spirocyclic ring, e.g.,
##STR00014##
[0159] In some embodiments, Cy.sup.1 is a monocyclic 5- to 9-membered heteroaryl or a bicyclic 7- to 10-membered heteroaryl which is unsubstituted or substituted with one, two or three R.sup.3a, wherein R.sup.3a is selected from deuterium, alkoxy, alkyl, halogen, R.sup.3bSO.sub.2, cycloalkyl, cyano, R.sup.3bC(O)N(R.sup.3c), N(R.sup.3bR.sup.3c)C(O), N(R.sup.3bR.sup.3c), R.sup.3bOC(O), oxo, phenyl or heterocyclyl, wherein said alkyl moiety in the group alkyl or alkoxy is unsubstituted or substituted with halogen, alkoxy, hydroxy, or cyano; said cycloalkyl or heterocyclyl is unsubstituted or substituted with alkoxy, alkyl, halogen, or hydroxy, wherein R.sup.3b and R.sup.3c are each independently hydrogen or alkyl; preferably, R.sup.3a is selected from deuterium, alkoxy, halogen-substituted alkoxy, alkoxyalkyl-, alkyl, halogen-substituted alkyl, halogen, oxo, hydroxyalkyl-, cyano, cyano-substituted alkyl, N(R.sup.3bR.sup.3c)C(O), N(R.sup.3bR.sup.3c), R.sup.3bOC(O), oxo, phenyl or heterocyclyl, said cycloalkyl or heterocyclyl is unsubstituted or substituted with alkoxy, alkyl, halogen, or hydroxy, wherein R.sup.3b and R.sup.3c are each independently hydrogen or alkyl; more preferably R.sup.3a is selected from deuterium, fluoro, bromo, chloro, methyl, difluoromethyl, trifluoromethyl, trifluoromethoxy, methoxy, methoxymethyl, methylsulfonyl, ethoxy, isopropoxy, isopropyl, tert-butyl, cyclopropyl, 1-hydroxyethyl, ethyl, 1,1-difluoroethyl, cyano, cyclopropyl-, acetamido, 1-methoxyethyl, cyanomethyl, carbamoyl, methoxycarbonyl, methylcarbamoyl, dimethylcarbamoyl, (difluoromethoxy)methyl, amino, isopropylamino, (difluoromethoxy)ethyl, azetidin-1-yl, 2-methoxypropan-2-yl, 1-methoxycyclopropyl, oxetan-3-yl, 1-methylazetidin-3-yl, 1-hydroxyazetidin-3-yl, phenyl or oxo.
[0160] In some embodiments, said monocyclic 5- to 9-membered heteroaryl is pyrazolyl, imidazolyl, thiazolyl, pyridinyl, pyrimidinyl, pyrazinyl or pyridazinyl, each of which is unsubstituted or substituted with one, two or three R.sup.3a as disclosed herein. In some embodiments, said monocyclic 5- to 9-membered heteroaryl is 1H-pyrazol-1-yl, 1H-pyrazol-3-yl, 1H-pyrazol-4-yl, thiazol-4-yl, thiazol-5-yl, 1H-pyrazol-5-yl, 1H-imidazol-2-yl, 1H-imidazol-4-yl, 1H-imidazol-5-yl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, 1H-indol-2-yl, 1H-indol-3-yl, 1H-indol-4-yl, 1H-indol-5-yl, 1H-indol-6-yl, 1H-indol-7-yl, pyrazin-2-yl, or pyridazin-4-yl, each of which is unsubstituted or substituted with one, two or three R.sup.3a as disclosed herein. In some embodiments, R.sup.3a is selected from deuterium, fluoro, bromo, chloro, methyl, difluoromethyl, trifluoromethyl, trifluoromethoxy, methoxy, ethoxy, isopropoxy, isopropyl, tert-butyl, carbamoyl, methylcarbamoyl, dimethylcarbamoyl, isopropylamino, azetidin-1-yl, phenyl or oxo.
[0161] In some embodiments, said bicyclic 7- to 10-membered heteroaryl is indolyl, triazolopyridinyl, imidazopyridinyl, benzooxazolyl, benzo[d]thiazolyl, quinolinyl, isoquinolinyl, naphthyridinyl, dioxinopyridineyl, quinoxalinyl, benzo[d]imidazolyl, benzofuranyl, benzoxazinyl, imidazo[4,5-b]pyridinyl, thiazolo[5,4-b]pyridinyl, thiazolo[4,5-b]pyridinyl, thieno[2,3-b]pyridinyl, dihydrodioxinopyridinyl, dioxinopyridinyl, dihydro-2H-pyranopyridinyl, 2H-pyranopyridinyl, dihydrofuropyridinyl, furopyridinyl, benzo[d]thiazolyl, isoindolinyl or thieno[3,2-b]pyridinyl, each of which is unsubstituted or substituted with one, two or three R.sup.3a as disclosed herein. In some embodiments, said bicyclic 7- to 10-membered heteroaryl is quinoxalin-6-yl, quinoxalin-2-yl, isoquinolin-3-yl, isoquinolin-7-yl, quinolin-6-yl, 1H-benzo[d]imidazol-5-yl, benzofuran-6-yl, benzofuran-5-yl, benzo[e][1,3]oxazin-6-yl, 2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl, 2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-7-yl, 3,4-dihydro-2H-pyrano[3,2-b]pyridin-6-yl, 3,4-dihydro-2H-pyrano[3,2-b]pyridin-8-yl, 2,3-dihydrofuro[2,3-b]pyridin-6-yl, 2,3-dihydrofuro[3,2-b]pyridin-5-yl, benzo[d]thiazol-5-yl, benzo[d]thiazol-6-yl, isoindolin-5-yl, 1H-benzo[d]imidazol-2-yl, 1H-benzo[d]imidazol-4-yl, 1H-benzo[d]imidazol-6-yl, 1H-benzo[d]imidazol-7-yl, [1,2,4]triazolo[1,5-a]pyridin-2-yl, [1,2,4]triazolo[1,5-a]pyridin-5-yl, [1,2,4]triazolo[1,5-a]pyridin-6-yl, [1,2,4]triazolo[1,5-a]pyridin-7-yl, [1,2,4]triazolo[1,5-a]pyridin-8-yl, 3H-imidazo[4,5-b]pyridine-2-yl, 3H-imidazo[4,5-b]pyridine-5-yl, 3H-imidazo[4,5-b]pyridine-6-yl, 3H-imidazo[4,5-b]pyridine-7-yl, 1H-imidazo[4,5-b]pyridin-2-yl, 1H-imidazo[4,5-b]pyridin-5-yl, 1H-imidazo[4,5-b]pyridin-6-yl, 1H-imidazo[4,5-b]pyridin-7-yl, benzo[d]oxazol-2-yl, benzo[d]oxazol-4-yl, benzo[d]oxazol-5-yl, benzo[d]oxazol-6-yl, benzo[d]oxazol-7-yl, benzo[d]thiazol-2-yl, benzo[d]thiazol-4-yl, benzo[d]thiazol-7-yl, quinolin-2-yl, quinolin-3-yl, quinolin-4-yl, quinolin-5-yl, quinolin-7-yl, isoquinolin-1-yl, isoquinolin-4-yl, isoquinolin-5-yl, isoquinolin-6-yl, isoquinolin-8-yl, quinoxalin-3-yl, quinoxalin-4-yl, quinoxalin-5-yl, quinoxalin-7-yl, quinoxalin-8-yl, 1,8-naphthyridin-2-yl, 1,8-naphthyridin-3-yl, 1,8-naphthyridin-4-yl, 2,3-dihydro-[1,4]dioxino[2,3-b]pyridine-6-yl, 2,3-dihydro-[1,4]dioxino[2,3-b]pyridine-7-yl, 2,3-dihydro-[1,4]dioxino[2,3-b]pyridine-8-yl, quinoxalin-6-yl-2,3-d2, 1H-indol-2-yl, 1-methyl-1H-benzo[d]imidazol-6-yl, 3H-imidazo[4,5-b]pyridin-2-yl, 4,5,6,7-tetrahydro-1H-benzo[d]imidazol-2-yl, thiazolo[5,4-b]pyridin-2-yl, thiazolo[5,4-b]pyridin-5-yl, thiazolo[5,4-b]pyridin-6-yl, thiazolo[5,4-b]pyridin-7-yl, thiazolo[4,5-b]pyridin-2-yl, thiazolo[4,5-b]pyridin-5-yl, thiazolo[4,5-b]pyridin-6-yl, thiazolo[4,5-b]pyridin-7-yl, thieno[2,3-b]pyridin-2-yl, thieno[2,3-b]pyridin-3-yl, thieno[2,3-b]pyridin-4-yl, thieno[2,3-b]pyridin-5-yl, thieno[2,3-b]pyridin-6-yl, thieno[3,2-b]pyridin-2-yl, thieno[3,2-b]pyridin-3-yl, thieno[3,2-b]pyridin-5-yl, thieno[3,2-b]pyridin-6-yl, thieno[3,2-b]pyridin-7-yl, each of which is unsubstituted or substituted with one, two or three R.sup.3a as disclosed herein.
[0162] In some embodiments, Cy.sup.1 is quinoxalinyl, e.g., quinoxalin-2-yl, quinoxalin-3-yl, quinoxalin-4-yl, quinoxalin-5-yl, quinoxalin-6-yl, quinoxalin-7-yl, quinoxalin-8-yl, preferably quinoxalin-6-yl, which is unsubstituted or substituted with one, two or three R.sup.3a, wherein R.sup.3a is selected from deuterium, alkoxy, alkyl, halogen, oxo, R.sup.3bSO.sub.2, cycloalkyl, cyano, R.sup.3bC(O)N(R.sup.3c), N(R.sup.3bR.sup.3c)C(O), N(R.sup.3bR.sup.3c), R.sup.3bOC(O), or heterocyclyl, wherein said alkyl moiety in the group alkyl or alkoxy is unsubstituted or substituted with halogen, alkoxy, hydroxy, or cyano; said cycloalkyl or heterocyclyl is unsubstituted or substituted with alkoxy, alkyl, halogen, or hydroxy, wherein R.sup.3b and R.sup.3c are each independently hydrogen or alkyl; preferably, R.sup.3a is selected from deuterium, alkoxy, halogen-substituted alkoxy, alkoxyalkyl-, alkyl, halogen-substituted alkyl, halogen, R.sup.3bSO.sub.2, cycloalkyl, hydroxyalkyl-, cyano, R.sup.3bC(O)N(R.sup.3c) cyano-substituted alkyl, N(R.sup.3bR.sup.3c)C(O), N(R.sup.3bR.sup.3c), R.sup.3bOC(O), or heterocyclyl, said cycloalkyl or heterocyclyl is unsubstituted or substituted with alkoxy, alkyl, halogen, or hydroxy, wherein R.sup.3b and R.sup.3c are each independently hydrogen or alkyl; more preferably R.sup.3a is selected from deuterium, fluoro, bromo, chloro, methyl, difluoromethyl, trifluoromethoxy, methoxy, methoxymethyl, trifluoromethyl, methylsulfonyl, difluoro, ethoxy, isopropoxy, trifluoromethoxy, difluoromethoxy, cyclopropyl, 1-hydroxyethyl, ethyl, 1,1-difluoroethyl, cyano, dimethoxy, dichloro, cyclopropyl-, acetamido, 1-methoxyethyl, cyanomethyl, carbamoyl, methoxycarbonyl, dimethylcarbamoyl, (difluoromethoxy)methyl, amino, 1-(difluoromethoxy)ethyl, azetidin-1-yl, 2-methoxypropan-2-yl, 1-methoxycyclopropyl, oxetan-3-yl, 1-methylazetidin-3-yl, or 1-hydroxyazetidin-3-yl.
[0163] In some embodiments, Cy.sup.1 is quinoxalin-2-yl or quinoxalin-6-yl, which is unsubstituted or substituted with one, two or three R.sup.3a, wherein R.sup.3a is deuterium, methyl, ethyl, isopropyl, cyanomethyl, 2-cyanoethyl, 2-cyanopropan-2-yl, hydroxymethyl, 2-hydroxyethyl, 1-hydroxyethyl, 3-hydroxypropyl, 2-hydroxypropan-2-yl, difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, 2-fluoropropan-2-yl, methoxy, isopropoxy, difluoromethoxy, fluoro, chloro, oxo, cyano, amino, cyclopropyl, 2,2-difluorocyclopropyl, 1-fluorocyclopropyl, 2-fluorocyclopropyl, or cyclobutyl; preferably deuterium, methyl, ethyl, trifluoromethyl, methoxy, isopropoxy, difluoromethoxy, fluoro, chloro, cyano, amino, or cyclopropyl. In some embodiments, Cy.sup.1 is quinoxalin-6-yl, 3-methylquinoxalin-6-yl, 3-(trifluoromethyl)quinoxalin-6-yl, or quinoxalin-2-yl.
[0164] In some embodiments, Cy.sup.1 is benzo[d]thiazol-5-yl, benzo[d]thiazol-6-yl, isoindolin-4-yl, indolin-5-yl, 1H-benzo[d]imidazol-5-yl, benzofuran-6-yl, benzofuran-5-yl, or benzo[e][1,3]oxazin-6-yl, isoquinolin-3-yl, isoquinolin-7-yl, quinolin-6-yl, 2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl, 2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-7-yl, 3-dihydro-[1,4]dioxino[2,3-c]pyridin-7-yl, 3,4-dihydro-2H-pyrano[3,2-b]pyridin-6-yl, 3,4-dihydro-2H-pyrano[3,2-b]pyridin-8-yl, 2,3-dihydrofuro[2,3-b]pyridin-6-yl, 2,3-dihydrofuro[3,2-b]pyridin-5-yl, each of which is unsubstituted or substituted with one, two or three R.sup.3a, wherein R.sup.3a is deuterium, methyl, ethyl, isopropyl, cyanomethyl, 2-cyanoethyl, 2-cyanopropan-2-yl, hydroxymethyl, 2-hydroxyethyl, 1-hydroxyethyl, 3-hydroxypropyl, 2-hydroxypropan-2-yl, difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, 2-fluoropropan-2-yl, methoxy, isopropoxy, difluoromethoxy, fluoro, chloro, bromo, oxo, cyano, amino, cyclopropyl, 2,2-difluorocyclopropyl, 1-fluorocyclopropyl, 2-fluorocyclopropyl, or cyclobutyl; preferably deuterium, methyl, ethyl, trifluoromethyl, methoxy, isopropoxy, difluoromethoxy, fluoro, chloro, cyano, amino, or cyclopropyl.
[0165] In some embodiments, Cy.sup.1 is [0166] 4-fluoro-2-methoxyphenyl, p-tolyl, 4-fluoro-2-(trifluoromethyl)phenyl, 2-(difluoromethoxy)-4-fluorophenyl, 4-fluoro-2-(trifluoromethoxy)phenyl, 4-fluoro-2-(methoxymethyl)phenyl, 4-fluoro-2-(1-methoxyethyl)phenyl, 1-(difluoromethoxy-4-fluorophenyl, 2-(difluoromethyl)-4-fluorophenyl, 2-(1,1-difluoroethoxy)-4-fluorophenyl, 3-(trifluoromethyl)phenyl, 3-isopropoxyphenyl, 3-(trifluoromethoxy)phenyl, 3-isopropylphenyl, 3-(tert-butyl)phenyl, 3-(2-hydroxypropan-2-yl)phenyl, 3-(difluoromethyl)-4-fluorophenyl, 4-fluoro-3-(trifluoromethyl)phenyl, 2-fluoro-5-(trifluoromethyl)phenyl, 4-methyl-3-(trifluoromethyl)phenyl, 4-methyl-2-(trifluoromethyl)phenyl, 3-cyclopropylphenyl, 3-(1-methylcyclopropyl)phenyl, 3-(oxetan-3-yl)phenyl, 4-fluoro-3-(methylcarbamoyl)phenyl, 3-(1-hydroxy-2-methylpropan-2-yl)phenyl, 3-(2-cyanopropan-2-yl)phenyl, 4-fluoro-3-(prop-1-en-2-yl)phenyl, 4-fluoro-3-(1-methylcyclopropyl)phenyl, 3-methyl-4-(trifluoromethyl)phenyl, 4-(morpholinomethyl)phenyl, 3-(2-oxa-6-azaspiro[3.3]heptan-6-yl)phenyl, 3-morpholinophenyl, 3-(morpholinomethyl)phenyl, 4-morpholinophenyl, 4-(trifluoromethyl)phenyl, 4-fluorophenyl, 3-methyl-2-(trifluoromethyl)phenyl, 3-(difluoromethyl)-5-fluorophenyl, 3-(1,1-difluoroethyl)-4-methoxyphenyl, m-tolyl, 3-methoxyphenyl, or 3-(difluoromethyl)phenyl; or [0167] 3,3-dimethyl-2,3-dihydro-1H-inden-5-yl; or [0168] 4,4-difluorochroman-6-yl, 2,2-dimethyl-4-oxochroman-7-yl, 3-methyl-4-oxo-4H-chromen-6-yl, 2,2-dimethylchroman-7-yl, or 2-methyl-4-oxo-4H-chromen-7-yl; or [0169] 2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 8-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 7-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 5-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 8-chloro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 2,2,3,3-tetrafluoro-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 3,3-difluoro-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 7-fluoro-2,2-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, or 7-chloro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl; or [0170] 2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 2,2-difluorobenzo[d][1,3]dioxol-5-yl, 2-ethyl-2-methylbenzo[d][1,3]dioxol-5-yl, spiro[benzo[d][1,3]dioxole-2,1-cyclobutan]-5-yl, 6-chloro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 7-chloro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 4-chloro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 7-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 4-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 4,6-difluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 2,2,7-trimethylbenzo[d][1,3]dioxol-5-yl, 7-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-4-yl, 2,2-dimethylbenzo[d][1,3]dioxol-4-yl, 2,2-difluorobenzo[d][1,3]dioxol-4-yl, 4-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, or 2,2-dimethylbenzo[d][1,3]dioxol-5-yl; or [0171] 2-(trifluoromethyl)pyridin-3-yl, 6-isopropoxypyridin-2-yl, 6-(trifluoromethyl)pyridin-2-yl, 2-(trifluoromethyl)pyridin-4-yl, 4-(trifluoromethyl)pyridin-2-yl, 2-(azetidin-1-yl)pyridin-4-yl, 2-(isopropylamino)pyridin-4-yl, 6-isopropylpyridin-2-yl, 2-isopropoxypyridin-4-yl, 6-(trifluoromethoxy)pyridin-2-yl, 2-(tert-butyl)pyridin-4-yl, 2-(methylcarbamoyl)pyridin-4-yl, 2-(2,2-dimethylcarbamoyl)pyridin-4-yl, 5,6-dimethylpyridin-2-yl, 2-phenylpyridin-4-yl, 5-(difluoromethyl)pyridin-2-yl, 5-methylpyridin-2-yl, 5-(trifluoromethyl)pyridin-2-yl, 2-methoxypyridin-4-yl, 1-methyl-6-oxo-1,6-dihydropyridin-3-yl, 1-methyl-2-oxo-1,2-dihydropyridin-4-yl, 2-(trifluoromethyl)thiazol-4-yl, or 1-methyl-5-(trifluoromethyl)-1H-pyrazol-4-yl; or [0172] quinoxalin-6-yl, 3-methylquinoxalin-6-yl, 3-(trifluoromethyl)quinoxalin-6-yl, or quinoxalin-2-yl; or [0173] isoquinolin-3-yl, 2-methyl-1-oxo-1,2-dihydroisoquinolin-7-yl, 2-methyl-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl, 1,2-dimethyl-4-oxo-1,4-dihydroquinolin-7-yl, 1,3-dimethyl-4-oxo-1,4-dihydroquinolin-6-yl, or 2-methylquinolin-7-yl; or [0174] 2-methyl-1H-benzo[d]imidazol-5-yl, 2,2-dimethyl-2,3-dihydrobenzofuran-6-yl, 2,2-dimethyl-3-oxo-2,3-dihydrobenzofuran-6-yl, 3,3-dimethyl-2,3-dihydrobenzofuran-5-yl, or 3-methyl-4-oxo-3,4-dihydro-2H-benzo[e][1,3]oxazin-6-yl; [0175] 2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl, 3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl, 4,4-difluoro-3,4-dihydro-2H-pyrano[3,2-b]pyridin-6-yl, 4,4-difluoro-3,4-dihydro-2H-pyrano[3,2-b]pyridin-8-yl, [0176] 7-chloro-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl, 7-fluoro-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl, 8-fluoro-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl, 2,2-dimethyl-2,3-dihydrofuro[2,3-b]pyridin-6-yl, 3,3-dimethyl-2,3-dihydrofuro[3,2-b]pyridin-5-yl, 3,3-dimethyl-2,3-dihydrofuro[3,2-b]pyridine-5-yl, 2,2-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-c]pyridin-7-yl, 2,2-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-7-yl, 3-methyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl, or 3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl; or [0177] 2-methylbenzo[d]thiazol-5-yl, 2-methylbenzo[d]thiazol-6-yl, 2-ethylbenzo[d]thiazol-5-yl, 2-isopropylbenzo[d]thiazol-5-yl, or 2-(trifluoromethyl)thiazol-5-yl; or [0178] 2-methyl-1,3-dioxoisoindolin-5-yl, 2-isopropyl-1,3-dioxoisoindolin-5-yl, 1-oxo-2-(2,2,2-trifluoroethyl)isoindolin-5-yl, 2,3,3-trimethyl-1-oxoisoindolin-5-yl, 2-methyl-3-oxoisoindolin-5-yl, or 2-methyl-1-oxoisoindolin-5-yl.
[0179] In some embodiments, the moiety m is
##STR00015##
wherein m is 0, 1, 2 or 3 and R.sup.3a is alkoxy, alkyl, halogen, and oxo.
[0180] In some embodiments, the moiety
##STR00016##
is 2,2-dimethylbenzo[d][1,3]dioxol-5-yl or 3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, wherein m is 0, 1, 2 or 3 and R.sup.3a is alkoxy, alkyl, halogen, and oxo. In some embodiments, the moiety
##STR00017##
is 2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 2,2-difluorobenzo[d][1,3]dioxol-5-yl, 2-ethyl-2-methylbenzo[d][1,3]dioxol-5-yl, spiro[benzo[d][1,3]dioxole-2,1-cyclobutan]-5-yl, 6-chloro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 7-chloro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 4-chloro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 7-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 4-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 4,6-difluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 2,2,7-trimethylbenzo[d][1,3]dioxol-5-yl, 2,2-difluorobenzo[d][1,3]dioxol-4-yl, 4-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl, 2,2-dimethylbenzo[d][1,3]dioxol-5-yl; 2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 8-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 7-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 5-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 8-chloro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 2,2,3,3-tetrafluoro-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 3,3-difluoro-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, 7-fluoro-2,2-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl, or 7-chloro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl.
General Synthetic Schemes
[0181] Compounds disclosed herein, including salts thereof, can be prepared using known organic synthesis techniques and can be synthesized according to any of numerous possible synthetic routes.
[0182] The reaction for preparing compounds disclosed herein can be carried out in suitable solvents which can be readily selected by one of skill in the art of organic synthesis. Suitable solvents can be substantially non-reactive with the starting materials, the intermediates, or products at the temperatures at which the reactions are carried out, e.g., temperatures which can range from room temperature to the solvent's boiling temperature. A given reaction can be carried out in one solvent or mixture of solvents.
[0183] The selection of appropriate protecting group, can be readily determined by one skilled in the art.
[0184] Reactions can be monitored according to any suitable method known in the art, such as NMR, UV, HPLC, LC-MS and TLC. Compounds can be purified by a variety of methods, including HPLC and normal phase silica flash column chromatography.
[0185] Chiral analytic HPLC was used for the retention time analysis of different chiral examples, the conditions were divided into the methods as below according to the column, mobile phase, solvent ration used. Preparation of homochiral examples may be carried out by techniques known to one skilled in the art. The absolute stereochemistry was not assigned at the newly formed carbon-nitrogen bond.
TABLE-US-00001 Abbreviations: Ac Acetyl AcOH Acetic acid AcONa Sodium acetate AIBN (E)-2,2-(diazene-1,2-diyl)bis(2-methylpropanenitrile) Aq Aqueous BINAP 2,2-bis(diphenylphosphaneyl)-1,1-binaphthalene Brine Saturated aqueous sodium chloride solution Bn Benzyl Boc Tert-butyloxycarbonyl (Boc).sub.2O Di-tert-butyl dicarbonate BOP Benzotriazol-1-yloxytris-(dimethylamino)phosphonium hexafluorophosphate DBU 1,8-Diazabicyclo[5.4.0]undec-7-ene DMF N,N-Dimethylformamide DBU 1,8-diazabicyclo[5.4.0]undec-7-ene DCE 1,2-dichloroethane DCM Dichloromethane DIBAL-H N,N-diisobutylaluminum hydride DIPEA N,N-diisopropylethylamine DMAc N,N-dimethylacetamide DMAP 4-N,N-dimethylaminopyridine DMF N,N-dimethylformamide DMSO Dimethyl sulfoxide EtOAc Ethyl acetate Eq Equivalent EtI Iodoethane EtOH Ethanol Et.sub.2Zn Diethylzinc Et.sub.3N Triethylamine g Grams HATU O-(7-Azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate HCl Hydrochloric acid HPLC High-performance liquid flash column chromatography IPA Isopropyl alcohol LiHMDS Lithium bis(trimethylsilyl)amide LDA Lithium diisopropylamide mg Milligrams mL Milliliters mmol Millimole MeCN Acetonitrile MeI Iodomethane MeOH Methanol Min Minutes MeMgBr Methylmagnesium bromide MS Mass spectrum MsCl Methanesulfonyl chloride n-Bu.sub.3SnH Tributylstannane n-BuLi Butyllithium n-BuOH Butan-1-ol n-Bu.sub.4NF Tetrabutylammonium fluoride NMR Nuclear magnetic resonance NCS N-Chlorosuccinimide NBS N-Bromosuccinimide Pd/C Palladium on carbon Pd(dppf)Cl.sub.2 [1,1-Bis(diphenylphosphino)ferrocene]dichloropalladium Pd(OAc).sub.2 Palladium diacetate Pd(PPh.sub.3).sub.2Cl.sub.2 Dichlorobis(triphenylphosphine)palladium Pd(PPh.sub.3).sub.4 Tetrakis(triphenylphosphine)palladium Pd.sub.2(dba).sub.3 Tris(dibenzylideneacetone)dipalladium PE Petroleum ether PPh.sub.3 Triphenylphosphane RT room temperature Selectfluor 1-(chloromethyl)-4-fluoro-1,4-diazabicyclo[2.2.2]octane-1,4-diium tetrafluoroborate TBAF Tetrabutylammonium fluoride TBSCl tert-Butyldimethylsilyl chloride tBuXPhos Bis(1,1-dimethylethyl)[2,4,6-tris(1-methylethyl)[1,1-biphenyl]-2- yl]phosphine tBuXPhos Pd G3 Methanesulfonato(2-di-t-butylphosphino-2,4,6-tri-i-propyl-1,1- biphenyl)(2-amino-1,1-biphenyl-2-yl)palladium(II) t-BuOK Potassium 2-methylpropan-2-olate TEA Triethanolamine TFA Trifluoroacetic acid THF Tetrahydrofuran THP Tetrahydropyran3,4-dihydro-2H-pyran TLC Thin layer flash column chromatography TMSCN Trimethylsilanecarbonitrile TrtCl Triphenylmethyl Chloride XPhos 2-Dicyclohexylphosphino-2,4,6-triisopropylbiphenyl
General Synthetic Schemes
[0186] Compounds disclosed herein, including salts thereof, can be prepared using known organic synthesis techniques and can be synthesized according to any of numerous possible synthetic routes.
[0187] The reaction for preparing compounds disclosed herein can be carried out in suitable solvents which can be readily selected by one of skill in the art of organic synthesis. Suitable solvents can be substantially non-reactive with the starting materials, the intermediates, or products at the temperatures at which the reactions are carried out, e.g., temperatures which can range from room temperature to the solvent's boiling temperature. A given reaction can be carried out in one solvent or mixture of solvents.
[0188] The selection of appropriate protecting group, can be readily determined by one skilled in the art.
[0189] Reactions can be monitored according to any suitable method known in the art, such as NMR, UV, HPLC, LC-MS and TLC. Compounds can be purified by a variety of methods, including HPLC and normal phase silica flash column chromatography.
[0190] Chiral analytic HPLC was used for the retention time analysis of different chiral examples, the conditions were divided into the methods as below according to the column, mobile phase, solvent ration used. Preparation of homochiral examples may be carried out by techniques known to one skilled in the art. The absolute stereochemistry was not assigned at the newly formed carbon-nitrogen bond.
[0191] The compounds disclosed herein can be prepared by following Scheme I to V.
##STR00018## [0192] wherein the substitutions from R.sup.1 to R.sup.9 and R.sup.8L (corresponding to L.sup.1-Cy.sup.1) are as defined as in Formula (I).
[0193] In scheme I, a commercially available Compound 1 (X.sub.1 is CH or N) is reacted with the appropriate chiral secondary amine by nucleophilic aromatic substitution reaction to give Compound 2. Compound 2 is reacted with appropriate R.sub.4X under basic condition (such as K.sub.2CO.sub.3, Cs.sub.2CO.sub.3) to give Compound 3. Compound 3 can be used to produce Compound 4 by normally Pd-catalyzed CO coupling reaction with appropriate Pd catalyst and ligand under a basic condition (such as KOH). Compound 4 is reacted with appropriate R.sub.1X under a basic condition (such as K.sub.2CO.sub.3, Cs.sub.2CO.sub.3) to give Compound 5. Compound 5 is reacted with formylation reagent (such as DMF) to give Compound 6 under a basic condition (such as n-BuLi or LDA). Reduction of Compound 6 to give Compound 7 as a alcohol using reducing agent (such as NaBH.sub.4). Compound 7 wherein contains a hydroxyl group may be chlorinated by treatment with a chlorination agent such as (SOCl.sub.2) to give Compound 8. Compound 8 as a benzyl halide may be converted into corresponding Compound 9 by treatment with a cyanation agent (such as TMSCN) under a basic condition (such as Cs.sub.2CO.sub.3 or n-Bu.sub.4NF). Compound 9 is deprotected using an acid condition (such as TFA or 4M solution of HCl in 1,4-dioxane) to give Compound 10. Tertiary amines Compound 11 is prepared by N-alkylation of secondary amines compound 10 by treatment with reductive alkylation with aldehydes or ketones, the most frequently used procedures via a phosphonium salt mediated alkylation of amines with corresponding alcohols (Florencio Zaragoza and Henrik Stephensen, J. Org. Chem. 2001, 66, 2518-2521).
[0194] Compound 7 disclosed herein can also be prepared by the following Scheme II.
##STR00019##
[0195] In Scheme II, Compound 3 may be converted into corresponding Compound 1a by treatment with a strong base (such as n-BuLi) and methyl chloroformate. Compound 1a wherein contains a chlorine atom may be converted into a corresponding compound containing a hydroxyl group by normally Pd-catalyzed CO coupling reaction with appropriate Pd catalyst and ligand under a basic condition (such as KOH), meanwhile, the ester group of Compound 1a is hydrolyzed to carboxylic acid of Compound 2a. Compound 2a is reacted with appropriate R.sub.1X under a basic condition (such as K.sub.2CO.sub.3, Cs.sub.2CO.sub.3) to give Compound 3a. Reduction of Compound 3a to give Compound 7 as a alcohol using reducing agent (such as NaBH.sub.4).
[0196] Compound 7 disclosed herein can also be prepared by the following Scheme III.
##STR00020##
[0197] In Scheme III, the Reduction of Compound 1a to give Compound 1b as a alcohol using a reducing agent (such as NaBH.sub.4). Compound 1b may be converted into corresponding Compound 2b by treatment with an organic base (such as imidazole) and TBSCl. Compound 2b wherein contains a chlorine atom may be converted into corresponding Compound 3b containing a hydroxyl group by normally Pd-catalyzed CO coupling reaction with appropriate Pd catalyst and ligand under a basic condition (such as KOH). Compound 3b is reacted with appropriate R.sub.1X under basic condition (such as K.sub.2CO.sub.3, Cs.sub.2CO.sub.3) to give Compound 4b. Compound 4b is deprotected using a reagent containing fluorine (such as tetrabutylammonium fluoride) to give Compound 7.
[0198] The compound of Formula 12c disclosed herein can be prepared by following Scheme IV.
##STR00021##
[0199] The halogenation of Compound 6c with electrophilic halogenating reagents (such as NBS, NCS, or Selectfluor) to afford a Compound 7c. Reduction of Compound 7c to give Compound 8c as a alcohol using a reductive agent (such as NaBH.sub.4). Compound 8c wherein contains a hydroxyl group may be chlorinated by treatment with a chlorination agent such as (SOCl.sub.2 or MsCl) to give Compound 9c. Compound 9c as a benzyl halide may be converted into corresponding Compound 10c by treatment with a cyanation agent (such as TMSCN) under a basic condition (such as Cs.sub.2CO.sub.3 or n-Bu.sub.4NF). Compound 10c is deprotected using an acid condition (such as TFA or 4M solution of HCl in 1,4-dioxane) to give Compound 11c. Tertiary amines Compound 12c is prepared by N-alkylation of secondary amines compound 11c by treatment with reductive alkylation with aldehydes or ketones, the most frequently used procedures via a phosphonium salt mediated alkylation of amines with corresponding alcohols (Florencio Zaragoza and Henrik Stephensen, J. Org. Chem. 2001, 66, 2518-2521).
[0200] The compound of Formula 11d disclosed herein can also be prepared by following Scheme V.
##STR00022##
[0201] In scheme V, Compound 2d is prepared by N-alkylation of secondary amines Compound 1d by treatment with alkylating agents (such as alkyl halides or sulfonates) or reductive alkylation with aldehydes or ketones, the most frequently used procedures via a phosphonium salt mediated alkylation of amines with corresponding alcohols. Compound 2d is deprotected using acid conditions (such as TFA or 4M solution of HCl in 1,4-dioxane) to give Compound 3d. Compound 1d was synthesized according to the method described in the literature (Shaker Youssef and Wolfgang Pfleiderer, J. Heterocyclic Chem. 35, 949-954, 1998). Compound 1d is reacted with the appropriate acid via cyclization reaction to give Compound 2d. Compound 2d is reacted with the appropriate chiral secondary amine (corresponding to Compound 3d) via phosphonium-mediated S.sub.NAr reactions to give Compound 3d in the presence of phosphonium reagents (such as BOP) under a basic condition (such as DBU, DIPEA). Compound 3d is deprotected using an acid condition (such as TFA) to give Compound 4d. Compound 4d wherein contains a hydroxyl group may be chlorinated by treatment with a chlorination agent such as (SOCl.sub.2) to give Compound 5d. Compound 5d as a benzyl halide may be converted into corresponding Compound 11d by treatment with a cyanation agent (such as TMSCN) under a basic condition (such as Cs.sub.2CO.sub.3 or n-Bu.sub.4NF).
Examples
[0202] The examples below are intended to be purely exemplary and should not be considered to be limiting in any way. Unless otherwise specified, the experimental methods in the Examples described below are conventional methods. Unless otherwise specified, the reagents and materials are all commercially available. All solvents and chemicals employed are of analytical grade or chemical purity. Solvents are all redistilled before use. Anhydrous solvents are all prepared according to standard methods or reference methods. Silica gel (100-200 meshes) for flash column chromatography and silica gel (GF254) for thin-layer flash column chromatography (TLC) are commercially available from Tsingdao Haiyang Chemical Co., Ltd. or Yantai Chemical Co., Ltd. of China; all are eluted with petroleum ether (60-90 C.)/ethyl acetate (v/v), and visualized by iodine or the solution of molybdphosphoric acid in ethanol unless otherwise specified. All extraction solvents, unless otherwise specified, are dried over anhydrous Na.sub.2SO.sub.4. .sup.1H NMR spectra are recorded on Bruck-400 nuclear magnetic resonance spectrometer with TMS (tetramethylsilane) as the internal standard. LC/MS data are recorded by using Agilent1100 High Performance Liquid Flash column chromatography-Ion Trap Mass Spectrometer (LC-MSD Trap) equipped with a diode array detector (DAD) detected at 214 nm and 254 nm, and an ion trap (ESI source). All compound names except the reagents were generated by ChemDraw.
Synthesis
TABLE-US-00002 Preparative HPLC Conditions (Method A) Column Phenomenex Gemini NX-C18, 150 21.2 mm, 5 m Column Temp. R.T. Detection Wavelength DAD, UV = 214/254 nm Run Time 17.0 min Flow Rate 20.0 mL/min Mobile Phase A 0.1% FA-H.sub.2O (v/v) Mobile Phase B 0.1% FA-CH.sub.3CN (v/v)
TABLE-US-00003 Preparative HPLC Conditions (Method B) Column Waters XSelect CSH C18, 150 19 mm, 5 m Column Temp. R.T. Detection Wavelength DAD, UV = 214/254 nm Run Time 17.0 min Flow Rate 17 mL/min Mobile Phase A 0.03% NH.sub.3H.sub.2OH.sub.2O (v/v) Mobile Phase B CH.sub.3CN
Intermediate 1: tert-butyl (2S,5R)-2,5-diethylpiperazine-1-carboxylate
##STR00023##
Step A: methyl (R)-2-(benzylamino)butanoate
##STR00024##
[0203] To a solution of methyl (R)-2-aminobutanoate (100.0 g, 0.85 mol) in CH.sub.3CN (1000 mL) was added benzyl bromide (146.1 g, 0.85 mol) at 0 C. under N.sub.2 atmosphere. The reaction was stirred at room temperature overnight and concentrated under reduced pressure. The resulting residue was dissolved in EtOAc (1000 mL) and washed with water (1000 mL3). The organic layers were concentrated under reduced pressure. The resulting residue was purified by flash column chromatography (PE:EtOAc=10:1) to give the titled compound (106 g, 600%). MS: M/e 208 (M+1).sup.+.
Step B: methyl (R)-2-((S)N-benzyl-2-((tert-butoxycarbonyl)amino)butanamido) butanoate
##STR00025##
[0204] To a solution of methyl (R)-2-(benzylamino) butanoate (117.0 g, 0.56 mol), (S)-2-((tert-butoxycarbonyl)amino)butanoic acid (170.5 g, 0.84 mmol) and 4-Methylmorpholine (113.1 g, 1.12 mmol) in DCM (2000 mL) was added HATU (319.0 g, 0.84 mmol) at 0 C. The reaction was stirred at room temperature overnight and quenched by water and washed with water (1500 mL2). The organic layers were concentrated under reduced pressure. The resulting residue was purified by flash column chromatography (PE:EtOAc=10:1) to give the titled compound (178 g, 80%). MS: Me 393 (M+1)+.
Step C: methyl (R)-2-((S)-2-amino-N-benzylbutanamido)butanoate
##STR00026##
[0205] To a solution of methyl (R)-2-((S)N-benzyl-2-((tert-butoxycarbonyl)amino)butanamido) butanoate (178 g, 0.45 mol) in 1,4-dioxane (100 mL) was added HCl (400 mL, 4 M in 1,4-dioxane) at room temperature. The resulting mixture was stirred at room temperature for another 2 hours and concentrated under vacuum to give the titled compound (200 g, crude). MS: M/e 293 (M+1)+.
Step D: (3S,6R)-1-benzyl-3,6-diethylpiperazine-2,5-dione
##STR00027##
[0206] To a solution of methyl (R)-2-((S)-2-amino-N-benzylbutanamido)butanoate (200 g, crude) in EtOAc (1000 mL) was added aq. NaHCO.sub.3 (300 mL) at room temperature. The reaction mixture was stirred at room temperature for another 2 hours. The organic layers were concentrated under reduced pressure. The resulting residue was triturated with MTBE to give the titled compound (61 g, 52% for 2 steps, ee: 97%). MS: M/e 261 (M+1).sup.+.
Step E: (2R,5S)-1-benzyl-2,5-diethylpiperazine
##STR00028##
[0207] To a solution of LiAlH.sub.4 (26.5 g, 0.69 mol) in THE (1000 mL) was added slowly (3S,6R)-1-benzyl-3,6-diethylpiperazine-2,5-dione (61.0 g, 0.23 mol) in THE (500 mL) at 0 C. The resulting mixture was stirred at room temperature for 2 hours, then stirred at 80 C. overnight. The reaction was quenched by water (27 mL) slowly at 0 C. Then, 1N aqueous NaOH solution (54 mL) and water (81 mL) was added sequentially. The resulting mixture was stirred for 2 hours. The white precipitates that formed was removed by filtration. The filter cake was washed with EtOAc (500 mL). The combined filtrates were evaporated. The resulting residue was dissolved in toluene. The solvent was removed under vacuum to dryness to afford the titled compound (51 g, 95%). MS: M/e 233 (M+1).sup.+.
Step F: tert-butyl (2S,5R)-4-benzyl-2,5-diethylpiperazine-1-carboxylate
##STR00029##
[0208] To a solution of (2R,5S)-1-benzyl-2,5-diethylpiperazine (46.4 g, 200 mmol) and Boc.sub.2O (48 g, 220 mmol) in DCM (800 mL) was added Et.sub.3N (30.3 g, 300 mmol). The reaction mixture was stirred at room temperature for 12 hours. The reaction was quenched with saturated NaCl (400 mL) at room temperature. The resulting mixture was extracted with DCM (800 mL2). The combined organic layers were dried over Na.sub.2SO.sub.4 and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography to give the titled compound (43.2 g, 65%). MS: M/e 333 (M+1).sup.+.
Step G: tert-butyl (2S,5R)-2,5-diethylpiperazine-1-carboxylate
##STR00030##
[0209] To a solution of tert-butyl (2S,5R)-4-benzyl-2,5-diethylpiperazine-1-carboxylate (43.2 g, 130 mmol) and Pd/C (6.6 g, 10% in water) in MeOH (240 mL) at room temperature was added AcOH (5 mL). The resulting mixture was degassed 3 times under H.sub.2 atmosphere and stirred at room temperature under H.sub.2 atmosphere for 12 hours. After filtration, the combined organic layers were concentrated under reduced pressure to give the crude product. The crude product was basified by Na.sub.2CO.sub.3 (4M) to pH 10 and extracted with EA (200 mL3). The combined organic layers were washed with brine (20 mL3), dried over Na.sub.2SO.sub.4 and concentrated under reduced pressure to give the titled compound (22 g, 70%). .sup.1H NMR (400 MHz, CDCl.sub.3) 3.96-3.84 (m, 1H), 3.74 (d, J=13.2 Hz, 1H), 3.12-3.00 (m, 2H), 2.72-2.60 (m, 1H), 2.52 (dd, J=1.2 Hz, 12.8 Hz, 1H), 1.86-1.71 (m, 1H), 1.69-1.49 (m, 4H), 1.43 (s, 9H), 0.95-0.80 (m, 6H) ppm. MS: M/e 243 (M+1).sup.+.
Intermediate 2: tert-butyl (2R,5S)-2,5-diethylpiperazine-1-carboxylate
##STR00031##
Step A: methyl (S)-2-(benzylamino)butanoate
##STR00032##
[0210] To a solution of compound 1 (250 g, 1.6 mol) in DCE (3500 mL) was added TEA (164 g, 1.6 mol), PhCHO (172 g, 1.6 mol) and NaBH(OAc).sub.3 (516 g, 2.4 mol) at ice-bath. The reaction mixture was stirred at rt for 5 h. The reaction mixture was quenched with Na.sub.2CO.sub.3 (1000 mL) and extracted with DCM (1500 mL2). The combined organic layers were washed with brine (1000 mL) and concentrated. The residue was purified by chromatography on silica gel (PE:EA=20:1) to afford titled compound (315 g, 93.8%) as colorless oil. MS: M/e 208 (M+1).sup.+
Step B: methyl (S)-2-((R)N-benzyl-2-((tert-butoxycarbonyl)amino)butanamido)butanoate
##STR00033##
[0211] To a solution of methyl (S)-2-(benzylamino)butanoate (244 g, 1.2 mol) in DCM (7000 mL) was added (R)-2-((tert-butoxycarbonyl)amino)butanoic acid (285 g, 1.4 mol) and EDCI (113 g, 0.16 mol) at ice-bath. The reaction mixture was stirred for 30 min at rt. EDCI (113 g, 0.16 mol) was added. The reaction mixture was stirred for an additional hour and additional EDCI (113 g, 0.16 mol) was added. After the reaction mixture was stirred for 2 days at rt. The reaction mixture was poured into H.sub.2O (2500 mL), and extracted with DCM (1500 mL2). The combined organic layers were washed with brine (1000 mL) and concentrated. The residue was purified by chromatography on silica gel (PE:EA=10:1) to afford titled compound (152 g, 33%) as colorless oil. MS: M/e 393 (M+1).sup.+
Step C: (3R,6S)-1-benzyl-3,6-diethylpiperazine-2,5-dione
##STR00034##
[0212] The reaction mixture of methyl (S)-2-((R)N-benzyl-2-((tert-butoxycarbonyl)amino)butanamido)butanoate (143 g, 0.36 mol) in 4 N HCl/Dioxane (1500 mL) was stirred at rt for 5 h. Then the reaction mixture was basified to adjust pH 9.0 with sat. NaHCO.sub.3 and stirred for 2 h. The mixture was extracted with DCM (1500 mL2). The combined organic layers were washed with brine (2 L) and concentrated. The residue was purified by chromatography on silica gel (PE:EA=2:1) to afford titled compound (80 g, 85%) as an off-white solid. .sup.1H NMR (DMSO-d.sub.6, 400 MHz): 8.23 (s, 1H), 7.36-7.23 (m, 5H), 5.10 (d, J=15.2 Hz, 1H), 4.15-4.13 (m, 1H), 4.08 (d, J=14.8 Hz 1H), 3.65-3.63 (m, 1H), 1.93-1.82 (m, 4H), 0.84-0.79 (m, 6H) ppm. MS: M/e 261 (M+1).sup.+.
Step D: (2S,5R)-1-benzyl-2,5-diethylpiperazine
##STR00035##
[0213] To a solution of (3R,6S)-1-benzyl-3,6-diethylpiperazine-2,5-dione (70 g, 0.27 mol) in THF (1000 mL) was added a solution of LiAlH.sub.4 (51 g, 1.35 mol) at ice-bath. The reaction mixture was stirred at 70 C. for 4 h. The reaction mixture was cooled, added water (51.15 mL), 10% NaOH (51.15 mL) and water (153.45 mL). The mixture was stirred for 4 h. The mixture was filtered. The filtrate was concentrated to afford titled compound (62 g, 99.9%) which was used to the next step without further purification. MS: M/e 233 (M+1).sup.+.
Step E: tert-butyl (2R,5S)-4-benzyl-2,5-diethylpiperazine-1-carboxylate
##STR00036##
[0214] To a solution of (2S,5R)-1-benzyl-2,5-diethylpiperazine (62 g, 0.27 mol) in THF (1000 mL) was added Boc.sub.2O (64 g, 0.29 mol) and DIEA (50 g, 0.39 mol) at ice-bath. The reaction mixture was stirred at rt overnight. The reaction mixture was poured into H.sub.2O (300 mL), and extracted with DCM (1000 mL2). The combined organic layers were washed with brine (1 L) and concentrated. The residue was purified by chromatography on silica gel (PE:EA=25:1) to afford titled compound (74 g, 88.7%) as colorless oil. .sup.1H NMR (DMSO-d.sub.6, 400 MHz): 7.34-7.23 (m, 5H), 3.88-3.84 (m, 2H), 3.67 (d, J=18.4 Hz, 1H), 3.50 (d, J=18.0 Hz, 1H), 3.06-3.04 (m, 1H), 2.58-2.50 (m, 1H), 2.27 (d, J=17.2 Hz, 1H), 1.82-1.75 (m, 1H), 1.59-1.45 (m, 13H), 0.88 (t, J=9.6 Hz, 3H), 0.74 (t, J=9.6 Hz, 3H) ppm. MS: M/e 333 (M+1).sup.+.
Step F: tert-butyl (2R,5S)-2,5-diethylpiperazine-1-carboxylate
##STR00037##
[0215] To a solution of tert-butyl (2R,5S)-4-benzyl-2,5-diethylpiperazine-1-carboxylate (70 g, 0.1 mol) in MeOH (500 mL) was added Pd/C (14 g) at rt. The reaction mixture was stirred at 30 C. overnight under H.sub.2 (1 atm). The resulting mixture was filtered and concentrated to afford titled compound (50 g, 98%) as colorless oil. .sup.1H NMR (DMSO-d.sub.6, 400 MHz): 3.76-3.74 (m, 1H), 3.63 (d, J=17.6 Hz, 1H), 3.00 (dd, J=5.2, 17.6 Hz, 1H), 2.89 (dd, J=6.0, 17.2 Hz, 1H), 2.57-2.56 (m, 1H), 2.41-2.24 (m, 1H), 1.79-1.72 (m, 1H), 1.57-1.46 (m, 3H), 1.38 (s, 9H), 0.88-0.77 (m, 6H) ppm. LCMS [column: Xbridge C18; column size: 4.6*50 mm, 3.5 um; mobile phase: from 95% water (0.1% TFA) and 5% CH.sub.3CN to 5% water (0.1% TFA) and 95% CH.sub.3CN in 6.5 min, finally under these conditions for 0.5 min.] purity is >95%, Rt=3.131 min; MS: M/e 243 (M+1).sup.+.
Intermediate 3: tert-butyl (2R,5S)-2-ethyl-5-methylpiperazine-1-carboxylate
##STR00038##
Step A: methyl benzyl-L-alaninate
##STR00039##
[0216] To a suspension of methyl L-alaninate hydrogen chloride (50 g, 0.36 mol) and K.sub.2CO.sub.3 (150 g, 1.08 mol) in MeCN (500 mL) was added BnBr (80 g, 0.47 mol) in drops at room temperature and the resulted mixture was stirred at room temperature for 16 hours. The mixture was filtered. The filter cake was washed with EtOAc (200 mL3). The organics were combined and concentrated. The resulted residue was diluted with EtOAc (500 mL), washed with saturated aqueous solution of NaHCO.sub.3 (200 mL), brine (200 mL3), dried and concentrated. The resulted oil was purified by flash column chromatography to give the titled compound (28.0 g, 40%). MS: M/e 194 (M+1).sup.+.
Step B: methyl N-benzyl-N((R)-2-((tert-butoxycarbonyl)amino)butanoyl)-L-alaninate
##STR00040##
[0217] To a mixture of methyl benzyl-L-alaninate (27.0 g, 140 mmol), (R)-2-((tert-butoxycarbonyl)amino)butanoic acid (31.2 g, 154 mmol) and DIPEA (36.5 g, 280 mmol) in CH.sub.2Cl.sub.2 (300 mL) was added HATU (60.0 g, 158 mmol) in portions at 0 C. The resulted mixture was stirred at room temperature for 20 hours. The mixture was washed with saturated aqueous solution of NaHCO.sub.3 (100 mL2), brine (100 mL2), dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by flash column chromatography to give the titled compound (45 g, 85%). MS: M/e 379 (M+1).sup.+.
Step C: methyl N((R)-2-aminobutanoyl)-N-benzyl-L-alaninate
##STR00041##
[0218] To a solution of methyl N-benzyl-N((R)-2-((tert-butoxycarbonyl)amino)butanoyl)-L-alaninate (41.0 g, 108.5 mmol) in EtOAc (300 mL) was added HCl (100 mL, 4M in Dioxane) at room temperature and the mixture was stirred at room temperature for 3 days. The mixture was concentrated to dryness to give the titled compound (38.0 g, crude) which was used for the next step directly. MS: M/e 279 (M+1).sup.+.
Step D: (3R,6S)-1-benzyl-3-ethyl-6-methylpiperazine-2,5-dione
##STR00042##
[0219] To a mixture of methyl N((R)-2-aminobutanoyl)-N-benzyl-L-alaninate hydrogen chloride (38.0 g, crude) in a mixed solvent EA/H.sub.2O (200 mL/50 mL) was added NaHCO.sub.3 (55 g, 650 mmol) in portions at room temperature. The resulted mixture was stirred at room temperature for 5 hours. Layers were separated. The aqueous layer was extracted with EtOAc (100 mL3). The combined organic layers was washed with brine (200 mL3), dried over Na.sub.2SO.sub.4 and concentrated. The resulting residue was purified by flash column chromatography to give the titled compound (24 g, 88% for 2 steps). MS: M/e 247 (M+1).sup.+.
Step E: (2S,5R)-1-benzyl-5-ethyl-2-methylpiperazine
##STR00043##
[0220] A solution of borane in THF (1M, 480 mL) was slowly added to (3R,6S)-1-benzyl-3-ethyl-6-methylpiperazine-2,5-dione (23.6 g, 95.9 mmol) over 15 min. The resulted mixture was stirred at 70 C. for 72 hours. The mixture was cooled to 0 C., MeOH (100 mL) was added slowly and followed by HCl (5M, 40 mL) slowly. Large amount of bubble was found. After stirring for 30 min, the mixture was heated at 70 C. for 2 hours. The mixture was concentrated to dryness and purified by flash column chromatography to give the titled compound (21.5 g, crude) which was used for the next step directly. MS: M/e 219 (M+1).sup.+.
Step F: tert-butyl (2R,5S)-4-benzyl-2-ethyl-5-methylpiperazine-1-carboxylate
##STR00044##
[0221] To a solution of (2S,5R)-1-benzyl-5-ethyl-2-methylpiperazine (19.5 g, crude) in THF (150 mL) was added K.sub.2CO.sub.3 (24.5 g, 177 mmol) and H.sub.2O (50 mL) at room temperature and followed by di-tert-butyl decarbonate (20.0 g, 91.7 mmol) in drops. The resulted mixture was stirred at room temperature for 16 hours. The mixture was diluted with of EtOAc (200 mL), washed with brine (100 mL3), dried over Na.sub.2SO.sub.4, concentrated and purified by flash column chromatography to give the titled compound (23.8 g, 86% for 2 steps). MS: M/e 319 (M+1).sup.+.
Step G: tert-butyl (2R,5S)-2-ethyl-5-methylpiperazine-1-carboxylate
##STR00045##
[0222] A mixture of tert-butyl (2R,5S)-4-benzyl-2-ethyl-5-methylpiperazine-1-carboxylate (23.8 g, 74.8 mmol), Pd/C (3.0 g), AcOH (2 mL) in MeOH (400 mL) was stirred at room temperature under H.sub.2 for 16 hours. The mixture was filtered and the filtrate was concentrated. The resulted oil was treated with aqueous solution of NaOH (5M, 50 mL), extracted with CH.sub.2Cl.sub.2 (100 mL3). The combined organic layers was washed with brine (100 mL3), dried over Na.sub.2SO.sub.4, concentrated to dryness to give the titled compound (14.3 g, 84%). .sup.1H NMR (400 MHz, DMSO-d6) 3.42-3.23 (m, 1H), 3.04 (d, J=12.0 Hz, 1H), 2.64-2.46 (m, 3H), 2.15-2.05 (m, 1H), 1.99 (dd, J=12.8, 1.6 Hz, 1H), 1.41-1.24 (m, 1H), 1.16-1.01 (m, 1H), 0.97 (s, 9H), 0.60 (d, J=6.8 Hz, 3H), 0.36 (t, J=7.6 Hz, 3H). MS: M/e 229 (M+1).sup.+.
Intermediate 4: tert-butyl (2S,5R)-5-ethyl-2-methylpiperazine-1-carboxylate
##STR00046##
Step A: methyl (R)-2-(benzylamino)butanoate
##STR00047##
[0223] To a solution of methyl (R)-2-aminobutanoate hydrogen chloride (100 g, 651 mmol) and K.sub.2CO.sub.3 (225 g, 1.628 mol) in CH.sub.3CN (700 mL) was added (bromomethyl)benzene (122.5 g, 716 mmol) dropwise at 0 C. The reaction mixture was stirred at room temperature for 12 hours. After filtration, the reaction solution was concentrated under reduced pressure. The resulting residue was purified by flash column chromatography to give the titled compound (70 g, 52%). MS: M/e 208 (M+1).sup.+.
Step B: methyl (R)-2-((S)N-benzyl-2-((tert-butoxycarbonyl)amino)propanamido)butanoate
##STR00048##
[0224] To a solution of methyl (R)-2-(benzylamino)butanoate (35 g, 169 mmol), (tert-butoxycarbonyl)-L-alanine (48 g, 254 mmol) and HATU (116 g, 304 mmol) in DCM (400 mL) was added NMM (43 g, 422 mmol). The reaction mixture solution was stirred at room temperature for 24 hours. The reaction was quenched with saturated NaCl (100 mL) at room temperature. The resulting mixture was extracted with DCM (300 mL2). The combined organic layers were dried over Na.sub.2SO.sub.4 and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography to give the titled compound (45 g, 70%). MS: M/e 379 (M+1).sup.+.
Step C: methyl (R)-2-((S)-2-amino-N-benzylpropanamido)butanoate hydrogen chloride
##STR00049##
[0225] To a solution of methyl (R)-2-((S)N-benzyl-2-((tert-butoxycarbonyl)amino)propanamido)butanoate (45 g, 119 mmol) in DCM (250 mL) at room temperature was added HCl (119 mL, 4 M in 1,4-dioxane). The reaction mixture was stirred at room temperature for 4 hours, concentrated under reduced pressure to give the titled compound (32 g, 86%). MS: M/e 279 (M+1).sup.+.
Step D: (3S,6R)-1-benzyl-6-ethyl-3-methylpiperazine-2,5-dione
##STR00050##
[0226] To a solution of methyl (R)-2-((S)-2-amino-N-benzylpropanamido)butanoate hydrogen chloride (32 g, 102 mmol) in NaHCO.sub.3 (150 mL, 4M) was added EA (150 mL). The reaction mixture was stirred at room temperature for 2 hours. The combined organic layers were dried over Na.sub.2SO.sub.4 and concentrated under reduced pressure to give the titled compound (22 g, 87%). MS: M/e 247 (M+1).sup.+.
Step E: (2R,5S)-1-benzyl-2-ethyl-5-methylpiperazine
##STR00051##
[0227] To a solution of (3S,6R)-1-benzyl-6-ethyl-3-methylpiperazine-2,5-dione (20 g, 81.3 mmol) in THE (300 mL) at 0 C. was added LiAlH.sub.4 (6.2 g, 163 mmol) in portion. The reaction mixture was stirred at 70 C. for 36 hours and added H.sub.2O (6.2 mL), followed NaOH (6.2 mL, 20%) and H.sub.2O (12.4 mL) at 0 C. to quench the reaction solution. After filtration, the combined organic layers were dried over Na.sub.2SO.sub.4 and concentrated under reduced pressure to give the titled compound (11 g, 61%). MS: M/e 219 (M+1).sup.+.
Step F: tert-butyl (2S,5R)-4-benzyl-5-ethyl-2-methylpiperazine-1-carboxylate
##STR00052##
[0228] To a solution of (2R,5S)-1-benzyl-2-ethyl-5-methylpiperazine (11 g, 50 mmol) and Boc.sub.2O (12 g, 55 mmol) in DCM (200 mL) was added Et.sub.3N (7.6 g, 75 mmol). The reaction mixture was stirred at room temperature for 12 hours. The reaction was quenched with saturated NaCl (100 mL) at room temperature. The resulting mixture was extracted with DCM (200 mL2). The combined organic layers were dried over Na.sub.2SO.sub.4 and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography to give the titled compound (14 g, 69%). MS: M/e 319 (M+1).sup.+.
Step G: tert-butyl (2S,5R)-5-ethyl-2-methylpiperazine-1-carboxylate
##STR00053##
[0229] To a solution of tert-butyl (2S,5R)-4-benzyl-5-ethyl-2-methylpiperazine-1-carboxylate (10 g, 31.4 mmol) and Pd/C (2.5 g, 10% in water) in MeOH (60 mL) at room temperature was added AcOH (2 mL). The resulting mixture was degassed 3 times under H.sub.2 atmosphere, and stirred at room temperature under H.sub.2 atmosphere for 12 hours. After filtration, the combined organic layers were concentrated under reduced pressure to give the crude product (AcOH salt). The crude product was basified by Na.sub.2CO.sub.3 (4M) to pH 10 and extracted with EA (80 mL3). The combined organic layers were washed with brine (20 mL3), dried over Na.sub.2SO.sub.4 and concentrated under reduced pressure to give the titled compound (5.5 g, 77%). .sup.1H NMR (400 MHz, CDCl.sub.3) 4.21-4.09 (m, 1H), 3.72-3.69 (d, J=12.9 Hz, 1H), 3.24-3.19 (dd, J=13.7, 4.0 Hz, 1H), 3.14-3.10 (dd, J=12.8, 4.7 Hz, 1H), 2.86-2.75 (m, 1H), 2.53-2.49 (dd, J=12.8, 2.7 Hz, 1H), 2.33-2.12 (m, 1H), 1.65-1.54 (m, 2H), 1.46 (s, 9H), 1.25-1.24 (d, J=4.0 Hz, 3H), 0.98-0.91 (m, 3H) ppm. MS: M/e 229 (M+1).sup.+.
Intermediate 5: 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00054##
Step A: tert-butyl (2R,5S)-4-(2-chloro-9H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00055##
[0230] To a solution of 2,6-dichloro-9H-purine (200 g, 529 mmol) in EtOH (2400 mL) was added tert-butyl (2R,5S)-2,5-dimethylpiperazine-1-carboxylate (238 g, 555 mmol), DIPEA (204.6 g, 794 mmol). The reaction was stirred at 80 C. for 16 h. The mixture was concentrated in vacuo. The residue was added DCM and washed with H.sub.2O. The organic phase was dried over Na.sub.2SO.sub.4 and concentrated in vacuo to give the titled compound (387 g, 99%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.97 (s, 1H), 6.17-5.05 (m, 2H), 4.43 (d, J=20.5 Hz, 1H), 3.83 (d, J=13.6 Hz, 1H), 3.80-3.35 (m, 2H), 1.50 (d, J=3.6 Hz, 9H), 1.30 (d, J=6.8 Hz, 3H), 1.18 (d, J=6.8 Hz, 3H) ppm. MS: M/e 367 (M+1).sup.+.
Step B: tert-butyl(2R,5S)-4-(2-chloro-9-methyl-9H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00056##
[0231] To a solution of tert-butyl (2R,5S)-4-(2-chloro-9H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (387 g, 1.06 mol) in acetone (2500 mL) was added K.sub.2CO.sub.3 (295 g, 2.12 mol) and CH.sub.3I (181 g, 1.27 mol). The reaction was stirred at room temperature overnight. The mixture was filtered. The filter cake was washed with DCM. The filtrate was concentrated in vacuo. The residue was dissolved in DCM and washed by H.sub.2O. The organic phase was dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was added a solution of PE/EtOAc=2/1 and stirred for 15 mins. The mixture was filtered. The filter cake was dried in vacuo to give the titled compound (350 g, 87%). .sup.1H NMR (400 MHz, CD3OD) 7.95 (s, 1H), 6.19-4.92 (m, 2H), 4.43 (d, J=19.6 Hz, 1H), 3.82 (d, J=13.8 Hz, 1H), 3.75 (s, 3H), 3.70-3.34 (m, 2H), 1.49 (s, 9H), 1.30 (d, J=6.7 Hz, 3H), 1.17 (d, J=6.8 Hz, 3H) ppm. MS: M/e 381 (M+1).sup.+.
Step C: tert-butyl(2R,5S)-2,5-dimethyl-4-(9-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl) piperazine-1-carboxylate
##STR00057##
[0232] To a solution of tert-butyl (2R,5S)-4-(2-chloro-9-methyl-9H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (100 g, 263 mmol) in dioxane/H.sub.2O (1400 mL/170 mL) was added 2-Di-tert-butylphosphino-2,4,6-triisopropylbiphenyl (11.2 g, 26.3 mmol) KOH (44 g, 789 mmol) and Pd.sub.2(dba).sub.3 (2.41 g, 2.63 mmol). The reaction was stirred at 90 C. overnight. The mixture was cooled down to rt, added EtOAc (1.5 L) and separated. The H.sub.2O phase was filtered. The filtrate was adjusted pH 6-7 and extracted by DCM. The organic phase was dried over Na.sub.2SO.sub.4 and concentrated in vacuo to give the titled compound (100.8 g). .sup.1H NMR (400 MHz, CD3OD) 7.56 (s, 1H), 5.67-4.90 (m, 2H), 4.36 (d, J=22.3 Hz, 1H), 3.76 (d, J=13.5 Hz, 1H), 3.62 (s, 3H), 3.57-3.33 (m, 2H), 1.49 (d, J=4.2 Hz, 9H), 1.24 (d, J=7.0 Hz, 3H), 1.15 (d, J=6.8 Hz, 3H) ppm. MS: M/e 363 (M+1).sup.+.
Step D: tert-butyl (2R,5S)-4-(3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00058##
[0233] To a mixture of tert-butyl (2R,5S)-2,5-dimethyl-4-(9-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)piperazine-1-carboxylate (96 g, 265 mmol) in dioxane (2000 mL) and was added Cs.sub.2CO.sub.3 (432 g, 1.33 mol) and CH.sub.3I (376 g, 2.65 mol). The reaction was stirred at room temperature overnight and 70 C. for 2 h. The reaction mixture was filtered. The filtrate was removed under reduced pressure to dryness. The crude product dissolved in DCM and washed with H.sub.2O. The organic phase was dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was added PE/EtOAc=500 ml/500 ml and stirred for 0.5 h. The mixture was filtered. The filter cake was dried to give the titled compound (52 g, 53%). .sup.1H NMR (400 MHz, CDCl.sub.3) 7.26 (s, 1H), 5.71 (s, 0.5H), 5.52 (s, 0.5H), 4.51 (s, 0.5H), 4.30 (s, 0.5H), 3.96 (d, J=6.9 Hz, 3H), 3.91-3.55 (m, 5H), 3.31 (m, 2H), 1.47 (s, 9H), 1.35-1.23 (m, 3H), 1.15 (d, J=6.5 Hz, 3H) ppm. MS: M/e 376 (M+1).sup.+.
Step E: tert-butyl(2R,5S)-4-(8-formyl-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00059##
[0234] To a solution of tert-butyl (2R,5S)-4-(3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (20 g, 53.2 mmol) in THE (1.2 L) was added LDA (106 mL, 2M, 212 mmol) at below 60 C. The reaction was stirred at 65 C. for 1 hour, then added DMF (19.4 g, 266 mmol) at 78 C. The reaction was stirred at 60 C. for another 1 hour. The reaction solvent was quenched by saturated NH.sub.4Cl and extracted by EtOAc. The organic phase was dried over Na.sub.2SO.sub.4 and concentrated in vacuo to give the titled compound (23.1 g). .sup.1H NMR (400 MHz, CD.sub.3OD) 9.68 (s, 1H), 6.35-5.72 (m, 1H), 5.40-4.65 (m, 1H), 4.39 (s, 1H), 4.07 (s, 3H), 3.80 (s, 3H), 3.78-3.35 (m, 3H), 1.49 (s, 9H), 1.33-1.24 (m, 3H), 1.18-1.07 (m, 3H) ppm. MS: M/e 405 (M+1).sup.+.
Step F: tert-butyl (2R,5S)-4-(8-(hydroxymethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00060##
[0235] NaBH.sub.4 (1.5 g, 40 mol) was added to tert-butyl (2R,5S)-4-(8-formyl-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (23.1 g, 57 mmol) in MeOH (10 mL) at 0 C. for 5 min. The reaction solvent was quenched with water and extracted with DCM. The organic layer was purified by flash column chromatography (DCM/MeOH) to give the titled compound (15.6 g, 68%). MS: M/e 406 (M+1).sup.+.
Step G: tert-butyl (2R,5S)-4-(8-(chloromethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00061##
[0236] To a solution of tert-butyl (2R,5S)-4-(8-(hydroxymethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (12 g, 29.6 mmol) in DCM (150 mL) was added SOCl.sub.2 (4.2 g, 35.5 mmol) at 0 C. The reaction was stirred at rt for 5 min. The reaction solvent was quenched with water and washed with water, sat. NaHCO.sub.3 and sat. NaCl. The organic layer was dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The crude product was used in next step directly.
Step H: tert-butyl (2R,5S)-4-(8-(cyanomethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00062##
[0237] To the solution of tert-butyl (2R,5S)-4-(8-(chloromethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate from last step in MeCN (400 mL) were added (14.6 g, 148 mmol) and CS.sub.2CO.sub.3 (28.9 g, 88.8 mmol). The resulting mixture was stirred at 60 C. for 1 hours. The mixture was added H.sub.2O and extracted by EtOAc. The organic phase was dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The crude product was purified by flash column chromatography (DCM/MeOH) to give the titled compound (5.8 g, two steps 47%). .sup.1H NMR (400 MHz, CD.sub.3OD) 6.25-5.64 (m, 1H), 5.36-4.66 (m, 1H), 4.38 (s, 1H), 4.23 (s, 2H), 3.94 (s, 3H), 3.80 (s, 3H), 3.75-3.33 (m, 3H), 1.48 (s, 9H), 1.28 (dd, J=19.6, 6.3 Hz, 3H), 1.15 (d, J=6.7 Hz, 3H) ppm. MS: M/e 416 (M+1).sup.+.
Step I: 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00063##
[0238] To a stirred solution of tert-butyl (2R,5S)-4-(8-(cyanomethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (1.3 g, 3 mmol) in CH.sub.2Cl.sub.2 (15 mL) was added TFA (4 mL). After stirred for 3 hours, the reaction mixture was concentrated to give the residue, which was basified to pH=1012 with aq.K.sub.2CO.sub.3, extracted with CH.sub.2Cl.sub.2/IPA (3/1, 50 mL6). The combined organic layers were dried over Na.sub.2SO.sub.4, concentrated to give the titled compound (700 mg, 71%). MS: M/e 316 (M+1).sup.+.
Intermediate 6: 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00064##
Step A: tert-butyl (2R,5S)-4-(2-chloro-9-ethyl-9H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00065##
[0239] To a solution of tert-butyl (2R,5S)-4-(2-chloro-9H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (6.97 g, 19 mmol) in acetone (100 mL) was added K.sub.2CO.sub.3 (5.2 g, 38 mmol), followed by EtI (5.9 g, 38 mmol). After the addition, the reaction mixture was stirred overnight. The reaction mixture was filtered, and the filtrate was concentrated to give the residue, which was treated with CH.sub.2Cl.sub.2 (100 mL), washed with brine, dried over Na.sub.2SO.sub.4, concentrated to give the titled compound (6.5 g, 87%). MS: M/e 395 (M+1).sup.+.
Step B: tert-butyl (2R,5S)-4-(9-ethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00066##
[0240] To a solution of tert-butyl (2R,5S)-4-(2-chloro-9-ethyl-9H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (6.5 g, 16.4 mmol) in dioxane (80 mL) was added aq.KOH (3.68 g, 65.8 mmol in 15 mL H.sub.2O), Pd.sub.2(dba).sub.3 (751 mg, 0.82 mmol) and t-BuXphOS (695 mg, 16.4 mmol). After the addition, the reaction mixture was stirred for an hour at 100 C. The reaction mixture was diluted with H.sub.2O (80 mL), extracted with EtOAc (60 mL3). The organic layers were discarded and the aqueous layer was acidified to pH=67 with aq.citric acid and extracted with EtOAc (100 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated to give the titled compound (5.3 g, 86%). MS: M/e 377 (M+1).sup.+.
Step C: tert-butyl (2R,5S)-4-(9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00067##
[0241] To a stirred solution of tert-butyl (2R,5S)-4-(9-ethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (5.3 g, 14.1 mmol) in dioxane (500 mL) was added Cs.sub.2CO.sub.3 (13.7 g, 42 mmol), followed by Mel (20 g, 0.14 mmol). After the addition, the reaction mixture was stirred for 3 days at RT and stirred at 80 C. for 3 hours. The reaction mixture was filtered, and the filtrate was concentrated to give the residue, which was purified by flash column chromatography to give the titled compound (3.32 g, 60%). MS: M/e 391 (M+1).sup.+.
Step D: tert-butyl (2R,5S)-4-(9-ethyl-8-formyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00068##
[0242] To a stirred solution of tert-butyl (2R,5S)-4-(9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (3.32 g, 8.51 mmol) in THE (300 mL) was added dropwise LDA (2.0 M, 17 mL, 34 mmol) at 78 C. After stirred for an hour at that temperature, DMF (3.06 g, 42.6 mmol) was added dropwise, then the mixture was allowed warm to RT. The reaction was quenched with aq.NH.sub.4Cl, extracted with EtOAc (200 mL2). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated to give the titled compound (crude, 100%), which was directly used to the next step.
Step E: tert-butyl (2R,5S)-4-(9-ethyl-8-(hydroxymethyl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00069##
[0243] To a stirred solution of tert-butyl (2R,5S)-4-(9-ethyl-8-formyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (crude, 8.51 mmol) in MeOH (20 mL) was added NaBH.sub.4 (224 mg, 6 mmol). After stirred for 10 min, the reaction was quenched with H.sub.2O, extracted with CH.sub.2Cl.sub.2 (50 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated and purified by flash column chromatography to give the titled compound (2.9 g, 81%).
Step F: tert-butyl (2R,5S)-4-(9-ethyl-3-methyl-8-(((methylsulfonyl)oxy)methyl)-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00070##
[0244] To a stirred solution of tert-butyl (2R,5S)-4-(9-ethyl-8-(hydroxymethyl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (2.9 g, 6.9 mmol) in CH.sub.2Cl.sub.2 (30 mL) was added Et.sub.3N (1.4 g, 13.8 mmol), followed by MsCl (1.2 g, 10.4 mmol). After the addition, the reaction mixture was stirred for 2 hours, the reaction mixture was washed with H.sub.2O (20 mL), extracted with CH.sub.2Cl.sub.2 (30 mL2). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated to give the titled compound (2.4 g, 70%).
Step G: tert-butyl (2R,5S)-4-(8-(cyanomethyl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00071##
[0245] To a stirred solution of tert-butyl (2R,5S)-4-(9-ethyl-3-methyl-8-(((methylsulfonyl)oxy)methyl)-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (2.4 g, 4.8 mmol) in CH.sub.3CN (30 mL) was added Cs.sub.2CO.sub.3 (3.2 g, 9.6 mmol), followed by TMSCN (0.95 g, 9.6 mmol). After the addition, the reaction mixture was stirred at 70 C. for 2 hours. The reaction mixture was poured into H.sub.2O (50 mL), extracted with EtOAc (30 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated and purified by flash column chromatography to give the titled compound (1.3 g, 63%).
Step H: 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00072##
[0246] To a stirred solution of tert-butyl (2R,5S)-4-(8-(cyanomethyl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (1.3 g, 3 mmol) in CH.sub.2Cl.sub.2 (15 mL) was added TFA (4 mL). After stirred for 3 hours, the reaction mixture was concentrated to give the residue, which was basified to pH=1012 with aq.K.sub.2CO.sub.3, extracted with CH.sub.2Cl.sub.2/IPA (3/1, 50 mL6). The combined organic layers were dried over Na.sub.2SO.sub.4, concentrated to give the titled compound (700 mg, 71%). MS: M/e 330 (M+1).sup.+.
Compound A1: 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(quinoxalin-6-yl)ethyl)piperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00073##
Step A: tert-butyl (2R,5S)-4-(5-chloro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00074##
[0247] To a solution of 5,7-dichloro-3H-imidazo[4,5-b]pyridine (2 g, 10.6 mmol) in DMSO (20 mL) were added tert-butyl (2R,5S)-2,5-dimethylpiperazine-1-carboxylate (11.4 g, 53.2 mmol), DIPEA (6.8 g, 53.2 mmol). The resulting mixture was stirred at 160 C. for 30 hours in sealed tube. The mixture was diluted with EtOAc then washed with water. The reaction solvent was concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (3.2 g, 82%). MS: M/e 366 (M+1).sup.+.
Step B: tert-butyl (2R,5S)-4-(5-chloro-3-methyl-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00075##
[0248] To a solution of tert-butyl (2R,5S)-4-(5-chloro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (3.2 g, 8.22 mmol) in acetone (50 mL) were added K.sub.2CO.sub.3 (2.3 g, 16.4 mmol) and CH.sub.3I (2.3 g, 16.4 mmol). The resulting mixture was stirred at rt overnight. The organic layer was concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (2.5 g, 76%). MS: M/e 380 (M+1).sup.+.
Step C: methyl 7-((2S,5R)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin-1-yl)-5-chloro-3-methyl-3H-imidazo[4,5-b]pyridine-2-carboxylate
##STR00076##
[0249] To a solution of tert-butyl (2R,5S)-4-(5-chloro-3-methyl-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (1 g, 2.63 mmol) in THE (15 mL) were added n-BuLi (2.5 mL, 1.6M, 3.95 mmol) at 78 C. The resulting mixture was stirred at 78 C. for 1 h and warmed to 40 C. for another 1 h, then added methyl carbonochloridate (1.98 g, 21.1 mmol) at 78 C. The reaction solvent was stirred at 78 C. for another 10 min. The reaction solvent was quenched by saturated NH.sub.4Cl and extracted by EtOAc. The crude product was concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (700 mg, 61%). MS: M/e 438 (M+1).sup.+.
Step D: 7-((2S,5R)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridine-2-carboxylic acid
##STR00077##
[0250] To a solution of methyl 7-((2S,5R)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin-1-yl)-5-chloro-3-methyl-3H-imidazo[4,5-b]pyridine-2-carboxylate (0.9 g, 2.01 mmol) in dioxane (15 mL) were added t-BuXphos (88 mg, 0.2 mmol) KOH (346 mg, 6.2 mmol in 1.8 mL water) and Pd.sub.2(dba).sub.3 (94 mg, 0.1 mmol). The resulting mixture was stirred at 90 C. for 4 hours. The reaction solvent was poured into water and extracted by EtOAc (discard). The aqueous layer was adjusted to pH=3 with citric acid. The aqueous layer was extracted with EtOAc to give the titled compound (900 mg, crude). MS: M/e 406 (M+1).sup.+.
Step E: methyl 7-((2S,5R)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridine-2-carboxylate
##STR00078##
[0251] To a solution of 7-((2S,5R)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridine-2-carboxylic acid (700 mg, 1.73 mmol) in dioxane (20 mL) was added Cs.sub.2CO.sub.3 (1.69 g, 5.19 mmol) and CH.sub.3I (736 mg, 5.19 mmol). The resulting mixture was stirred at 90 C. overnight. The reaction solvent was removed under reduced pressure to dryness. The crude product was purified by flash column chromatography to give the titled compound (163 mg, 22%). MS: M/e 434 (M+1).sup.+.
Step F: tert-butyl (2R,5S)-4-(2-(hydroxymethyl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00079##
[0252] To a solution of methyl 7-((2S,5R)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridine-2-carboxylate (190 mg, 0.44 mmol) in THF (10 mL) were added NaBH.sub.4 (34 mg, 0.88 mmol). The resulting mixture was stirred at 50 C. overnight. Then added NaBH.sub.4 (34 mg, 0.88 mmol), the resulting mixture was stirred at 60 C. overnight. The reaction solvent was quenched by water and extracted with (DCM/IPA=4/1). The residue was purified by Prep-TLC (DCM:MeOH=20:1) to give the titled compound (130 mg, 73%) as a white solid. MS: M/e 406 (M+1).sup.+.
Step G: tert-butyl (2R,5S)-4-(2-(chloromethyl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00080##
[0253] To a solution of tert-butyl (2R,5S)-4-(2-(hydroxymethyl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (80 mg, 0.197 mmol) in DCM (20 mL) were added SOCl.sub.2 (90 mg). The resulting mixture was stirred at RT for 5 min. The reaction solvent was washed with water, saturated NaHCO.sub.3 and saturated NaCl. The organic layer was concentrated to give the titled compound which was used to next step directly without further purification.
Step H: tert-butyl (2R,5S)-4-(2-(cyanomethyl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00081##
[0254] To a solution of tert-butyl (2R,5S)-4-(2-(chloromethyl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (crude) in CH.sub.3CN (3 mL) were added TMSCN (80 mg, 0.79 mmol) Cs.sub.2CO.sub.3 (192 mg, 0.58 mmol). The resulting mixture was stirred at 70 C. for 30 min. The reaction solvent was removed under vacuum. The crude product was purified by Prep-TLC (DCM:MeOH=30:1) to give the titled compound (50 mg, two step 61% yield). MS: M/e 415 (M+1).sup.+.
Step I: 2-(7-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00082##
[0255] To a solution of tert-butyl (2R,5S)-4-(2-(cyanomethyl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (50 mg, 0.12 mmol) in DCM (5 mL) were added TFA (1 mL). The resulting mixture was stirred at rt for 3 hours. The reaction solvent was removed under vacuum. The crude product was dissolved with water and adjusted to pH=9 with saturated NaHCO.sub.3. The aqueous layer was extracted with (DCM/IPA=4/1) and purified by Prep-TLC (DCM:MeOH=30:1) to give the titled compound (20 mg, 52%). MS: Me 315 (M+1).sup.+.
Step J: 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(quinoxalin-6-yl)ethyl)piperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00083##
[0256] To a solution of 2-(7-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (10 mg, 0.032 mmol) in CH.sub.3CN (10 mL) was added 1-(quinoxalin-6-yl)ethan-1-ol (11 mg, 0.064 mmol), (cyanomethyl)trimethylphosphonium iodide (31 mg, 0.127 mmol) and DIPEA (41 mg, 0.32 mmol). The resulting mixture was stirred at 105 C. for 15 hours. The reaction solvent was removed under reduced pressure to dryness. The crude product was purified by Prep-TLC(DCM:MeOH=20:1) and further purified by Prep-HPLC(Method A) to give the titled compound (1 mg, 18%). .sup.1H NMR (400 MHz, CD.sub.3OD) 8.90-8.80 (m, 2H), 8.15-7.95 (m, 3H), 5.52 (s, 1H), 4.20 (s, 2H), 3.97 (s, 3H), 3.83 (s, 3H), 3.75-3.55 (m, 2H), 3.50-3.33 (m, 2H), 3.20-3.02 (m, 1H), 2.98-2.63 (m, 2H), 2.25-2.15 (m, 1H), 1.49-1.30 (m, 4H), 1.25-1.18 (m, 3H), 1.08-1.01 (m, 1H) ppm. MS: M/e 471 (M+1).sup.+.
[0257] Compound A1 also can be synthesized according to the following synthetic method.
Compound A1: 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(quinoxalin-6-yl)ethyl)piperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00084##
Step A: tert-butyl (2R,5S)-4-(5-chloro-2-(hydroxymethyl)-3-methyl-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00085##
[0258] To a solution of methyl 7-((2S,5R)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin-1-yl)-5-chloro-3-methyl-3H-imidazo[4,5-b]pyridine-2-carboxylate (1.6 g, 3.66 mmol) in THE (10 mL) were added NaBH.sub.4 (278 mg, 7.32 mmol). The resulting mixture was stirred at 70 C. for 24 hours. The reaction solvent was quenched by water and extracted with EtOAc. The residue was purified by flash column chromatography (DCM:MeOH=20:1) to give the titled compound (1.2 g, 80% yield) as a white solid. MS: M/e 410 (M+1).sup.+.
Step B: tert-butyl (2R,5S)-4-(2-(((tert-butyldimethylsilyl)oxy)methyl)-5-chloro-3-methyl-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00086##
[0259] To a mixture of tert-butyl (2R,5S)-4-(5-chloro-2-(hydroxymethyl)-3-methyl-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (1.1 g, 2.6 mmol) in DCM (15 mL) and was added TBSCl (0.58 g, 3.9 mmol) and imidazole (0.53 g, 7.78 mmol). The resulting mixture was stirred at RT overnight. The reaction solvent was washed with water and purified by flash column chromatography (PE/EtOAc) to give the titled compound (1.2 g, 85% yield) as a colourless oil. MS: M/e 524 (M+1).sup.+.
Step C tert-butyl (2R,5S)-4-(2-(((tert-butyldimethylsilyl)oxy)methyl)-3-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00087##
[0260] To a solution of tert-butyl (2R,5S)-4-(2-(((tert-butyldimethylsilyl)oxy)methyl)-5-chloro-3-methyl-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (1.2 g, 2.3 mmol) in 1,4-dioxane (20 mL) were added t-BuXphos (98 mg, 0.11 mmol), KOH (385 mg, 6.9 mmol in 2.5 mL water) and Pd.sub.2(dba).sub.3 (107 mg, 0.11 mmol). The resulting mixture was stirred at 80 C. for 3 hours. The reaction solvent was poured into water and extracted by EtOAc (discard). The aqueous layer was adjusted to pH=3 with citric acid. The aqueous layer was extracted with EtOAc and purified by flash column chromatography (PE/EtOAc) to give the titled compound (900 mg, crude) as a white solid. MS: M/e 506 (M+1).sup.+.
Step D tert-butyl (2R,5S)-4-(2-(((tert-butyldimethylsilyl)oxy)methyl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00088##
[0261] To a mixture of tert-butyl (2R,5S)-4-(2-(((tert-butyldimethylsilyl)oxy)methyl)-3-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (600 mg, 1.19 mmol) in dioxane (80 mL) and was added Cs.sub.2CO.sub.3 (2.3 g, 7.1 mmol) and CH.sub.3I (1 g, 7.1 mmol). The resulting mixture was stirred at rt overnight. The reaction solvent was removed under reduced pressure to dryness. The crude product was purified by flash column chromatography (PE/EtOAc and DCM/MeOH) to give the titled compound (380 mg). MS: M/e 520 (M+1).sup.+.
Step E: tert-butyl (2R,5S)-4-(2-(hydroxymethyl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00089##
[0262] To a solution of tert-butyl (2R,5S)-4-(2-(((tert-butyldimethylsilyl)oxy)methyl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (380 mg, 0.73 mmol) in THF (10 mL) were added TBAF (1.1 mL, 1.1 mmol). The resulting mixture was stirred at rt for 1 hour. The reaction solvent was concentrated to dryness. The resulting residue was purified by flash column chromatography (DCM/MeOH) to give the titled compound (300 mg, crude) as a yellow solid. MS: M/e 406 (M+1).sup.+.
Step F: tert-butyl (2R,5S)-4-(2-(chloromethyl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00090##
[0263] To a solution of tert-butyl (2R,5S)-4-(2-(hydroxymethyl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (150 mg, 0.37 mmol) in DCM (150 mL) were added SOCl.sub.2 (220 mg, 1.85 mmol). The resulting mixture was stirred at rt for 5 min. The reaction solvent was washed with water, NaHCO.sub.3 aq. and NaCl aq., The organic layer was concentrated to give the titled compound which was used to next step directly without further purification.
Step G: tert-butyl (2R,5S)-4-(2-(cyanomethyl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00091##
[0264] To the solution of tert-butyl (2R,5S)-4-(2-(chloromethyl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (crude) in CH.sub.3CN (20 mL) were added TMSCN (183 mg, 1.85 mmol) and TBAF (1.8 ml, 1.85 mmol). The resulting mixture was stirred at 0 C. for 30 min. The reaction solvent was removed under vacuum. The crude product was purified by Prep-TLC (DCM:MeOH=30:1) to give the titled compound (60 mg). MS: M/e 415 (M+1).sup.+.
Step H: 2-(7-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00092##
[0265] To a solution of tert-butyl (2R,5S)-4-(2-(cyanomethyl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (120 mg, 0.29 mmol) in DCM (10 mL) were added TFA (3 mL). The resulting mixture was stirred at rt for 1.5 hours. The reaction solvent was removed under vacuum. The crude product was dissolved with water and adjusted to pH=9 with NaHCO.sub.3 aq. The aqueous layer was extracted with (DCM:IPA=3:1) to give the titled compound (100 mg, crude) as a yellow solid. MS: M/e 315 (M+1).sup.+.
Step I: 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(quinoxalin-6-yl)ethyl)piperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00093##
[0266] To a mixture of 2-(7-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (100 mg, 0.32 mmol) in CH.sub.3CN (10 mL) and was added 1-(quinoxalin-6-yl)ethan-1-ol (83 mg, 0.48 mmol), (cyanomethyl)trimethylphosphonium iodide (387 mg, 1.6 mmol) and DIPEA (410 mg, 3.2 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure to dryness. The crude product was purified by Prep-TLC (DCM:MeOH=50:1) to give the titled Compound A1 which was further separated into Compound A1a (1 mg) and Compound A1b (1 mg) by Prep-HPLC(Method B).
[0267] Compound A1a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.90-8.8. (m, 2H), 8.20-7.98 (m, 3H), 5.52 (s, 1H), 5.09-4.95 (m, 1H), 4.55-4.30 (m, 1H), 4.00-3.90 (m, 4H), 3.83 (s, 3H), 3.50-3.34 (m, 1H), 3.10-3.02 (m, 1H), 2.98-2.63 (m, 4H), 1.49-1.30 (m, 6H), 1.08-1.01 (m, 3H) ppm. MS: M/e 471 (M+1).sup.+
[0268] Compound A1b (the later peak) .sup.1H NMR (400 MHz, CD.sub.3OD) 8.90-8.8. (m, 2H), 8.15-7.95 (m, 3H), 5.52 (s, 2H), 4.87-4.55 (m, 2H), 3.97 (s, 3H), 3.83-3.75 (m, 4H), 3.70-3.55 (m, 2H), 2.95-2.63 (m, 2H), 2.25-2.15 (m, 1H), 1.49-1.30 (m, 3H), 1.25-1.13 (m, 6H) ppm. MS: M/e 471 (M+1).sup.+
Compound A4: 2-(7-((2S,5R)-4-(1-(4-fluoro-2-methoxyphenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00094##
[0269] To a solution of 2-(7-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (30 mg, 0.096 mmol) in CH.sub.3CN (3 mL) was added 1-(4-fluoro-2-methoxyphenyl)ethan-1-ol (33 mg, 1.91 mmol), (cyanomethyl)trimethylphosphonium iodide (116 mg, 0.48 mmol) and DIPEA (123 mg, 0.96 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure to dryness. The crude product was purified by Prep-TLC (DCM:MeOH=20:1) to give the titled Compound A4, which was further separated into Compound A4a (0.7 mg) and Compound A4b (0.91 mg) by Prep-HPLC (Method A).
[0270] Compound A4a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.58-7.50 (m, 1H), 6.78-6.59 (m, 2H), 5.51 (s, 1H), 4.80-4.55 (m, 1H), 4.55-3.98 (m, 2H), 3.97 (s, 3H), 3.83 (s, 6H), 3.60-3.50 (m, 1H), 2.98-2.55 (m, 4H), 2.25-2.10 (m, 1H), 1.35-1.01 (m, 9H) ppm. MS: M/e 467 (M+1).sup.+.
[0271] Compound A4b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.55-7.45 (m, 1H), 6.68-6.56 (m, 2H), 5.51 (s, 1H), 4.57-4.10 (m, 2H), 4.05-3.85 (m, 4H), 3.85-3.55 (m, 5H), 3.50-3.34 (m, 2H), 2.99-2.60 (m, 3H), 2.15-1.98 (m, 1H), 1.60-1.10 (m, 5H), 1.08-0.60 (m, 4H) ppm. MS: M/e 467 (M+1).sup.+.
Compound A5: 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(p-tolyl)ethyl)piperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00095##
[0272] To a mixture of 2-(7-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (30 mg, 0.1 mmol) in CH.sub.3CN (3 mL) and was added 1-(p-tolyl)ethan-1-ol (39 mg, 0.29 mmol), (cyanomethyl)trimethylphosphonium iodide (116 mg, 0.5 mmol) and DIPEA (123 mg, 1 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure to dryness. The crude product was purified by Prep-TLC (DCM:MeOH=20:1) to give the titled Compound A5, which was further separated into Compound A5a (0.65 mg) and Compound A5b (0.48 mg) by Prep-HPLC (Method A).
[0273] Compound A5a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.25 (d, J=7.9 Hz, 2H), 7.11 (d, J=7.8 Hz, 2H), 5.51 (s, 1H), 4.78-4.65 (m, 1H), 4.21 (s, 1H), 3.97 (s, 3H), 3.97 (s, 3H), 3.66-3.41 (m, 2H), 2.80-2.61 (m, 4H), 2.31 (s, 3H), 2.26-2.19 (m, 1H), 1.34 (d, J=10.5 Hz, 3H), 1.18 (d, J=6.6 Hz, 3H), 1.13 (d, J=6.1 Hz, 3H) ppm. MS: M/e 433 (M+1).sup.+.
[0274] Compound A5b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.26 (d, J=7.9 Hz, 2H), 7.14 (d, J=7.8 Hz, 2H), 5.50 (s, 1H), 4.56-4.45 (m, 1H), 4.20 (s, 1H), 3.97 (s, 3H), 3.83 (s, 3H), 3.62 (s, 1H), 3.40-3.33 (m, 1H), 2.98-2.75 (m, 5H), 2.32 (s, 3H), 1.36 (d, J=6.5 Hz, 3H), 1.31 (d, J=6.5 Hz, 3H), 0.97 (d, J=6.5 Hz, 3H) ppm. MS: M/e 433 (M+1).sup.+.
Compound A6: 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(1-methyl-5-(trifluoromethyl)-1H-pyrazol-4-yl)ethyl)piperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00096##
Step A: 1-(1-methyl-5-(trifluoromethyl)-1H-pyrazol-4-yl)ethan-1-ol
##STR00097##
[0275] To a solution of 1-methyl-5-(trifluoromethyl)-1H-pyrazole-4-carbaldehyde (200 mg, 1.12 mmol) in THE (8 ml) at 60 C. under N.sub.2, was added MeMgBr (3M, 0.41 ml, 1.23 mmol) dropwise. The solution was warmed to RT naturally and stirred at RT overnight. After completed, the solution was quenched with H.sub.2O (10 ml) and then extracted with EtOAc (10 ml2). The organic layer was washed with aq. NaHCO.sub.3 (10 ml), brine (10 ml), dried over anhydrous Na.sub.2SO.sub.4 and then concentrated under reduced pressure to give the titled compound (217 mg, 100%), which was used directly for the next step without further purification. MS: M/e 195 (M+1).sup.+.
Step B: 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(1-methyl-5-(trifluoromethyl)-1H-pyrazol-4-yl)ethyl)piperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00098##
[0276] To a mixture of 2-(7-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (30 mg, 0.1 mmol) in CH.sub.3CN (3 mL) and was added 1-(1-methyl-5-(trifluoromethyl)-1H-pyrazol-4-yl)ethan-1-ol (56 mg, 0.29 mmol), (cyanomethyl)trimethylphosphonium iodide (116 mg, 0.5 mmol) and DIPEA (123 mg, 1 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure to dryness. The crude product was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled Compound A6, which was further separated into Compound A6a (0.77 mg) and Compound A6b (0.64 mg) by Prep-HPLC (Method A).
[0277] Compound A6a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.62 (s, 1H), 5.51 (s, 1H), 4.77-4.63 (m, 1H), 4.21 (s, 1H), 3.97 (s, 6H), 3.83 (s, 3H), 3.81-3.70 (m, 1H), 3.57-3.45 (m, 2H), 2.86-2.70 (m, 3H), 2.31 (d, J=12.5 Hz, 1H), 1.35 (d, J=6.6 Hz, 3H), 1.21 (d, J=6.6 Hz, 3H), 1.09 (d, J=6.5 Hz, 3H) ppm. MS: M/e 491 (M+1).sup.+.
[0278] Compound A6b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.61 (s, 1H), 5.51 (s, 1H), 4.75-4.63 (m, 1H), 4.23-4.17 (m, 1H), 3.98 (s, 3H), 3.97 (s, 3H), 3.83 (s, 3H), 3.39-3.33 (m, 2H), 3.04-2.90 (m, 2H), 2.85-2.61 (m, 3H), 1.33 (d, J=6.7 Hz, 3H), 1.29 (d, J=6.5 Hz, 3H), 1.00 (d, J=6.5 Hz, 3H) ppm. MS: M/e 491 (M+1).sup.+.
Compound A7: 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(2-(trifluoromethyl)pyridin-3-yl)ethyl)piperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00099##
Step A: 1-(2-(trifluoromethyl)pyridin-3-yl)ethan-1-ol
##STR00100##
[0279] To a solution of 2-(trifluoromethyl)nicotinaldehyde (200 mg, 1.1 mmol) in THE (10 mL) was added methylmagnesium bromide (0.7 mL, 2.2 mmol, 3M in Et.sub.2O) at 0 C. under N.sub.2 atmosphere. The reaction mixture was stirred at room temperature for 2 hours. The reaction was quenched by water (5 mL) and extracted with EtOAc (15 mL3). The organic phases were concentrated under reduced pressure. The resulting residue was purified by Prep-TLC (PE:EtOAc=2:1) to give the target compound (120 mg, 55% yield) as a clear oil. MS: M/e 192 (M+1).sup.+.
Step B: 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(2-(trifluoromethyl)pyridin-3-yl)ethyl)piperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00101##
[0280] To a mixture of 2-(7-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (30 mg, 0.1 mmol) in CH.sub.3CN (3 mL) and was added 1-(2-(trifluoromethyl)pyridin-3-yl)ethan-1-ol (56 mg, 0.29 mmol), (cyanomethyl)trimethylphosphonium iodide (116 mg, 0.5 mmol) and DIPEA (123 mg, 1 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure. The resulting residue was purified by Prep-TLC (DCM:MeOH=20:1) to give the titled Compound A7, which was further separated into Compound A7a (1.21 mg) and Compound A7b (0.81 mg) by Prep-HPLC (Method A).
[0281] Compound A7a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.55 (d, J=4.6 Hz, 1H), 8.48 (d, J=8.4 Hz, 1H), 7.68 (dd, J=7.9, 4.7 Hz, 1H), 5.51 (s, 1H), 4.20 (d, J=4.9 Hz, 1H), 3.99-3.91 (m, 4H), 3.83 (s, 3H), 3.72-3.53 (m, 3H), 2.85-2.55 (m, 3H), 1.93 (d, J=12.1 Hz, 1H), 1.36 (d, J=6.4 Hz, 3H), 1.21-1.11 (m, 6H) ppm. MS: M/e 488 (M+1).sup.+.
[0282] Compound A7b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.56 (d, J=4.2 Hz, 1H), 8.51 (d, J=7.9 Hz, 1H), 7.70 (dd, J=8.2, 4.4 Hz, 1H), 5.51 (s, 1H), 4.25-4.16 (m, 2H), 3.97 (s, 3H), 3.83 (s, 3H), 3.42-3.33 (m, 2H), 3.08-2.60 (m, 5H), 1.43 (d, J=6.6 Hz, 3H), 1.29 (d, J=6.4 Hz, 3H), 1.02 (d, J=6.5 Hz, 3H) ppm. MS: M/e 488 (M+1).sup.+.
Compound A9: 2-(7-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethyl piperazin-1-yl)-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00102##
Step A: tert-butyl (tert-butoxycarbonyl)(4,6-dichloro-3-nitropyridin-2-yl)carbamate
##STR00103##
[0283] To a solution of 4,6-dichloro-3-nitropyridin-2-amine (2 g, 9.66 mmol) in THE (20 ml) at 0 C. was added NaH (60%, 1.16 g, 29 mmol). Then (Boc).sub.2O (6.3 g, 28.90 mmol) was added. The mixture was stirred at rt overnight. The reaction was diluted with EtOAc (30 ml) and then washed with water (20 ml). The organic layer was concentrated under reduced pressure. The resulting residue was purified by flash column chromatography with 0-60% EtOAc in PE to give the titled compound (3.63 g, 92%). MS: M/e 408 (M+1).sup.+.
Step B: tert-butyl (tert-butoxycarbonyl)(6-chloro-4-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-nitropyridin-2-yl)carbamate
##STR00104##
[0284] A solution of tert-butyl (tert-butoxycarbonyl)(4,6-dichloro-3-nitropyridin-2-yl)carbamate (1.9 g, 4.67 mmol), (2R,5S)-1-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazine (1.5 g, 4.93 mmol) and DIPEA (3 g, 23.26 mmol) in DCM (30 ml) was stirred at RT overnight. The mixture was concentrated under reduced pressure. The resulting residue was purified by flash column chromatography with 0-15% EtOAc in PE to give the titled compound (1.8 g, 57%). MS: M/e 676 (M+1).sup.+.
Step C: tert-butyl (tert-butoxycarbonyl)(4-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-nitro-6-oxo-1,6-dihydropyridin-2-yl)carbamate
##STR00105##
[0285] A solution of tert-butyl (tert-butoxycarbonyl)(6-chloro-4-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-nitropyridin-2-yl)carbamate (0.6 g, 0.89 mmol), Pd.sub.2(dba).sub.3 (81.4 mg, 0.089 mmol), t-BuXPhos (75.6 mg, 0.18 mmol) and KOH (100 mg, 1.79 mmol) in dioxane (15 ml) and water (3 ml) was stirred at 80 C. for 3 h in a sealed tube under N.sub.2. The mixture was concentrated. The residue was diluted with EtOAc (20 ml) and washed with brine (10 ml). The organic layer was concentrated to dryness. The resulting residue was purified by flash column chromatography with 0-60% EtOAc in PE to give the titled compound (0.4 g, 68%). MS: M/e 658 (M+1).sup.+.
Step D: tert-butyl (tert-butoxycarbonyl)(4-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-1-methyl-3-nitro-6-oxo-1,6-dihydropyridin-2-yl)carbamate
##STR00106##
[0286] A solution of tert-butyl (tert-butoxycarbonyl)(4-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-nitro-6-oxo-1,6-dihydropyridin-2-yl)carbamate (1.2 g, 1.83 mmol), Mel (0.52 g, 3.66 mmol) and Cs.sub.2CO.sub.3 (1.8 g, 5.52 mmol) in dioxane (20 ml) was stirred at 80 C. overnight. The mixture was concentrated. The residue was diluted with EtOAc (30 ml) and washed with brine (15 ml). The organic layer was concentrated to dryness. The resulting residue was purified by flash column chromatography with 0-50% EtOAc in PE to give the titled compound (0.66 g, 54%). MS: M/e 672 (M+1).sup.+.
Step E: tert-butyl (3-amino-4-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-1-methyl-6-oxo-1,6-dihydropyridin-2-yl)(tert-butoxycarbonyl)carbamate
##STR00107##
[0287] A solution of tert-butyl (tert-butoxycarbonyl)(4-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-1-methyl-3-nitro-6-oxo-1,6-dihydropyridin-2-yl)carbamate (0.66 g, 0.98 mmol) and Pd/C (0.1 g) in MeOH (15 ml) under H.sub.2 was stirred at rt overnight. The mixture was filtered. The filtrate was concentrated to dryness to give the titled compound (0.62 g, 98%). MS: M/e 642 (M+1).sup.+.
Step F: 5,6-diamino-4-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-1-methylpyridin-2 (1H)-one
##STR00108##
[0288] A solution of tert-butyl (3-amino-4-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-1-methyl-6-oxo-1,6-dihydropyridin-2-yl)(tert-butoxycarbonyl)carbamate (310 mg, 0.48 mmol) and TFA (3 ml) in DCM (15 ml) was stirred at rt for 2 h. The mixture was washed with NaHCO.sub.3 (aq., 10 ml), brine (10 ml), dried over Na.sub.2SO.sub.4 and then concentrated to dryness to give the titled compound (200 mg, 93%). MS: M/e 442 (M+1).sup.+.
Step G: N-(3-amino-4-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-1-methyl-6-oxo-1,6-dihydropyridin-2-yl)-2-cyanoacetamide
##STR00109##
[0289] A solution of 5,6-diamino-4-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-1-methylpyridin-2 (1H)-one (200 mg, 0.45 mmol), 2-cyanoacetic acid (46 mg, 0.54 mmol), HATU (259 mg, 0.68 mmol) and DIPEA (176 mg, 1.36 mmol) in THE (10 ml) was stirred at rt for 30 min. The mixture was diluted with EtOAc (20 ml) and washed with brine (15 ml). The organic layer was concentrated. The resulting residue was purified by flash column chromatography with 0-10% MeOH in DCM to give the titled compound (130 mg, 56%). MS: M/e 509 (M+1).sup.+.
Step H: 2-(7-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00110##
[0290] A solution of N-(3-amino-4-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-1-methyl-6-oxo-1,6-dihydropyridin-2-yl)-2-cyanoacetamide (65 mg, 0.13 mmol) in AcOH (1 ml) was stirred at 80 C. overnight. The mixture was concentrated under reduced pressure. The resulting residue was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled Compound A9 (18 mg), which was further separated into Compound A9a (4 mg) and Compound A9b (7 mg) by Prep-HPLC (Method A).
[0291] Compound A9: .sup.1H NMR (400 MHz, DMSO-d6) 13.10-12.50 (m, 1H), 8.09-7.95 (m, 1H), 7.67-7.49 (m, 2H), 5.44-5.17 (m, 1H), 4.27 (s, 0.5H), 4.23 (s, 0.5H), 4.22-4.11 (m, 1H), 4.01 (s, 0.5H), 3.92-3.72 (m, 1H), 3.57 (s, 0.5H), 3.48-3.35 (m, 4H), 3.26-3.14 (m, 0.5H), 3.13-2.96 (m, 1H), 2.90 (s, 0.5H), 2.85 (d, J=11.4 Hz, 1H), 2.67 (s, 0.5H), 1.87 (d, J=11.5 Hz, 0.5H), 1.28 (d, J=6.2 Hz, 2H), 1.21 (dd, J=11.4, 6.5 Hz, 2.5H), 1.11-1.00 (m, 2H), 0.96-0.85 (m, 2.5H) ppm. MS: M/e 491 (M+1).sup.+.
[0292] Compound A9a (the earlier peak): .sup.1H NMR (400 MHz, DMSO-d6)) 13.21-12.27 (m, 1H), 8.04-7.95 (m, 1H), 7.63-7.50 (m, 2H), 5.32 (s, 1H), 4.19 (s, 2H), 3.82 (s, 1H), 3.58 (s, 1H), 3.47-3.43 (m, 1H), 3.41 (s, 3H), 3.31-3.25 (m, 1H), 3.10-2.90 (m, 1H), 2.81 (s, 1H), 1.87 (d, J=11.2 Hz, 1H), 1.28 (d, J=6.4 Hz, 3H), 1.07 (d, J=6.4 Hz, 3H), 0.94 (s, 3H) ppm. MS: M/e 491 (M+1).sup.+.
[0293] Compound A9b (the later peak): .sup.1H NMR (400 MHz, DMSO-d6)) 12.76 (s, 1H), 8.11-8.01 (m, 1H), 7.67-7.52 (m, 2H), 5.26 (s, 1H), 4.22 (s, 2H), 4.00 (s, 1H), 3.41 (s, 3H), 3.31-3.28 (m, 2H), 3.20 (d, J=9.2 Hz, 1H), 3.00 (s, 1H), 2.85 (d, J=11.0 Hz, 1H), 2.69 (s, 1H), 1.22 (t, J=7.6 Hz, 6H), 0.93 (d, J=6.4 Hz, 3H) ppm. MS: M/e 491 (M+1).sup.+.
Compound A10: 2-(6-((2R,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2-(methoxymethyl)-5-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00111##
Step A: 2,6-dichloro-9-methyl-9H-purine
##STR00112##
[0294] To a solution of 2,6-dichloro-9H-purine (20 g, 106 mmol) and K.sub.2CO.sub.3 (29.3 g, 212 mmol) in DMF (120 mL) was added CH.sub.3I (22.5 g, 158 mmol). The reaction was stirred at room temperature overnight. The reaction mixture was diluted with water, extracted with EtOAc (300 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (EtOAc:PE=1:1) to give the titled compound (14 g, 65%). MS: M/e 203 (M+1).sup.+.
Step B: tert-butyl (2R,5R)-4-(2-chloro-9-methyl-9H-purin-6-yl)-5-(hydroxymethyl)-2-methylpiperazine-1-carboxylate
##STR00113##
[0295] To a solution of 2,6-dichloro-9-methyl-9H-purine (2.0 g, 9.9 mmol) and tert-butyl (2R,5R)-5-(hydroxymethyl)-2-methylpiperazine-1-carboxylate (2.28 g, 9.9 mmol) in n-BuOH (40 mL) was added DIPEA (3.83 g, 29.7 mmol). The reaction was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure to give the title compound (3.3 g, yield: 84%). MS: M/e 397 (M+1).sup.+.
Step C: tert-butyl (2R,5R)-4-(2-chloro-9-methyl-9H-purin-6-yl)-5-(methoxymethyl)-2-methylpiperazine-1-carboxylate
##STR00114##
[0296] To a solution of tert-butyl (2R,5R)-4-(2-chloro-9-methyl-9H-purin-6-yl)-5-(hydroxymethyl)-2-methylpiperazine-1-carboxylate (3.3 g, 8.329 mmol) in DMF (50 mL) at 0 C. was added NaH (1.33 g, 60% contained, 33.3 mmol). The mixture was stirred at 0 C. for 0.5 hours. Then CH.sub.3I was added. The reaction was stirred at room temperature for 2 hours. The reaction mixture was quenched with water and extracted with EtOAc (120 mL2). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, filtered and concentrated to dryness. The resulting residue was purified by flash column chromatography (DCM:MeOH=10:1) to give the titled compound (2.4 g, 70%). MS: M/e 411 (M+1).sup.+.
Step D: tert-butyl (2R,5R)-4-(2-hydroxy-9-methyl-9H-purin-6-yl)-5-(methoxymethyl)-2-methylpiperazine-1-carboxylate
##STR00115##
[0297] To a solution of tert-butyl (2R,5R)-4-(2-chloro-9-methyl-9H-purin-6-yl)-5-(methoxymethyl)-2-methylpiperazine-1-carboxylate (2.4 g, 5.85 mmol), Pd.sub.2(dba).sub.3 (536 mg, 0.585 mmol), t-BuXphos (498 mg, 1.17 mmol) and KOH (5.85 mL, 3M, 17.55 mmol) in dioxane (30 mL) was degassed 3 times under N.sub.2 atmosphere. Then the mixture was stirred at 90 C. for 12 hours. The reaction mixture was diluted with water and extracted with EtOAc (60 mL2). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, filtered and concentrated to dryness. The resulting residue was purified by flash column chromatography (DCM:MeOH=10:1) to give the titled compound (1.7 g, 74%). MS: M/e 393 (M+1).sup.+.
Step E: tert-butyl (2R,5R)-4-(3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-5-(methoxymethyl)-2-methylpiperazine-1-carboxylate
##STR00116##
[0298] To a solution of tert-butyl (2R,5R)-4-(2-hydroxy-9-methyl-9H-purin-6-yl)-5-(methoxymethyl)-2-methylpiperazine-1-carboxylate (1.7 g, 4.33 mmol) and Cs.sub.2CO.sub.3 (2.82 g, 8.66 mmol) in dioxane (40 mL) was added CH.sub.3I (923 mg, 6.50 mmol). The reaction was stirred at room temperature overnight. The reaction mixture was diluted with water, extracted with EtOAc (80 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (DCM:MeOH=10:1) to give the titled compound (960 mg, 54%). MS: M/e 407 (M+1).sup.+.
Step F: tert-butyl (2R,5R)-4-(8-bromo-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-5-(methoxymethyl)-2-methylpiperazine-1-carboxylate
##STR00117##
[0299] To a solution of tert-butyl (2R,5R)-4-(3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-5-(methoxymethyl)-2-methylpiperazine-1-carboxylate (300 mg, 0.739 mmol) in CH.sub.3CN (10 mL) was added NBS (197 mg, 1.108 mmol). The reaction was stirred at room temperature for 4 hours. The reaction mixture was diluted with saturated NH.sub.4Cl, extracted with EtOAc (50 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (DCM:MeOH=10:1) to give the titled compound (350 mg, 98%). MS: M/e 485 (M+1).sup.+.
Step G: methyl 6-((2R,5R)-4-(tert-butoxycarbonyl)-2-(methoxymethyl)-5-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purine-8-carboxylate
##STR00118##
[0300] To a solution of tert-butyl (2R,5R)-4-(8-bromo-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-5-(methoxymethyl)-2-methylpiperazine-1-carboxylate (350 mg, 0.723 mmol), Pd(dppf)Cl.sub.2 and Et.sub.3N (146 mg, 1.446 mmol) in MeOH (10 mL) was degassed 3 times under CO atmosphere. Then the mixture was stirred at 90 C. for 12 hours under CO atmosphere. The reaction mixture was diluted with water and extracted with EtOAc (35 mL2). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, filtered and concentrated to dryness. The resulting residue was purified by flash column chromatography (DCM:MeOH=8:1) to give the titled compound (280 mg, 83%). MS: M/e 465 (M+1).sup.+.
Step H: tert-butyl (2R,5R)-4-(8-(hydroxymethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-5-(methoxymethyl)-2-methylpiperazine-1-carboxylate
##STR00119##
[0301] To a solution of methyl 6-((2R,5R)-4-(tert-butoxycarbonyl)-2-(methoxymethyl)-5-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purine-8-carboxylate (280 mg, 0.603 mmol) in MeOH (10 mL) was added NaBH.sub.4 (92 mg, 2.414 mmol). The reaction was stirred at room temperature for 2 hours. The reaction mixture was quenched with saturated NH.sub.4Cl, extracted with EtOAc (30 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (DCM:MeOH=8:1) to give the titled compound (200 mg, 76%). MS: M/e 437 (M+1).sup.+.
Step I: tert-butyl (2R,5R)-4-(8-(chloromethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-5-(methoxymethyl)-2-methylpiperazine-1-carboxylate
##STR00120##
[0302] To a solution of tert-butyl (2R,5R)-4-(8-(hydroxymethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-5-(methoxymethyl)-2-methylpiperazine-1-carboxylate (200 mg, 0.459 mmol) in DCM (8 mL) was added SOCl.sub.2 (218 mg, 1.835 mmol). The reaction was stirred at room temperature for 0.5 hours. The reaction mixture concentrated to give the titled compound (190 mg, 91%). MS: M/e 455 (M+1).sup.+.
Step J: tert-butyl (2R,5R)-4-(8-(cyanomethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-5-(methoxymethyl)-2-methylpiperazine-1-carboxylate
##STR00121##
[0303] To a solution of tert-butyl (2R,5R)-4-(8-(chloromethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-5-(methoxymethyl)-2-methylpiperazine-1-carboxylate (190 mg, 0.419 mmol) in DCM (15 mL) was added TBAF (1.67 mL, 1M, 1.674 mmol), followed TMSCN (166 mg, 1.674 mmol). The reaction was stirred at room temperature overnight. The reaction mixture was diluted with water, extracted with DCM (40 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (DCM:MeOH=8:1) to give the titled compound (150 mg, 81%). MS: M/e 446 (M+1).sup.+.
Step K: 2-(6-((2R,5R)-2-(methoxymethyl)-5-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00122##
[0304] To a stirred solution of tert-butyl (2R,5R)-4-(8-(cyanomethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-5-(methoxymethyl)-2-methylpiperazine-1-carboxylate (150 mg, 0.337 mmol) in CH.sub.2Cl.sub.2 (15 mL) was added TFA (3 mL). Then reaction mixture was stirred at room temperature for 4 hours, concentrated to give a residue, basified to pH=1011 with saturated NaHCO.sub.3 aq., and extracted with CH.sub.2Cl.sub.2/IPA (3/1, 30 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, and concentrated to give the titled compound (40 mg, 34%). MS: M/e 346 (M+1).sup.+.
Step L: 2-(6-((2R,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2-(methoxymethyl)-5-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00123##
[0305] A solution of 2-(6-((2R,5R)-2-(methoxymethyl)-5-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (40 mg, 0.116 mol), 1-(4-fluoro-2-(trifluoromethyl)phenyl)ethan-1-ol (36 mg, 0.174 mol), (cyanomethyl)trimethylphosphonium iodide (56 mg. 0.232 mol) and DIPEA (45 mg, 0.348 mol) in CH.sub.3CN (2 ml). The mixture solution was degassed 3 times under N.sub.2 atmosphere. Then the mixture solution was stirred at 105 C. for 24 hours. The reaction mixture was concentrated under reduced pressure. The crude product was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled Compound A10 (6 mg), which was further separated into Compound A10a (2 mg) and Compound A10b (1.5 mg) by Prep-HPLC (Method A).
[0306] Compound A10: .sup.1H NMR (400 MHz, CD.sub.3OD) 8.07-7.94 (m, 1H), 7.43-7.39 (m, 2H), 6.32-6.05 (m, 0.5H), 5.95-5.66 (m, 0.5H), 5.37-5.12 (m, 0.5H), 4.81-4.57 (m, 0.5H), 4.21 (dd, J=24.3, 4.1 Hz, 2H), 3.93 (t, J=5.6 Hz, 3H), 3.79 (s, 3H), 3.76-3.59 (m, 3H), 3.43 (d, J=4.0 Hz, 1H), 3.26-3.18 (m, 3H), 2.94-2.53 (m, 2H), 2.35-1.98 (m, 1H), 1.39-1.34 (m, 3H), 1.04-0.89 (m, 3H) ppm. MS: M/e 536 (M+1).sup.+
[0307] Compound A10a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.06-7.91 (m, 1H), 7.45-7.36 (m, 2H), 6.06-6.02 (m, 0.5H), 5.95-5.91 (m, 0.5H), 5.16-5.12 (m, 0.5H), 4.94-4.90 (m, 0.5H), 4.21 (d, J=19.3 Hz, 1H), 3.94 (s, 3H), 3.92-3.88 (m, 1H), 3.79 (d, J=2.7 Hz, 3H), 3.69-3.60 (m, 3H), 3.40-3.33 (m, 1H), 3.22 (d, J=10.8 Hz, 3H), 2.94-2.51 (m, 2H), 2.27 (d, J=12.3 Hz, 1H), 1.31 (d, J=6.2 Hz, 3H), 1.11-0.98 (m, 3H) ppm. MS: M/e 536 (M+1).sup.+
[0308] Compound A10b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.08-7.98 (m, 1H), 7.44-7.39 (m, 2H), 6.31 (s, 0.5H), 5.67 (d, J=13.6 Hz, 0.5H), 5.39 (s, 0.5H), 4.64-4.57 (m, 0.5H), 4.11-3.98 (m, 2H), 3.93 (d, J=11.3 Hz, 3H), 3.85-3.73 (m, 4H), 3.50-3.45 (m, 0.5H), 3.43 (d, J=3.9 Hz, 3H), 3.35-3.31 (m, 0.5H), 3.27-3.06 (m, 1.5H), 3.01-2.59 (m, 3.5H), 1.29-1.21 (m, 3H), 0.90 (d, J=6.5 Hz, 3H) ppm. MS: M/e 536 (M+1).sup.+
Compound A11: 2-(6-((2R,5R)-4-(1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2-(methoxymethyl)-5-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00124##
Step A: 1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol
##STR00125##
[0309] To a solution of 1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-one (720 mg, 4.1 mmol) in MeOH (20 mL) was added NaBH.sub.4 (228 mg, 6.0 mmol) at room temperature and the mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated to dryness. The resulting residue was treated with saturated NaHCO.sub.3 aq., extracted with EtOAc (20 mL3). The combined layers were washed with brine (20 mL3), dried over Na.sub.2SO.sub.4 and concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (360 mg, 50%). MS: M/e 181 (M+1).sup.+.
Step B: 2-(6-((2R,5R)-4-(1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2-(methoxymethyl)-5-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00126##
[0310] A solution of 2-(6-((2R,5R)-2-(methoxymethyl)-5-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.145 mol), 1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (52 mg, 0.290 mol), (cyanomethyl)trimethylphosphonium iodide (70 mg. 0.290 mol) and DIPEA (56 mg, 0.435 mol) in CH.sub.3CN (1 ml). The mixture solution was degassed 3 times under N.sub.2 atmosphere. Then the mixture solution was stirred at 105 C. for 24 hours. The reaction mixture was concentrated under reduced pressure. The crude product was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled Compound A11, which was further separated into Compound A11a (3 mg) and Compound A11b (4 mg) by Prep-HPLC (Method A).
[0311] Compound A11a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.85 (s, 1H), 6.81-6.74 (m, 2H), 5.98 (d, J=53.1 Hz, 1H), 5.16 (s, 0.5H), 4.89-4.84 (m, 0.5H), 4.30-4.19 (m, 4.5H), 3.94 (s, 3H), 3.79 (s, 3H), 3.76-3.65 (m, 1.5H), 3.63-3.58 (m, 2H), 3.49-3.40 (m, 1H), 3.36-3.32 (m, 1H), 3.26 (s, 3H), 2.83-2.58 (m, 3H), 1.31 (d, J=6.3 Hz, 3H), 1.04 (d, J=6.3 Hz, 3H) ppm. MS: M/e 508 (M+1).sup.+
[0312] Compound A11b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.84 (s, 1H), 6.83-6.75 (m, 2H), 6.24 (s, 0.5H), 5.69 (s, 0.5H), 5.32 (s, 0.5H), 4.59 (s, 0.5H), 4.30-4.21 (m, 4.5H), 3.96-3.82 (m, 4H), 3.80-3.75 (m, 4H), 3.70-3.48 (m, 2H), 3.39 (s, 3H), 3.19-3.06 (m, 1.5H), 2.95 (s, 1H), 2.88-2.64 (m, 2H), 1.29 (d, J=6.4 Hz, 3H), 0.88 (d, J=6.5 Hz, 3H) ppm. MS: M/e 508 (M+1).sup.+
Compound A12: 2-(6-((2R,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2-(methoxymethyl)-5-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00127##
Step A: 3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxine-6-carbonitrile
##STR00128##
[0313] To a solution of 4-fluoro-3-hydroxybenzonitrile (100 g, 0.73 mol) and Cs.sub.2CO.sub.3 (951 g, 2.92 mol) in DMAc (1000 mL) was added 2,2-dimethyloxirane (210 g, 2.92 mol) at room temperature. The reaction mixture was stirred at 100 C. for overnight. The reaction mixture was filtered. The filtrates were diluted with EtOAc (1000 mL) and washed with H.sub.2O (1000 mL). The aqueous was extracted with EtOAc (1000 mL2). The combined organics were concentrated. The residue was triturated by H.sub.2O (500 mL) to give the crude product (120 g), which was recrystallized by n-Hexane to give the titled compound (71 g).
Step B: 3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxine-6-carbaldehyde
##STR00129##
[0314] To a solution of 3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxine-6-carbonitrile (71 g, 0.37 mol) in THE (700 mL) was added DIBAL-H (550 mL, 0.55 mol, 1M in n-Hexane) slowly at 0 C. The reaction mixture was stirred at 0 C. for 2 hours. The reaction was monitored by HPLC (quenched by 3N HCl). After completely disappearance of 3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxine-6-carbonitrile, the reaction was quenched by 2N HCl slowly at 0 C. The resulting mixture was filtered. The filtrates were concentrated. The residue was dissolved in EtOAc (1000 mL) and washed with 1N HCl (1000 mL2). The organic phases were concentrated to dryness. The crude product was purification by flash column chromatography to give the titled compound (64 g).
Step C: 1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol
##STR00130##
[0315] To a solution of 3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxine-6-carbaldehyde (31 g, 0.16 mol) in THE (300 mL) was added MeMgBr (106 mL, 0.32 mmol, 3M in Et.sub.2O) slowly at 0 C. The resulting mixture was stirred at r.t for 2 hours. The reaction mixture was quenched with H.sub.2O slowly at 0 C., then filtered. The filtrates were extracted with EtOAc (500 mL2). The combined organic phases were concentrated to give the crude product (30 g, contained 1.9% BP 1). The crude product was dissolved in DCM (300 mL), TEA (29.1 g, 2 eq), TrtCl (16 g, 0.4 eq) were added successively. The reaction was stirred at r.t for 2 hours. The reaction was monitored by LCMS. After completely disappearance of BP 1, the reaction was diluted with DCM (300 mL) and washed with H.sub.2O (300 mL). The organic phases were concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (26 g).
Step D: 2-(6-((2R,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2-(methoxymethyl)-5-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00131##
[0316] A solution of 2-(6-((2R,5R)-2-(methoxymethyl)-5-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (250 mg, 0.725 mol), 1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (302 mg, 1.449 mol), (cyanomethyl)trimethylphosphonium iodide (352 mg. 1.449 mol) and DIPEA (281 mg, 2.175 mol) in CH.sub.3CN (3 ml). The mixture solution was degassed 3 times under N.sub.2 atmosphere. Then the mixture solution was stirred at 105 C. for 24 hours. The reaction mixture was concentrated under reduced pressure. The crude product was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled Compound A12 (240 mg), which was further separated into Compound A12a (70 mg) and Compound A12b (68 mg) by chiral Prep-HPLC. The chiral separation conditions are shown below.
TABLE-US-00004 Column CHIRALPAK ID Column Size 2 cm 25 cm, 5 um Mobile Phase A Hex:DCM(0.5% 2M NH3MeOH) Mobile Phase B MeOH Flow Rate 20 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-HPLC Equipment Prep-HPLC-Gilson
[0317] Compound A12a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.84-6.73 (m, 3H), 6.24 (s, 0.5H), 5.67-5.63 (m, 0.5H), 5.35-5.31 (m, 0.5H), 4.63-4.59 (m, 0.5H), 3.93 (s, 3H), 3.88 (s, 3H), 3.80-3.75 (m, 4H), 3.50-3.43 (m, 1H), 3.39 (s, 3H), 3.30 (s, 2H), 3.15-3.07 (m, 2H), 2.90 (s, 1H), 2.83-2.75 (m, 1H), 1.32 (s, 6H), 1.25 (d, J=6.4 Hz, 3H), 0.85 (d, J=6.5 Hz, 3H) ppm. MS: M/e 536 (M+1).sup.+
[0318] Compound A12b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.81-6.72 (m, 3H), 6.03 (s, 0.5H), 5.89 (d, J=13.2 Hz, 0.5H), 5.13 (s, 0.5H), 4.84-4.79 (m, 0.5H), 3.93 (s, 3H), 3.87 (s, 2H), 3.78 (s, 3H), 3.75-3.68 (m, 1H), 3.66-3.56 (m, 2H), 3.54-3.49 (m, 1H), 3.40-3.30 (m, 4H), 3.29-3.18 (m, 3H), 2.60-2.40 (m, 1H), 1.31 (s, 6H), 1.27 (d, J=6.4 Hz, 3H), 0.99 (d, J=6.2 Hz, 3H) ppm. MS: M/e 536 (M+1).sup.+
Compound A13: 2-(6-((2R,5R)-2-(methoxymethyl)-5-methyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00132##
Step A: 7-bromo-2-methylquinoxaline
##STR00133##
[0319] 7-bromo-2-chloroquinoxaline (243 g, 1 mol) and Ferric acetylacetonate (17.63 g, 50 mmol) were dissolved in dry THE (1.5 L). A methyl magnesium bromide (3M in ethyl ether) solution (370 ml, 1.1 mol) was added dropwise at 0 C. After stirred for 1 hour, the reaction mixture was quenched with 1M aqueous HCl solution (1 L). The organic was removed under reduced pressure. DCM (1 L) was added to dilute the solid and filtered to separate the undissolved substance. The organic layer was washed with brine, dried over MgSO.sub.4, filtered and the solvent was removed under vacuum. The residue (230 g) was slurried from ethanol (920 ml) for 2 hours and filtered. The solid (150 g) was dissolved in EtOAc (2 L) and decolorized with active carbon (70 g). The mixture was filtered and concentrated to give the titled compound (120 g, 54%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) 8.89 (s, 1H), 8.22 (d, J=2.0 Hz, 1H), 8.00 (d, J=8.9 Hz, 1H), 7.91 (dd, J=8.9, 2.0 Hz, 1H), 2.72 (s, 3H) ppm. MS: M/e 223 (M+1).sup.+.
Step B: 1-(3-methylquinoxalin-6-yl)ethan-1-one
##STR00134##
[0320] To a solution of 7-bromo-2-methylquinoxaline (66 g, 0.3 mol) and Cs.sub.2CO.sub.3 (147 g, 0.45 mol) in DMF/H.sub.2O (8/1, 1.3 L) was added 1-(vinyloxy)butane (240 g, 2.4 mol), Pd(OAc).sub.2 (5.4 g, 0.024 mol) and DPPP (29.6 g, 0.072 mol). The mixture was stirred at 116 C. under N.sub.2 for 16 h. The reaction was cooled to 010 C., treated with 2N HCl (600 mL) drop wise, stirred at 010 C. for one hour. The mixture was diluted with water (500 mL), a suspension was formed. The mixture was filtered through a pad of Celtic, washed with water (500 mL). The filtrate was extracted with EtOAc (2 L), washed with brine, dried over Na.sub.2SO.sub.4, filtered, concentrated. The water layer and brine layer were combined, extracted with DCM (1 L2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, concentrated to dryness. The combined residue was purified by flash column chromatography to give titled compound (35 g, 62%). .sup.1HNMR (400 MHz, CDCl.sub.3) 8.83 (s, 1H), 8.60 (d, J=1.6 Hz, 1H), 8.28 (dd, J=1.6 Hz, 8.8 Hz, 1H), 8.13 (d, J=9.2 Hz, 1H), 2.82 (s, 3H), 2.76 (s, 3H) ppm. MS: M/e 187 (M+1).sup.+.
Step C: 1-(3-methylquinoxalin-6-yl)ethan-1-ol
##STR00135##
[0321] To a solution of 1-(3-methylquinoxalin-6-yl)ethan-1-one (55.4 g, 298 mmol) in EtOH (500 ml) at 0 C. was added NaBH.sub.4 (6.8 g, 179 mmol) in some portions. After addition, the reaction was stirred at 0 C. for 1 hour. The reaction was quenched by adding water, most of EtOH was concentrated off under reduced pressure. The residue was diluted with water, extracted with DCM (500 mL3), washed with brine, dried over Na.sub.2SO.sub.4, filtered, concentrated. The resulting residue was purified by flash column chromatography to give the titled compound (47.6 g, 85%). .sup.1H NMR (400 MHz, CDCl.sub.3) 8.71 (s, 1H), 8.04 (d, J=8.8 Hz, 1H), 7.98 (d, J=0.8 Hz, 1H), 7.75 (dd, J=1.6 Hz, 8.4 Hz, 1H), 5.17-5.09 (m, 1H), 2.77 (s, 3H), 1.60 (d, J=6.8 Hz, 3H) ppm. MS: M/e 189 (M+1).sup.+.
Step D: 2-(6-((2R,5R)-2-(methoxymethyl)-5-methyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00136##
[0322] A solution of 2-(6-((2R,5R)-2-(methoxymethyl)-5-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (43 mg, 0.125 mol), 1-(3-methylquinoxalin-6-yl)ethan-1-ol (47 mg, 0.249 mol), (cyanomethyl)trimethylphosphonium iodide (61 mg, 0.249 mol) and DIPEA (48 mg, 0.375 mol) in CH.sub.3CN (1 ml). The mixture solution was degassed 3 times under N.sub.2 atmosphere. Then the mixture solution was stirred at 105 C. for 24 hours. The reaction mixture was concentrated under reduced pressure to give the titled Compound A13 (crude), which was further separated into Compound A13a (6 mg) and Compound A13b (4 mg) by Prep-HPLC(Method A).
[0323] Compound A13a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.78 (s, 1H), 8.03 (d, J=8.6 Hz, 1H), 7.94 (s, 1H), 7.89 (dd, J=8.7, 1.5 Hz, 1H), 6.16-5.85 (m, 1H), 5.13 (s, 0.5H), 4.96-4.91 (m, 0.5H), 4.19 (d, J=25.6 Hz, 1H), 3.94 (s, 3H), 3.79 (s, 3H), 3.76 (d, J=6.6 Hz, 1H), 3.72-3.63 (m, 3.5H), 3.45-3.39 (m, 0.5H), 3.19 (s, 3H), 2.83-2.77 (m, 0.5H), 2.76 (s, 3H), 2.66-2.59 (m, 1.5H), 2.45 (d, J=12.1 Hz, 1H), 1.43 (d, J=6.5 Hz, 3H), 1.09 (d, J=6.5 Hz, 3H) ppm. MS: M/e 516 (M+1).sup.+.
[0324] Compound A13b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.79 (s, 1H), 8.05 (d, J=8.6 Hz, 1H), 7.96 (s, 1H), 7.90 (dd, J=8.7, 1.6 Hz, 1H), 6.31 (s, 0.5H), 5.68-5.62 (m, 0.5H), 5.40 (s, 0.5H), 4.63-4.59 (m, 0.5H), 4.19 (d, J=33.4 Hz, 1H), 4.08-3.86 (m, 5H), 3.89-3.78 (m, 1H), 3.82 (s, 3H), 3.56-3.48 (m, 0.5H), 3.41 (s, 3H), 3.23-3.14 (m, 1.5H), 2.95-2.82 (m, 2.5H), 2.77-2.63 (m, 3.5H), 1.42 (d, J=6.5 Hz, 3H), 0.94 (d, J=6.5 Hz, 3H) ppm. MS: M/e 516 (M+1).sup.+.
Compound A14: 2-(6-fluoro-7-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00137##
Step A: tert-butyl(2R,5S)-4-(6-fluoro-2-formyl-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00138##
[0325] To a solution of tert-butyl (2R,5S)-4-(2-formyl-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (5.5 g, 13.7 mmol) in THE (30 mL) were added 1-Chloromethyl-4-fluoro-1,4-diazoniabicyclo[2.2.2]octane bis(tetrafluoroborate) (7.2 g, 20.5 mmol). The resulting mixture was stirred at 40 C. overnight. The reaction solvent was collected by filtration and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography to give the titled compound (2.5 g, 43%). MS: M/e 422 (M+1).sup.+.
Step B: tert-butyl(2R,5S)-4-(6-fluoro-2-(hydroxymethyl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00139##
[0326] To a solution of tert-butyl(2R,5S)-4-(6-fluoro-2-formyl-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (3 g, 7.1 mmol) in MeOH (10 mL) were added NaBH.sub.4 (270 mg, 7.1 mmol). The resulting mixture was stirred at 0 C. for 5 min. The reaction solvent was diluted with DCM and washed with water, the organic layer was concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (2.4 g, 73%). MS: M/e 424 (M+1).sup.+.
Step C: tert-butyl(2R,5S)-4-(2-(chloromethyl)-6-fluoro-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00140##
[0327] To a solution of SOCl.sub.2 (1.4 g, 11.8 mmol) in DCM (600 mL) were added tert-butyl(2R,5S)-4-(6-fluoro-2-(hydroxymethyl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (1 g, 2.36 mmol). The resulting mixture was stirred at RT for 10 min. The reaction solvent was washed with water, aq. NaHCO.sub.3 and brine. The organic layer was concentrated to give the titled compound which was used to next step directly without further purification.
Step D: tert-butyl(2R,5S)-4-(2-(cyanomethyl)-6-fluoro-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00141##
[0328] To the solution of tert-butyl(2R,5S)-4-(2-(chloromethyl)-6-fluoro-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (crude) in CH.sub.3CN (30 mL) were added TMSCN (1.17 g, 11.8 mmol) Cs.sub.2CO.sub.3 (3.8 g, 11.8 mmol). The resulting mixture was stirred at 60 C. for 30 min. The reaction solvent was removed under vacuum. The crude product was purified by flash column chromatography to give the titled compound (400 mg). MS: M/e 433 (M+1).sup.+.
Step E: 2-(7-((2S,5R)-2,5-dimethylpiperazin-1-yl)-6-fluoro-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00142##
[0329] To a solution of tert-butyl(2R,5S)-4-(2-(cyanomethyl)-6-fluoro-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (400 mg, 0.93 mmol) in DCM (10 mL) were added TFA (3 mL). The resulting mixture was stirred at rt for 2 hours. The reaction solvent was removed under vacuum. The crude product was dissolved with water and adjusted to pH=9 with aq. NaHCO.sub.3. The aqueous layer was extracted with (DCM/IPA=3/1) to give the titled compound (400 mg, crude). MS: M/e 333 (M+1).sup.+.
Step F: 2-(6-fluoro-7-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00143##
[0330] To a solution of 2-(7-((2S,5R)-2,5-dimethylpiperazin-1-yl)-6-fluoro-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (30 mg, 0.09 mmol) in CH.sub.3CN (3 mL) and was added 1-(4-fluoro-2-(trifluoromethyl)phenyl)ethan-1-ol (57 mg, 0.27 mmol), (cyanomethyl)trimethylphosphonium iodide (110 mg, 0.45 mmol) and DIPEA (117 mg, 0.9 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure. The resulting residue was purified by Prep-TLC (DCM:MeOH=20:1) to give the titled Compound 14, which was further separated into Compound A14a (2 mg) and Compound A14b (1 mg) by Prep-HPLC(Method A).
[0331] Compound A14a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.10-8.02 (m, 1H), 7.39 (t, J=8.2 Hz, 2H), 4.66 (br s, 1H), 4.23-4.09 (m, 2H), 4.02-3.93 (m, 5H), 3.89 (s, 3H), 3.56 (s, 1H), 2.95-2.65 (m, 2H), 1.95 (d, J=12.2 Hz, 1H), 1.31 (d, J=6.4 Hz, 3H), 1.26 (d, J=6.7 Hz, 3H), 1.15 (d, J=6.5 Hz, 3H) ppm. MS: M/e 523 (M+1).sup.+.
[0332] Compound A14b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.17-8.06 (m, 1H), 7.47-7.35 (m, 2H), 4.95 (br s, 1H), 4.24-3.95 (m, 2H), 3.98 (s, 3H), 3.89 (s, 3H), 3.80 (q, J=13.1 Hz, 2H), 3.17-3.07 (m, 1H), 2.89 (d, J=10.9 Hz, 1H), 2.75-2.66 (m, 2H), 1.52 (d, J=6.6 Hz, 3H), 1.25 (d, J=6.4 Hz, 3H), 1.00 (d, J=6.5 Hz, 3H) ppm. MS: M/e 523 (M+1).sup.+.
Compound A16: 2-(6-fluoro-7-((2S,5R)-4-(1-(4-fluoro-2-methoxyphenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00144##
[0333] To a solution of 2-(7-((2S,5R)-2,5-dimethylpiperazin-1-yl)-6-fluoro-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (30 mg, 0.09 mmol) in CH.sub.3CN (3 mL) and was added 1-(4-fluoro-2-methoxyphenyl)ethan-1-ol (46 mg, 0.27 mmol), (cyanomethyl)trimethylphosphonium iodide (110 mg, 0.45 mmol) and DIPEA (117 mg, 0.9 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure. The resulting residue was purified by Prep-HPLC (Method A) to give the titled compound (4 mg, 8%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.56-7.42 (m, 1H), 6.95-6.67 (m, 2H), 4.68-4.42 (m, 1H), 4.37 (d, J=3.8 Hz, 2H), 4.11-3.90 (m, 1H), 3.80 (s, 3H), 3.80 (d, J=2.9 Hz, 3H), 3.76 (s, 3H), 3.68-3.51 (m, 1H), 3.36-3.34 (m, 1H), 2.96-2.55 (m, 2H), 2.07 (d, J=11.4 Hz, 1H), 1.30 (d, J=6.4 Hz, 2H), 1.24-1.12 (m, 4H), 1.04 (d, J=6.4 Hz, 1H), 0.91 (d, J=6.4 Hz, 2H) ppm. MS: M/e 485 (M+1).sup.+.
Compound A18: 2-(7-((2S,5R)-4-(1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-6-fluoro-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00145##
[0334] To a solution of 2-(7-((2S,5R)-2,5-dimethylpiperazin-1-yl)-6-fluoro-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (30 mg, 0.09 mmol) in CH.sub.3CN (3 mL) and was added 1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (49 mg, 0.27 mmol), (cyanomethyl)trimethylphosphonium iodide (110 mg, 0.45 mmol) and DIPEA (117 mg, 0.9 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure to give the titled Compound A18, which was further separated into Compound A18a (1 mg) and Compound A18b (2 mg) by Prep-HPLC (Method A).
[0335] Compound A18a (the earlier peak): .sup.1H NMR (400 MHz, DMSO-d.sub.6) 6.82 (d, J=7.1 Hz, 1H), 6.81 (s, 2H), 4.56 (s, 1H), 4.38 (s, 2H), 4.23 (s, 4H), 3.88 (s, 3H), 3.76 (s, 3H), 3.64-3.42 (m, 3H), 2.91 (d, J=8.6 Hz, 1H), 2.68 (s, 1H), 2.58-2.56 (m, 1H), 1.26 (d, J=6.4 Hz, 3H), 1.21 (d, J=6.5 Hz, 3H), 0.92 (d, J=6.4 Hz, 3H) ppm. MS: M/e 495 (M+1).sup.+.
[0336] Compound A18b (the later peak): .sup.1H NMR (400 MHz, DMSO-d.sub.6) 6.84 (s, 1H), 6.79 (t, J=8.3 Hz, 2H), 4.40 (s, 1H), 4.37 (s, 2H), 4.21 (s, 4H), 3.87 (s, 3H), 3.76 (s, 5H), 3.44 (d, J=6.6 Hz, 1H), 3.33-3.31 (m, 1H), 2.64 (d, J=8.0 Hz, 1H), 2.10 (d, J=9.1 Hz, 1H), 1.20 (d, J=6.3 Hz, 3H), 1.13 (d, J=6.4 Hz, 3H), 1.03 (d, J=6.4 Hz, 3H) ppm. MS: M/e 495 (M+1).sup.+.
Compound A19: 2-(7-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-6-fluoro-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00146##
[0337] A solution of 2-(7-((2S,5R)-2,5-dimethylpiperazin-1-yl)-6-fluoro-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (250 mg, 0.753 mol), 1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (313 mg, 1.506 mol), (cyanomethyl)trimethylphosphonium iodide (366 mg. 1.506 mol) and DIPEA (292 mg, 2.259 mol) in CH.sub.3CN (3 ml). The mixture solution was degassed 3 times under N.sub.2 atmosphere. Then the mixture solution was stirred at 105 C. for 24 hours. The reaction mixture was concentrated under reduced pressure. The crude product was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled Compound A19 (60 mg), which was further separated into Compound A19a (18 mg) and Compound A19b (18 mg) by chiral Prep-HPLC. The chiral separation conditions are shown below.
TABLE-US-00005 Column CHIRALPAK IH Column Size 2 cm 25 cm, 5 um Mobile Phase A MtBE Mobile Phase B MeOH:DCM Flow Rate 20 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-HPLC Equipment Prep-HPLC-Gilson
[0338] Compound A19a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.83-6.77 (m, 3H), 4.67 (s, 1H), 4.58 (s, 1H), 3.98 (s, 3H), 3.93 (s, 3H), 3.89 (s, 2H), 3.72-3.61 (m, 3H), 3.04 (dd, J=11.5, 3.3 Hz, 1H), 2.85-2.75 (m, 1H), 2.62 (dd, J=11.5, 3.7 Hz, 1H), 1.36 (d, J=6.4 Hz, 3H), 1.33 (s, 6H), 1.30 (d, J=6.5 Hz, 4H), 1.01 (d, J=6.4 Hz, 3H) ppm. MS: M/e 523 (M+1).sup.+.
[0339] Compound A19b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.82 (d, J=6.6 Hz, 2H), 6.77 (d, J=8.8 Hz, 1H), 4.60-4.53 (m, 2H), 4.06-3.95 (m, 1H), 3.98 (s, 3H), 3.93-3.89 (m, 1H), 3.90 (s, 3H), 3.87 (s, 2H), 3.5-3.39 (m, 2H), 2.76 (dd, J=11.7, 3.2 Hz, 1H), 2.21 (d, J=11.3 Hz, 1H), 1.31 (d, J=2.7 Hz, 6H), 1.28 (d, J=6.5 Hz, 4H), 1.24 (d, J=6.5 Hz, 3H), 1.11 (d, J=6.4 Hz, 3H) ppm. MS: M/e 523 (M+1).sup.+.
Compound A20: 2-(7-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-ethyl-6-fluoro-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00147##
Step A: tert-butyl (2R,5S)-4-(5-chloro-3-ethyl-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00148##
[0340] To a solution of tert-butyl (2R,5S)-4-(5-chloro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (49 g, 134.2 mmol) and K.sub.2CO.sub.3 (55.6 g, 403 mmol) in acetone (300 mL) was added CH.sub.3CH.sub.2I (41.9 g, 268.4 mmol). The reaction was stirred at room temperature for 16 hours. After filtration, the reaction mixture was diluted with water, extracted with EtOAc (300 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (PE:EtOAc=1:1) to give the titled compound (23.5 g, 45%). MS: M/e 394 (M+1).sup.+.
Step B: tert-butyl (2R,5S)-4-(3-ethyl-5-hydroxy-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00149##
[0341] To a solution of tert-butyl (2R,5S)-4-(5-chloro-3-ethyl-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (23.5 g, 60 mmol), Pd.sub.2(dba).sub.3 (5.47 g, 6 mmol), t-BuXphos (5.11 g, 12 mmol) and KOH (60 mL, 3M, 180 mmol) in dioxane (120 mL) was degassed 3 times under N.sub.2 atmosphere. Then the mixture was stirred at 90 C. for 12 hours. The reaction mixture was diluted with water and extracted with EtOAc (150 mL2). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, filtered and concentrated to dryness. The resulting residue was purified by flash column chromatography (DCM:MeOH=10:1) to give the titled compound (15.6 g, 69%). MS: M/e 376 (M+1).sup.+.
Step C: tert-butyl (2R,5S)-4-(3-ethyl-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00150##
[0342] To a solution of tert-butyl (2R,5S)-4-(3-ethyl-5-hydroxy-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (7.5 g, 20 mmol) and Cs.sub.2CO.sub.3 (22.8 g, 60 mmol) in dioxane (210 mL) was added CH.sub.3I (28.4 g, 200 mmol). The reaction was stirred at room temperature for 16 hours. After filtration, the reaction mixture was diluted with water, extracted with EtOAc (150 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (DCM:MeOH=20:1) to give the titled compound (5 g, 64%). MS: M/e 390 (M+1).sup.+.
Step D: tert-butyl (2R,5S)-4-(3-ethyl-2-formyl-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00151##
[0343] To a solution of tert-butyl (2R,5S)-4-(3-ethyl-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (5.0 g, 12.853 mmol) in THE (150 mL) was degassed 3 times under N.sub.2 atmosphere. Then added LDA (19.3 mL, 2M, 38.560 mmol) dropwise at 78 C. The mixture was stirred at 78 C. for 1 hours. Added DMF (9.39 g, 128.53 mmol) to quench the reaction solution. The reaction mixture was stirred for 1 hour and diluted with saturated NH.sub.4Cl, extracted with EtOAc (150 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (DCM:MeOH=20:1) to give the titled compound (5.0 g, 93%). MS: M/e 418 (M+1).sup.+.
Step E: tert-butyl (2R,5S)-4-(3-ethyl-6-fluoro-2-formyl-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00152##
[0344] To a solution of tert-butyl (2R,5S)-4-(3-ethyl-2-formyl-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (1.0 g, 2.40 mmol) in THE (60 mL) was added Selectfluor (2.55 g, 7.20 mmol). The reaction was stirred at 60 C. for 6 hours. After filtration, the reaction mixture was concentrated to dryness. The resulting residue was purified by flash column chromatography (PE:EtOAc=1:1) to give the titled compound (450 mg, 43%). MS: M/e 436 (M+1).sup.+.
Step F: tert-butyl (2R,5S)-4-(3-ethyl-6-fluoro-2-(hydroxymethyl)-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00153##
[0345] To a solution of tert-butyl (2R,5S)-4-(3-ethyl-6-fluoro-2-formyl-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (450 mg, 1.034 mmol) in MeOH (10 mL) was added NaBH.sub.4 (39 mg, 1.034 mmol). The reaction was stirred at room temperature for 1 hours. The reaction mixture was quenched with saturated NH.sub.4Cl, extracted with EtOAc (30 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (DCM:MeOH=10:1) to give the titled compound (350 mg, 77%). MS: M/e 438 (M+1).sup.+.
Step G: tert-butyl (2R,5S)-4-(2-(chloromethyl)-3-ethyl-6-fluoro-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00154##
[0346] To a solution of tert-butyl (2R,5S)-4-(3-ethyl-6-fluoro-2-(hydroxymethyl)-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (350 mg, 0.801 mmol) in DCM (20 mL) was added SOCl.sub.2 (100 mg, 0.841 mmol). The reaction was stirred at room temperature for 0.5 hours. The reaction mixture concentrated to give the titled compound (350 mg, 96%). MS: M/e 456 (M+1).sup.+.
Step H: tert-butyl (2R,5S)-4-(2-(cyanomethyl)-3-ethyl-6-fluoro-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00155##
[0347] To a solution of tert-butyl (2R,5S)-4-(2-(chloromethyl)-3-ethyl-6-fluoro-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (350 mg, 0.769 mmol) in DCM (20 mL) was added TBAF (1.54 mL, 1M, 1.538 mmol), followed TMSCN (153 mg, 1.538 mmol). The reaction was stirred at room temperature for 16 hours. The reaction mixture was diluted with water, extracted with DCM (40 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (PE:EtOAc=1:5) to give the titled compound (300 mg, 87%). MS: M/e 447 (M+1).sup.+.
Step I: 2-(7-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3-ethyl-6-fluoro-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00156##
[0348] To a stirred solution of tert-butyl (2R,5S)-4-(2-(cyanomethyl)-3-ethyl-6-fluoro-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (300 mg, 0.673 mmol) in CH.sub.2Cl.sub.2 (20 mL) was added TFA (5 mL). Then the reaction mixture was stirred at room temperature for 4 hours, concentrated to give a residue, basified to pH=1011 with saturated NaHCO.sub.3 aq., and extracted with CH.sub.2Cl.sub.2/IPA (3/1, 50 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, and concentrated to give the titled compound (180 mg, 78%). MS: M/e 347 (M+1).sup.+.
Step J: 2-(7-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-ethyl-6-fluoro-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00157##
[0349] A solution of 2-(7-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3-ethyl-6-fluoro-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (180 mg, 0.520 mmol), 1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (216 mg, 1.040 mmol), (cyanomethyl) trimethyl phosphonium iodide (253 mg. 1.040 mmol) and DIPEA (201 mg, 1.560 mol) in CH.sub.3CN (2 ml). The mixture solution was degassed 3 times under N.sub.2 atmosphere. Then the mixture solution was stirred at 105 C. for 24 hours. The reaction mixture was concentrated under reduced pressure. The crude product was purified by Prep-TLC (DCM:MeOH=15:1) and Prep-HPLC(Method A) to give the titled compound (16 mg, 6%). .sup.1H NMR (400 MHz, CD.sub.3OD) 6.88-6.84 (m, 3H), 4.59 (s, 1H), 4.42 (d, J=7.2 Hz, 2H), 4.26-4.21 (m, 0.5H), 4.08-3.99 (m, 0.5H), 3.89 (d, J=11.0 Hz, 6H), 3.76-3.49 (m, 2H), 3.13-3.04 (m, 2H), 3.03-2.31 (m, 2H), 1.44 (dd, J=17.7, 10.6 Hz, 6H), 1.33 (d, J=5.8 Hz, 8H), 1.21 (t, J=17.9 Hz, 4H) ppm. MS: M/e 537 (M+1).sup.+
Compound A21: 2-(7-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-ethyl-6-fluoro-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00158##
Step A: 2-chloro-6-(hydroxymethyl)pyridin-3-ol
##STR00159##
[0350] A solution of 2-chloropyridin-3-ol (2 g, 15.50 mmol), formaldehyde (37%, 12.6 g, 155 mmol), NaHCO.sub.3 (3.9 g, 46.5 mmol) in water (50 mL) was stirred at 90 C. overnight. The reaction mixture was quenched with con. HCl and extracted with CH.sub.2Cl.sub.2/IPA (3/1, 10 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated to give the titled compound (2.1 g, 85%) as colorless oil. MS: M/e 160 (M+1).sup.+.
Step B: 1-((2-chloro-6-(hydroxymethyl)pyridin-3-yl)oxy)propan-2-one
##STR00160##
[0351] A solution of 2-chloro-6-(hydroxymethyl)pyridin-3-ol (2.1 g, 13.21 mmol), 1-chloropropan-2-one (1.46 g, 15.85 mmol), KI (219 mg, 1.32 mmol) and K.sub.2CO.sub.3 (3.65 g, 26.42 mmol) in ACN (100 mL) was stirred at 60 C. for 1 h. The mixture was diluted with CH.sub.2Cl.sub.2 (20 mL), then washed with H.sub.2O, brine, dried over Na.sub.2SO.sub.4, concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (2.7 g, 95%) as colorless oil. MS: M/e 216 (M+1).sup.+.
Step C: 6-chloro-5-(2-oxopropoxy)picolinaldehyde
##STR00161##
[0352] A mixture of 1-((2-chloro-6-(hydroxymethyl)pyridin-3-yl)oxy)propan-2-one (2.7 g, 12.56 mmol) and Dess-Martin Periodinane (6.4 g, 15.07 mmol) in DCM (30 mL) was stirred at r.t for 3 h. The reaction mixture was diluted with DCM and washed with water, concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (2.6 g, 97%). MS: M/e 214 (M+1).sup.+.
Step D: 1-((2-chloro-6-(1-hydroxyethyl)pyridin-3-yl)oxy)-2-methylpropan-2-ol
##STR00162##
[0353] To a stirred solution of 6-chloro-5-(2-oxopropoxy)picolinaldehyde (2.6 g, 12.21 mmol) in dry THE (200 mL) was added dropwise MeMgBr (1.0 M, 30.5 mL, 30.5 mmol) at 0 C. After then, the mixture was stirred for half an hour. The reaction was quenched with aq.NH.sub.4Cl, extracted with EtOAc (100 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (2.6 g, 87%) as colorless oil. MS: M/e 246 (M+1).sup.+.
Step E: 1-(3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethan-1-ol
##STR00163##
[0354] A mixture of 1-((2-chloro-6-(1-hydroxyethyl)pyridin-3-yl)oxy)-2-methylpropan-2-ol (1 g, 4.08 mmol), Pd(OAc).sub.2 (91 mg, 0.41 mmol) BINAP (254 mg, 0.41 mmol) and Cs.sub.2CO.sub.3 (2.66 g, 8.16 mmol) in toluene (30 mL) was stirred at 100 C. under N.sub.2 overnight. The reaction mixture was extracted with EtOAc (100 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (550 mg, 64%) as colorless oil. MS: M/e 210 (M+1).sup.+.
Step F: 2-(7-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-ethyl-6-fluoro-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00164##
[0355] A solution of 2-(7-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3-ethyl-6-fluoro-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (120 mg, 0.347 mmol), 1-(3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethan-1-ol (218 mg, 1.040 mmol), (cyanomethyl)trimethylphosphonium iodide (337 mg. 1.388 mmol) and DIPEA (224 mg, 1.735 mol) in CH.sub.3CN (2 ml). The mixture solution was degassed 3 times under N.sub.2 atmosphere. Then the mixture solution was stirred at 105 C. for 24 hours. The reaction mixture was concentrated under reduced pressure. The crude product was purified by Prep-TLC (DCM:MeOH=15:1) and Prep-HPLC(Method A) to give the titled compound (11 mg, 6%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.33 (t, J=6.1 Hz, 1H), 7.12 (t, J=8.0 Hz, 1H), 4.65 (d, J=34.1 Hz, 1H), 4.43 (dd, J=13.1, 6.0 Hz, 2H), 4.26-4.05 (m, 2H), 4.03-3.97 (m, 0.5H), 3.98 (s, 2H), 3.89 (s, 3H), 3.79-3.62 (m, 2H), 3.38-3.34 (m, 1.5H), 3.23-3.00 (m, 1H), 2.88-2.50 (m, 1H), 1.46 (t, J=7.3 Hz, 6H), 1.41 (s, 6H), 1.32 (d, J=6.4 Hz, 4H), 1.21 (d, J=6.3 Hz, 2H) ppm. MS: M/e 538 (M+1).sup.+
Compound A22: 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(quinoxalin-6-yl)ethyl)piperazin-1-yl)-6-fluoro-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00165##
[0356] To a solution of 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(quinoxalin-6-yl)ethyl)piperazin-1-yl)-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (20 mg, 0.0.04 mmol) in CH.sub.3CN (0.2 mL) and THE (2 mL) and was added 1-Chloromethyl-4-fluoro-1,4-diazoniabicyclo[2.2.2]octane bis(tetrafluoroborate) (23 mg, 0.06 mmol). The resulting mixture was stirred for 10 min at rt. The reaction solvent was removed under reduced pressure. The resulting residue was purified by Prep-TLC to give the titled Compound A22, which was further separated into Compound A22a (0.63 mg) and Compound A22b (0.3 mg, crude) by Prep-HPLC(Method A).
[0357] Compound A22a (the earlier peak): .sup.1H NMR (400 MHz, DMSO-d.sub.6) 8.93 (t, J=4.2 Hz, 2H), 8.18-7.90 (m, 3H), 4.37 (s, 2H), 4.01 (d, J=6.5 Hz, 1H), 3.93-3.83 (m, 4H), 3.76 (s, 3H), 3.69-3.61 (m, 1H), 3.51 (d, J=13.5 Hz, 1H), 3.06-2.93 (m, 2H), 2.68 (d, J=7.9 Hz, 1H), 1.42-1.29 (m, 5H), 1.20-1.08 (m, 2H), 0.97 (t, J=11.8 Hz, 2H) ppm. MS: M/e 489 (M+1).sup.+.
Compound A23: 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-6-fluoro-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00166##
[0358] A solution of 2-(7-((2S,5R)-2,5-dimethylpiperazin-1-yl)-6-fluoro-3,4-dimethyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (40 mg, 0.093 mol), 1-(3-methylquinoxalin-6-yl)ethan-1-ol (35 mg, 0.185 mol), (cyanomethyl)trimethylphosphonium iodide (45 mg. 0.185 mol) and DIPEA (36 mg, 0.279 mol) in CH.sub.3CN (1 ml). The mixture solution was degassed 3 times under N.sub.2 atmosphere. Then the mixture solution was stirred at 105 C. for 24 hours. The reaction mixture was concentrated under reduced pressure. The crude product was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled Compound A23, which was further separated into Compound A23a (7 mg) and Compound A23b (4 mg) by Prep-HPLC(Method A).
[0359] Compound A23a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.79 (s, 1H), 8.05 (d, J=8.6 Hz, 1H), 7.98 (d, J=1.4 Hz, 1H), 7.93 (dd, J=8.7, 1.7 Hz, 1H), 4.84-4.76 (m, 2H), 4.23-4.18 (m, 1H), 4.07-3.98 (m, 1H), 3.96 (s, 3H), 3.89 (s, 3H), 3.85-3.75 (m, 2H), 3.17 (dd, J=11.5, 3.7 Hz, 1H), 2.84-2.78 (m, 2H), 2.77 (s, 3H), 1.48 (dd, J=6.5 Hz, 3H), 1.45 (dd, J=6.5 Hz, 3H), 1.07 (d, J=6.5 Hz, 3H) ppm. MS: M/e 503 (M+1).sup.+.
[0360] Compound A23b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.78 (s, 1H), 8.02 (d, J=8.6 Hz, 1H), 7.98 (s, 1H), 7.95 (dd, J=8.6, 1.7 Hz, 1H), 4.84-4.80 (m, 1H), 4.63-4.54 (m, 1H), 4.25-1.28 (m, 1H), 4.10-3.99 (m, 2H), 3.98 (s, 3H), 3.90 (s, 3H), 3.88-3.83 (m, 1H), 3.61-3.50 (m, 1H), 2.90 (dd, J=11.9, 3.8 Hz, 1H), 2.76 (s, 3H), 2.16 (dd, J=11.8, 2.5 Hz, 1H), 1.44 (d, J=6.6 Hz, 3H), 1.27 (d, J=6.5 Hz, 3H), 1.20 (d, J=6.5 Hz, 3H) ppm. MS: M/e 503 (M+1).sup.+.
Compound A24: 2-(6-((2S,5R)-4-(1-(4-fluoro-2-methoxyphenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00167##
[0361] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.159 mol), 1-(4-fluoro-2-methoxyphenyl)ethan-1-ol (40 mg, 0.238 mol), (cyanomethyl)trimethylphosphonium iodide (77 mg. 0.318 mol) and DIPEA (62 mg, 0.477 mol) in CH.sub.3CN (1 ml). The mixture solution was degassed 3 times under N.sub.2 atmosphere. Then the mixture solution was stirred at 105 C. for 24 hours. The reaction mixture was concentrated under reduced pressure. The crude product was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled Compound A24, which was further separated into Compound A24a (5 mg) and Compound A24b (3 mg) by Prep-HPLC (Method A).
[0362] Compound A24a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.56 (s, 1H), 6.90-6.64 (m, 2H), 5.99-5.94 (m, 0.5H), 5.08-5.04 (m, 0.5H), 4.85-4.41 (m, 1H), 4.29-4.03 (m, 2H), 3.93 (s, 3H), 3.83 (s, 3H), 3.79 (s, 3H), 3.70-3.58 (m, 2H), 2.95-2.64 (m, 2H), 2.56-2.20 (m, 1H), 1.43-1.35 (m, 3H), 1.30 (d, J=5.7 Hz, 3H), 1.18-1.01 (m, 3H) ppm. MS: M/e 468 (M+1).sup.+.
[0363] Compound A24b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.52 (dd, J=8.4, 7.0 Hz, 1H), 6.80 (dd, J=11.2, 2.2 Hz, 1H), 6.74 (td, J=8.4, 2.3 Hz, 1H), 6.19-5.02 (m, 2H), 4.38-3.34 (m, 1H), 4.21 (s, 1H), 3.93 (s, 3H), 3.85 (s, 3H), 3.80 (s, 3H), 3.53-3.38 (m, 1.5H), 3.12-2.91 (m, 2H), 2.92-2.70 (m, 1.5H), 1.41 (d, J=6.3 Hz, 3H), 1.33 (d, J=6.5 Hz, 3H), 1.00 (d, J=6.1 Hz, 3H) ppm. MS: M/e 468 (M+1).sup.+.
Compound A25: 2-(6-((2S,5R)-4-(1-(2-(difluoromethoxy)-4-fluorophenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00168##
[0364] A mixture of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.158 mmol), 1-(2-(difluoromethoxy)-4-fluorophenyl)ethan-1-ol (65.4 mg, 0.317 mmol), (cyanomethyl)trimethylphosphonium iodide (115 mg, 0.474 mmol) and DIPEA (204 mg, 1.58 mmol) in CH.sub.3CN (4 mL) was stirred overnight at 100 C. in a sealed tube. The reaction mixture was diluted with EtOAc (20 mL), washed with H.sub.2O, brine, dried over Na.sub.2SO.sub.4, concentrated to give the residue, which was purified by Pre-TLC (CH.sub.2Cl.sub.2/MeOH=10:1) to give the titled compound (18 mg, 23%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.81-7.65 (m, 1H), 7.13-6.68 (m, 3H), 6.04-4.45 (m, 2H), 4.21 (s, 2H), 4.16-3.97 (m, 1H), 3.93 (s, 3H), 3.78 (s, 3H), 3.65-3.46 (m, 1H), 3.04-2.60 (m, 2.5H), 2.28-2.07 (m, 0.5H), 1.50-1.22 (m, 6H), 1.13-0.85 (m, 3H) ppm. MS: M/e 504 (M+1).sup.+.
Compound A26: 2-(6-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethoxy)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00169##
[0365] A mixture of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.158 mmol), 1-(4-fluoro-2-(trifluoromethoxy)phenyl)ethan-1-ol (71 mg, 0.317 mmol), (cyanomethyl)trimethylphosphonium iodide (115 mg, 0.474 mmol) and DIPEA (204 mg, 1.58 mmol) in CH.sub.3CN (4 mL) was stirred overnight at 100 C. in a sealed tube. The reaction mixture was diluted with EtOAc (20 mL), washed with H.sub.2O, brine, dried over Na.sub.2SO.sub.4, concentrated to give the residue, which was purified by Pre-TLC (CH.sub.2Cl.sub.2/MeOH=10:1) to give the titled compound (4 mg). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.47-7.34 (m, 1H), 7.23-7.07 (m, 2H), 6.32-4.99 (m, 2H), 4.27-4.06 (m, 3H), 3.93 (s, 3H), 3.78 (s, 3H), 3.62-3.54 (m, 1H), 2.94-2.71 (m, 2H), 2.15-2.04 (m, 1H), 1.54-1.18 (m, 6H), 1.04-0.89 (m, 3H) ppm. MS: M/e 522 (M+1).sup.+.
Compound A27: 2-(6-((2S,5R)-4-(1-(4-fluoro-2-(methoxymethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00170##
[0366] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.159 mol), 1-(4-fluoro-2-(methoxymethyl)phenyl)ethan-1-ol (44 mg, 0.238 mol), (cyanomethyl)trimethylphosphonium iodide (77 mg. 0.318 mol) and DIPEA (62 mg, 0.477 mol) in CH.sub.3CN (1 ml). The mixture solution was degassed 3 times under N.sub.2 atmosphere. Then the mixture solution was stirred at 105 C. for 24 hours. The reaction mixture was concentrated under reduced pressure. The crude product was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled Compound A27, which was further separated into Compound A27a (2 mg) and Compound A27b (1.5 mg) by Prep-HPLC (Method A).
[0367] Compound A27a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.74-7.64 (m, 1H), 7.04 (dd, J=13.8, 5.8 Hz, 2H), 6.00-5.89 (m, 1H), 5.02-4.96 (m, 0.5H), 4.83-4.71 (m, 0.5H), 4.68-4.38 (m, 2H), 4.21 (s, 1H), 3.95-3.91 (m, 3H), 3.82-3.75 (m, 4H), 3.67-3.62 (m, 2H), 3.37 (s, 3H), 2.96-2.56 (m, 2H), 2.12 (d, J=11.6 Hz, 1H), 1.30 (d, J=6.5 Hz, 3H), 1.24 (d, J=6.3 Hz, 3H), 1.05 (d, J=6.5 Hz, 3H) ppm. MS: M/e 482 (M+1).sup.+.
[0368] Compound A27b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.61 (dd, J=8.6, 5.9 Hz, 1H), 7.11 (dd, J=9.9, 2.6 Hz, 1H), 7.04 (dd, J=9.7, 7.1 Hz, 1H), 6.17-6.13 (m, 0.5H), 5.65 (d, J=12.5 Hz, 0.5H), 5.25 (s, 0.5H), 4.63-4.51 (m, 2.5H), 4.22-4.17 (m, 1H), 4.06-3.91 (m, 4H), 3.78 (s, 3H), 3.55-3.49 (m, 1H), 3.40 (s, 3H), 3.15-3.10 (m, 1H), 2.92-2.75 (m, 3H), 1.45 (d, J=17.2 Hz, 3H), 1.27 (d, J=6.4 Hz, 3H), 0.88 (d, J=6.5 Hz, 3H) ppm. MS: M/e 482 (M+1).sup.+.
Compound A28: 2-(6-((2S,5R)-4-(1-(4-fluoro-2-(1-methoxyethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00171##
[0369] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.16 mmol), 1-(4-fluoro-2-(1-methoxyethyl)phenyl)ethan-1-ol (31 mg, 0.16 mmol), (cyanomethyl)trimethylphosphonium iodide (115 mg, 0.48 mmol) and DIPEA (102 mg, 0.79 mmol) in MeCN (3 ml) was stirred at 100 C. overnight. After completed, the solution was concentrated under reduced pressure to dryness. The resulting residue was purified by Prep-HPLC (Method A) to give the titled compound (12 mg, 15%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.74-7.59 (m, 1H), 7.16-6.96 (m, 2H), 6.00-4.80 (m, 3H), 4.57 (s, 3H), 3.93 (s, 3H), 3.78 (s, 3H), 3.60-3.22 (m, 2H), 2.96-2.70 (m, 4H), 2.23-1.99 (m, 1H), 1.60-1.41 (m, 6H), 1.26-1.03 (m, 3H), 0.90 (t, J=7.5 Hz, 3H) ppm. MS: M/e 496 (M+1).sup.+
Compound A29: 2-(6-((2S,5R)-4-(1-(2-(1-(difluoromethoxy)ethyl)-4-fluorophenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00172##
Step A: 1-bromo-2-(1-(difluoromethoxy)ethyl)-4-fluorobenzene
##STR00173##
[0370] To a solution of 1-(2-bromo-5-fluorophenyl)ethan-1-ol (657 mg, 3 mmol) in DCM (11.8 ml) and H.sub.2O (1.8 ml) were added (bromodifluoromethyl)trimethylsilane (1.8 g, 9 mmol) and AcOK (1.79 g, 18 mmol). The mixture was stirred at RT for 4 hours. The reaction was diluted with DCM and washed with water. The organic layer was separated, dried by Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (260 mg, 32%). MS: M/e 269 (M+1).sup.+.
Step B: 1-(2-(1-(difluoromethoxy)ethyl)-4-fluorophenyl)ethan-1-one
##STR00174##
[0371] To a solution of 1-bromo-2-(1-(difluoromethoxy)ethyl)-4-fluorobenzene (268 mg, 1 mol) in toluene (10 mL) was added tributyl(1-ethoxyvinyl)stannane (430 mg, 1.2 mol) and Pd(PPh.sub.3).sub.4 (140 mg, 0.2 mmol). The reaction mixture was protected by N.sub.2 atmosphere and stirred at 100 C. for overnight. The mixture was cooled down to rt, added HCl/Dioxane (4M, 10 ml) and stirred at room temperature for 30 mins. The mixture was concentrated in vacuo. The residue was added H.sub.2O and adjusted pH 7-8 by NaHCO.sub.3 aqueous solution. The resulting mixture was extracted with EtOAc, and then concentrated by using a rotary evaporator, to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (190 mg, 82%). MS: M/e 233 (M+1).sup.+.
Step C: 1-(2-(1-(difluoromethoxy)ethyl)-4-fluorophenyl)ethan-1-ol
##STR00175##
[0372] NaBH.sub.4 (38 mg, 1 mmol) was added to 1-(2-(1-(difluoromethoxy)ethyl)-4-fluorophenyl)ethan-1-one (190 mg, 0.8 mol) in EtOH (200 ml) at 0 C. for 1 hour. The reaction was quenched by adding water. The mixture was extracted with EtOAc and washed with brine. The organic layer was separated, dried by Na.sub.2SO.sub.4, filtered and concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (50 mg, 26%). MS: M/e 235 (M+1).sup.+.
Step D: 2-(6-((2S,5R)-4-(1-(2-(1-(difluoromethoxy)ethyl)-4-fluorophenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00176##
[0373] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (40 mg, 0.12 mmol) and (cyanomethyl)trimethylphosphonium iodide (120 mg, 1 mmol) in CH.sub.3CN (5 mL) was added DIPEA (250 mg, 2 mmol). The mixture solution was degassed 3 times under N.sub.2 atmosphere. Then the mixture solution was stirred at 105 C. for 24 hours. The reaction was quenched with saturated NH.sub.4Cl (20 mL) at room temperature. The resulting mixture was extracted with EtOAc (35 mL2). The combined organic layers were dried over Na.sub.2SO.sub.4 and concentrated under reduced pressure. The resulting residue was purified by Prep-HPLC(Method A) to give the titled Compound A29a (0.51 mg) and Compound A29b (0.83 mg).
[0374] Compound A29a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.72 (dd, J=8.8, 6.0 Hz, 1H), 7.14 (dd, J=10.3, 2.7 Hz, 1H), 7.04 (dd, J=8.5, 5.6 Hz, 1H), 6.55-6.17 (m, 1H), 5.66 (d, J=6.4 Hz, 1H), 5.34 (s, 1H), 4.56 (s, 2H), 3.94 (s, 3H), 3.84 (d, J=6.5 Hz, 1H), 3.78 (s, 3H), 3.59 (s, 1H), 2.77 (s, 1H), 2.68 (d, J=12.3 Hz, 1H), 2.21-2.15 (m, 1H), 2.03 (m, 1H), 1.50 (d, J=6.5 Hz, 3H), 1.30 (s, 3H), 1.22 (d, J=6.6 Hz, 3H), 1.07 (d, J=6.6 Hz, 3H) ppm. MS: M/e 532 (M+1).sup.+.
[0375] Compound A29b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.62-7.54 (m, 1H), 7.21 (dd, J=10.4, 2.7 Hz, 1H), 7.04 (td, J=8.4, 2.6 Hz, 1H), 6.58-6.20 (m, 1H), 5.90 (d, J=6.6 Hz, 1H), 5.65 (d, J=13.2 Hz, 1H), 5.26 (s, 1H), 4.56 (s, 2H), 3.90 (dd, J=21.3, 5.6 Hz, 4H), 3.78 (s, 3H), 3.55-3.48 (m, 1H), 2.84 (s, 2H), 2.03 (m, 1H), 1.51 (d, J=6.3 Hz, 3H), 1.30 (d, J=6.5 Hz, 6H), 0.91 (d, J=6.5 Hz, 3H) ppm. MS: M/e 532 (M+1).sup.+.
Compound A30: 2-(6-((2S,5R)-4-(1-(2-(difluoromethyl)-4-fluorophenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00177##
Step A: 1-(2-(difluoromethyl)-4-fluorophenyl)ethan-1-one
##STR00178##
[0376] A mixture of 1-bromo-2-(difluoromethyl)-4-fluorobenzene (1.12 g, 5 mmol), tributyl(1-ethoxyvinyl)stannane (2.17 g, 6 mmol) and Pd(PPh.sub.3).sub.2Cl.sub.2 (350 mg, 0.5 mmol) in toluene (10 mL) was stirred at 100 C. under N.sub.2 overnight. Then to the solution was added HCl (4 mL, 4 M in 1,4-dioxane) in drops and the mixture was stirred at room temperature for 0.5 hour. The reaction mixture was diluted with EtOAc (50 mL), treated with saturated NaHCO.sub.3 aq. to pH8, washed with brine (20 mL3), dried over Na.sub.2SO.sub.4 and concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (800 mg, 85%). .sup.1H NMR (400 MHz, CD.sub.3OD) 8.10 (dd, J=8.2, 5.9 Hz, 1H), 7.54-7.18 (m, 3H), 2.61 (d, J=6.9 Hz, 3H) ppm. MS: M/e 189 (M+1).sup.+.
Step B: 1-(2-(difluoromethyl)-4-fluorophenyl)ethan-1-ol
##STR00179##
[0377] To a solution of 1-(2-(difluoromethyl)-4-fluorophenyl)ethan-1-one (400 mg, 2.13 mmol) in MeOH (5 mL) was added NaBH.sub.4 (65 mg, 1.7 mmol) at C. and the resulting mixture was stirred at room temperature for 1 hour. The resulting residue was treated with water, extracted with EtOAc (20 mL3). The combined organic layers were washed with brine (20 mL3), dried over Na.sub.2SO.sub.4 and concentrated to dryness. The resulting residue was purified by flash column chromatography (EtOAc:PE=0-100% in 20 minutes) to give the titled compound (190 mg, 47%). .sup.1H NMR (400 MHz, DMSO-d6) 7.70-7.62 (m, 1H), 7.51-7.19 (m, 3H), 5.44 (d, J=4.0 Hz, 1H), 5.10-5.02 (m, 1H), 1.32 (d, J=6.4 Hz, 3H) ppm. MS: M/e 191 (M+1).sup.+.
Step C: 2-(6-((2S,5R)-4-(1-(2-(difluoromethyl)-4-fluorophenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00180##
[0378] A mixture of 1-(2-(difluoromethyl)-4-fluorophenyl)ethan-1-ol (57 mg, 0.3 mmol), 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3, 9-dihydro-2H-purin-8-yl)acetonitrile (63 mg, 0.2 mmol), (cyanomethyl)trimethylphosphonium iodide (97 mg, 0.4 mmol) and DIPEA (103 mg, 0.8 mmol) in MeCN (2 mL) was stirred at 100 C. for overnight in a sealed tube. The mixture was treated with water, extracted with EtOAc. The combined organic layers were washed with brine (20 mL2), dried over Na.sub.2SO.sub.4, concentrated to dryness. The resulting residue was purified by Prep-TLC (CH.sub.2Cl.sub.2:MeOH=13:1) to give the titled compound (5 mg, 5%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.81-7.10 (m, 4H), 6.29-5.22 (m, 2H), 4.22 (t, J=13.1 Hz, 1H), 3.93 (d, J=6.6 Hz, 3H), 3.86-3.76 (m, 3H), 3.67-3.50 (m, 1H), 3.35-3.20 (m, 2H), 2.92-2.69 (m, 2H), 2.09-1.98 (m, 1H), 1.48-1.22 (m, 6H), 1.05-0.87 (m, 3H) ppm. MS: M/e 488 (M+1).sup.+.
Compound A31: 2-(6-((2S,5R)-4-(1-(2-(1,1-difluoroethoxy)-4-fluorophenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00181##
[0379] A mixture of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.158 mmol), 1-(2-(1,1-difluoroethoxy)-4-fluorophenyl)ethan-1-ol (64 mg, 0.317 mmol), (cyanomethyl)trimethylphosphonium iodide (115 mg, 0.474 mmol) and DIPEA (204 mg, 1.58 mmol) in CH.sub.3CN (4 mL) was stirred overnight at 100 C. in a sealed tube. The reaction mixture was diluted with EtOAc (20 mL), washed with H.sub.2O, brine, dried over Na.sub.2SO.sub.4, concentrated to give the residue, which was purified by Pre-TLC (CH.sub.2Cl.sub.2/MeOH=10:1) to the desired compound, which was further purified by Prep-HPLC (Method A) to give the titled compound (0.5 mg). .sup.1H NMR (400 MHz, CD.sub.3OD) 8.06-7.95 (m, 1H), 7.28-7.16 (m, 2H), 6.21-5.21 (m, 2H), 4.28-4.14 (m, 1H), 4.03-3.97 (m, 1H), 3.96-3.90 (m, 3H), 3.78 (s, 3H), 3.63 (s, 1H), 2.98-2.66 (m, 4H), 2.08-1.91 (m, 3H), 1.56-1.28 (m, 3H), 1.28-1.18 (m, 3H), 1.08-0.88 (m, 3H) ppm. MS: M/e 518 (M+1).sup.+.
Compound A33: 2-(6-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00182##
[0380] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.159 mol), 1-(4-fluoro-2-(trifluoromethyl)phenyl)ethan-1-ol (66 mg, 0.317 mol), (cyanomethyl)trimethylphosphonium iodide (77 mg. 0.317 mol) and DIPEA (62 mg, 0.477 mol) in CH.sub.3CN (1 ml). The mixture solution was degassed 3 times under N.sub.2 atmosphere. Then the mixture solution was stirred at 105 C. for 24 hours. The reaction mixture was concentrated under reduced pressure. The crude product was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled Compound A33, which was further separated into Compound A33a (4 mg) and Compound A33b (3 mg) by Prep-HPLC (Method A).
[0381] Compound A33a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.10-8.00 (m, 1H), 7.40 (t, J=8.6 Hz, 2H), 5.94-5.89 (m, 1H), 5.07-5.03 (m, 0.5H), 4.86-4.82 (m, 0.5H), 4.23-4.18 (m, 1.5H), 3.93 (s, 3H), 3.92-3.87 (m, 1H), 3.79-3.74 (m, 3.5H), 3.64 (s, 1H), 3.46-3.42 (m, 0.5H), 2.73-2.64 (m, 1.5H), 1.99 (d, J=12.2 Hz, 1H), 1.32 (d, J=6.3 Hz, 3H), 1.23 (dd, J=17.0, 6.3 Hz, 3H), 1.04 (d, J=6.4 Hz, 3H) ppm. MS: M/e 506 (M+1).sup.+.
[0382] Compound A33b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.18-8.05 (m, 1H), 7.44 (t, J=8.8 Hz, 2H), 6.23-6.18 (m, 0.5H), 5.66 (d, J=13.3 Hz, 0.5H), 5.29 (s, 0.5H), 4.60 (d, J=13.1 Hz, 0.5H), 4.08-4.02 (m, 1H), 3.95 (d, J=14.6 Hz, 3H), 3.83-3.76 (m, 3.5H), 3.57-3.52 (m, 0.5H), 3.35-3.31 (m, 1.5H), 3.25-3.20 (m, 0.5H), 2.97-2.84 (m, 2H), 2.83-2.77 (m, 1H), 1.52 (dd, J=26.0, 6.6 Hz, 3H), 1.30-1.21 (m, 3H), 0.99-0.83 (m, 3H) ppm. MS: M/e 506 (M+1).sup.+.
Compound A34: 2-(6-((2S,5R)-2,5-diethyl-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl) piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00183##
Step A: 2-chloro-6-((2S,5R)-2,5-diethyl-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl) piperazin-1-yl)-9-methyl-9H-purine
##STR00184##
[0383] To a solution of 2,6-dichloro-9-methyl-9H-purine (311 mg, 1.54 mmol) and (2R,5S)-2,5-diethyl-1-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)piperazine (450 mg, 1.40 mmol) in n-BuOH (20 mL) was added DIPEA (542 mg, 4.20 mmol). The reaction was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure to give the title compound (650 mg, 84%). MS: M/e 499 (M+1).sup.+.
Step B: 6-((2S,5R)-2,5-diethyl-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-9-methyl-9H-purin-2-ol
##STR00185##
[0384] To a solution of 2-chloro-6-((2S,5R)-2,5-diethyl-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-9-methyl-9H-purine (600 mg, 1.204 mmol), Pd.sub.2(dba).sub.3 (55 mg, 0.06 mmol), t-BuXphos (51 mg, 0.12 mmol) and KOH (1.2 mL, 3M, 3.6 mmol) in dioxane (10 mL) was degassed 3 times under N.sub.2 atmosphere. Then the mixture was stirred at 90 C. for 12 hours. The reaction mixture was diluted with water and extracted with EtOAc (30 mL2). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, filtered and concentrated to dryness. The resulting residue was purified by flash column chromatography (DCM:MeOH=10:1) to give the titled compound (470 mg, 82%). MS: M/e 481 (M+1).sup.+.
Step C: 6-((2S,5R)-2,5-diethyl-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-3,9-dimethyl-3,9-dihydro-2H-purin-2-one
##STR00186##
[0385] To a solution of 6-((2S,5R)-2,5-diethyl-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-9-methyl-9H-purin-2-ol (470 mg, 0.979 mmol) and Cs.sub.2CO.sub.3 (638 mg, 1.958 mmol) in dioxane (30 mL) was added CH.sub.3I (208 mg, 1.468 mmol). The reaction was stirred at room temperature overnight. The reaction mixture was diluted with water, extracted with EtOAc (60 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (DCM:MeOH=10:1) to give the titled compound (270 mg, 56%). MS: M/e 495 (M+1).sup.+.
Step D: 6-((2S,5R)-2,5-diethyl-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purine-8-carbaldehyde
##STR00187##
[0386] To a solution of 6-((2S,5R)-2,5-diethyl-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-3,9-dimethyl-3,9-dihydro-2H-purin-2-one (240 mg, 0.485 mmol) in THE (15 mL) was degassed 3 times under N.sub.2 atmosphere. Then added LDA (0.97 mL, 2M, 1.940 mmol) at 78 C. The mixture was stirred at 78 C. for 1 hour, DMF (106 mg, 1.455 mmol) was added. The reaction solution was stirred to room temperature for 2 hours. The reaction mixture was diluted with water, extracted with EtOAc (45 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (DCM:MeOH=10:1) to give the titled compound (180 mg, 71%). MS: M/e 523 (M+1).sup.+.
Step E: 6-((2S,5R)-2,5-diethyl-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-8-(hydroxymethyl)-3,9-dimethyl-3,9-dihydro-2H-purin-2-one
##STR00188##
[0387] To a solution of 6-((2S,5R)-2,5-diethyl-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3, 9-dihydro-2H-purine-8-carbaldehyde (180 mg, 0.345 mmol) in MeOH (10 mL) was added NaBH.sub.4 (20 mg, 0.517 mmol). The reaction was stirred at room temperature for 1 hour. The reaction mixture was quenched with saturated NH.sub.4Cl, extracted with EtOAc (30 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (DCM:MeOH=8:1) to give the titled compound (150 mg, 83%). MS: M/e 525 (M+1).sup.+.
Step F: 8-(chloromethyl)-6-((2S,5R)-2,5-diethyl-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl) piperazin-1-yl)-3,9-dimethyl-3,9-dihydro-2H-purin-2-one
##STR00189##
[0388] To a solution of 6-((2S,5R)-2,5-diethyl-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-8-(hydroxymethyl)-3,9-dimethyl-3,9-dihydro-2H-purin-2-one (150 mg, 0.286 mmol) in DCM (5 mL) was added SOCl.sub.2 (136 mg, 1.145 mmol). The reaction was stirred at room temperature for 0.5 hours. The reaction mixture concentrated to give the titled compound (150 mg, 97%). MS: M/e 543 (M+1).sup.+.
Step G: 2-(6-((2S,5R)-2,5-diethyl-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00190##
[0389] To a solution of 8-(chloromethyl)-6-((2S,5R)-2,5-diethyl-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-3,9-dimethyl-3,9-dihydro-2H-purin-2-one (150 mg, 0.277 mmol) and Cs.sub.2CO.sub.3 (361 mg, 1.107 mmol) in CH.sub.3CN (10 mL) was added TMSCN (110 mg, 1.107 mmol). The reaction was stirred at room temperature for 4 hours. The reaction mixture was diluted with water, extracted with DCM (30 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled Compound A34, which was further separated into Compound A34a (19 mg) and Compound A34b (10 mg) by Prep-HPLC (Method A).
[0390] Compound A34a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.09-7.96 (m, 1H), 7.40 (t, J=7.9 Hz, 2H), 6.14 (d, J=13.7 Hz, 0.5H), 5.78-5.74 (m, 0.5H), 5.12 (d, J=13.2 Hz, 0.5H), 4.91-4.87 (m, 0.5H), 4.21 (d, J=24.8 Hz, 2H), 4.02-3.93 (m, 1H), 3.92 (d, J=4.3 Hz, 3H), 3.79 (t, J=3.9 Hz, 3H), 3.62 (d, J=13.7 Hz, 0.5H), 3.25-3.19 (m, 1.5H), 2.68-2.51 (m, 1H), 2.11 (d, J=12.2 Hz, 1H), 1.84-1.66 (m, 2H), 1.54-1.48 (m, 2H), 1.28 (d, J=6.4 Hz, 3H), 1.04-0.99 (m, 3H), 0.66-0.61 (m, 3H) ppm. MS: M/e 534 (M+1).sup.+.
[0391] Compound A34b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.06 (dd, J=8.2, 6.0 Hz, 1H), 7.46-7.40 (m, 2H), 6.05-6.01 (m, 0.5H), 5.88 (d, J=13.5 Hz, 0.5H), 5.18-5.13 (m, 0.5H), 4.87-4.83 (m, 0.5H), 4.30-4.09 (m, 3H), 3.92 (d, J=10.3 Hz, 3H), 3.78 (d, J=2.1 Hz, 3H), 3.37 (d, J=11.7 Hz, 0.5H), 3.08-2.96 (m, 1.5H), 2.88-2.71 (m, 1H), 2.28 (t, J=10.4 Hz, 1H), 2.18-1.96 (m, 2H), 1.51-1.35 (m, 2H), 1.26 (dd, J=6.2, 2.4 Hz, 3H), 0.99-0.93 (m, 3H), 0.79-0.64 (m, 3H) ppm. MS: M/e 534 (M+1).sup.+.
Compound A35: 2-(9-ethyl-6-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00191##
[0392] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.15 mmol), 1-(4-fluoro-2-(trifluoromethyl)phenyl)ethan-1-ol (32 mg, 0.15 mmol), (cyanomethyl)trimethylphosphonium iodide (110 mg, 0.46 mmol) and DIPEA (98 mg, 0.76 mmol) in MeCN (3 ml) was stirred at 100 C. overnight. After completed, the solution was concentrated under reduced pressure. The resulting residue was purified by Prep-HPLC (Method A) to give the titled Compound A35a (16 mg) and Compound A35b (9 mg).
[0393] Compound A35a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.11-8.00 (m, 1H), 7.39 (t, J=8.9 Hz, 2H), 5.92 (d, J=13.2 Hz, 1H), 5.05 (s, 1H), 4.37 (q, J=7.2 Hz, 2H), 4.26-4.20 (m, 1H), 3.88 (s, 1H), 3.76 (s, 3H), 3.65 (s, 1H), 3.48 (s, 1H), 2.84-2.56 (m, 2H), 1.99 (d, J=12.5 Hz, 1H), 1.46 (t, J=7.2 Hz, 3H), 1.32 (d, J=6.3 Hz, 3H), 1.23 (dd, J=18.9, 6.8 Hz, 3H), 1.06 (d, J=6.6 Hz, 3H) ppm. MS: M/e 520 (M+1).sup.+
[0394] Compound A35b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.16-8.05 (m, 1H), 7.42 (t, J=9.0 Hz, 2H), 6.34-5.04 (m, 2H), 4.37 (dd, J=15.2, 7.5 Hz, 2H), 4.05 (s, 1H), 3.76 (s, 3H), 3.61-3.36 (m, 1H), 3.26-3.10 (m, 1H), 2.99-2.88 (m, 2H), 2.78 (s, 2H), 1.57-1.41 (m, 6H), 1.25 (d, J=5.8 Hz, 3H), 0.90 (t, J=6.7 Hz, 3H) ppm. MS: M/e 520 (M+1).sup.+
Compound A36: 2-(6-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-bis(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00192##
Step A: 2,6-dichloro-9-(methyl-d3)-9H-purine
##STR00193##
[0395] To a stirred solution of 2,6-dichloro-9H-purine (3.78 g, 20 mmol) in DMF (30 mL) was added K.sub.2CO.sub.3 (5.52 g, 40 mmol), followed by CD3I (4.35 g, 30 mmol). After the addition, the reaction mixture was stirred overnight. The reaction mixture was treated with H.sub.2O (50 mL) and extracted with CH.sub.2Cl.sub.2 (50 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated and purified by flash column chromatography to give the titled compound (2.52 g, 61%). MS: M/e 206 (M+1).sup.+.
Step B: tert-butyl (2R,5S)-4-(2-chloro-9-(methyl-d3)-9H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00194##
[0396] A mixture of 2,6-dichloro-9-(methyl-d3)-9H-purine (2.52 g, 12.2 mmol), tert-butyl (2R,5S)-2,5-dimethylpiperazine-1-carboxylate (2.61 g, 12.2 mmol) and DIPEA (2.37 g, 18.3 mmol) in n-BuOH (25 mL) was stirred for 3 days. The reaction mixture was concentrated to give the titled (4.3 g, 92%). MS: M/e 384 (M+1).sup.+.
Step C: tert-butyl (2R,5S)-4-(2-hydroxy-9-(methyl-d3)-9H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00195##
[0397] To a stirred solution of tert-butyl (2R,5S)-4-(2-chloro-9-(methyl-d3)-9H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (3.51 g, 9.2 mmol) in dioxane (50 mL) was added Pd.sub.2(dba).sub.3 (390 mg, 0.92 mmol), t-BuXPhos (390 mg, 0.92 mmol) and aq.KOH (1.55 g, 27.6 mmol in 5 mL H.sub.2O). After the addition, the reaction mixture was stirred at 100 C. for 3 hours under N.sub.2. The mixture was poured into H.sub.2O (100 mL), extracted with EtOAc (50 mL2). The combined organic layers were discarded and the aqueous layer was acidified to pH=34 with aq.citric acid, then extracted with CH.sub.2Cl.sub.2 (50 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated to give the titled compound (4.15 g, 100%). MS: M/e 366 (M+1).sup.+.
Step D: tert-butyl (2R,5S)-4-(3,9-bis(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00196##
[0398] To a stirred mixture of tert-butyl (2R,5S)-4-(2-hydroxy-9-(methyl-d3)-9H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (2 g, 5.46 mmol) in dioxane (50 mL) was added Cs.sub.2CO.sub.3 (5.3 g, 16.38 mmol), followed by CD3I (1.58 g, 10.9 mmol). After then, the mixture was stirred overnight at 80 C. in a sealed tube. The mixture was poured into H.sub.2O (150 mL), extracted with EtOAc (50 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (1 g, 48%). MS: M/e 383 (M+1).sup.+.
Step E: tert-butyl (2R,5S)-4-(8-formyl-3,9-bis(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00197##
[0399] To a stirred solution of tert-butyl (2R,5S)-4-(3,9-bis(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (500 mg, 1.3 mmol) in THE (10 mL) was added LDA (2.0 M, 1.73 mL, 5.2 mmol) dropwise at 78 C. After stirred for an hour, DMF (379 mg, 5.2 mmol) was added and the mixture was allowed warm to room temperature and stirred for an hour. The mixture was quenched with aq.NH.sub.4Cl, extracted with EtOAc (10 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated and directly used to the next step without further purification. MS: M/e 411 (M+1).sup.+.
Step F: tert-butyl (2R,5S)-4-(8-(hydroxymethyl)-3,9-bis(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00198##
[0400] To a stirred solution of tert-butyl (2R,5S)-4-(8-formyl-3,9-bis(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (crude, 1.3 mmol) in MeOH (15 mL) was added NaBH.sub.4 (74.1 mg, 1.96 mmol), then stirred for 10 min. The reaction mixture was poured into H.sub.2O (30 mL), extracted with EtOAc (15 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated and purified by flash column chromatography to give the titled compound (240 mg, 45%). MS: M/e 413 (M+1).sup.+.
Step G: tert-butyl (2R,5S)-4-(8-(chloromethyl)-3,9-bis(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00199##
[0401] To a stirred solution of tert-butyl (2R,5S)-4-(8-(hydroxymethyl)-3,9-bis(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (240 mg, 0.58 mmol) in CH.sub.2Cl.sub.2 (10 mL) was added SOCl.sub.2 (138 mg, 1.16 mmol). After then, the reaction mixture was stirred for 10 min. The reaction mixture was concentrated to give the titled compound, which was directly used to the next step. MS: M/e 431 (M+1).sup.+
Step H: tert-butyl (2R,5S)-4-(8-(cyanomethyl)-3,9-bis(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00200##
[0402] To a stirred solution of tert-butyl (2R,5S)-4-(8-(chloromethyl)-3,9-bis(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (crude, 0.58 mmol) in CH.sub.2Cl.sub.2 (10 mL) was added TBAF (1.0 M, 2.32 mL, 2.32 mmol), followed by TMSCN (232 mg, 2.32 mmol). After the addition, the reaction mixture was stirred for 5 hours. The reaction mixture was poured into H.sub.2O (30 mL) and extracted with EtOAc (15 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated and purified by flash column chromatography to give the titled compound (crude, 100%). MS: M/e 422 (M+1).sup.+
Step I: 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-bis(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00201##
[0403] To a stirred solution of tert-butyl (2R,5S)-4-(8-(cyanomethyl)-3,9-bis(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (crude, 0.58 mmol) in CH.sub.2Cl.sub.2 (10 mL) was added TFA (2 mL). After the addition, the reaction mixture was stirred for an hour. The reaction mixture was concentrated to give the residue, which was treated with EtOAc/H.sub.2O (20 mL/20 mL), most TBAF was extracted, and the organic layers were discarded. The aqueous layer was basified to pH=1012 with aq.Na.sub.2CO.sub.3 and extracted with CH.sub.2Cl.sub.2/IPA (3/1, 10 mL4). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated to give the titled compound (120 mg, 64%). MS: M/e 322 (M+1).sup.+
Step J: 2-(6-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-bis(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00202##
[0404] A mixture of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-bis(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (32.1 mg, 0.1 mmol), 1-(4-fluoro-2-(trifluoromethyl)phenyl)ethan-1-ol (41.6 mg, 0.2 mmol), (cyanomethyl)trimethylphosphonium iodide (73 mg, 0.3 mmol) and DIPEA (129 mg, 1 mmol) in CH.sub.3CN (4 mL) was stirred overnight at 100 C. in a sealed tube. The reaction mixture was diluted with EtOAc (20 mL), washed with H.sub.2O, brine, dried over Na.sub.2SO.sub.4, concentrated to give the residue, which was purified by Pre-TLC (CH.sub.2Cl.sub.2/MeOH=10:1) to give the titled compound (15 mg). .sup.1H NMR (400 MHz, CD.sub.3OD) 8.14-8.02 (m, 1H), 7.46-7.34 (m, 2H), 6.24-5.01 (m, 2H), 4.29-4.15 (m, 2H), 4.10-3.41 (m, 3H), 2.99-2.62 (m, 3H), 1.57-1.46 (m, 1.5H), 1.34-1.21 (m, 4.5H), 1.07-1.04 (m, 1.5H), 0.93-0.86 (m, 1.5H) ppm. MS: M/e 512 (M+1).sup.+.
Compound A37: 2-(6-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-9-(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00203##
Step A: tert-butyl (2R,5S)-2,5-dimethyl-4-(3-methyl-9-(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-6-yl)piperazine-1-carboxylate
##STR00204##
[0405] To a stirred mixture of tert-butyl (2R,5S)-4-(2-hydroxy-9-(methyl-d3)-9H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (2 g, 5.46 mmol) in dioxane (50 mL) was added Cs.sub.2CO.sub.3 (5.3 g, 16.38 mmol), followed by Mel (1.54 g, 10.9 mmol). After then, the mixture was stirred overnight at 80 C. in a sealed tube. The mixture was poured into H.sub.2O (150 mL), extracted with EtOAc (15 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (860 mg, 41%). MS: M/e 380 (M+1).sup.+.
Step B: tert-butyl (2R,5S)-4-(8-formyl-3-methyl-9-(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00205##
[0406] To a stirred solution of tert-butyl (2R,5S)-2,5-dimethyl-4-(3-methyl-9-(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-6-yl)piperazine-1-carboxylate (860 mg, 2.25 mmol) in THE (15 mL) was added LDA (2.0 M, 4.5 mL, 9 mmol) dropwise at 78 C. After stirred for an hour, DMF (657 mg, 9 mmol) was added, and the mixture was stirred for an hour. The mixture was quenched with aq.NH.sub.4Cl, extracted with EtOAc (20 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated to dryness. The resulting residue was directly used to the next step without further purification. MS: M/e 408 (M+1).sup.+.
Step C: tert-butyl (2R,5S)-4-(8-(hydroxymethyl)-3-methyl-9-(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00206##
[0407] To a stirred solution of tert-butyl (2R,5S)-4-(8-formyl-3-methyl-9-(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (crude, 2.25 mmol) in MeOH (15 mL) was added NaBH.sub.4 (128 mg, 3.37 mmol), then stirred for 10 min. The reaction mixture was poured into H.sub.2O (30 mL), extracted with EtOAc (15 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated and purified by flash column chromatography to give the titled compound (420 mg, 45%) as a white solid. MS: M/e 410 (M+1).sup.+.
Step D: tert-butyl (2R,5S)-4-(8-(chloromethyl)-3-methyl-9-(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00207##
[0408] To a stirred solution of tert-butyl (2R,5S)-4-(8-(hydroxymethyl)-3-methyl-9-(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (420 mg, 1.02 mmol) in CH.sub.2Cl.sub.2 (10 mL) was added SOCl.sub.2 (243 mg, 2.04 mmol). After then, the reaction mixture was stirred for 10 min. The reaction mixture was concentrated to give the titled compound, which was directly used to the next step. MS: M/e 428 (M+1).sup.+
Step E: tert-butyl (2R,5S)-4-(8-(cyanomethyl)-3-methyl-9-(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00208##
[0409] To a stirred solution of tert-butyl (2R,5S)-4-(8-(chloromethyl)-3-methyl-9-(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (crude, 1.02 mmol) in CH.sub.2Cl.sub.2 (15 mL) was added TBAF (1.0 M, 4 mL, 4.08 mmol), followed by TMSCN (408 mg, 4.08 mmol). After the addition, the reaction mixture was stirred for 4 hours. The reaction mixture was poured into H.sub.2O (30 mL) and extracted with EtOAc (15 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (240 mg, 56%). MS: M/e 419 (M+1).sup.+
Step F: 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3-methyl-9-(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00209##
[0410] To a stirred solution of tert-butyl (2R,5S)-4-(8-(cyanomethyl)-3-methyl-9-(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (240 mg, 0.57 mmol) in CH.sub.2Cl.sub.2 (10 mL) was added TFA (2 mL). After the addition, the reaction mixture was stirred for an hour. The reaction mixture was concentrated to give the residue, which was basified to pH=1012 with aq.Na.sub.2CO.sub.3 and extracted with CH.sub.2Cl.sub.2/IPA (3/1, 30 mL4). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated to give the titled compound (120 mg, 66%). MS: M/e 319 (M+1).sup.+
Step G: 2-(6-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-9-(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00210##
[0411] A mixture of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3-methyl-9-(methyl-d3)-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (31.8 mg, 0.1 mmol), 1-(4-fluoro-2-(trifluoromethyl)phenyl)ethan-1-ol (41.6 mg, 0.2 mmol), (cyanomethyl)trimethylphosphonium iodide (73 mg, 0.3 mmol) and DIPEA (129 mg, 1 mmol) in CH.sub.3CN (4 mL) was stirred overnight at 100 C. in a sealed tube. The reaction mixture was diluted with EtOAc (20 mL), washed with H.sub.2O, brine, dried over Na.sub.2SO.sub.4, concentrated to give the residue, which was purified by Pre-TLC (EtOAc, then CH.sub.2Cl.sub.2/MeOH=10:1) to give the titled compound (5 mg). .sup.1H NMR (400 MHz, CD.sub.3OD) 8.12-8.03 (m, 1H), 7.46-7.36 (m, 2H), 6.23-5.00 (m, 2H), 4.25-4.15 (m, 2H), 4.10-3.85 (m, 1H), 3.78 (s, 3H), 3.68-3.41 (m, 1H), 3.00-2.63 (m, 2.5H), 1.99 (d, J=12.4 Hz, 0.5H), 1.56-1.19 (m, 6H), 1.07-0.88 (m, 3H) ppm. MS: M/e 509 (M+1).sup.+.
Compound A41: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00211##
Step A: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00212##
[0412] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (100 mg, 0.31 mmol) in CH.sub.3CN (2 mL) and was added 1-(3-(trifluoromethyl)phenyl)ethan-1-ol (180 mg, 0.95 mmol), (cyanomethyl)trimethylphosphonium iodide (308 mg, 1.27 mmol) and DIPEA (0.4 g, 3.1 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure and purified by Prep-TLC (DCM/MeOH=30/1) to give the titled compound (crude), which was further purified to give the titled compound (20 mg) by Prep-HPLC (Method A). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.78-7.46 (m, 4H), 6.01-4.95 (m, 2H), 4.21 (s, 2H), 3.93 (s, 3H), 3.78 (s, 3H), 3.74-3.45 (m, 2H), 3.05-2.75 (m, 2H), 2.73-2.03 (m, 1H), 1.98-1.21 (m, 6H), 1.15-0.83 (m, 3H) ppm. MS: M/e 488 (M+1).sup.+.
Compound A42: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00213##
[0413] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (300 mg, 0.9 mmol) in CH.sub.3CN (8 mL) and was added 1-(3-(trifluoromethyl)phenyl)ethan-1-ol (520 mg, 2.7 mmol), (cyanomethyl)trimethylphosphonium iodide (875 mg, 3.6 mmol) and DIPEA (1.16 g, 9 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure and purified by Prep-TLC (DCM/MeOH=30/1) to give the titled Compound A42, which was further separated into Compound A42a (68.23 mg) and Compound A42b (76.15 mg) by chiral Prep-HPLC. The chiral separation conditions are shown below.
TABLE-US-00006 Column CHIRALPAK IH Column Size 2 cm 25 cm, 5 um Mobile Phase A MtBE Mobile Phase B MeOH:DCM Flow Rate 20 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-HPLC Equipment Prep-HPLC-Gilson
[0414] Compound A42: .sup.1H NMR (400 MHz, CD.sub.3OD) 7.78-7.60 (m, 2H), 7.60-7.46 (m, 2H), 6.25-5.52 (m, 1H), 5.43-5.01 (m, 0.5H), 4.73-4.62 (m, 0.5H), 4.37 (q, J=7.0 Hz, 2H), 4.23 (s, 2H), 3.85-3.72 (m, 4H), 3.67-3.40 (m, 1.5H), 2.94-2.54 (m, 2H), 2.13 (d, J=12.2 Hz, 0.5H), 1.46 (t, J=6.9 Hz, 4.5H), 1.39-1.31 (m, 3H), 1.26 (d, J=5.2 Hz, 1.5H), 1.06 (d, J=6.3 Hz, 1.5H), 0.92 (d, J=6.4 Hz, 1.5H) ppm. MS: M/e 502 (M+1).sup.+
[0415] Compound A42a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.73-7.63 (m, 2H), 7.60-7.49 (m, 2H), 6.35-4.51 (m, 2H), 4.37 (d, J=7.1 Hz, 2H), 4.22 (s, 2H), 3.76 (s, 3H), 3.73 (s, 1H), 3.65-3.36 (m, 1H), 2.85 (d, J=20.8 Hz, 3H), 1.52-1.40 (m, 6H), 1.33 (d, J=6.5 Hz, 3H), 0.91 (d, J=6.4 Hz, 3H) ppm. MS: M/e 502 (M+1).sup.+.
[0416] Compound A42b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.75 (s, 1H), 7.63 (d, J=7.1 Hz, 1H), 7.60-7.47 (m, 2H), 6.05-4.95 (m, 2H), 4.37 (q, J=7.2 Hz, 2H), 4.23 (s, 2H), 3.76 (s, 3H), 3.56 (dd, J=30.0, 23.5 Hz, 3H), 2.68 (s, 1H), 2.13 (d, J=12.1 Hz, 1H), 1.46 (t, J=7.2 Hz, 3H), 1.35 (d, J=6.5 Hz, 3H), 1.27 (s, 3H), 1.06 (d, J=6.3 Hz, 3H) ppm. MS: M/e 502 (M+1).sup.+.
Compound A43: 2-(9-ethyl-6-((2S,5R)-4-(1-(3-isopropoxyphenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00214##
Step A: 1-(3-isopropoxyphenyl)ethan-1-ol
##STR00215##
[0417] To a solution of 3-isopropoxybenzaldehyde (500 mg, 3.045 mmol) in THE (30 mL) was degassed 3 times under N.sub.2 atmosphere. Then added CH.sub.3MgBr (2 mL, 3M, 6.09 mmol) dropwise at 0 C. The mixture was stirred at 0 C. for 4 hours and diluted with saturated NH.sub.4Cl, extracted with EtOAc (150 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (PE:EtOAc=4:1) to give the titled compound (500 mg, 91%). .sup.1H NMR (400 MHz, CDCl.sub.3) 7.29-7.21 (m, 1H), 6.92 (d, J=6.7 Hz, 2H), 6.85-6.73 (m, 1H), 4.86 (q, J=6.4 Hz, 1H), 4.57 (dt, J=12.1, 6.0 Hz, 1H), 1.79 (s, 1H), 1.48 (d, J=6.4 Hz, 3H), 1.34 (d, J=6.1 Hz, 6H) ppm.
Step B: 2-(9-ethyl-6-((2S,5R)-4-(1-(3-isopropoxyphenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00216##
[0418] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (120 mg, 0.365 mmol), 1-(3-isopropoxyphenyl)ethan-1-ol (197 mg, 1.094 mmol), (cyanomethyl)trimethylphosphonium iodide (177 mg. 0.730 mmol) and DIPEA (141 mg, 1.094 mol) in CH.sub.3CN (2 ml). The mixture solution was degassed 3 times under N.sub.2 atmosphere. Then the mixture solution was stirred at 105 C. for 24 hours. The reaction mixture was concentrated under reduced pressure. The crude product was purified by Prep-TLC (DCM:MeOH=15:1) and Prep-HPLC (Method A) to give the titled compound (58 mg, 32%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.26-7.14 (m, 1H), 6.93 (dd, J=19.2, 6.1 Hz, 2H), 6.79 (t, J=9.2 Hz, 1H), 6.23-5.53 (m, 1H), 5.26-5.01 (m, 0.5H), 4.85-4.71 (m, 0.5H), 4.59 (dd, J=11.9, 6.0 Hz, 1H), 4.37 (q, J=6.9 Hz, 2H), 4.22 (s, 1H), 3.76 (s, 3H), 3.61 (s, 1H), 3.48 (s, 2H), 3.10-2.58 (m, 2.5H), 2.29 (d, J=11.7 Hz, 0.5H), 1.46 (t, J=6.9 Hz, 5H), 1.31 (t, J=6.1 Hz, 10H), 0.98 (dd, J=55.1, 6.2 Hz, 3H) ppm. MS: M/e 492 (M+1).sup.+
Compound A44: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-(trifluoromethoxy)phenyl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00217##
[0419] To a solution of 1-(3-(trifluoromethoxy)phenyl)ethan-1-ol (60 mg, 0.29 mmol), 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (60 mg, 0.19 mmol) and (cyanomethyl) trimethyl phosphonium iodide (68 mg, 0.28 mmol) in CH.sub.3CN (5 mL) was added DIPEA (116 mg, 0.9 mmol). The mixture was sealed in a bottle and heated at 100 C. for 16 hours. The mixture was cooled to room temperature, diluted with water, extracted with EtOAc (10 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled compound (19 mg, 19%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.45-7.34 (m, 3H), 7.17-7.12 (m, 1H), 5.97-5.91 (m, 1H), 5.31-5.24 (m, 1H), 4.38-4.34 (m, 2H), 4.22 (s, 2H), 3.76 (s, 3H), 3.59-3.48 (m, 2H), 2.88-2.84 (m, 2H), 2.70-2.66 (m, 0.5H), 2.18-2.15 (m, 0.5H), 1.49-1.44 (m, 4H), 1.35-1.29 (m, 4H), 1.06-1.02 (m, 2H), 0.92-0.90 (m, 2H) ppm. MS: M/e 518 (M+1).sup.+.
Compound A48: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(quinoxalin-6-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00218##
[0420] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl) acetonitrile (50 mg, 0.117 mol), 1-(quinoxalin-6-yl)ethan-1-ol (41 mg, 0.233 mol), (cyanomethyl)trimethylphosphonium iodide (57 mg. 0.233 mol) and DIPEA (45 mg, 0.351 mol) in CH.sub.3CN (2 ml). The mixture solution was degassed 3 times under N.sub.2 atmosphere. Then the mixture solution was stirred at 105 C. for 24 hours. The reaction mixture was concentrated under reduced pressure. The resulting residue was purified by Prep-HPLC (Method A) to give the titled compound (7 mg, 13%). .sup.1H NMR (400 MHz, CD.sub.3OD) 8.87 (t, J=3.3 Hz, 2H), 8.13-8.02 (m, 3H), 5.90-4.65 (m, 2H), 4.26-4.15 (m, 2H), 4.00-3.96 (m, 1H), 3.94 (s, 3H), 3.78 (s, 3H), 3.68-3.49 (m, 1H), 3.15-2.00 (m, 3H), 1.54-1.49 (m, 1.5H), 1.44 (dd, J=12.1, 6.5 Hz, 3H), 1.31-1.27 (m, 1.5H), 1.11 (d, J=6.5 Hz, 1.5H), 0.96 (d, J=6.3 Hz, 1.5H) ppm. MS: M/e 472 (M+1).sup.+.
Compound A49: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00219##
[0421] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (315 mg, 1 mol), 1-(3-methylquinoxalin-6-yl)ethan-1-ol (376 mg, 2 mol), (cyanomethyl)trimethylphosphonium iodide (486 mg. 2 mol) and DIPEA (387 mg, 3 mol) in CH.sub.3CN (3 ml). The mixture solution was degassed 3 times under N.sub.2 atmosphere. Then the mixture solution was stirred at 105 C. for 24 hours. The reaction mixture was concentrated under reduced pressure. The crude product was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled Compound A49 (80 mg), which was further separated into Compound A49a (32 mg) and Compound A49b (35 mg) by Prep-HPLC (Method A).
[0422] Compound A49: .sup.1H NMR (400 MHz, CD.sub.3OD) 8.79 (d, J=4.7 Hz, 1H), 8.09-7.90 (m, 3H), 6.27-4.55 (m, 2H), 4.20 (s, 2H), 3.96-3.86 (m, 4H), 3.78 (s, 3H), 3.76-3.60 (m, 2H), 2.97-2.87 (m, 1.5H), 2.77 (d, J=1.6 Hz, 3H), 2.20 (d, J=11.5 Hz, 0.5H), 1.45-1.39 (m, 3H), 1.36-1.22 (m, 3H), 1.10-0.94 (m, 3H) ppm. MS: M/e 486 (M+1).sup.+.
[0423] Compound A49a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.79 (s, 1H), 8.06 (d, J=8.6 Hz, 1H), 7.98 (s, 1H), 7.93 (dd, J=8.7, 1.6 Hz, 1H), 6.29-5.16 (m, 2H), 4.58 (s, 1H), 4.30-4.16 (m, 2H), 3.96-3.89 (m, 4H), 3.79 (s, 3H), 3.00-2.85 (m, 3H), 2.77 (s, 3H), 1.50 (s, 3H), 1.41 (t, J=8.9 Hz, 3H), 0.94 (t, J=8.4 Hz, 3H) ppm. MS: M/e 486 (M+1).sup.+.
[0424] Compound A49b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.78 (s, 1H), 8.03 (d, J=8.6 Hz, 1H), 7.96 (s, 1H), 7.92 (dd, J=8.7, 1.6 Hz, 1H), 5.96-5.68 (m, 1H), 5.08-4.92 (m, 1H), 4.58 (s, 1H), 4.20 (s, 2H), 3.94 (s, 3H), 3.80-3.73 (m, 4H), 3.67-3.61 (m, 1H), 2.79-2.73 (m, 4H), 2.20 (d, J=11.5 Hz, 1H), 1.45 (d, J=6.5 Hz, 3H), 1.28 (d, J=5.6 Hz, 3H), 1.10 (d, J=6.5 Hz, 3H) ppm. MS: M/e 486 (M+1).sup.+.
Compound A50: 2-(6-((2S,5R)-5-ethyl-2-methyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00220##
Step A: tert-butyl (2S,5R)-5-ethyl-2-methyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazine-1-carboxylate
##STR00221##
[0425] A solution of 1-(3-methylquinoxalin-6-yl)ethan-1-ol (500 mg, 2.66 mmol), tert-butyl (2S,5R)-5-ethyl-2-methylpiperazine-1-carboxylate (758 mg, 3.32 mmol), (cyanomethyl)trimethylphosphonium iodide (1.29 g, 5.31 mmol) and DIPEA (2.74 g, 21.24 mmol) in MeCN (6 ml) was stirred at 100 C. overnight. The reaction was diluted with EtOAc (20 ml) and washed with brine (10 ml). The organic layer was concentrated to dryness. The resulting residue was purified by flash column chromatography with 0-20% EtOAc in PE to give the titled compound product (1 g, 94%). MS: M/e 399 (M+1).sup.+.
Step B: 7-(1-((2R,5S)-2-ethyl-5-methylpiperazin-1-yl)ethyl)-2-methylquinoxaline
##STR00222##
[0426] A solution of tert-butyl (2S,5R)-5-ethyl-2-methyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazine-1-carboxylate (1 g, 2.51 mmol) and TFA (4 ml) in DCM (16 ml) was stirred at rt for 60 min. The reaction was concentrated under reduced pressure. The residue was diluted with EtOAc (20 ml) and washed with aq. NaHCO.sub.3 (10 ml2). The organic layer was dried over Na.sub.2SO.sub.4 and concentrated to dryness. The resulting residue was purified by flash column chromatography with 0-5% MeOH in DCM to give the titled compound product (745 mg, 100%). MS: M/e 299 (M+1).sup.+.
Step C: 8-((benzyloxy)methyl)-6-((2S,5R)-5-ethyl-2-methyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-3,9-dihydro-2H-purin-2-one
##STR00223##
[0427] A solution of 7-(1-((2R,5S)-2-ethyl-5-methylpiperazin-1-yl)ethyl)-2-methylquinoxaline (645 mg, 2.16 mmol), 8-((benzyloxy)methyl)-6-hydroxy-3,9-dimethyl-3,9-dihydro-2H-purin-2-one (649 mg, 2.16 mmol), BOP (1.44 g, 3.25 mmol) and DBU (993 mg, 6.49 mmol) in CH.sub.3CN (15 ml) was stirred at rt for 24 h. The reaction was concentrated to dryness. The resulting residue was diluted with EtOAc (20 ml) and washed with brine (10 ml). The organic layer was dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by flash column chromatography with 0-10% MeOH in DCM to give the titled compound (crude), which was used directly for the next step without further purification. MS: M/e 581 (M+1).sup.+.
Step D: 6-((2S,5R)-5-ethyl-2-methyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-8-(hydroxymethyl)-3,9-dimethyl-3,9-dihydro-2H-purin-2-one
##STR00224##
[0428] A solution of 8-((benzyloxy)methyl)-6-((2S,5R)-5-ethyl-2-methyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-3,9-dihydro-2H-purin-2-one (crude) in TFA (20 ml) was stirred at 66 C. for 2 days. The solution was concentrated to dryness. The residue was diluted with EtOAc (20 ml), washed with aq. NaHCO.sub.3 (10 ml2) and brine (10 ml), dried over Na.sub.2SO.sub.4 and concentrated. The resulting residue was purified by flash column chromatography with 0-15% MeOH in DCM to give the titled compound (580 mg) as an off-white solid. MS: M/e 491 (M+1).sup.+.
Step E: 8-(chloromethyl)-6-((2S,5R)-5-ethyl-2-methyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-3,9-dihydro-2H-purin-2-one
##STR00225##
[0429] To a solution of 6-((2S,5R)-5-ethyl-2-methyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-8-(hydroxymethyl)-3,9-dimethyl-3,9-dihydro-2H-purin-2-one (580 mg, 1.18 mmol) in DCM (15 ml) at 0 C., was added SOCl.sub.2 (423 mg, 3.55 mmol) and stirred at 0 C. for 20 min. The reaction was washed with aq. NaHCO.sub.3 (10 ml) and brine (10 ml), dried over Na.sub.2SO.sub.4 and concentrated to dryness to give the titled compound (601 mg, 100%), which was used directly for the next step. MS: M/e 509 (M+1).sup.+.
Step F: 2-(6-((2S,5R)-5-ethyl-2-methyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00226##
[0430] A solution of 8-(chloromethyl)-6-((2S,5R)-5-ethyl-2-methyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-3,9-dihydro-2H-purin-2-one (601 mg, 1.18 mmol), TMSCN (351 mg, 3.55 mmol) and Cs.sub.2CO.sub.3 (1.15 g, 3.53 mmol) in MeCN (15 ml) was stirred at 66 C. for 60 min. The reaction mixture was poured into water (15 ml) and then extracted with EtOAc (20 ml2). The organic layer was washed with brine (10 ml), dried over Na.sub.2SO.sub.4 and concentrated to dryness. The resulting residue was purified by flash column chromatography with 0-10% MeOH in DCM to give the titled Compound A50, which was separated into Compound A50a (40 mg) and Compound A50b (60 mg) by chiral Prep-HPLC. The chiral separation conditions are shown below.
TABLE-US-00007 Column CHIRALPAK IA Column Size 2 cm 25 cm, 5 um Mobile Phase A MtBE Mobile Phase B MeOH:DCM Flow Rate 20 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-HPLC Equipment Prep-HPLC-Gilson
[0431] Compound A50a (the earlier peak): .sup.1H NMR (400 MHz, DMSO-d6) 8.83 (s, 1H), 8.05 (d, J=8.6 Hz, 1H), 7.92 (s, 1H), 7.85 (d, J=8.6 Hz, 1H), 5.97 (s, 0.5H), 5.70 (d, J=14.0 Hz, 0.5H), 5.05 (s, 0.5H), 4.64 (d, J=14.0 Hz, 0.5H), 4.36 (d, J=33.8 Hz, 2H), 3.98 (q, J=6.5 Hz, 1H), 3.82 (d, J=23.8 Hz, 3H), 3.64 (s, 3H), 3.31 (s, 0.5H), 3.00 (s, 0.5H), 2.82 (d, J=25.2 Hz, 2H), 2.70 (s, 3H), 2.34 (s, 1H), 1.56-1.47 (m, 1H), 1.45-1.39 (m, 1H), 1.38-1.30 (m, 6H), 0.63 (t, J=7.2 Hz, 3H) ppm. MS: M/e 500 (M+1).sup.+.
[0432] Compound A50b (the later peak): .sup.1H NMR (400 MHz, DMSO-d6) 8.81 (s, 1H), 8.02 (d, J=8.7 Hz, 1H), 7.95 (s, 1H), 7.86 (d, J=8.6 Hz, 1H), 5.97 (d, J=13.2 Hz, 0.5H), 5.73 (s, 0.5H), 4.98-4.80 (m, 1H), 4.37 (d, J=37.3 Hz, 2H), 3.83 (s, 4H), 3.64 (s, 3H), 3.59 (s, 0.5H), 3.32-3.19 (m, 0.5H), 3.11 (s, 1H), 2.70 (s, 3H), 2.55 (d, J=9.0 Hz, 1H), 2.16 (d, J=11.8 Hz, 1H), 1.51-1.41 (m, 2H), 1.34 (d, J=6.2 Hz, 3H), 1.21-1.10 (m, 3H), 0.99 (t, J=9.2 Hz, 3H) ppm. MS: M/e 500 (M+1).sup.+.
Compound A51: 2-(6-((2S,5R)-2,5-diethyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00227##
Step A: tert-butyl (2S,5R)-2,5-diethyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazine-1-carboxylate
##STR00228##
[0433] A solution of 1-(3-methylquinoxalin-6-yl)ethan-1-ol (500 mg, 2.66 mmol), tert-butyl (2S,5R)-2,5-diethylpiperazine-1-carboxylate (805 mg, 3.33 mmol), (cyanomethyl)trimethylphosphonium iodide (1.29 g, 5.31 mmol) and DIPEA (2.74 g, 21.24 mmol) in MeCN (6 ml) was stirred at 100 C. overnight. The reaction was diluted with EtOAc (20 ml) and washed with brine (10 ml). The organic layer was concentrated to dryness. The resulting residue was purified by flash column chromatography with 0-20% EtOAc in PE to give the titled compound (1 g, 91%). MS: M/e 413 (M+1).sup.+.
Step B: 7-(1-((2R,5S)-2,5-diethylpiperazin-1-yl)ethyl)-2-methylquinoxaline
##STR00229##
[0434] A solution of tert-butyl (2S,5R)-2,5-diethyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazine-1-carboxylate (1 g, 2.43 mmol) and TFA (4 ml) in DCM (16 ml) was stirred at rt for 30 min. The reaction was concentrated under reduced pressure. The residue was diluted with EtOAc (20 ml) and washed with aq. NaHCO.sub.3 (10 ml2). The organic layer was dried over Na.sub.2SO.sub.4 and concentrated to dryness. The resulting residue was purified by flash column chromatography with 0-5% MeOH in DCM to give the titled compound (757 mg, 100%). MS: M/e 313 (M+1).sup.+.
Step C: 8-((benzyloxy)methyl)-6-((2S,5R)-2,5-diethyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-3,9-dihydro-2H-purin-2-one
##STR00230##
[0435] A solution of 7-(1-((2R,5S)-2,5-diethylpiperazin-1-yl)ethyl)-2-methylquinoxaline (400 mg, 1.28 mmol), 8-((benzyloxy)methyl)-6-hydroxy-3,9-dimethyl-3,9-dihydro-2H-purin-2-one (423 mg, 1.41 mmol), BOP (737 mg, 1.67 mmol) and DBU (392 mg, 2.56 mmol) in MeCN (12 ml) was stirred at rt for 24 h and then 50 C. overnight. The reaction was concentrated to dryness. The resulting residue was diluted with EtOAc (20 ml) and washed with brine (10 ml). The organic layer was dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by flash column chromatography with 0-10% MeOH in DCM to give the titled compound (crude), which was used directly for the next step without further purification. MS: M/e 595 (M+1).sup.+.
Step D: 6-((2S,5R)-2,5-diethyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-8-(hydroxymethyl)-3,9-dimethyl-3,9-dihydro-2H-purin-2-one
##STR00231##
[0436] A solution of 8-((benzyloxy)methyl)-6-((2S,5R)-2,5-diethyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-3,9-dihydro-2H-purin-2-one (crude) in TFA (10 ml) was stirred at 66 C. for 3 days. The solution was concentrated to dryness. The residue was diluted with EtOAc (20 ml), washed with aq. NaHCO.sub.3 (10 ml2) and brine (10 ml), dried over Na.sub.2SO.sub.4 and concentrated. The resulting residue was purified by flash column chromatography with 0-15% MeOH in EtOAc to give the titled compound (250 mg). MS: M/e 505 (M+1).sup.+.
Step E: 8-(chloromethyl)-6-((2S,5R)-2,5-diethyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-3,9-dihydro-2H-purin-2-one
##STR00232##
[0437] To a solution of 6-((2S,5R)-2,5-diethyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-8-(hydroxymethyl)-3,9-dimethyl-3,9-dihydro-2H-purin-2-one (250 mg, 0.50 mmol) in DCM (15 ml) at 0 C., was added SOCl.sub.2 (177 mg, 1.49 mmol) and stirred at 0 C. for 25 min. The reaction was washed with aq. NaHCO.sub.3 (10 ml) and brine (10 ml), dried over Na.sub.2SO.sub.4 and concentrated to dryness to give the titled compound (259 mg, 100%), which was used directly for the next step. MS: M/e 523 (M+1).sup.+.
Step F: 2-(6-((2S,5R)-2,5-diethyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00233##
[0438] A solution of 8-(chloromethyl)-6-((2S,5R)-2,5-diethyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-3,9-dihydro-2H-purin-2-one (259 mg, 0.50 mmol), TMSCN (148 mg, 1.49 mmol) and Cs.sub.2CO.sub.3 (485 mg, 1.49 mmol) in MeCN (10 ml) was stirred at 70 C. for 60 min. The reaction mixture was poured into water (15 ml) and then extracted with EtOAc (15 ml2). The organic layer was washed with brine (10 ml), dried over Na.sub.2SO.sub.4 and concentrated to dryness. The resulting residue was purified by Prep-TLC with EtOAc:MeOH (13:1) to give the titled Compound A51, which was separated into Compound A51a (21 mg) and Compound A51b (31 mg) by chiral Prep-HPLC. The chiral separation conditions are shown below.
TABLE-US-00008 Column CHIRALPAK IA Column Size 2 cm 25 cm, 5 um Mobile Phase A MtBE Mobile Phase B MeOH:DCM Flow Rate 20 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-HPLC Equipment Prep-HPLC-Gilson
[0439] Compound A51a (the earlier peak): .sup.1H NMR (400 MHz, DMSO-d6) 8.82 (s, 1H), 8.04 (d, J=8.5 Hz, 1H), 7.91 (s, 1H), 7.84 (d, J=8.8 Hz, 1H), 5.85 (s, 0.5H), 5.74 (d, J=13.0 Hz, 0.5H), 4.98 (s, 0.5H), 4.72 (d, J=13.1 Hz, 0.5H), 4.40 (s, 1H), 4.31 (s, 1H), 3.97 (q, J=6.1 Hz, 1H), 3.81 (d, J=17.1 Hz, 3H), 3.63 (s, 3H), 3.30-3.28 (m, 0.5H), 2.94 (q, J=22.9, 1.5H), 2.78 (d, J=12.4 Hz, 0.5H), 2.70 (s, 3H), 2.67 (s, 0.5H), 2.32 (dd, J=23.0, 11.0 Hz, 1H), 2.02-1.87 (m, 2H), 1.50 (s, 1H), 1.36 (d, J=5.2 Hz, 3H), 1.33-1.26 (m, 1H), 0.87 (dt, J=14.7, 7.3 Hz, 3H), 0.65-0.54 (m, 3H) ppm. MS: M/e 514 (M+1).sup.+.
[0440] Compound A51b (the later peak): .sup.1H NMR (400 MHz, DMSO-d6) 8.81 (s, 1H), 8.02 (d, J=8.6 Hz, 1H), 7.93 (s, 1H), 7.85 (d, J=8.6 Hz, 1H), 6.01 (d, J=13.8 Hz, 0.5H), 5.57 (s, 0.5H), 4.98 (d, J=12.5 Hz, 0.5H), 4.74 (s, 0.5H), 4.41 (s, 1H), 4.31 (s, 1H), 3.82 (d, J=13.9 Hz, 4H), 3.64 (s, 3H), 3.55 (d, J=13.5 Hz, 0.5H), 3.12 (dd, J=20.6, 10.0 Hz, 1.5H), 2.70 (s, 3H), 2.59 (d, J=8.9 Hz, 0.5H), 2.47 (s, 0.5H), 2.23 (d, J=12.4 Hz, 1H), 1.84-1.64 (m, 2H), 1.44 (s, 2H), 1.33 (d, J=5.6 Hz, 3H), 0.97 (s, 3H), 0.54 (t, J=7.3 Hz, 3H) ppm. MS: M/e 514 (M+1).sup.+.
Compound A52: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00234##
[0441] A mixture of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (165 mg, 0.5 mmol), 1-(3-methylquinoxalin-6-yl)ethan-1-ol (122.2 mg, 0.65 mmol) and (cyanomethyl)trimethylphosphonium iodide (243 mg, 1 mmol) in CH.sub.3CN (5 mL) was added DIPEA (645 mg, 5 mmol). The mixture was stirred at 100 C. overnight in a sealed tube. The reaction mixture was diluted with EtOAc (20 mL), washed with brine, dried over Na.sub.2SO.sub.4, concentrated. The resulting residue was purified by flash column chromatography to give the titled Compound A52 (65 mg), which was separated into Compound A52a (23 mg) and Compound A52b (28 mg) by chiral Prep-HPLC. The chiral separation conditions are shown below.
TABLE-US-00009 Column CHIRALPAK IH Column Size 2 cm 25 cm, 5 um Mobile Phase A MtBE Mobile Phase B MeOH:DCM Flow Rate 20 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-HPLC Equipment Prep-HPLC-Gilson
[0442] Compound A52a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.79 (s, 1H), 8.06 (d, J=8.4 Hz, 1H), 8.00-7.90 (m, 2H), 6.33-5.16 (m, 2H), 4.43-4.29 (m, 2H), 4.20 (s, 2H), 3.96-3.89 (m, 1H), 3.76 (s, 3H), 3.67-3.42 (m, 1H), 3.04-2.85 (m, 3H), 2.77 (s, 3H), 1.62-1.37 (m, 9H), 0.95 (d, J=6.4 Hz, 3H) ppm. MS: M/e 500 (M+1).sup.+.
[0443] Compound A52b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.77 (s, 1H), 8.07-7.85 (m, 3H), 6.11-4.94 (m, 2H), 4.37 (q, J=7.2 Hz, 2H), 4.22 (s, 1H), 3.76 (s, 4H), 3.71-3.42 (m, 2H), 3.30-3.10 (m, 1H), 2.76 (s, 4H), 2.20 (d, J=12 Hz, 1H), 1.54-1.36 (m, 6H), 1.37-1.22 (m, 3H), 1.10 (d, J=6.4 Hz, 3H) ppm. MS: M/e 500 (M+1).sup.+.
Compound A53: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-3-ethyl-9-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00235##
Step A: tert-butyl (2R,5S)-4-(3-ethyl-9-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00236##
[0444] To a mixture of tert-butyl (2R,5S)-2,5-dimethyl-4-(9-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)piperazine-1-carboxylate (5 g, 13.8 mmol) in 1,4-dioxane (700 mL) and was added Cs.sub.2CO.sub.3 (22.5 g, 69 mmol) and CH.sub.3CH.sub.2I (10.7 g, 69 mmol). The reaction was stirred at room temperature overnight. The reaction mixture was filtered. The filtrate was removed under reduced pressure to dryness. The crude product was purified by flash column chromatography to give the titled compound (1.7 g, 31%). MS: M/e 391 (M+1).sup.+.
Step B: tert-butyl (2R,5S)-4-(3-ethyl-8-formyl-9-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00237##
[0445] To a solution of tert-butyl (2R,5S)-4-(3-ethyl-9-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (1.7 g, 4.4 mmol) in THE (100 mL) was added LDA (6.1 mL, 2M, 13.2 mmol) at below 60 C. The reaction was stirred at 65 C. for 1 hour, then added DMF (1.56 g, 22 mmol) at 78 C. The reaction was stirred at 60 C. for another 1 hour. The reaction solvent was quenched by saturated NH.sub.4Cl and extracted by EtOAc. The crude product was concentrated to dryness. The resulting residue was purified by flash column chromatography (DCM/MeOH) to give the titled compound (1.2 g, 66%). MS: M/e 419 (M+1).sup.+.
Step C: tert-butyl (2R,5S)-4-(3-ethyl-8-(hydroxymethyl)-9-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00238##
[0446] NaBH.sub.4 (106 mg, 2.38 mol) was added to tert-butyl (2R,5S)-4-(3-ethyl-8-formyl-9-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (1.2 g, 2.8 mmol) in MeOH (15 mL) at 0 C. for 5 min. The reaction solvent was quenched with water and extracted with DCM. The organic layer was purified by flash column chromatography (DCM/MeOH) to give the titled compound (800 mg, 66%). MS: M/e 421 (M+1).sup.+.
Step D: tert-butyl (2R,5S)-4-(8-(chloromethyl)-3-ethyl-9-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00239##
[0447] To a solution of SOCl.sub.2 (340 mg, 2.8 mmol) in DCM (400 mL) was added tert-butyl (2R,5S)-4-(3-ethyl-8-(hydroxymethyl)-9-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (800 mg, 1.9 mmol) at 10 C. The reaction was stirred at rt for 5 min. The reaction solvent was quenched with water and washed with water, sat. NaHCO.sub.3 and sat. NaCl. The organic layer was dried over Na.sub.2SO.sub.4 and used the next step directly.
Step E: tert-butyl (2R,5S)-4-(8-(cyanomethyl)-3-ethyl-9-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00240##
[0448] To the solution of tert-butyl (2R,5S)-4-(8-(chloromethyl)-3-ethyl-9-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate in DCM (40 mL) were added TMSCN (564 mg, 5.7 mmol) and TBAF (1M, 5.7 mL, 5.7 mmol). The resulting mixture was stirred at room temperature for 3 hours. The reaction solvent was washed by water and dried over Na.sub.2SO.sub.4, removed under vacuum. The crude product was purified by flash column chromatography (DCM/MeOH) to give the titled compound (500 mg, two step 61%). MS: M/e 430 (M+1).sup.+.
Step F: 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3-ethyl-9-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00241##
[0449] To a solution of tert-butyl (2R,5S)-4-(8-(cyanomethyl)-3-ethyl-9-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (500 mg, 1.17 mmol) in DCM (10 mL) were added TFA (3 mL). The resulting mixture was stirred at rt for 3 hours. The reaction solvent was removed under vacuum. The crude product was dissolved with water and adjusted to pH=9 with saturated NaHCO.sub.3. The aqueous layer was extracted with (DCM/IPA=4/1) to give the titled compound by lyophilization (400 mg, crude). MS: M/e 330 (M+1).sup.+.
Step G: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-methylquinoxalin-6-yl)ethyl)piperazin-1-yl)-3-ethyl-9-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00242##
[0450] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3-ethyl-9-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (200 mg, 0.6 mmol) in CH.sub.3CN (5 mL) and was added 1-(3-methylquinoxalin-6-yl)ethan-1-ol (339 mg, 1.8 mmol), (cyanomethyl)trimethylphosphonium iodide (583 mg, 2.4 mmol) and DIPEA (0.74 g, 6 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure and purified by Prep-TLC (DCM:MeOH=30: 1) to give the titled Compound A53 (50 mg), which was further separated into Compound A53a (20 mg) and Compound A53b (16 mg) by chiral Prep-SFC. The chiral separation conditions are shown below.
TABLE-US-00010 Column CHIRALPAK IG Column Size 3 cm 25 cm, 5 um Mobile Phase A CO2 Mobile Phase B MeOH Flow Rate 100 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-SFC Equipment Prep-SFC-150
[0451] Compound A53a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.79 (s, 1H), 8.06 (d, J=8.6 Hz, 1H), 8.01-7.86 (m, 2H), 6.20-5.14 (m, 2H), 4.25-4.10 (m, 4H), 3.89 (s, 4H), 3.55 (d, J=61.5 Hz, 1H), 2.90 (d, J=27.4 Hz, 3H), 2.77 (s, 3H), 1.50 (s, 3H), 1.44-1.29 (m, 6H), 0.95 (d, J=6.3 Hz, 3H) ppm. MS: M/e 500 (M+1).sup.+.
[0452] Compound A53b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.78 (s, 1H), 8.03 (d, J=8.6 Hz, 1H), 7.99-7.87 (m, 2H), 6.01-4.98 (m, 2H), 4.32-4.26 (m, 2H), 4.21 (s, 1H), 3.97-3.82 (m, 3H), 3.82-3.47 (m, 3H), 2.76 (s, 3H), 2.71-2.66 (m, 2H), 2.19 (d, J=11.9 Hz, 1H), 1.47-1.41 (m, 3H), 1.36 (t, J=7.0 Hz, 3H), 1.29 (s, 3H), 1.10 (d, J=6.4 Hz, 3H) ppm. MS: M/e 500 (M+1).sup.+.
Compound A54: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-(trifluoromethyl)quinoxalin-6-yl)ethyl) piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00243##
Step A: 7-bromo-2-(trifluoromethyl)quinoxaline
##STR00244##
[0453] To a solution of 3,3-dibromo-1,1,1-trifluoropropan-2-one (5.77 g, 21.386 mmol) and AcONa (8.44 g, 106.9 mmol) in CH.sub.30H (60 mL) and H.sub.2O (60 mL) was stirred at 90 C. for 30 mins. Then 4-bromobenzene-1,2-diamine (2.0 g, 10.7 mmol) was added. The mixture was stirred at room temperature for 12 hours. After filtered, the mixture was extracted with EtOAc (35 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (PE:EtOAc=5:1) to give the crude compound (3.2 g). The crude product (1.6 g) was further separated into the titled compound (680 mg, MS: M/e 277 (M+1).sup.+) and another position isomer (620 mg, MS: M/e 277 (M+1).sup.+) by chiral Prep-SFC. The chiral separation conditions are shown below.
TABLE-US-00011 Column CHIRALPAK IG Column Size 3 cm 25 cm, 5 um Mobile Phase A CO.sub.2 Mobile Phase B MeOH Flow Rate 100 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-SFC Equipment Prep-SFC-150
Step B: 1-(3-(trifluoromethyl)quinoxalin-6-yl)ethan-1-one
##STR00245##
[0454] To a solution of 6-bromo-2-(trifluoromethyl)quinoxaline (360 mg, 1.304 mmol), tributyl(1-ethoxyvinyl)stannane (942 mg, 2.609 mmol), Pd(PPh.sub.3).sub.2Cl.sub.2 (183 mg, 0.261 mmol) in toluene (20 mL) was stirred at 90 C. under N.sub.2 for 12 hours. The reaction mixture was quenched with saturated NaHCO.sub.3 aq. (30 mL), extracted with EtOAc (50 mL2), combined, washed brine (30 mL2), dried and concentrated to dryness. The resulting oil was diluted with THE (30 mL). Then to the solution was added HCl (3 mL, 4 M in 1,4-dioxane) in drops and the mixture was stirred at room temperature for 1 hour. The reaction mixture was diluted with EtOAc (30 mL), treated with saturated NaHCO.sub.3 aq. to pH8, washed with brine (30 mL3), dried over Na.sub.2SO.sub.4 and concentrated to dryness. The resulting residue was purified by flash column chromatography (PE:EtOAc=6:1) to give the titled compound (300 mg, 96%). MS: M/e 241 (M+1).sup.+
Step C: 1-(3-(trifluoromethyl)quinoxalin-6-yl)ethan-1-ol
##STR00246##
[0455] To a solution of 1-(3-(trifluoromethyl)quinoxalin-6-yl)ethan-1-one (300 mg, 1.25 mmol) in CH.sub.30H (15 mL) was added NaBH.sub.4 (48 mg, 1.25 mmol). The reaction was stirred at room temperature for 1 hours. The reaction mixture was quenched with saturated NH.sub.4Cl, extracted with EtOAc (35 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (PE:EtOAc=5:1) to give the titled compound (290 mg, 96%). MS: M/e 243 (M+1).sup.+
Step D: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-(trifluoromethyl)quinoxalin-6-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00247##
[0456] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (70 mg, 0.213 mmol), 1-(3-(trifluoromethyl)quinoxalin-6-yl)ethan-1-ol (77 mg, 0.319 mmol), (cyanomethyl)trimethylphosphonium iodide (104 mg. 0.426 mmol) and DIPEA (82 mg, 0.639 mol) in CH.sub.3CN (1 ml). The mixture solution was degassed 3 times under N.sub.2 atmosphere. Then the mixture solution was stirred at 100 C. for 24 hours. The reaction mixture was concentrated under reduced pressure. The crude product was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled compound (56 mg, 48%). .sup.1H NMR (400 MHz, CD.sub.3OD) 9.23 (d, J=4.1 Hz, 1H), 8.27-8.12 (m, 3H), 6.25-5.56 (m, 1H), 5.32-5.01 (m, 0.5H), 4.78-4.56 (m, 0.5H), 4.37 (q, J=7.1 Hz, 2H), 4.21 (s, 1H), 3.99 (q, J=6.3 Hz, 0.5H), 3.84 (q, J=6.3 Hz, 0.5H), 3.76 (s, 3H), 3.72-3.47 (m, 1H), 3.30-3.17 (m, 1H), 3.06-2.65 (m, 2.5H), 2.19 (d, J=12.1 Hz, 0.5H), 1.45 (dd, J=13.2, 6.5 Hz, 7H), 1.28 (s, 2H), 1.12 (d, J=6.4 Hz, 1.5H), 0.97 (d, J=6.3 Hz, 1.5H) ppm. MS: M/e 554 (M+1).sup.+
Compound A55: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-methylbenzo[d]thiazol-5-yl)ethyl) piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00248##
[0457] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (100 mg, 0.3 mmol) in CH.sub.3CN (2 mL) and was added 1-(2-methylbenzo[d]thiazol-5-yl)ethan-1-ol (64 mg, 0.33 mmol), (cyanomethyl)trimethylphosphonium iodide (111 mg, 0.46 mmol) and DIPEA (156 mg, 1.2 mmol). The resulting mixture was stirred at 100 C. overnight. The reaction solvent was removed under reduced pressure. The resulting residue was purified by Prep-TLC (DCM:MeOH=13:1) to give the titled compound (36 mg, 24%). .sup.1H NMR (400 MHz, CDCl.sub.3) 7.94 (d, J=17.7 Hz, 1H), 7.77 (t, J=8.6 Hz, 1H), 7.42 (t, J=7.8 Hz, 1H), 5.75-4.95 (m, 2H), 4.30 (d, J=7.3 Hz, 2H), 3.96-3.67 (m, 6H), 3.55 (d, J=6.1 Hz, 2H), 2.84 (d, J=3.1 Hz, 4H), 2.21-2.05 (m, 1H), 1.51 (d, J=6.4 Hz, 4H), 1.38-1.22 (m, 5H), 1.06-0.84 (m, 3H) ppm. MS: M/e 460 (M+1).sup.+.
Compound A56: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-methylbenzo[d]thiazol-6-yl)ethyl) piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00249##
Step A: 1-(2-methylbenzo[d]thiazol-6-yl)ethan-1-ol
##STR00250##
[0458] To a solution of 1-(2-methylbenzo[d]thiazol-6-yl)ethan-1-one (150 mg, 0.79 mmol) in MeOH (15 mL) was added NaBH.sub.4 (30 mg, 0.79 mmol) at room temperature and the resulting mixture was stirred at room temperature for 15 mins. The reaction mixture was diluted with DCM (200 mL). The organic layer was washed with water, dried over Na.sub.2SO.sub.4 and concentrated to give the titled compound (130 mg, 86%). MS: M/e 194 (M+1).sup.+.
Step B: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-methylbenzo[d]thiazol-6-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00251##
[0459] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.15 mmol) in CH.sub.3CN (3 mL) and was added 1-(2-methylbenzo[d]thiazol-6-yl)ethan-1-ol (29 mg, 0.15 mmol), (cyanomethyl)trimethylphosphonium iodide (110 mg, 0.46 mmol) and DIPEA (98 mg, 0.76 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure to dryness. The resulting residue was purified by Prep-HPLC (Method A) to give the titled Compound A56a (26 mg) and Compound A56b (24 mg).
[0460] Compound A56a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.92 (s, 2H), 7.62 (s, 1H), 6.38-4.98 (m, 2H), 4.37 (d, J=7.2 Hz, 2H), 4.24 (s, 2H), 4.11-3.83 (m, 1H), 3.77 (s, 3H), 3.73-3.32 (m, 2H), 3.29-3.05 (m, 2H), 2.84 (s, 3H), 1.93-1.41 (m, 6H), 1.40-1.04 (m, 6H) ppm. MS: M/e 505 (M+1).sup.+
[0461] Compound A56b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.41-7.83 (m, 2H), 7.81-7.52 (m, 1H), 6.74-5.95 (m, 1H), 5.95-5.17 (m, 1H), 4.84-4.48 (m, 2H), 4.36 (s, 2H), 4.26 (s, 2H), 3.78 (s, 3H), 3.73-3.36 (m, 2H), 3.10-2.95 (m, 1H), 2.87 (s, 3H), 1.82 (s, 3H), 1.66-1.35 (m, 6H), 1.35-1.13 (m, 3H) ppm. MS: M/e 505 (M+1).sup.+
Compound A57: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-methyl-1H-benzo[d]imidazol-5-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00252##
Step A: mixture of 6-bromo-2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazole and 5-bromo-2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazole
##STR00253##
[0462] To a solution of 5-bromo-2-methyl-1H-benzo[d]imidazole (1.05 g, 5 mmol) in THE (20 mL) was added NaH (60%, 400 mg, 10 mmol) at 0 C. After 30 minutes, (2-(chloromethoxy)ethyl)trimethylsilane (1 g, 6 mmol) in THE (5 ml) was added to the reaction and the mixture was stirred at RT for 2 hours. The reaction mixture was quenched with H.sub.2O and extracted with EtOAc. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (DCM:MeOH=10:1) to give the titled compound (1.5 g, 88%). MS: M/e 341 (M+1).sup.+.
Step B: mixture of 1-(2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-6-yl)ethan-1-one and 1-(2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-5-yl)ethan-1-one
##STR00254##
[0463] A mixture of 6-bromo-2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazole and 5-bromo-2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazole (1.5 g, 4.4 mmol), tributyl(1-ethoxyvinyl)stannane (1.9 g, 5.3 mmol) and Pd(PPh.sub.3).sub.2Cl.sub.2 (308 mg, 0.44 mmol) in toluene (20 mL) was stirred at 100 C. under N.sub.2 overnight. The reaction mixture was concentrated under reduced pressure. The residue was diluted with THE (20 mL) followed by addition of TFA (1.5 mL) and stirred for 30 minutes. The resulting mixture was washed with saturated NaHCO.sub.3 aq., extracted with EtOAc, dried and concentrated to dryness. The resulting oil was purified by flash column chromatography (DCM:MeOH=10:1) to give the titled compound (500 mg, 37%) as a mixture. MS: M/e 305 (M+1).sup.+.
Step C: mixture of 1-(2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-6-yl)ethan-1-ol and 1-(2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-5-yl)ethan-1-ol
##STR00255##
[0464] To a solution of the mixture of 1-(2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-6-yl)ethan-1-one and 1-(2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-5-yl)ethan-1-one (200 mg, 0.67 mmol) in EtOH (4 mL) was added NaBH.sub.4 (25 mg, 0.67 mmol). The mixture was stirred at room temperature for 1 hour. The mixture was treated with water, extracted with DCM. The combined organic layers were dried over MgSO.sub.4 and concentrated to dryness. The resulting residue (200 mg, crude) was used in the next step without further purification. MS: M/e 307 (M+1).sup.+.
Step D: mixture of 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-5-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile and 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-6-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00256##
[0465] To a solution of the mixture of 1-(2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-6-yl)ethan-1-ol and 1-(2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-5-yl)ethan-1-ol (100 mg, 0.33 mmol), 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (100 mg, 0.3 mmol), (cyanomethyl)trimethylphosphonium iodide (145 mg, 0.6 mmol) and DIPEA (155 mg, 1.2 mmol) in MeCN (3 mL) was stirred at 100 C. overnight in a sealed tube. The mixture was diluted with EA (20 mL), washed with brine (10 mL3), dried, concentrated. The resulting residue was purified by Prep-TLC (CH.sub.2Cl.sub.2:MeOH=13:1) to give the titled compound (20 mg, 11%) as a mixture. MS: M/e 618 (M+1).sup.+.
Step E: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-methyl-1H-benzo[d]imidazol-5-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00257##
[0466] To a stirred mixture of 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-5-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile and 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-6-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (20 mg) in DCM (1 mL) was added TFA (1 mL). The resulting mixture was stirred at room temperature overnight. The reaction mixture was concentrated to dryness. The resulting residue was purified by Prep-HPLC(Method A) to give the titled compound (2 mg, 12%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.49-7.40 (m, 2H), 7.26 (t, J=8.6 Hz, 1H), 5.90 (s, 2H), 4.37 (d, J=7.0 Hz, 2H), 3.76 (s, 3H), 3.68-3.57 (m, 2H), 2.90 (s, 2H), 2.65 (s, 2H), 2.55 (d, J=3.6 Hz, 3H), 2.25 (d, J=12.0 Hz, 1H), 1.46 (t, J=6.9 Hz, 4H), 1.37 (dd, J=10.3, 6.5 Hz, 3H), 1.30-1.14 (m, 2H), 1.06-0.88 (m, 3H) ppm. MS: M/e 488 (M+1).sup.+.
Compound A58: 2-(6-((2S,5R)-4-(1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethyl piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00258##
[0467] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.159 mol), 1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (43 mg, 0.238 mol), (cyanomethyl)trimethylphosphonium iodide (77 mg. 0.318 mol) and DIPEA (62 mg, 0.477 mol) in CH.sub.3CN (1 ml). The mixture solution was degassed 3 times under N.sub.2 atmosphere. Then the mixture solution was stirred at 105 C. for 24 hours. The reaction mixture was concentrated under reduced pressure. The resulting residue was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled Compound A58 (30 mg), which was further separated into Compound A58a (12 mg) and Compound A58b (12 mg) by Prep-HPLC (Method A).
[0468] Compound A58: .sup.1H NMR (400 MHz, CDCl.sub.3) 7.69-7.44 (m, 0.5H), 7.30-7.27 (m, 1H), 6.91-6.68 (m, 1.5H), 6.21-4.42 (m, 2H), 4.29-4.23 (m, 4H), 4.06-3.86 (m, 4H), 3.80 (d, J=9.6 Hz, 3H), 3.55-3.40 (m, 2H), 3.35-3.04 (m, 0.5H), 3.01-2.52 (m, 1.5H), 1.95-1.72 (m, 2H), 1.47-1.32 (m, 3H), 1.25 (s, 3H), 1.10-0.75 (m, 3H) ppm. MS: M/e 478 (M+1).sup.+
[0469] Compound A58a (the earlier peak): .sup.1H NMR (400 MHz, CDCl.sub.3) 7.53-7.33 (m, 1H), 7.22-6.70 (m, 2H), 6.28-5.17 (m, 2H), 4.32-4.23 (m, 4H), 3.91 (d, J=27.8 Hz, 3H), 3.79 (s, 3H), 3.56-3.51 (m, 2H), 3.23-2.62 (m, 2H), 2.10-1.95 (m, 2H), 1.48-1.36 (m, 3H), 1.25 (s, 3H), 1.20-0.93 (m, 3H) ppm. MS: M/e 478 (M+1).sup.+
[0470] Compound A58b (the later peak): .sup.1H NMR (400 MHz, CDCl.sub.3) 7.52-7.34 (m, 1H), 7.10-6.78 (m, 2H), 6.15-5.80 (m, 0.5H), 5.53-5.13 (m, 1H), 4.59-4.54 (m, 0.5H), 4.29-4.24 (m, 4H), 4.05-3.90 (m, 4H), 3.79 (s, 3H), 3.59-3.18 (m, 2H), 3.07-2.42 (m, 2H), 2.09-1.82 (m, 2H), 1.54-1.37 (m, 3H), 1.31 (s, 3H), 1.26-0.82 (m, 3H) ppm. MS: M/e 478 (M+1).sup.+.
Compound A59: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00259##
[0471] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (60 mg, 0.190 mol), 1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (80 mg, 0.381 mol), (cyanomethyl)trimethylphosphonium iodide (93 mg. 0.381 mol) and DIPEA (147 mg, 1.143 mol) in CH.sub.3CN (1 ml). The mixture solution was degassed 3 times under N.sub.2 atmosphere. Then the mixture solution was stirred at 105 C. for 24 hours. The reaction mixture was concentrated under reduced pressure. The resulting residue was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled Compound A59 (30 mg), which was further separated into Compound A59a (8 mg) and Compound A59b (8 mg) by Prep-HPLC (Method A).
[0472] Compound A59a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.92-6.71 (m, 3H), 5.93-5.76 (m, 1H), 5.10-5.05 (m, 0.5H), 4.57-4.50 (m, 0.5H), 4.23-4.16 (m, 0.5H), 3.93 (s, 3H), 3.88 (s, 2H), 3.77 (s, 3H), 3.75-3.53 (m, 1.5H), 3.49-3.43 (m, 2H), 2.93-2.54 (m, 2H), 2.33 (d, J=12.3 Hz, 1H), 1.34 (d, J=6.6 Hz, 3H), 1.31 (d, J=1.5 Hz, 6H), 1.26 (d, J=6.6 Hz, 3H), 1.05 (d, J=6.4 Hz, 3H) ppm. MS: M/e 506 (M+1).sup.+
[0473] Compound A59b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.89-6.76 (m, 3H), 6.10-5.47 (m, 1H), 5.35-4.56 (m, 1H), 4.20 (d, J=5.0 Hz, 0.5H), 3.93 (s, 3H), 3.89 (s, 2H), 3.78 (s, 3H), 3.69-3.60 (m, 1H), 3.48-3.34 (m, 1.5H), 3.13-2.64 (m, 4H), 1.43 (d, J=6.4 Hz, 3H), 1.34 (d, J=8.8 Hz, 9H), 0.95 (d, J=6.4 Hz, 3H) ppm. MS: M/e 506 (M+1).sup.+
Compound A60: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00260##
[0474] A mixture of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (165 mg, 0.5 mmol), 1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (270 mg, 1.3 mmol) and (cyanomethyl)trimethylphosphonium iodide (243 mg, 1 mmol) in CH.sub.3CN (5 mL) was added DIPEA (645 mg, 5 mmol). The mixture was stirred at 100 C. overnight in a sealed tube. The reaction mixture was diluted with EtOAc (20 mL), washed with brine, dried over Na.sub.2SO.sub.4, concentrated. The resulting residue was purified by flash column chromatography (DCM:MeOH=15:1) to give the titled Compound A60 (30 mg), which was separated into Compound A60a (9 mg) and Compound A60b (12 mg) by chiral Prep-HPLC. The chiral separation conditions are shown below.
TABLE-US-00012 Column CHIRALPAK IH Column Size 2 cm 25 cm, 5 um Mobile Phase A MtBE Mobile Phase B MeOH:DCM Flow Rate 20 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-HPLC Equipment Prep-HPLC-Gilson
[0475] Compound A60a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.84-6.74 (m, J=6.4 Hz, 3H), 6.13-5.04 (m, 2H), 4.60-4.52 (m, 0.5H), 4.36 (q, J=7.2 Hz, 2H), 4.25-4.17 (m, 0.5H), 3.89 (s, 2H), 3.76 (s, 3H), 3.67-3.41 (m, 2H), 3.06-2.67 (m, 4H), 1.50-1.39 (m, 6H), 1.35-1.23 (m, 9H), 0.93 (d, J=6 Hz, 3H) ppm. MS: M/e 520 (M+1).sup.+.
[0476] Compound A60b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.84-6.74 (m, 3H), 6.02-5.00 (m, 2H), 4.37 (q, J=7.2 Hz, 2H), 3.87 (s, 2H), 3.76 (s, 3H), 3.70-3.31 (m, 4H), 3.09-2.52 (m, 2H), 2.26 (d, J=11.6 Hz, 1H), 1.46 (t, J=7.2 Hz, 3H), 1.34-1.21 (m, 12H), 1.02 (d, J=6 Hz, 3H) ppm. MS: M/e 500 (M+1).sup.+.
Compound A61: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-ethyl-9-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00261##
[0477] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3-ethyl-9-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (330 mg, 1 mmol) in CH.sub.3CN (5 mL) and was added 1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (624 mg, 3 mmol), (cyanomethyl)trimethylphosphonium iodide (972 mg, 4 mmol) and DIPEA (1.3 g, 10 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure and purified by Prep-TLC (DCM:MeOH=30:1) to give the titled Compound A61 (120 mg), which was further separated into Compound A61a (55 mg) and Compound A61b (40 mg) by chiral Prep-SFC. The chiral separation conditions are shown below.
TABLE-US-00013 Column CHIRALPAK IG Column Size 3 cm 25 cm, 5 um Mobile Phase A CO2 Mobile Phase B MeOH Flow Rate 100 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-SFC Equipment Prep-SFC-150
[0478] Compound A61a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.80 (d, J=3.0 Hz, 3H), 6.25-5.14 (m, 2H), 4.28 (q, J=6.9 Hz, 2H), 3.89 (d, J=3.7 Hz, 5H), 3.54-3.33 (m, 3H), 2.95-2.90 (m, 2H), 2.89-2.80 (m, 2H), 1.43 (d, J=5.9 Hz, 3H), 1.39-1.30 (m, 9H), 1.26 (d, J=6.4 Hz, 3H), 0.87 (d, J=6.5 Hz, 3H) ppm. MS: M/e 520 (M+1).sup.+.
[0479] Compound A61b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.85-6.70 (m, 3H), 6.01-4.99 (m, 2H), 4.28 (q, J=7.2 Hz, 2H), 3.93-3.86 (m, 5H), 3.75-3.70 (m, 1H), 3.51 (d, J=25.8 Hz, 1H), 3.35-3.33 (m, 2H), 2.79 (s, 1H), 2.58 (s, 1H), 2.25 (d, J=12.3 Hz, 1H), 1.41-1.16 (m, 15H), 1.01 (d, J=6.4 Hz, 3H) ppm. MS: M/e 520 (M+1).sup.+.
Compound A62: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-diethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00262##
Step A: 1-ethyl-6-(ethylamino)pyrimidine-2,4 (1H,3H)-dione
##STR00263##
[0480] To a stirred mixture of 6-chloro-1-methylpyrimidine-2,4 (1H,3H)-dione (35 g, 0.2 mol) in EtOH (200 mL) was added aq. CH.sub.3CH.sub.2NH.sub.2 (w/w=50%, 36 g, 0.4 mol). After the addition, the reaction mixture was stirred at 65 C. overnight. The reaction mixture was cooled to room temperature and filtered. The caked was collected and dried to give the titled compound (35 g, 96%) as a white solid. MS: M/e 184 (M+1).sup.+.
Step B: 1-ethyl-6-(ethylamino)-5-nitrosopyrimidine-2,4 (1H,3H)-dione
##STR00264##
[0481] To a stirred mixture of 1-ethyl-6-(ethylamino)pyrimidine-2,4 (1H,3H)-dione (35 g, 0.19 mol) in H.sub.2O (400 mL) was added NaNO.sub.2 (14.5 g, 0.21 mol) at 05 C. Then AcOH (34.2 g, 0.57 mol) was added dropwise at that temperature. After then, the mixture was stirred for an hour. The reaction mixture was filtered. The resulting cake was collected and dried to give the titled compound, which was directly used to the next step. MS: M/e 213 (M+1).sup.+.
Step C: 5-amino-1-ethyl-6-(ethylamino)-4-hydroxypyrimidin-2 (1H)-one
##STR00265##
[0482] To a stirred mixture of 1-ethyl-6-(ethylamino)-5-nitrosopyrimidine-2,4 (1H,3H)-dione (crude, 0.19 mol) in H.sub.2O (400 mL) was added Na.sub.2S.sub.2O.sub.4 (99.2 g, 0.57 mmol) portionwise at 2030 C. After the addition, the reaction mixture was stirred for 30 minutes and cooled in an ice-water bath for several hours and filtered. The cake was collected, dried to give the titled compound (17 g, 45%) as a white solid. MS: M/e 199 (M+1).sup.+.
Step D: 8-((benzyloxy)methyl)-3,9-diethyl-6-hydroxy-3,9-dihydro-2H-purin-2-one
##STR00266##
[0483] A mixture of 5-amino-1-ethyl-6-(ethylamino)-4-hydroxypyrimidin-2 (1H)-one (17 g, 86 mmol) in 2-(benzyloxy)acetic acid (15 g, 90 mmol) was stirred at 240 C. for 1.5 hour. The mixture was allowed cool to room temperature and dissolved in aq.NaOH (2.0 M, 100 mL), then acidified to pH=56 with aq.HCl and filtered. The cake was collected, dried to give the titled compound (17 g, 60%) as a light-yellow solid. MS: M/e 329 (M+1).sup.+.
Step E: tert-butyl (2R,5S)-4-(8-((benzyloxy)methyl)-3,9-diethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00267##
[0484] To a stirred mixture of 8-((benzyloxy)methyl)-3,9-diethyl-6-hydroxy-3,9-dihydro-2H-purin-2-one (4.28 g, 20 mmol) in CH.sub.3CN (50 mL) was added BOP (3.06 g, 20 mmol), followed by DBU (8.8 g, 20 mmol). After the addition, the reaction was stirred overnight. The reaction mixture was concentrated to give the residue, which was dissolved in EtOAc (150 mL), washed with H.sub.2O (20 mL), brine, dried over Na.sub.2SO.sub.4, concentrated. The resulting residue was purified by flash column chromatography to give the titled compound (1.8 g, 34%) as a white solid. MS: M/e 525 (M+1).sup.+.
Step F: tert-butyl (2R,5S)-4-(3,9-diethyl-8-(hydroxymethyl)-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00268##
[0485] To a stirred solution of tert-butyl (2R,5S)-4-(8-((benzyloxy)methyl)-3,9-diethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (1.8 g, 3.43 mmol) in MeOH/AcOH (100 mL/2 mL) was added Pd/C (500 mg). After the addition, the reaction mixture was stirred for 2 days under H.sub.2 (1 atm). The reaction mixture was filtered, and filtrate was concentrated to give the titled compound (1.5 g, 100%) as a whited solid. MS: M/e 435 (M+1).sup.+.
Step G: tert-butyl (2R,5S)-4-(8-(cyanomethyl)-3,9-diethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00269##
[0486] To a stirred solution of tert-butyl (2R,5S)-4-(3,9-diethyl-8-(hydroxymethyl)-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (crude, 3.24 mmol) in CH.sub.2Cl.sub.2 (15 mL) was added SOCl.sub.2 (0.41 g, 3.4 mmol). After stirred for 10 min, the reaction mixture was concentrated and dissolved in CH.sub.2Cl.sub.2 (10 mL), TBAF (1.0 M, 6.48 mL, 6.48 mmol) was added followed by TMSCN (0.64 g, 6.48 mmol). After the addition, the reaction mixture was stirred overnight. The reaction mixture was concentrated to give the residue, treated with H.sub.2O and extracted with EtOAc (40 mL2). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (0.9 g, 63%) as a white solid. MS: M/e 444 (M+1).sup.+.
Step H: 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-diethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00270##
[0487] To a stirred solution of tert-butyl (2R,5S)-4-(8-(cyanomethyl)-3,9-diethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (0.9 g, 2.02 mmol) in CH.sub.2Cl.sub.2 (10 mL) was added TFA (2 mL). After then, the reaction mixture was stirred for 2 hours. The reaction mixture was concentrated to give the residue, which was basified to pH=910 with K.sub.2CO.sub.3 and extracted with CH.sub.2Cl.sub.2/IPA (3/1, 40 mL5). The combined organic layers were dried over Na.sub.2SO.sub.4, concentrated, further lyophilized to give the titled compound (0.9 g, 100%), which was directly used to the next step. MS: M/e 342 (M+1).sup.+.
Step I: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-diethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00271##
[0488] A mixture of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-diethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (1 g, 2.92 mmol), 1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (0.91 g, 4.4 mmol), (cyanomethyl)trimethylphosphonium iodide (1.42 g, 5.84 mmol) and DIPEA (1.9 g, 14.6 mmol) in CH.sub.3CN (20 mL) was stirred overnight at 100 C. in a sealed tube. The reaction mixture was concentrated to give the residue, treated with H.sub.2O and extracted with EtOAc (30 mL2). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated. The resulting residue was purified by flash column chromatography to give the titled compound (230 mg, 15%). .sup.1H NMR (400 MHz, CD3OD) 6.90-6.75 (m, 3H), 6.21-4.98 (m, 2H), 4.33-4.17 (m, 6H), 3.88 (d, J=2.4 Hz, 2H), 3.78-3.68 (m, 1H), 3.63-3.41 (m, 1.5H), 3.27-3.19 (m, 1H), 3.06-2.76 (m, 1.5H), 1.46 (t, J=7.2 Hz, 3H), 1.40-1.34 (m, 6H), 1.34-1.29 (m, 9H), 1.08-0.84 (m, 3H) ppm. MS: M/e 534 (M+1).sup.+.
Compound A63: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00272##
Step A: tert-butyl (2R,5S)-4-(2-chloro-9H-purin-6-yl)-2-ethyl-5-methylpiperazine-1-carboxylate
##STR00273##
[0489] To a solution of 2,6-dichloro-9H-purine (8.0 g, 42.33 mmol) and tert-butyl (2R,5S)-2-ethyl-5-methylpiperazine-1-carboxylate (10.6 g, 46.56 mmol) in EtOH (130 mL) was added DIPEA (10.92 g, 84.66 mmol). The reaction was stirred at 80 C. for 12 hours. The reaction mixture was concentrated under reduced pressure to give the title compound (16.0 g, 99%). MS: M/e 381 (M+1).sup.+.
Step B: tert-butyl (2R,5S)-4-(2-chloro-9-methyl-9H-purin-6-yl)-2-ethyl-5-methylpiperazine-1-carboxylate
##STR00274##
[0490] To a solution of tert-butyl (2R,5S)-4-(2-chloro-9H-purin-6-yl)-2-ethyl-5-methylpiperazine-1-carboxylate (16.0 g, 42.11 mmol) and K.sub.2CO.sub.3 (11.62 g, 84.21 mmol) in acetone (150 mL) was added CH.sub.3I (12.0 g, 84.21 mmol). The reaction was stirred at room temperature for 16 hours. The reaction mixture was diluted with water, extracted with EtOAc (300 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (EtOAc:PE=1:1) to give the titled compound (14 g, 84%). MS: M/e 395 (M+1).sup.+.
Step C: tert-butyl (2R,5S)-2-ethyl-4-(2-hydroxy-9-methyl-9H-purin-6-yl)-5-methylpiperazine-1-carboxylate
##STR00275##
[0491] To a solution of tert-butyl (2R,5S)-4-(2-chloro-9-methyl-9H-purin-6-yl)-2-ethyl-5-methylpiperazine-1-carboxylate (14.0 g, 35.53 mmol), Pd.sub.2(dba).sub.3 (3.26 g, 3.56 mmol), t-BuXphos (3.03 g, 7.11 mmol) and KOH (35.6 mL, 3M, 106.06 mmol) in dioxane (120 mL) was degassed 3 times under N.sub.2 atmosphere. Then the mixture was stirred at 90 C. for 12 hours. The reaction mixture was diluted with water and extracted with EtOAc (90 mL2). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, filtered and concentrated to dryness. The resulting residue was purified by flash column chromatography (DCM:MeOH=10:1) to give the titled compound (13 g, 97%). MS: M/e 377 (M+1).sup.+.
Step D: tert-butyl (2R,5S)-4-(3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2-ethyl-5-methylpiperazine-1-carboxylate
##STR00276##
[0492] To a solution of tert-butyl (2R,5S)-2-ethyl-4-(2-hydroxy-9-methyl-9H-purin-6-yl)-5-methylpiperazine-1-carboxylate (13 g, 34.6 mmol) and Cs.sub.2CO.sub.3 (33.8 g, 103.7 mmol) in dioxane (180 mL) was added CH.sub.3I (14.7 g, 103.7 mmol). The reaction was stirred at room temperature for 16 hours. The reaction mixture was diluted with water, extracted with EtOAc (150 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (DCM:MeOH=20:1) to give the titled compound (9.5 g, 70%). MS: M/e 391 (M+1).sup.+.
Step E: tert-butyl (2R,5S)-2-ethyl-4-(8-formyl-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-5-methylpiperazine-1-carboxylate
##STR00277##
[0493] To a solution of tert-butyl (2R,5S)-4-(3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2-ethyl-5-methylpiperazine-1-carboxylate (9.5 g, 24.35 mmol) in THE (150 mL) was degassed 3 times under N.sub.2 atmosphere. Then added LDA (49 mL, 2M, 97.44 mmol) dropwise at 78 C. The mixture was stirred at 78 C. for 1 hours. Added DMF (8.9 g, 121.75 mmol) to quench the reaction solution. The reaction mixture was stirred for 1 hour and diluted with saturated NH.sub.4Cl, extracted with EtOAc (100 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (DCM:MeOH=20:1) to give the titled compound (6.8 g, 67%). MS: M/e 419 (M+1).sup.+.
Step F: tert-butyl (2R,5S)-2-ethyl-4-(8-(hydroxymethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-5-methylpiperazine-1-carboxylate
##STR00278##
[0494] To a solution of tert-butyl (2R,5S)-2-ethyl-4-(8-formyl-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-5-methylpiperazine-1-carboxylate (3.7 g, 8.831 mmol) in MeOH (30 mL) was added NaBH.sub.4 (671 mg, 17.66 mmol). The reaction was stirred at room temperature for 1 hours. The reaction mixture was quenched with saturated NH.sub.4Cl, extracted with EtOAc (50 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (DCM:MeOH=10:1) to give the titled compound (2.8 g, 75%). MS: M/e 421 (M+1).sup.+.
Step G: tert-butyl (2R,5S)-4-(8-(chloromethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2-ethyl-5-methylpiperazine-1-carboxylate
##STR00279##
[0495] To a solution of tert-butyl (2R,5S)-2-ethyl-4-(8-(hydroxymethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-5-methylpiperazine-1-carboxylate (1.6 g, 3.80 mmol) in DCM (20 mL) was added SOCl.sub.2 (497 mg, 4.181 mmol). The reaction was stirred at room temperature for 0.5 hours. The reaction mixture concentrated to give the titled compound (1.62 g, 97%). MS: M/e 439 (M+1).sup.+.
Step H: tert-butyl (2R,5S)-4-(8-(cyanomethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2-ethyl-5-methylpiperazine-1-carboxylate
##STR00280##
[0496] To a solution of tert-butyl (2R,5S)-4-(8-(chloromethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2-ethyl-5-methylpiperazine-1-carboxylate (1.62 g, 3.7 mmol) in DCM (25 mL) was added TBAF (7.4 mL, 1M, 7.4 mmol), followed TMSCN (734 mg, 7.4 mmol). The reaction was stirred at room temperature for 16 hours. The reaction mixture was diluted with water, extracted with DCM (40 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (PE:EtOAc=1:5) to give the titled compound (1.3 g, 82%). MS: M/e 430 (M+1).sup.+.
Step I: 2-(6-((2S,5R)-5-ethyl-2-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00281##
[0497] To a stirred solution of tert-butyl (2R,5S)-4-(8-(cyanomethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2-ethyl-5-methylpiperazine-1-carboxylate (1.3 g, 3.03 mmol) in CH.sub.2Cl.sub.2 (30 mL) was added TFA (6 mL). Then reaction mixture was stirred at room temperature for 4 hours, concentrated to give a residue, basified to pH=1011 with saturated NaHCO.sub.3 aq., and extracted with CH.sub.2Cl.sub.2/IPA (3/1, 50 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, and concentrated to give the titled compound (800 mg, 80%). MS: M/e 330 (M+1).sup.+.
Step J: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00282##
[0498] A solution of 2-(6-((2S,5R)-5-ethyl-2-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (420 mg, 1.276 mmol), 1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (531 mg, 2.552 mmol), (cyanomethyl)trimethylphosphonium iodide (620 mg. 2.552 mmol) and DIPEA (494 mg, 3.828 mol) in CH.sub.3CN (2 ml). The mixture solution was degassed 3 times under N.sub.2 atmosphere. Then the mixture solution was stirred at 105 C. for 24 hours. The reaction mixture was concentrated under reduced pressure. The resulting residue was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled Compound A63 (260 mg), which was further separated into Compound A63a (87 mg) and Compound A63b (68 mg) by Prep-HPLC (Method A).
[0499] Compound A63a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.85-6.72 (m, 3H), 6.10-5.73 (m, 1H), 5.18-5.12 (m, 0.5H), 4.83-4.77 (m, 0.5H), 3.91 (s, 2H), 3.85 (s, 3H), 3.78 (s, 3H), 3.63-3.54 (m, 1H), 3.42-3.34 (m, 0.5H), 3.30-3.24 (m, 2H), 3.12-3.06 (m, 0.5H), 2.80 (s, 2H), 2.53-2.46 (m, 1H), 1.48-1.36 (m, 4H), 1.32 (d, J=3.1 Hz, 6H), 1.29 (d, J=6.3 Hz, 4H), 0.72 (t, J=7.3 Hz, 3H) ppm. MS: M/e 520 (M+1).sup.+
[0500] Compound A63b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.86-6.66 (m, 3H), 6.16-5.76 (m, 1H), 5.04-4.95 (m, 1H), 3.92 (s, 3H), 3.87 (s, 2H), 3.78 (s, 3H), 3.69-3.57 (m, 0.5H), 3.48-3.40 (m, 1H), 3.30-3.24 (m, 2H), 3.14-3.05 (m, 1H), 2.85-2.45 (m, 1.5H), 2.27 (d, J=12.1 Hz, 1H), 1.51-1.39 (m, 2H), 1.31 (d, J=1.5 Hz, 6H), 1.25-1.19 (m, 6H), 1.00 (t, J=7.3 Hz, 3H) ppm. MS: M/e 520 (M+1).sup.+
Compound A64: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-diethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00283##
Step A: tert-butyl (2R,5S)-4-(8-((benzyloxy)methyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-diethylpiperazine-1-carboxylate
##STR00284##
[0501] To a solution of 8-((benzyloxy)methyl)-6-hydroxy-3,9-dimethyl-3,9-dihydro-2H-purin-2-one (3.85 g, 12.83 mmol), tert-butyl (2R,5S)-2,5-diethylpiperazine-1-carboxylate (6.21 g, 25.66 mmol) and BOP (11.35 g, 25.66 mmol) in CH.sub.3CN (100 mL) was added DBU (3.91 g, 25.66 mmol). The reaction mixture was diluted with water, extracted with EtOAc (150 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (PE:EtOAc=3:1) to give the titled compound (4.7 g, 70%). MS: M/e 525 (M+1).sup.+.
Step B: tert-butyl (2R,5S)-2,5-diethyl-4-(8-(hydroxymethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)piperazine-1-carboxylate
##STR00285##
[0502] To a solution of tert-butyl (2R,5S)-4-(8-((benzyloxy)methyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-diethylpiperazine-1-carboxylate (4.7 g, 8.97 mmol) and Pd/C (800 mg, 10% in water) in MeOH (60 mL) at room temperature was added AcOH (2 mL). The resulting mixture was degassed 3 times under H.sub.2 atmosphere and stirred at room temperature under H.sub.2 atmosphere for 36 hours. After filtration, the combined organic layers were concentrated under reduced pressure to give the titled compound (3.2 g, 82%). MS: M/e 435 (M+1).sup.+.
Step C: tert-butyl (2R,5S)-4-(8-(chloromethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-diethylpiperazine-1-carboxylate
##STR00286##
[0503] To a solution of tert-butyl (2R,5S)-2,5-diethyl-4-(8-(hydroxymethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)piperazine-1-carboxylate (1.84 g, 4.24 mmol) in DCM (30 mL) was added SOCl.sub.2 (555 mg, 4.66 mmol). The reaction was stirred at room temperature for 0.5 hours. The reaction mixture concentrated to give the titled compound (1.9 g, 99%). MS: M/e 453 (M+1).sup.+.
Step D: tert-butyl (2R,5S)-4-(8-(cyanomethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-diethylpiperazine-1-carboxylate
##STR00287##
[0504] To a solution of tert-butyl (2R,5S)-4-(8-(chloromethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-diethylpiperazine-1-carboxylate (1.90 g, 4.20 mmol) in DCM (40 mL) was added TBAF (8.4 mL, 1M, 8.4 mmol), followed TMSCN (834 mg, 8.4 mmol). The reaction was stirred at room temperature for 16 hours. The reaction mixture was diluted with water, extracted with DCM (40 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (PE:EtOAc=1:5) to give the titled compound (1.3 g, 70%). MS: M/e 444 (M+1).sup.+.
Step E: 2-(6-((2S,5R)-2,5-diethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00288##
[0505] To a stirred solution of tert-butyl (2R,5S)-4-(8-(cyanomethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-diethylpiperazine-1-carboxylate (1.3 g, 2.935 mmol) in CH.sub.2Cl.sub.2 (30 mL) was added TFA (5 mL). Then reaction mixture was stirred at room temperature for 4 hours, concentrated to give a residue, basified to pH=1011 with saturated NaHCO.sub.3 aq., and extracted with CH.sub.2Cl.sub.2/IPA (3/1, 50 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, and concentrated to give the titled compound (820 mg, 82%). MS: M/e 344 (M+1).sup.+.
Step F: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-diethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00289##
[0506] A solution of 2-(6-((2S,5R)-2,5-diethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (450 mg, 1.312 mmol), 1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (546 mg, 2.624 mmol), (cyanomethyl)trimethylphosphonium iodide (638 mg. 2.624 mmol) and DIPEA (508 mg, 3.936 mol) in CH.sub.3CN (2 ml). The mixture solution was degassed 3 times under N.sub.2 atmosphere. Then the mixture solution was stirred at 105 C. for 24 hours. The reaction mixture was concentrated under reduced pressure. The resulting residue was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled Compound A64 (240 mg), which was further separated into Compound A64a (68 mg) and Compound A64b (54 mg) by Prep-HPLC (Method A).
[0507] Compound A64a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.87-6.70 (m, 3H), 5.93 (s, 0.5H), 5.84 (d, J=13.3 Hz, 0.5H), 5.07 (s, 0.5H), 4.83 (s, 0.5H), 3.91 (d, J=7.7 Hz, 3H), 3.87 (s, 2H), 3.77 (s, 3H), 3.60-3.53 (m, 1H), 3.40-3.32 (m, 1.5H), 3.06-2.89 (m, 1.5H), 2.84-2.65 (m, 1H), 2.49-2.36 (m, 1H), 2.10-1.87 (m, 2H), 1.49-1.38 (m, 1H), 1.32 (d, J=3.6 Hz, 7H), 1.28 (d, J=6.4 Hz, 4H), 0.92 (t, J=7.3 Hz, 3H), 0.68 (dt, J=13.9, 7.0 Hz, 3H) ppm. MS: M/e 534 (M+1).sup.+
[0508] Compound A64b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.82 (s, 1H), 6.80-6.74 (m, 2H), 6.09 (d, J=13.5 Hz, 0.5H), 5.69 (s, 0.5H), 5.08 (d, J=13.3 Hz, 0.5H), 4.81 (s, 0.5H), 3.92 (d, J=3.7 Hz, 3H), 3.87 (d, J=1.6 Hz, 2H), 3.77 (s, 3H), 3.59-3.52 (m, 0.5H), 3.47-3.38 (m, 1H), 3.20 (d, J=13.1 Hz, 0.5H), 3.12-3.03 (m, 1H), 2.53-3.35 (m, 2H), 1.89-1.70 (m, 2H), 1.51-1.38 (m, 2H), 1.31 (d, J=8.0 Hz, 8H), 1.25 (d, J=6.7 Hz, 3H), 0.98 (t, J=5.8 Hz, 3H), 0.67 (dd, J=8.6, 5.4 Hz, 3H) ppm. MS: M/e 534 (M+1).sup.+
Compound A65: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00290##
[0509] A solution of 2-(9-ethyl-6-((2S,5R)-5-ethyl-2-methylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.15 mmol), 1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (30 mg, 0.15 mmol), (cyanomethyl)trimethylphosphonium iodide (106 mg, 0.44 mmol) and DIPEA (94 mg, 0.73 mmol) in MeCN (3 ml) was stirred at 100 C. overnight. After completed, the solution was concentrated under reduced pressure to dryness. The resulting residue was purified by Prep-HPLC (Method A) to give the titled Compound A65 (50 mg, 50%). .sup.1H NMR (400 MHz, CD.sub.3OD) 6.91-6.72 (m, 3H), 6.42-5.58 (m, 1H), 5.33-4.88 (m, 1H), 4.36 (q, J=7.0 Hz, 2H), 4.23 (s, 1H), 3.89 (d, J=8.7 Hz, 2H), 3.76 (s, 3H), 3.71-3.48 (m, 1H), 3.47-3.31 (m, 1H), 3.25-3.08 (m, 1H), 3.02-2.87 (m, 1H), 2.79-2.33 (m, 2H), 1.52-1.34 (m, 8H), 1.34-1.20 (m, 9H), 1.04-0.75 (m, 3H) ppm. MS: M/e 534 (M+1).sup.+
Compound A66: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-3-ethyl-9-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00291##
Step A: 1-ethyl-6-(methylamino)pyrimidine-2,4 (1H,3H)-dione
##STR00292##
[0510] To a stirred mixture of 6-chloro-1-methylpyrimidine-2,4 (1H,3H)-dione (10 g, 57.5 mmol) in EtOH (120 mL) was added aq. CH.sub.3NH.sub.2 (w/w=33%, 16 g, 0.174 mol). After the addition, the reaction mixture was stirred at 65 C. overnight. The reaction mixture was cooled to room temperature and filtered. The caked was collected and dried to give the titled compound (10 g) as a white solid, which was directly used to the next step. MS: M/e 170 (M+1).sup.+.
Step B: 1-ethyl-6-(methylamino)-5-nitrosopyrimidine-2,4 (1H,3H)-dione
##STR00293##
[0511] To a stirred mixture of 1-ethyl-6-(methylamino)pyrimidine-2,4 (1H,3H)-dione (10 g, 0.059 mol) in H.sub.2O (100 mL) was added NaNO.sub.2 (4.5 g, 0.065 mol) at 05 C. Then AcOH (10.6 g, 0.177 mol) was added dropwise at that temperature. After then, the mixture was stirred for 1.5 hour. The reaction mixture was filtered. The resulting cake was collected and dried to give the titled compound (7.2 g, 84%), which was directly used to the next step. MS: M/e 199 (M+1).sup.+.
Step C: 5-amino-1-ethyl-4-hydroxy-6-(methylamino)pyrimidin-2 (1H)-one
##STR00294##
[0512] To a stirred mixture of 1-ethyl-6-(ethylamino)-5-nitrosopyrimidine-2,4 (1H,3H)-dione (7.2, 0.036 mol) in H.sub.2O (70 mL) was added Na.sub.2S.sub.2O.sub.4 (18.7 g, 0.107 mol) portionwise at 20-30 C. After the addition, the reaction mixture was stirred for 30 minutes and cooled in an ice-water bath for several hours and filtered. The cake was collected, dried to give the titled compound (17 g, 45%) as a white solid, which was directly used to the next step. MS: M/e 1 (M+1).sup.+.
Step D: 8-((benzyloxy)methyl)-3-ethyl-6-hydroxy-9-methyl-3,9-dihydro-2H-purin-2-one
##STR00295##
[0513] A mixture of 5-amino-1-ethyl-6-(ethylamino)-4-hydroxypyrimidin-2 (1H)-one (2.5 g, 13.5 mmol) in 2-(benzyloxy)acetic acid (2.5 g, 14.9 mmol) was stirred at 240 C. for 1.5 hour. The mixture was allowed cool to room temperature and dissolved in aq.NaOH (2.0 M, 40 mL), then acidified to pH=56 with aq.HCl and filtered. The cake was collected, dried to give the titled compound (2.8 g, 66%) as a light-yellow solid, which was directly used to the next step. MS: M/e 315 (M+1).sup.+.
Step E: tert-butyl (2R,5S)-4-(8-((benzyloxy)methyl)-3-ethyl-9-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2-ethyl-5-methylpiperazine-1-carboxylate
##STR00296##
[0514] To a stirred mixture of 8-((benzyloxy)methyl)-3-ethyl-6-hydroxy-9-methyl-3,9-dihydro-2H-purin-2-one (1 g, 3.2 mmol) in CH.sub.3CN (10 mL) was added tert-butyl tert-butyl (2R,5S)-2-ethyl-5-methylpiperazine-1-carboxylate (1.36 g, 6.4 mmol), BOP (2.8 g, 6.4 mmol), followed by DBU (1 g, 6.4 mmol). After the addition, the reaction was stirred overnight. The reaction mixture was concentrated to give the residue, which was dissolved in EtOAc (50 mL), washed with H.sub.2O (20 mL), brine, dried over Na.sub.2SO.sub.4, concentrated. The resulting residue was purified by flash column chromatography to give the titled compound (1.6 g, 95%) as a white solid. MS: M/e 525 (M+1).sup.+.
Step F: tert-butyl (2R,5S)-2-ethyl-4-(3-ethyl-8-(hydroxymethyl)-9-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-5-methylpiperazine-1-carboxylate
##STR00297##
[0515] To a stirred solution of tert-butyl (2R,5S)-4-(8-((benzyloxy)methyl)-3-ethyl-9-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2-ethyl-5-methylpiperazine-1-carboxylate (1.6 g, 3.1 mmol) in MeOH/AcOH (15 mL/0.5 mL) was added Pd/C (500 mg). After the addition, the reaction mixture was stirred for 2 days under H.sub.2 (1 atm). The reaction mixture was filtered. The resulting filtrate was concentrated to give the titled compound (1.3 g, 81%) as a whited solid, which was directly used to the next step. MS: M/e 435 (M+1).sup.+.
Step G: tert-butyl (2R,5S)-4-(8-(cyanomethyl)-3-ethyl-9-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2-ethyl-5-methylpiperazine-1-carboxylate
##STR00298##
[0516] To a stirred solution of tert-butyl (2R,5S)-2-ethyl-4-(3-ethyl-8-(hydroxymethyl)-9-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-5-methylpiperazine-1-carboxylate (3.3 g, 7.6 mmol) in CH.sub.2Cl.sub.2 (15 mL) was added SOCl.sub.2 (1.35 g, 11.4 mmol). After stirred for 10 min, the reaction mixture was concentrated and dissolved in CH.sub.2Cl.sub.2 (100 mL), TBAF (1.0 M, 38 mL, 38 mmol) was added followed by TMSCN (3.7 g, 38 mmol). After the addition, the reaction mixture was stirred overnight. The reaction mixture was concentrated to give the residue, treated with H.sub.2O and extracted with EtOAc (100 mL2). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated. The resulting residue was purified by flash column chromatography to give the titled compound (1.5 g, 45%) as a white solid. MS: M/e 444 (M+1).sup.+.
Step H: 2-(3-ethyl-6-((2S,5R)-5-ethyl-2-methylpiperazin-1-yl)-9-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00299##
[0517] To a stirred solution of tert-butyl (2R,5S)-4-(8-(cyanomethyl)-3-ethyl-9-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2-ethyl-5-methylpiperazine-1-carboxylate (0.4 g, 0.9 mmol) in CH.sub.2Cl.sub.2 (10 mL) was added TFA (2 mL). After then, the reaction mixture was stirred for 2 hours. The reaction mixture was concentrated to give the residue, which was basified to pH=910 with K.sub.2CO.sub.3 (water 5 mL) and extracted with CH.sub.2Cl.sub.2/IPA (3/1, 100 mL3). The combined organic layers were dried over Na.sub.2SO.sub.4, concentrated, further lyophilized to give the titled compound (0.38 g, crude), which was directly used to the next step. MS: M/e 342 (M+1).sup.+.
Step I: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-3-ethyl-9-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00300##
[0518] A mixture of 2-(3-ethyl-6-((2S,5R)-5-ethyl-2-methylpiperazin-1-yl)-9-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (0.38 g, 1.1 mmol), 1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (0.7 g, 3.3 mmol), (cyanomethyl)trimethylphosphonium iodide (1.1 g, 4.43 mmol) and DIPEA (1.43 g, 11 mmol) in CH.sub.3CN (8 mL) was stirred overnight at 105 C. in a sealed tube. The reaction mixture was concentrated to give the residue, treated with H.sub.2O and extracted with EtOAc (30 mL2). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated. The reaction solvent was removed under reduced pressure to give the titled Compound A66, which was further separated into Compound A66a (25 mg) and Compound A66b (10 mg) by Prep-HPLC (Method A).
[0519] Compound A66a (the earlier peak): 1H NMR (400 MHz, DMSO-d6) 6.87-6.73 (m, 3H), 6.21-4.93 (m, 2H), 4.28 (q, J=7.0 Hz, 2H), 4.23-4.18 (m, 1H), 3.88 (d, J=3.9 Hz, 5H), 3.68-3.38 (m, 2H), 3.13 (s, 2H), 2.70-2.45 (m, 1H), 2.35 (s, 1H), 1.49 (d, J=7.5 Hz, 2H), 1.41-1.15 (m, 15H), 1.01 (t, J=7.3 Hz, 3H) ppm. MS: M/e 534 (M+1).sup.+.
[0520] Compound A66b (the later peak): 1H NMR (400 MHz, DMSO-d6) 6.90-6.70 (m, 3H), 6.10-5.05 (m, 2H), 4.28 (q, J=7.0 Hz, 2H), 4.21 (s, 1H), 3.89 (d, J=5.8 Hz, 5H), 3.75-3.35 (m, 2H), 3.12-2.61 (m, 3H), 2.52 (s, 1H), 1.53-1.22 (m, 17H), 0.74 (t, J=7.2 Hz, 3H) ppm. MS: M/e 534 (M+1).sup.+.
Compound A67: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-diethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00301##
[0521] A solution of 2-(6-((2S,5R)-2,5-diethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.14 mmol), 1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (29 mg, 0.14 mmol), (cyanomethyl)trimethylphosphonium iodide (102 mg, 0.42 mmol) and DIPEA (90 mg, 0.70 mmol) in CH.sub.3CN (3 ml) was stirred at 100 C. overnight. After completed, the solution was concentrated under reduced pressure to dryness. The resulting residue was purified by Prep-HPLC (Method A) to give the titled compound (42 mg, 58%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.01-6.75 (m, 3H), 6.42-4.93 (m, 2H), 4.41-4.20 (m, 4H), 3.91 (d, J=6.7 Hz, 2H), 3.77 (s, 3H), 3.48 (s, 2H), 2.89-2.49 (m, 2H), 2.14-1.67 (m, 3H), 1.66-1.36 (m, 8H), 1.32 (d, J=7.3 Hz, 6H), 1.09-0.93 (m, 3H), 0.85-0.60 (m, 3H) ppm. MS: M/e 548 (M+1).sup.+
Compound A68: 2-(6-((2S,5R)-4-(1-(8-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00302##
Step A: 3,4-difluoro-5-hydroxybenzonitrile
##STR00303##
[0522] A mixture of 5-bromo-2,3-difluorophenol (3 g, 14 mmol), tBuXPhos Pd G3 (222 mg, 0.28 mmol) and Zn(CN).sub.2 (3.27 g, 28 mmol) in THE (15 mL) and H.sub.2O (15 mL) was stirred at 40 C. for overnight. The reaction mixture was extracted with EtOAc (50 mL2). The combined organics were concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (1.7 g, 76%). MS: M/e 156 (M+1).sup.+.
Step B: 8-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxine-6-carbonitrile
##STR00304##
[0523] To a solution of 3,4-difluoro-5-hydroxybenzonitrile (1.7 g, 10.9 mmol) and Cs.sub.2CO.sub.3 (10.7 g, 32.9 mmol) in DMAc (50 mL) was added 2,2-dimethyloxirane (6.3 g, 87.2 mol) at room temperature. The reaction mixture was stirred at 100 C. for overnight. The reaction mixture was filtered. H.sub.2O (50 mL) was added to the filtrates. The mixture was extracted with PE (100 mL3). The organic phases were concentrated and purified by silica gel chromatography (PE:EtOAc=10:1) to give the titled compound (1.3 g, 57%). MS: M/e 208 (M+1).sup.+.
Step C: 8-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxine-6-carbaldehyde
##STR00305##
[0524] To a solution of 8-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxine-6-carbonitrile (1.3 g, 6.2 mmol) in THE (20 mL) was added Dibal-H (9 mL, 9 mmol, 1M in n-Hexane) slowly at 0 C. The reaction mixture was stirred at 0 C. for 2 hours. The reaction was quenched by 2 N HCl (30 mL). The resulting mixture was stirred at room temperature for 1 hours. The mixture was extracted with EtOAc (50 mL3). The organic phases were concentrated and purified by combi flash to give the titled compound (0.9 g, 69%). MS: M/e 211 (M+1).sup.+.
Step D: 1-(8-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol
##STR00306##
[0525] To a solution of 8-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxine-6-carbaldehyde (0.9 g, 4.3 mmol) in THE (10 mL) was added MeMgBr (2.7 mL, 8.6 mmol, 3M in Et.sub.2O) slowly at 0 C. The resulting mixture was stirred at 0 C. for 2 hours. The reaction mixture was quenched with H.sub.2O (10 mL) and extracted with EtOAc (30 mL3). The combined organic phases were concentrated and purified by combi flash to give the titled compound (550 mg, 57% yield). MS: M/e 227 (M+1).sup.+.
Step E: 2-(6-((2S,5R)-4-(1-(8-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00307##
[0526] To a solution of 1-(8-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol
[0527] (64 mg, 0.28 mmol), 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (60 mg, 0.19 mmol) and (cyanomethyl) trimethyl phosphonium iodide (68 mg, 0.28 mmol) in CH.sub.3CN (5 mL) was added DIPEA (116 mg, 0.9 mmol). The mixture was sealed in a bottle and heated at 100 C. for 16 hours. The mixture was cooled to room temperature, diluted with water, extracted with EtOAc (10 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled Compound A68 (23 mg, 23%), which was separated into Compound A68a (5 mg) and Compound A68b (4 mg) by chiral Prep-HPLC. The chiral separation conditions are shown below.
TABLE-US-00014 Column CHIRALPAK IH Column Size 2 cm 25 cm, 5 um Mobile Phase A MtBE (0.5% 2 mM NH3MeOH) Mobile Phase B MeOH:DCM Flow Rate 20 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-HPLC Equipment Prep-HPLC-Gilson
[0528] Compound A68: .sup.1H NMR (400 MHz, CD.sub.3OD) 6.73-6.65 (m, 2H), 6.01-5.95 (m, 1H), 5.50-5.03 (m, 1H), 4.21 (s, 2H), 3.95-3.91 (m, 4H), 3.74-3.71 (m, 4H), 3.29-3.25 (m, 2H), 2.83-2.12 (m, 3H), 1.44-1.26 (m, 12H), 1.05-0.90 (m, 3H) ppm. MS: M/e 524 (M+1).sup.+.
[0529] Compound A68a (the earlier peak) .sup.1H NMR (400 MHz, CD.sub.3OD) 6.71 (d, J=11.3 Hz, 1H), 6.64 (s, 1H), 6.19-5.95 (m, 1H), 5.50-5.25 (m, 1H), 4.20 (s, 1H), 3.93 (d, J=3.4 Hz, 4H), 3.78 (s, 3H), 3.50-3.10 (m, 2H), 2.90-2.75 (m, 5H), 1.46-1.35 (m, 6H), 1.29-1.24 (m, 5H), 0.89-0.88 (m, 3H) ppm. MS: M/e 524 (M+1).sup.+.
[0530] Compound A68b (the later peak) .sup.1H NMR (400 MHz, CD.sub.3OD) 6.71 (d, J=11.3 Hz, 1H), 6.63 (s, 1H), 6.01-5.75 (m, 1H), 5.23-5.03 (m, 1H), 4.21 (s, 1H), 3.95-3.93 (m, 5H), 3.78 (s, 3H), 3.51-3.47 (m, 1H), 2.83-2.51 (m, 4H), 2.25-2.22 (m, 1H), 1.40-1.26 (m, 12H), 1.05-0.95 (m, 3H) ppm. MS: M/e 524 (M+1).sup.+.
Compound A69: 2-(6-((2S,5R)-4-(1-(7-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,4,5,9-tetrahydro-2H-purin-8-yl)acetonitrile
##STR00308##
Step A: 2,4-difluoro-5-hydroxybenzonitrile
##STR00309##
[0531] To a solution of 5-bromo-2,4-difluorophenol (1.0 g, 4.785 mmol), t-BuXphos Pd G3 (190 mg, 0.239 mmol), t-BuXphos (204 mg, 0.48 mmol) and Zn(CN).sub.2 (1.12 g, 9.570 mmol) in THE (20 mL) and H.sub.2O (10 mL) was degassed 3 times under N.sub.2 atmosphere. Then the mixture was stirred at 70 C. for 12 hours. After filtration, the reaction mixture was diluted with water and extracted with EtOAc (50 mL2). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, filtered and concentrated to dryness. The resulting residue was purified by flash column chromatography (PE:EtOAc=5:1) to give the titled compound (730 mg, 98%).
Step B: 7-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxine-6-carbonitrile
##STR00310##
[0532] To a solution of 2,4-difluoro-5-hydroxybenzonitrile (700 mg, 4.516 mmol) and Cs.sub.2CO.sub.3 (5.15 g, 13.548 mmol) in DMAc (25 mL) was added 2,2-dimethyloxirane (4.885 g, 67.74 mmol). The reaction was stirred at 90 C. for 12 hours. After filtration, the reaction mixture was diluted with water, extracted with PE (60 mL3), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (PE:EtOAc=5:1) to give the titled compound (220 mg, 24%). .sup.1H NMR (400 MHz, CDCl.sub.3) 7.09 (d, J=6.1 Hz, 1H), 6.66 (d, J=9.7 Hz, 1H), 3.88 (s, 2H), 1.36 (s, 6H) ppm.
Step C: 7-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxine-6-carbaldehyde
##STR00311##
[0533] To a solution of 7-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxine-6-carbaldehyde (200 mg, 0.966 mmol) in THE (15 mL) was degassed 3 times under N.sub.2 atmosphere. Then added DIBAL-H (1.45 mL, 1M, 1.45 mmol) dropwise at 0 C. The mixture was stirred at 0 C. for 4 hours. The reaction solution was quenched by 1M HCl. After filtration, the reaction mixture was diluted with water, extracted with EtOAc (50 mL3), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (PE:EtOAc=5:1) to give the titled compound (120 mg, 54%).
Step D: 1-(7-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol
##STR00312##
[0534] To a solution of 7-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxine-6-carbaldehyde (110 mg, 0.523 mmol) in THE (10 mL) was degassed 3 times under N.sub.2 atmosphere. Then added CH.sub.3MgBr (0.35 mL, 3M, 1.048 mmol) dropwise at 0 C. The mixture was stirred at 0 C. for 4 hours. The reaction solution was quenched with saturated NH.sub.4Cl, extracted with EtOAc (50 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (PE:EtOAc=5:1) to give the titled compound (86 mg, 73%).
Step E: 2-(6-((2S,5R)-4-(1-(7-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,4,5,9-tetrahydro-2H-purin-8-yl)acetonitrile
##STR00313##
[0535] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (80 mg, 0.254 mmol), 1-(7-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (86 mg, 0.381 mmol), (cyanomethyl)trimethylphosphonium iodide (123 mg. 0.508 mmol) and DIPEA (98 mg, 0.762 mol) in CH.sub.3CN (1 ml). The mixture solution was degassed 3 times under N.sub.2 atmosphere. Then the mixture solution was stirred at 100 C. for 24 hours. The reaction mixture was concentrated under reduced pressure. The resulting residue was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled Compound A69 (29 mg), which was further separated into Compound A69a (8 mg) and Compound A69b (6 mg) by Prep-HPLC (Method A).
[0536] Compound A69: .sup.1H NMR (400 MHz, CD.sub.3OD) 7.08-6.90 (m, 1H), 6.54 (dt, J=24.0, 11.8 Hz, 1H), 6.15-5.52 (m, 1H), 5.37-4.96 (m, 0.5H), 4.72-4.49 (m, 0.5H), 4.21 (s, 1H), 3.93 (s, 3H), 3.88 (d, J=6.0 Hz, 2H), 3.80 (s, 3H), 3.76-3.67 (m, 1.5H), 3.58-3.47 (m, 0.5H), 3.23 (dd, J=14.8, 7.4 Hz, 1H), 2.95-2.87 (m, 1H), 2.80-2.55 (m, 1.5H), 2.32-2.19 (m, 0.5H), 1.37 (d, J=5.2 Hz, 3H), 1.32 (d, J=5.9 Hz, 6H), 1.29 (s, 3H), 1.02 (d, J=5.4 Hz, 1.5H), 0.91 (d, J=5.6 Hz, 1.5H) ppm. MS: M/e 524 (M+1).sup.+
[0537] Compound A69a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.95 (d, J=6.9 Hz, 1H), 6.56 (d, J=10.7 Hz, 1H), 6.20-6.00 (m, 0.5H), 5.78-5.46 (m, 0.5H), 5.26-5.05 (m, 0.5H), 4.76-4.51 (m, 0.5H), 4.21 (s, 1H), 3.93 (s, 3H), 3.90 (d, J=4.4 Hz, 2H), 3.78 (s, 3H), 3.58-3.47 (m, 1H), 2.95-2.87 (m, 2H), 2.83-2.65 (m, 2H), 2.32-1.98 (m, 1H), 1.42 (d, J=4.4 Hz, 3H), 1.32 (d, J=4.0 Hz, 6H), 1.28 (d, J=6.3 Hz, 3H), 0.90 (d, J=5.9 Hz, 3H) ppm. MS: M/e 524 (M+1).sup.+
[0538] Compound A69b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.05 (d, J=6.9 Hz, 1H), 6.49 (d, J=11.0 Hz, 1H), 6.02-5.78 (m, 1H), 5.14-5.01 (m, 0.5H), 4.84-4.71 (m, 0.5H), 4.21 (s, 1H), 3.93 (s, 3H), 3.88 (s, 2H), 3.78 (s, 3H), 3.54-3.31 (m, 2H), 2.74-2.53 (m, 2H), 2.34-1.98 (m, 2H), 1.31 (s, 6H), 1.30 (s, 6H), 1.02 (d, J=6.2 Hz, 3H) ppm. MS: M/e 524 (M+1).sup.+
Compound A70: 2-(6-((2S,5R)-4-(1-(5-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00314##
Step A: 2,4-difluoro-3-hydroxybenzonitrile
##STR00315##
[0539] To a solution of 3-bromo-2,6-difluorophenol (1.0 g, 4.785 mmol), t-BuXphos Pd G3 (190 mg, 0.239 mmol), t-BuXphos (204 mg, 0.48 mmol) and Zn(CN).sub.2 (1.12 g, 9.570 mmol) in THE (20 mL) and H.sub.2O (10 mL) was degassed 3 times under N.sub.2 atmosphere. Then the mixture was stirred at 70 C. for 12 hours. After filtration, the reaction mixture was diluted with water and extracted with EtOAc (50 mL2). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, filtered and concentrated to dryness. The resulting residue was purified by flash column chromatography (PE:EtOAc=5:1) to give the titled compound (690 mg, 93%). MS: M/e 156 (M+1).sup.+
Step B: 5-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxine-6-carbonitrile
##STR00316##
[0540] To a solution of 2,4-difluoro-3-hydroxybenzonitrile (690 mg, 4.452 mmol) and Cs.sub.2CO.sub.3 (5.08 g, 13.355 mmol) in DMAc (25 mL) was added 2,2-dimethyloxirane (4.815 g, 66.78 mmol). The reaction was stirred at 90 C. for 12 hours. After filtration, the reaction mixture was diluted with water, extracted with PE (60 mL3), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (PE:EtOAc=5:1) to give the titled compound (500 mg, 54%). MS: M/e 208 (M+1).sup.+
Step C: 5-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxine-6-carbaldehyde
##STR00317##
[0541] To a solution of 5-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxine-6-carbonitrile (500 mg, 2.415 mmol) in THE (50 mL) was degassed 3 times under N.sub.2 atmosphere. Then added DIBAL-H (4.8 mL, 1M, 4.831 mmol) dropwise at 0 C. The mixture was stirred at 0 C. for 4 hours. The reaction solution was quenched by 1M HCl. After filtration, the reaction mixture was diluted with water, extracted with EtOAc (50 mL3), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (PE:EtOAc=5:1) to give the titled compound (400 mg, 79%). MS: M/e 211 (M+1).sup.+
Step D: 1-(5-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol
##STR00318##
[0542] To a solution of 5-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxine-6-carbaldehyde (400 mg, 1.905 mmol) in THE (20 mL) was degassed 3 times under N.sub.2 atmosphere. Then added CH.sub.3MgBr (1.3 mL, 3M, 3.810 mmol) dropwise at 0 C. The mixture was stirred at 0 C. for 4 hours. The reaction solution was quenched with saturated NH.sub.4Cl, extracted with EtOAc (50 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography (PE:EtOAc=5:1) to give the titled compound (320 mg, 74%). .sup.1H NMR (400 MHz, CDCl.sub.3) 6.84 (dd, J=13.2, 6.3 Hz, 1H), 6.68 (dd, J=20.2, 9.8 Hz, 1H), 5.04 (dd, J=23.0, 6.2 Hz, 1H), 3.95 (s, 2H), 1.49 (dd, J=8.5, 3.5 Hz, 3H), 1.40 (s, 6H) ppm.
Step E: 2-(6-((2S,5R)-4-(1-(5-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00319##
[0543] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (80 mg, 0.254 mmol), 1-(5-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (86 mg, 0.381 mmol), (cyanomethyl)trimethylphosphonium iodide (123 mg. 0.508 mmol) and DIPEA (98 mg, 0.762 mol) in CH.sub.3CN (1 ml). The mixture solution was degassed 3 times under N.sub.2 atmosphere. Then the mixture solution was stirred at 100 C. for 24 hours. The reaction mixture was concentrated under reduced pressure. The resulting residue was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled compound (38 mg, 29%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.16-6.89 (m, 1H), 6.78-6.56 (m, 1H), 6.04-5.45 (m, 1H), 5.35-5.08 (m, 0.5H), 4.73-4.54 (m, 0.5H), 4.27-4.08 (m, 1.5H), 4.01-3.96 (m, 2.5H), 3.86 (s, 3H), 3.79 (s, 3H), 3.76-3.66 (m, 0.5H), 3.57-3.36 (m, 1H), 3.25-3.21 (m, 0.5H), 2.99-2.52 (m, 2.5H), 2.26 (t, J=11.3 Hz, 0.5H), 1.39 (d, J=8.0 Hz, 2H), 1.34 (d, J=6.2 Hz, 6H), 1.27 (d, J=8.5 Hz, 4H), 1.06-0.85 (m, 3H) ppm. MS: M/e 524 (M+1).sup.+
Compound A71: 2-(9-ethyl-6-((2S,5R)-4-(1-(5-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00320##
[0544] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (80 mg, 0.243 mmol), 1-(5-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (82 mg, 0.365 mmol), (cyanomethyl)trimethylphosphonium iodide (119 mg. 0.486 mmol) and DIPEA (94 mg, 0.729 mol) in CH.sub.3CN (1 ml). The mixture solution was degassed 3 times under N.sub.2 atmosphere. Then the mixture solution was stirred at 100 C. for 24 hours. The reaction mixture was concentrated under reduced pressure. The resulting residue was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled compound (46 mg, 35%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.14-6.95 (m, 1H), 6.67 (q, J=9.9 Hz, 1H), 6.17-5.39 (m, 1H), 5.35-4.94 (m, 0.5H), 4.65-4.58 (m, 0.5H), 4.45-4.29 (m, 2H), 4.21-4.06 (m, 1H), 3.92 (d, J=11.1 Hz, 2H), 3.76 (s, 3H), 3.58-3.38 (m, 1H), 3.30-3.05 (m, 2H), 2.93-2.55 (m, 2H), 2.35-1.95 (m, 1H), 1.46 (t, J=7.1 Hz, 3H), 1.37-1.32 (m, 6H), 1.26 (d, J=17.2 Hz, 6H), 1.03 (d, J=4.7 Hz, 1.5H), 0.92 (d, J=5.6 Hz, 1.5H) ppm. MS: M/e 538 (M+1).sup.+
Compound A72: 2-(6-((2S,5R)-4-(1-(8-chloro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00321##
Step A: 3-chloro-4-fluoro-5-hydroxybenzonitrile
##STR00322##
[0545] A mixture of 5-bromo-3-chloro-2-fluorophenol (1 g, 4.4 mmol), tBuXPhos Pd G3 (71 mg, 0.09 mmol) and Zn(CN).sub.2 (1.0 g, 8.8 mmol) in THE (5 mL) and H.sub.2O (5 mL) was stirred at 40 C. for overnight. The reaction mixture was extracted with EtOAc (20 mL2). The combined organics were concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (210 mg, 28%). MS: M/e 172 (M+1).sup.+.
Step B: 8-chloro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxine-6-carbonitrile
##STR00323##
[0546] To a solution of 3-chloro-4-fluoro-5-hydroxybenzonitrile (210 mg, 1.2 mmol) and Cs.sub.2CO.sub.3 (1.2 g, 3.6 mmol) in DMAc (5 mL) was added 2,2-dimethyloxirane (690 mg, 9.6 mol) at room temperature. The reaction mixture was stirred at 100 C. for overnight. The reaction mixture was filtered. H.sub.2O (10 mL) was added to the filtrates. The mixture was extracted with PE (50 mL3). The organic phases were concentrated to dryness. The resulting residue was purified by flash column chromatography (PE:EtOAc=10:1) to give the titled compound (170 mg, 62%). MS: M/e 224 (M+1).sup.+.
Step C: 1-(8-chloro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-one
##STR00324##
[0547] To a solution of 8-chloro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxine-6-carbonitrile (170 mg, 0.76 mmol) in THE (2 mL) was added MeMgBr (1.5 mL, 1.5 mmol, 1M in THF) slowly at 0 C. The reaction mixture was stirred at 70 C. for overnight. The reaction was quenched by 2 N HCl (10 mL). The resulting mixture was stirred at room temperature for 1 hours. The mixture was extracted with EtOAc (50 mL3). The organic phases were concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (150 mg, 82%). MS: M/e 241 (M+1).sup.+.
Step D: 1-(8-chloro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol
##STR00325##
[0548] To a solution of 1-(8-chloro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-one (150 mg, 0.63 mmol) in MeOH (5 mL) was added NaBH.sub.4 (26 mg, 0.7 mmol) slowly at 0 C. The resulting mixture was stirred at room temperature for 2 hours. The reaction mixture was quenched with H.sub.2O (10 mL) and extracted with EtOAc (30 mL3). The combined organic phases were concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (120 mg, 80% yield). MS: M/e 243 (M+1).sup.+.
Step E: 2-(6-((2S,5R)-4-(1-(8-chloro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00326##
[0549] To a solution of 1-(8-chloro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (60 mg, 0.28 mmol), 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (60 mg, 0.19 mmol) and (cyanomethyl) trimethyl phosphonium iodide (68 mg, 0.28 mmol) in CH.sub.3CN (5 mL) was added DIPEA (116 mg, 0.9 mmol). The mixture was sealed in a bottle and heated at 100 C. for 16 hours. The mixture was cooled to room temperature, diluted with water, extracted with EtOAc (10 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled compound (18 mg, 18%). .sup.1H NMR (400 MHz, CD.sub.3OD) 6.96 (s, 1H), 6.79 (s, 1H), 6.01-5.75 (m, 1H), 5.28-5.01 (m, 1H), 4.38-4.36 (m, 2H), 4.23 (s, 2H), 3.99 (s, 2H) 3.76 (s, 3H), 3.75-3.30 (m, 2H), 2.90-2.75 (m, 3H), 1.49-1.44 (m, 3H), 1.34-1.24 (m, 12H), 1.05-0.90 (m, 3H) ppm. MS: M/e 554 (M+1).sup.+.
Compound A74: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00327##
Step A: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00328##
[0550] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (300 mg, 0.91 mmol), 1-(3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethan-1-ol (190 mg, 0.91 mmol), (cyanomethyl)trimethylphosphonium iodide (662 mg, 2.74 mmol) and DIPEA (588 mg, 4.56 mmol) in MeCN (15 ml) was stirred at 100 C. overnight. After completed, the solution was concentrated under reduced pressure, The resulting residue was purified by Prep-HPLC (Method A) to give the titled Compound A74 (121 mg, 26%), which was further purified by Prep-HPLC (Method A) to give the titled Compound A74a (40 mg) and Compound A74b (29 mg).
[0551] Compound A74: .sup.1H NMR (400 MHz, CD.sub.3OD) 7.31 (dd, J=8.0, 5.2 Hz, 1H), 7.12 (dd, J=8.1, 2.7 Hz, 1H), 6.50-4.50 (m, 2H), 4.37 (q, J=6.9 Hz, 2H), 4.22 (s, 1H), 3.96 (s, 2H), 3.76 (s, 3H), 3.73-3.52 (m, 2H), 3.50-3.31 (m, 2H), 2.95-2.77 (m, 1.5H), 2.20-2.10 (m, 0.5H), 1.46 (t, J=7.0 Hz, 6H), 1.41 (s, 6H), 1.30 (d, J=6.6 Hz, 3H), 1.11-0.91 (m, 3H) ppm. MS: M/e 521 (M+1).sup.+
[0552] Compound A74a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.32 (d, J=8.1 Hz, 1H), 7.12 (d, J=8.0 Hz, 1H), 6.32-5.45 (m, 1H), 5.39-4.89 (m, 1H), 4.36 (d, J=7.2 Hz, 2H), 4.30-4.20 (m, 1H), 3.96 (s, 2H), 3.76 (s, 3H), 3.64 (d, J=5.6 Hz, 1H), 3.42-3.31 (m, 2H), 2.90 (s, 2H), 2.81 (d, J=11.9 Hz, 1H), 1.46 (t, J=7.1 Hz, 6H), 1.41 (s, 6H), 1.29 (d, J=6.5 Hz, 3H), 0.93 (d, J=6.3 Hz, 3H) ppm. MS: M/e 521 (M+1).sup.+
[0553] Compound A74b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.30 (d, J=8.0 Hz, 1H), 7.14 (d, J=8.1 Hz, 1H), 6.18-5.63 (m, 1H), 5.27-4.90 (m, 1H), 4.37 (q, J=7.0 Hz, 2H), 3.95 (s, 2H), 3.76 (s, 3H 3.59-3.51 (m, 1H), 3.49-3.39 (m, 1H), 3.30 (s, 3H), 2.72 (s, 1H), 2.18 (d, J=11.8 Hz, 1H), 1.46 (t, J=7.2 Hz, 3H), 1.40 (s, 6H), 1.31 (d, J=6.3 Hz, 6H), 1.02 (d, J=5.2 Hz, 3H) ppm. MS: M/e 521 (M+1).sup.+
Compound A77: 2-(6-((2S,5R)-4-(1-(4,4-difluorochroman-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00329##
Step A: 6-bromospiro[chromane-4,2-[1,3] dithiolane]
##STR00330##
[0554] To a stirred solution of 6-bromo-chroman-4-one (2 g, 8.8 mmol) in DCM (20 mL) at 0 C. are added 1,2-ethanedithiol (1.7 g, 17.6 mmol) and boron trifluoride etherate (640 mg, 4.4 mmol). The reaction mixture is stirred at room temperature for overnight before it is poured into 1.0 N NaOH solution. The mixture is extracted with DCM (50 mL) and the organic layers are separated, concentrated and purified by combi flash to give the titled compound (2.3 g, 96%) MS: M/e 303 (M+1).sup.+.
Step B: 6-bromo-4,4-difluorochromane
##STR00331##
[0555] A suspension of N-iodosuccinimide (3.3 g, 15 mmol) in DCM (40 mL) is cooled down to 70 C. Hydrogen fluoride pyridine complex (2.5 g, 30 mmol) is added dropwise. A solution of 6-bromospiro[chromane-4,2-[1,3] dithiolane](2.3 g, 7.6 mmol) in DCM is added dropwise and the mixture is stirred at 70 C. for 30 min. Then the reaction solution is poured into a mixture of hexane (10 mL) and DCM (10 mL) and the resulting mixture is filtered through a short pad of silica gel. The filtrate is concentrated and purified by combi flash to give the title compound (1.3 g, 69%). MS: M/e 249 (M+1).sup.+.
Step C: 1-(4,4-difluorochroman-6-yl)ethan-1-one
##STR00332##
[0556] A mixture of 6-bromo-4,4-difluorochromane (500 mg, 2.0 mmol), tributyl(1-ethoxyvinyl) stannane (1.1 g, 3.0 mmol) and Pd(PPh.sub.3).sub.2Cl.sub.2 (140 mg, 0.2 mmol) in toluene (5 mL) was stirred at 100 C. under N.sub.2 for 4 hours. The reaction mixture was quenched with saturated NaHCO.sub.3 aq. (10 mL), extracted with EtOAc (25 mL3), combined, washed brine (10 mL3), dried and concentrated to dryness. The resulting oil was diluted with THE (5 mL). Then to the solution was added HCl (2 mL, 4 M in 1,4-dioxane) in drops and the mixture was stirred at room temperature for 1 hour. The reaction mixture was diluted with EtOAc (20 mL), treated with saturated NaHCO.sub.3 aq. to pH8, washed with brine (20 mL3), dried over Na.sub.2SO.sub.4 and concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (320 mg, 75%). MS: M/e 213 (M+1).sup.+.
Step D: 1-(4,4-difluorochroman-6-yl)ethan-1-ol
##STR00333##
[0557] To a solution of 1-(4,4-difluorochroman-6-yl)ethan-1-one (320 mg, 1.5 mmol) in MeOH (10 mL) was added NaBH.sub.4 (66 mg, 1.8 mmol) at room temperature and the resulting mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated to dryness. The resulting residue was treated with saturated NaHCO.sub.3 aq., extracted with EtOAc (10 mL3). The combined organic layers were washed with brine (10 mL3), dried over Na.sub.2SO.sub.4 and concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (280 mg, 88%). MS: M/e 215 (M+1).sup.+.
Step E: 2-(6-((2S,5R)-4-(1-(4,4-difluorochroman-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00334##
[0558] To a solution of 1-(4,4-difluorochroman-6-yl)ethan-1-ol (60 mg, 0.28 mmol), 2-(6-((2S, 5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl) acetonitrile (60 mg, 0.19 mmol) and (cyanomethyl) trimethyl phosphonium iodide (68 mg, 0.28 mmol) in CH.sub.3CN (5 mL) was added DIPEA (116 mg, 0.9 mmol). The mixture was sealed in a bottle and heated at 100 C. for 16 hours. The mixture was cooled to room temperature, diluted with water, extracted with EtOAc (10 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled compound (8 mg, 9%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.60-7.45 (m, 2H), 6.92 (s, 1H), 6.02-5.75 (m, 1H), 5.25-5.01 (m, 1H), 4.34 (s, 2H), 4.21 (s, 2H), 3.93 (s, 3H), 3.69 (s, 3H), 3.68-3.60 (m, 1H), 3.32-3.28 (m, 1H), 2.81-2.75 (m, 3H), 2.51-2.48 (m, 2H), 1.38-1.29 (m, 9H) ppm. MS: M/e 512 (M+1).sup.+.
Compound A78: 2-(6-((2S,5R)-4-(1-(4,4-difluorochroman-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00335##
[0559] To a solution of 1-(4,4-difluorochroman-6-yl)ethan-1-ol (60 mg, 0.27 mmol), 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (60 mg, 0.18 mmol) and (cyanomethyl) trimethyl phosphonium iodide (66 mg, 0.27 mmol) in CH.sub.3CN (5 mL) was added DIPEA (116 mg, 0.9 mmol). The mixture was sealed in a bottle and heated at 100 C. for 16 hours. The mixture was cooled to room temperature, diluted with water, extracted with EtOAc (10 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by Prep-TLC (DCM:MeOH=15:1) to give the titled compound (9 mg, 10%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.61-7.58 (m, 1H), 7.44-7.38 (m, 1H), 6.92-6.86 (m, 1H), 6.01-5.70 (m, 1H), 5.25-4.99 (m, 1H), 4.38-4.33 (m, 4H), 4.23 (s, 2H), 3.71 (s, 3H), 3.68-3.63 (m, 1H), 3.50-3.30 (m, 1H), 3.23-3.13 (m, 1H), 3.00-2.50 (m, 2H), 2.48-2.45 (m, 2H), 1.46-1.28 (m, 9H), 0.99-0.67 (m, 3H) ppm. MS: M/e 526 (M+1).sup.+.
Compound A79: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2,2,3,3-tetrafluoro-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00336##
Step A: 1-(2,2,3,3-tetrafluoro-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-one
##STR00337##
[0560] A solution of 6-bromo-2,2,3,3-tetrafluoro-2,3-dihydrobenzo[b][1,4]dioxine (450 mg, 1.57 mmol), tributyl(1-ethoxyvinyl)stannane (849 mg, 2.35 mmol) and Pd(PPh.sub.3).sub.2Cl.sub.2 (110 mg, 0.16 mmol) in toluene (5 ml) was stirred at 100 C. under N.sub.2 overnight. After completed, the mixture was cooled to rt. HCl (4M in dioxane, 2 ml) was added and stirred at rt for 20 min. The reaction mixture was diluted with EtOAc (20 ml), washed with brine (10 ml), dried over Na.sub.2SO.sub.4 and then concentrated under reduced pressure. The residue was purified by flash column chromatography with 0-10% EtOAc in PE to give the titled compound (390 mg, 99%) as a colorless liquid. .sup.1H NMR (400 MHz, CDCl.sub.3) 7.80 (dd, J=8.7 Hz, 1.7 Hz, 1H), 7.77 (d, J=1.7 Hz, 1H), 7.24 (d, J=8.7 Hz, 1H), 2.60 (s, 3H) ppm.
Step B: 1-(2,2,3,3-tetrafluoro-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol
##STR00338##
[0561] To a solution of 1-(2,2,3,3-tetrafluoro-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-one (390 mg, 1.56 mmol) in MeOH (4 ml) at rt, was added NaBH.sub.4 (59 mg, 1.56 mmol). The solution was stirred at rt for 15 min. The reaction solution was diluted with EtOAc (15 ml), washed with brine (10 ml), dried over Na.sub.2SO.sub.4 and then evaporated to dryness to give the titled compound (390 mg, 100%) as a light-yellow oil. .sup.1H NMR (400 MHz, CDCl.sub.3) 7.20 (d, J=1.7 Hz, 1H), 7.17 (dd, J=8.4 Hz, 1.7 Hz, 1H), 7.11 (d, J=8.4 Hz, 1H), 4.92 (q, J=6.2 Hz, 1H), 1.49 (d, J=6.4 Hz, 3H) ppm.
Step C: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2,2,3,3-tetrafluoro-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00339##
[0562] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (370 mg, 1.17 mmol), 1-(2,2,3,3-tetrafluoro-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (290 mg, 1.15 mmol), (cyanomethyl)trimethylphosphonium iodide (856 mg, 3.52 mmol) and DIPEA (1.52 g, 11.78 mmol) in CH.sub.3CN (6 ml) was stirred at 100 C. overnight. The reaction was diluted with EtOAc (20 ml) and washed with brine (10 ml). The organic layer was concentrated under reduced pressure. The resulting residue was purified by flash column chromatography with 0-5% MeOH in DCM to give the titled Compound A79 (140 mg), which was further separated into Compound A79a (49 mg) and Compound A79b (61 mg) by chiral Prep-HPLC. The chiral separation conditions are shown below.
TABLE-US-00015 Column CHIRALPAK ID Column Size 2 cm 25 cm, 5 um Mobile Phase A Hex(0.5% 2 mM NH3MeOH) Mobile Phase B EtOH Flow Rate 20 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-HPLC Equipment Prep-HPLC-Gilson
[0563] Compound A79: .sup.1H NMR (400 MHz, DMSO-d6) 7.49-7.42 (m, 2H), 7.37 (t, J=8.4 Hz, 1H), 6.18-4.45 (m, 2H), 4.45-4.30 (m, 2H), 3.84 (s, 3H), 3.67 (q, J=6.3 Hz, 0.5H), 3.64 (s, 3H), 3.54-3.47 (m, 1H), 3.34-3.31 (m, 1.5H), 2.84-2.69 (m, 1.5H), 2.08 (d, J=12.0 Hz, 0.5H), 1.34 (s, 1.5H), 1.24 (dd, J=9.9, 6.5 Hz, 3H), 1.18 (d, J=6.5 Hz, 1.5H), 0.94 (d, J=6.1 Hz, 1.5H), 0.83 (d, J=6.5 Hz, 1.5H) ppm. MS: M/e 550 (M+1).sup.+.
[0564] Compound A79a (the earlier peak): .sup.1H NMR (400 MHz, DMSO-d6) 7.49-7.42 (m, 2H), 7.38 (d, J=8.5 Hz, 1H), 6.10-4.40 (m, 2H), 4.41-4.30 (m, 2H), 3.83 (s, 3H), 3.67 (q, J=6.0 Hz, 1H), 3.64 (s, 3H), 3.31-3.28 (m, 1H), 2.85-2.70 (m, 3H), 1.34 (s, 3H), 1.23 (d, J=6.4 Hz, 3H), 0.83 (d, J=6.3 Hz, 3H) ppm. MS: M/e 550 (M+1).sup.+.
[0565] Compound A79b (the later peak): .sup.1H NMR (400 MHz, DMSO-d6) 7.46-7.41 (m, 2H), 7.36 (d, J=8.3 Hz, 1H), 6.04-5.48 (m, 1H), 5.08-4.50 (m, 1H), 4.42-4.31 (m, 2H), 3.84 (s, 3H), 3.64 (s, 3H), 3.54-3.45 (m, 2H), 3.32-3.28 (m, 2H), 2.08 (d, J=11.8 Hz, 1H), 1.25 (d, J=6.3 Hz, 3H), 1.17 (d, J=5.8 Hz, 3H), 0.94 (d, J=6.1 Hz, 3H) ppm. MS: M/e 550 (M+1).sup.+.
Compound A80: 2-(6-((2S,5R)-4-(1-(3,3-difluoro-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00340##
##STR00341##
[0566] A solution of 5-bromo-2-fluorophenol (10 g, 0.052 mol), methyl 2-bromo-2,2-difluoroacetate (14.8 g, 0.078 mol), DBU (15.9 g, 0.10 mol) in DMF (60 ml) was stirred at 70 C. overnight. The solution was poured into water (50 ml) and then extracted with EtOAc (50 ml2). The organic layer was washed with brine and concentrated. The residue was purified by flash column chromatography with 0-1% EtOAc in PE to give the titled compound (8.4 g, 53%). .sup.1H NMR (400 MHz, CDCl.sub.3) 7.48 (d, J=6.3 Hz, 1H), 7.42-7.35 (m, 1H), 7.08 (t, J=9.2 Hz, 1H), 3.98 (s, 3H) ppm
Step B: 2-(5-bromo-2-fluorophenoxy)-2,2-difluoroethan-1-ol
##STR00342##
[0567] To a solution of methyl 2-(5-bromo-2-fluorophenoxy)-2,2-difluoroacetate (8.4 g, 0.028 mol) in THE (80 ml), was added LiAlH.sub.4 (1M in THF, 56 ml, 56 mmol) dropwise. The solution was stirred at RT overnight. The solution was quenched with water (50 ml) and separated. The organic layer was concentrated. The residue was purified by flash column chromatography with 0-10% EtOAc in PE to give the title compound (5 g, 65%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.34 (t, J=7.7 Hz, 1H), 7.28-7.16 (m, 2H), 3.94 (t, J=9.8 Hz, 2H) ppm
Step C: 3-(1,1-difluoro-2-hydroxyethoxy)-4-fluorobenzonitrile
##STR00343##
[0568] A solution of 2-(5-bromo-2-fluorophenoxy)-2,2-difluoroethan-1-ol (2.8 g, 0.010 mol), Pd(PPh.sub.3).sub.4 (1.2 g, 0.0010 mol) and Zn(CN).sub.2 (2.4 g, 0.021 mol) in DMF (20 ml) was stirred at 120 C. overnight under N.sub.2. The solution was poured into water (40 ml) and extracted with EtOAc (30 ml2). The organic layer was washed with brine (20 ml), dried and concentrated. The residue was purified by flash column chromatography with 0-20% EtOAc in PE to give the titled compound (390 mg, 17%). MS: M/e 218 (M+1).sup.+.
Step D: 3,3-difluoro-2,3-dihydrobenzo[b][1,4]dioxine-6-carbonitrile
##STR00344##
[0569] A solution of 3-(1,1-difluoro-2-hydroxyethoxy)-4-fluorobenzonitrile (260 mg, 1.20 mmol) and NaH (60%, 72 mg, 1.8 mmol) in THE (10 ml) was stirred at RT for 30 min. The mixture was diluted with EtOAc (10 ml), washed with brine (10 ml), dried and concentrated. The residue was purified by flash column chromatography with 0-15% EtOAc in PE to give the titled compound (150 mg, 63%). .sup.1H NMR (400 MHz, CDCl.sub.3) 7.36 (d, J=6.9 Hz, 2H), 7.11 (d, J=8.9 Hz, 1H), 4.33 (t, J=5.9 Hz, 2H) ppm
Step E: 1-(3,3-difluoro-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-one
##STR00345##
[0570] A solution of 3,3-difluoro-2,3-dihydrobenzo[b][1,4]dioxine-6-carbonitrile (200 mg, 1.1 mmol), MeMgBr (1M in THF, 1.52 ml, 1.52 mmol) and CuBr (10.9 mg, 0.076 mmol) in THF (6 ml) was stirred at 60 C. overnight. The mixture was cooled to RT, HCl (3N, 1 ml) was added and stirred for 30 min. The mixture was diluted with EtOAc (15 ml), washed with brine (10 ml), dried and concentrated. The residue was purified by flash column chromatography with 0-15% EtOAc in PE to give the titled compound (170 mg, 82%). .sup.1H NMR (400 MHz, CDCl.sub.3) 7.67 (d, J=8.4 Hz, 2H), 7.08 (d, J=8.4 Hz, 1H), 4.32 (t, J=5.9 Hz, 2H), 2.57 (s, 3H) ppm
Step F: 1-(3,3-difluoro-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol
##STR00346##
[0571] A solution of 1-(3,3-difluoro-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-one (170 mg, 0.79 mmol) and NaBH.sub.4 (30 mg, 0.79 mmol) in MeOH (4 ml) was stirred at RT for 40 min. The solution was diluted with EtOAc (10 ml), washed with brine (10 ml), dried and evaporated to dryness to give the titled compound (170 mg, 100%), which was used directly for the next step without further purification. MS: M/e 217 (M+1).sup.+.
Step G: 2-(6-((2S,5R)-4-(1-(3,3-difluoro-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00347##
[0572] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (261 mg, 0.79 mmol), 1-(3,3-difluoro-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (170 mg, 0.79 mmol), (cyanomethyl)trimethylphosphonium iodide (579 mg, 2.38 mmol) and DIPEA (1.02 g, 7.91 mmol) in MeCN (4 ml) was stirred at 100 C. overnight. The reaction was diluted with EtOAc (10 ml) and washed with brine (10 ml). The organic layer was concentrated under reduced pressure. The resulting residue was purified by flash column chromatography with 0-5% MeOH in DCM to give the titled Compound A80 (100 mg), which was further separated into Compound A80a (29 mg) and Compound A80b (32 mg) by chiral prep-HPLC. The chiral separation conditions are shown below.
TABLE-US-00016 Column CHIRALPAK IH Column Size 2 cm 25 cm, 5 um Mobile Phase A MtBE Mobile Phase B MeOH:DCM Flow Rate 20 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-HPLC Equipment Prep-HPLC-Gilson
Prep-HPLC Equipment Prep-HPLC-Gilson
[0573] Compound A80: .sup.1H NMR (400 MHz, CD.sub.3OD) 7.17-7.07 (m, 2H), 7.06-6.97 (m, 1H), 6.30-4.95 (m, 2H), 4.40-4.32 (m, 4H), 4.23 (s, 2H), 3.80-3.74 (m, 3.5H), 3.74-3.48 (m, 2H), 3.07-2.66 (m, 2H), 2.29 (d, J=12 Hz, 0.5H), 1.46 (t, J=7.1 Hz, 4.5H), 1.39-1.32 (m, 3H), 1.27 (d, J=5.3 Hz, 1.5H), 1.07 (d, J=6.0 Hz, 1.5H), 0.96 (d, J=4.8 Hz, 1.5H) ppm. MS: M/e 528 (M+1).sup.+.
[0574] Compound A80a (the earlier peak): .sup.1H NMR (400 MHz, DMSO-d6) [0888]7.16 (s, 1H), 7.10 (q, J=5.5 Hz, 2H), 6.10-5.40 (m, 1H), 5.25-4.92 & 4.51-4.44 (m, 1H), 4.62-4.53 (m, 2H), 4.44-4.33 (m, 2H), 4.32-4.16 (m, 2H), 3.61 (s, 3H), 3.56 (q, J=5.3 Hz, 1H), 3.33-3.28 (m, 2H), 2.80-2.72 (m, 2H), 1.45-1.27 (m, 6H), 1.21 (d, J=6.1 Hz, 3H), 0.82 (d, J=5.4 Hz, 3H) ppm. MS: M/e 528 (M+1).sup.+.
[0575] Compound A80b (the later peak): .sup.1H NMR (400 MHz, DMSO-d6) 7.18-7.04 (m, 3H), 5.93-5.63 (m, 1H), 5.00-4.65 (m, 1H), 4.56 (t, J=5.7 Hz, 2H), 4.47-4.35 (m, 2H), 4.33-4.20 (m, 2H), 3.61 (s, 3H), 3.53-3.40 (m, 2H), 3.33-3.29 (m, 2H), 2.14 (d, J=9.7 Hz, 1H), 1.34 (t, J=5.7 Hz, 3H), 1.23 (d, J=5.7 Hz, 3H), 1.21-1.12 (m, 3H), 1.00-0.88 (m, 3H) ppm. MS: M/e 528 (M+1).sup.+.
Compound A81: 2-(6-((2S,5R)-4-(1-(2,2-dimethyl-4-oxochroman-7-yl)ethyl)-2,5-dimethyl piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00348##
Step A: 7-acetyl-2,2-dimethylchroman-4-one
##STR00349##
[0576] A mixture of 7-bromo-2,2-dimethylchroman-4-one (510 mg, 2.0 mmol), tributyl(1-ethoxyvinyl)stannane (1.45 g, 4.0 mmol) and Pd(PPh.sub.3).sub.2Cl.sub.2 (70 mg, 0.1 mmol) in toluene (5 mL) was stirred at 100 C. under N.sub.2 for 16 hrs. The mixture was cooled and diluted with EtOAc (10 mL), washed with brine (10 mL2). The organic layer was added HCl/Dioxane (4M, 5 mL), stirred for 5 min and washed with brine (10 mL2), NaHCO.sub.3 (10 mL2), brine (10 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated to dryness. The resulted residue was purified by flash column chromatography to give the titled compound (458 mg, 95%). .sup.1H NMR (400 MHz, CDCl.sub.3) 7.94 (d, J=8.0 Hz, 1H), 7.56-7.46 (m, 2H), 2.77 (s, 2H), 2.61 (s, 3H), 1.48 (s, 6H) ppm.
Step B: 7-(1-hydroxyethyl)-2,2-dimethylchroman-4-one
##STR00350##
[0577] To a solution of 7-acetyl-2,2-dimethylchroman-4-one (458 mg, 2.0 mmol) in MeOH (5 mL) was added NaBH.sub.4 (38 mg, 1.0 mmol) at room temperature and the mixture was stirred at rt for 2 hours. The mixture was treated with NaHCO.sub.3 (15 mL), extracted with CH.sub.2Cl.sub.2 (10 mL5). The combined extracts was washed with brine (10 mL), dried over Na.sub.2SO.sub.4 and concentrated to dryness and purified by flash column chromatography to give the title compound (307 mg, 67%). .sup.1H NMR (400 MHz, DMSO-d6) 7.66 (d, J=8.0 Hz, 1H), 6.99 (d, J=8.0 Hz, 1H), 6.92 (s, 1H), 5.31 (d, J=4.4 Hz, 1H), 4.75-4.63 (m, 1H), 2.75 (s, 2H), 1.38 (s, 6H), 1.30 (d, J=6.4 Hz, 3H) ppm.
Step C: 2-(6-((2S,5R)-4-(1-(2,2-dimethyl-4-oxochroman-7-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00351##
[0578] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (100 mg, 0.3 mmol), 7-(1-hydroxyethyl)-2,2-dimethylchroman-4-one (134 mg, 0.61 mmol) and (cyanomethyl)trimethylphosphonium iodide (150 mg, 0.61 mmol) in CH.sub.3CN (2 mL) was added DIPEA (155 mg, 1.2 mmol). The mixture was stirred at 100 C. in a sealed tube for 16 hours. The mixture was cooled and diluted with EtOAc (20 mL), washed with brine (10 mL3), dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography and further purified by Prep-HPLC (Method A) to give the titled compound (68 mg, 43%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.82-7.71 (m, 1H), 7.12-7.03 (m, 1H), 7.00 (s, 1H), 6.38-5.45 (m, 1H), 5.41-4.50 (m, 1H), 4.37 (q, J=7.2 Hz, 2H), 4.23 (s, 2H), 3.76 (s, 3H), 3.68-3.37 (m, 2H), 2.96-2.15 (m, 5H), 1.48-1.27 (m, 15H), 1.08-0.88 (m, 3H) ppm. MS: M/e 532 (M+1).sup.+.
Compound A83: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-methyl-1,3-dioxoisoindolin-5-yl)ethyl) piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00352##
Step A: 5-bromo-2-methylisoindoline-1,3-dione
##STR00353##
[0579] To a 0 C. solution of 5-bromoisoindoline-1,3-dione (2.26 g, 10 mmol) in DMF (40 mL) was added NaH (600 mg, 15 mmol) in portions. The resulted mixture was stirred at 0 C. for 10 min. Iodomethane (2.13 g, 15 mmol) was added and the mixture was stirred at room temperature for 2 hours. The mixture was quenched with aqueous solution of NH.sub.4Cl (100 mL). EtOAc (100 mL) was added and the mixture was washed with brine (100 mL3), dried over Na.sub.2SO.sub.4, filtered and concentrated to dryness. The resulted residue was purified by column chromatography to give the titled compound (1.51 g, 63%). .sup.1H NMR (400 MHz, DMSO-d6) 8.06 (s, 1H), 8.02 (d, J=8.0 Hz, 1H), 7.79 (d, J=8.0 Hz, 1H), 3.03 (s, 3H) ppm.
Step B: 5-acetyl-2-methylisoindoline-1,3-dione
##STR00354##
[0580] A mixture of 5-bromo-2-methylisoindoline-1,3-dione (1.0 g, 4.16 mmol), tributyl(1-ethoxyvinyl)stannane (3.0 g, 8.33 mmol) and Pd(PPh.sub.3).sub.2Cl.sub.2 (140 mg, 0.2 mmol) in toluene (10 mL) was stirred at 100 C. under N.sub.2 for 16 hrs. The mixture was cooled and diluted with EtOAc (50 mL), washed with brine (20 mL2). The mixture was filtered through a celite pad and the organic layer was treated with HCl/Dioxane (4M, 5 mL), and washed with brine (20 mL2), NaHCO.sub.3 (20 mL2), dried over Na.sub.2SO.sub.4, filtered and concentrated to dryness. The resulted residue was purified by flash column chromatography to give the titled compound (800 mg, 93%). .sup.1H NMR (400 MHz, CDCl.sub.3) 8.38 (s, 1H), 8.32 (d, J=7.6 Hz, 1H), 7.95 (d, J=8.0 Hz, 1H), 3.22 (s, 3H), 2.70 (s, 3H) ppm.
Step C: 5-(1-hydroxyethyl)-2-methylisoindoline-1,3-dione
##STR00355##
[0581] To a solution of 5-acetyl-2-methylisoindoline-1,3-dione (800 mg, 3.9 mmol) in MeOH (10 mL) was added NaBH.sub.4 (80 mg, 2.1 mmol) at room temperature and the mixture was stirred at rt for 2 hours. The mixture was treated with NaHCO.sub.3 (50 mL), extracted with EtOAc (50 mL2). The combined extracts was washed with brine (50 mL2), dried over Na.sub.2SO.sub.4 and concentrated to dryness and purified by flash column chromatography to give the title compound (300 mg, 37%). .sup.1H NMR (400 MHz, DMSO-d6) 7.88-7.80 (m, 3H), 5.50 (s, 1H), 4.89 (q, J=6.0 Hz, 1H), 3.03 (s, 3H), 1.36 (d, J=6.4 Hz, 3H) ppm.
Step D: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-methyl-1,3-dioxoisoindolin-5-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00356##
[0582] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (82 mg, 0.25 mmol), 5-(1-hydroxyethyl)-2-methylisoindoline-1,3-dione (100 mg, 0.49 mmol) and (cyanomethyl)trimethylphosphonium iodide (121 mg, 0.50 mmol) in CH.sub.3CN (2 mL) was added DIPEA (129 mg, 1.0 mmol). The mixture was stirred at 100 C. in a sealed tube for 16 hours. The mixture was cooled and diluted with EtOAc (20 mL), washed with brine (10 mL3), dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography and further purified by Prep-HPLC(Method A) to give the titled compound (18 mg, 14%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.92 (s, 1H), 7.86-7.74 (m, 2H), 6.16-4.50 (m, 2H), 4.44-4.30 (m, 2H), 4.29-4.08 (m, 2H), 3.89-3.57 (m, 5H), 3.12 (s, 3H), 3.00-2.57 (m, 2.5H), 2.23-2.08 (m, 0.5H), 1.55-1.20 (m, 9H), 1.11-0.88 (m, 3H) ppm. MS: M/e 517 (M+1).sup.+.
Compound A85: 2-(6-((2S,5R)-4-(1-(2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethyl piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00357##
[0583] To a solution of 1-(2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethan-1-ol (116 mg, 0.6 mmol), 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (99 mg, 0.3 mmol) and (cyanomethyl)trimethylphosphonium iodide (145 mg, 0.6 mmol) in CH.sub.3CN (3 mL) was added DIPEA (155 mg, 1.2 mmol). The mixture was sealed in a bottle and heated at 100 C. overnight. Then the mixture was cooled to room temperature, diluted with water, extracted with EtOAc (50 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by Prep-TLC (DCM:MeOH=13:1) to give the titled Compound A85 (25 mg), which was further separated into Compound A85a (11 mg) and Compound A85b (10 mg) by chiral Prep-HPLC. The chiral separation conditions are shown below.
TABLE-US-00017 Column CHIRALPAK IF Column Size 2 cm 25 cm, 5 um Mobile Phase A MtBE Mobile Phase B ACN Flow Rate 20 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-HPLC Equipment Prep-HPLC-Gilson
[0584] Compound A85: .sup.1HNMR (400 MHz, CDCl.sub.3) 6.82 (d, J=9.4 Hz, 1H), 6.75-6.69 (m, 1H), 6.63 (d, J=7.9 Hz, 1H), 6.00-4.50 (m, 2H), 4.30 (d, J=6.7 Hz, 2H), 3.87 (d, J=23.4 Hz, 2H), 3.77 (s, 3H), 3.58-3.18 (m, 3H), 2.95-2.74 (m, 1H), 2.58-2.27 (m, 1H), 1.69 (d, J=8.6 Hz, 6H), 1.49 (dd, J=26.0, 18.9 Hz, 5H), 1.26 (dd, J=15.2, 6.4 Hz, 4H), 1.05-0.84 (m, 3H) ppm. MS: M/e 506 (M+1).sup.+.
[0585] Compound A85a (the earlier peak) .sup.1H NMR (400 MHz, DMSO-d6) 6.82 (s, 1H), 6.74 (q, J=7.9 Hz, 2H), 5.94-5.60 (m, 1H), 4.96-4.58 (m, 1H), 4.40 (s, 2H), 4.26 (d, J=7.2 Hz, 2H), 3.61 (s, 4H), 3.46 (s, 1H), 3.34 (s, 1H), 2.49-2.39 (m, 1H), 2.18 (d, J=12.3 Hz, 1H), 1.62 (s, 6H), 1.34 (t, J=7.0 Hz, 3H), 1.22 (d, J=6.5 Hz, 3H), 1.16 (s, 3H), 0.93 (s, 3H) ppm. MS: M/e 506 (M+1).sup.+.
[0586] Compound A85b (the later peak) .sup.1H NMR (400 MHz, DMSO-d6) 6.84 (s, 1H), 6.75 (q, J=8.0 Hz, 2H), 6.1-5.40 (m, 1H), 5.15-4.45 (m, 1H), 4.38 (s, 2H), 4.25 (s, 2H), 3.61 (s, 3H), 3.47 (d, J=6.3 Hz, 1H), 3.30-2.95 (m, 1H), 2.90-2.62 (m, 3H), 1.63 (d, J=5.0 Hz, 7H), 1.34 (s, 6H), 1.19 (d, J=6.2 Hz, 3H), 0.81 (d, J=5.9 Hz, 3H) ppm. MS: M/e 506 (M+1).sup.+.
Compound A86: 2-(9-cyclopropyl-6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00358##
Step A: 6-(cyclopropylamino)-1-methylpyrimidine-2,4 (1H,3H)-dione
##STR00359##
[0587] To a stirred mixture of 6-chloro-1-methylpyrimidine-2,4 (1H,3H)-dione (4.8 g, 30 mmol) in EtOH (50 mL) was added cyclopropanamine (3.42 g, 60 mmol). After the addition, the reaction mixture was stirred at 60 C. overnight. The reaction mixture was cooled to room temperature and filtered. The caked was collected and dried to give the titled compound (4.2 g, 77%). MS: M/e 182 (M+1).sup.+.
Step B: 6-(cyclopropylamino)-1-methyl-5-nitrosopyrimidine-2,4 (1H,3H)-dione
##STR00360##
[0588] To a stirred mixture of 6-(cyclopropylamino)-1-methylpyrimidine-2,4 (1H,3H)-dione (4.2 g, 23.2 mmol) in H.sub.2O (50 mL) was added NaNO.sub.2 (1.76 g, 25.5 mmol) at 05 C. Then AcOH (4.2 g, 69.6 mmol) was added dropwise at that temperature. After then, the mixture was stirred for 30 minutes. The reaction mixture was filtered and the cake was collected, dried to give the titled compound (3.3 g, 68%). MS: M/e 211 (M+1).sup.+.
Step C: 5-amino-6-(cyclopropylamino)-1-methylpyrimidine-2,4 (1H,3H)-dione
##STR00361##
[0589] To a stirred mixture of 6-(cyclopropylamino)-1-methyl-5-nitrosopyrimidine-2,4 (1H,3H)-dione (3.3 g, 15.7 mmol) in H.sub.2O (50 mL) was added Na.sub.2S.sub.2O.sub.4 (8.2 g, 47.1 mmol) portionwise at 20-30 C. After the addition, the reaction mixture was stirred for 30 minutes. The reaction mixture was cooled in a icebox for several hours and filtered. The cake was collected, dried to give the titled compound (2 g, 65%). MS: M/e 197 (M+1).sup.+.
Step D: 8-((benzyloxy)methyl)-9-cyclopropyl-3-methyl-3,9-dihydro-1H-purine-2,6-dione
##STR00362##
[0590] A mixture of 5-amino-6-(cyclopropylamino)-1-methylpyrimidine-2,4 (1H,3H)-dione (2 g, 10.2 mmol) in 2-(benzyloxy)acetic acid (1.86 g, 11.2 mmol) was stirred at 240 C. for 1.5 hour. The mixture was allowed cool to room temperature and dissolved in aq.NaOH (2.0 M, 20 mL), then acidified to pH=56 with aq.HCl and filtered. The cake was collected, dried to give the titled compound (2 g, 61%). MS: M/e 327 (M+1).sup.+.
Step E: tert-butyl (2R,5S)-4-(8-((benzyloxy)methyl)-9-cyclopropyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00363##
[0591] To a stirred mixture of 8-((benzyloxy)methyl)-9-cyclopropyl-3-methyl-3,9-dihydro-1H-purine-2,6-dione (978 mg, 3 mmol) and tert-butyl (2R,5S)-2,5-dimethylpiperazine-1-carboxylate (1.28 g, 6 mmol) in CH.sub.3CN (15 mL) was added BOP (2.65 g, 6 mmol), followed by DBU (918 mg, 6 mmol). After the addition, the reaction mixture was concentrated to give the residue, which was dissolved in EtOAc (50 mL), washed with H.sub.2O (20 mL), brine, dried over Na.sub.2SO.sub.4, concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (0.84 g, 54%). MS: M/e 523 (M+1).sup.+.
Step F: tert-butyl (2R,5S)-4-(9-cyclopropyl-8-(hydroxymethyl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00364##
[0592] To a stirred solution of tert-butyl (2R,5S)-4-(8-((benzyloxy)methyl)-9-cyclopropyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (0.84 g, 1.6 mmol) in MeOH/AcOH (80 mL/2 mL) was added Pd/C (200 mg). After the addition, the reaction mixture was stirred over a weekend under H.sub.2 (1 atm). The reaction mixture was filtered, and filtrate was concentrated to give the titled compound (crude, 100%). MS: M/e 433 (M+1).sup.+.
Step G: tert-butyl (2R,5S)-4-(8-(cyanomethyl)-9-cyclopropyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00365##
[0593] To a stirred solution of tert-butyl (2R,5S)-4-(9-cyclopropyl-8-(hydroxymethyl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (crude, 1.6 mmol) in CH.sub.2Cl.sub.2 (10 mL) was added SOCl.sub.2 (0.21 g, 1.77 mmol). After stirred for 10 min, the reaction mixture was quenched with aq.NaHCO.sub.3, washed with brine, dried over Na.sub.2SO.sub.4, concentrated to give the intermediate, dissolved in CH.sub.2Cl.sub.2 (10 mL), TBAF (1.0 M, 3.2 mL, 3.2 mmol) was added followed by TMSCN (0.32 g, 3.2 mmol). After the addition, the reaction mixture was stirred for 3 hours. The reaction mixture was concentrated to give the residue, treated with H.sub.2O and extracted with EtOAc (15 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (330 mg, 47%). MS: M/e 442 (M+1).sup.+.
Step H: 2-(9-cyclopropyl-6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00366##
[0594] To a stirred solution of tert-butyl (2R,5S)-4-(8-(cyanomethyl)-9-cyclopropyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (330 mg, 0.748 mmol) in EtOAc (10 mL) was added dioxane/HCl (g) (4.0 M, 4 mL). After the addition, the reaction mixture was stirred for 4 hours. The reaction mixture was concentrated to give the residue, which was treated with CH.sub.3CN/aq.NaHCO.sub.3 (20 mL/3 mL), stirred for 10 min and filtered. The filtrate was concentrated, dried to give the titled compound (240 mg, 97%), which was directly used to the next step. MS: M/e 342 (M+1).sup.+.
Step I: 2-(9-cyclopropyl-6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00367##
[0595] A mixture of 2-(9-cyclopropyl-6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (240 mg, 0.7 mmol), 1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (439 mg, 2.1 mmol), (cyanomethyl)trimethylphosphonium iodide (680 mg, 2.8 mmol) and DIPEA (903 mg, 7 mmol) in CH.sub.3CN (6 mL) was stirred overnight at 100 C. in a sealed tube. The reaction mixture was diluted with EtOAc (20 mL), washed with H.sub.2O, brine, dried over Na.sub.2SO.sub.4, concentrated to dryness. The resulting residue was purified by Pre-TLC (CH.sub.2Cl.sub.2/MeOH=10:1) to give the desired compound, and further purified by Prep-HPLC (Method A) to give the titled compound (60 mg, 18%). .sup.1H NMR (400 MHz, CD.sub.3OD) 6.86-6.73 (m, 3H), 6.25-4.97 (m, 2H), 4.31-4.19 (m, 1H), 3.88 (d, J=3.2 Hz, 2H), 3.83 (s, 3H), 3.64-3.39 (m, 3.5H), 3.04-2.82 (m, 1.5H), 2.65 (s, 1H), 2.31 (d, J=12.4 Hz, 0.5H), 2.04-1.94 (m, 0.5H), 1.48-1.34 (m, 3H), 1.34-1.28 (m, 9H), 1.27-1.13 (m, 4H), 1.05-0.88 (m, 3H) ppm. MS: M/e 532 (M+1).sup.+.
Compound A87: 2-(9-(2,2-difluoroethyl)-6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00368##
Step A: 6-((2,2-difluoroethyl)amino)-1-methylpyrimidine-2,4 (1H,3H)-dione
##STR00369##
[0596] To a stirred mixture of 6-chloro-1-methylpyrimidine-2,4 (1H,3H)-dione (4.8 g, 30 mmol) in EtOH (50 mL) was added 2,2-difluoroethan-1-amine (4.8 g, 60 mmol). After the addition, the reaction mixture was stirred at 60 C. overnight. The reaction mixture was cooled to room temperature and filtered. The caked was collected and dried to give the titled compound (6.2 g, 100%). MS: M/e 206 (M+1).sup.+.
Step B: 6-((2,2-difluoroethyl)amino)-1-methyl-5-nitrosopyrimidine-2,4 (1H,3H)-dione
##STR00370##
[0597] To a stirred mixture of 6-((2,2-difluoroethyl)amino)-1-methylpyrimidine-2,4 (1H,3H)-dione (6.1 g, 29.7 mmol) in H.sub.2O (80 mL) was added NaNO.sub.2 (2.3 g, 32.7 mmol) at 05 C. Then AcOH (5.3 g, 89.1 mmol) was added dropwise at that temperature. After then, the mixture was stirred for 30 minutes. The reaction mixture was filtered, and the cake was collected, dried to give the titled compound (7.6 g, 100%). MS: M/e 235 (M+1).sup.+.
Step C: 5-amino-6-((2,2-difluoroethyl)amino)-1-methylpyrimidine-2,4 (1H,3H)-dione
##STR00371##
[0598] To a stirred mixture of 6-((2,2-difluoroethyl)amino)-1-methyl-5-nitrosopyrimidine-2,4 (1H,3H)-dione (7.6 g, 32.5 mmol) in H.sub.2O (80 mL) was added Na.sub.2S.sub.2O.sub.4 (17 g, 97.4 mmol) portionwise at 2030 C. After the addition, the reaction mixture was stirred for 30 minutes. The reaction mixture was cooled in a icebox for several hours and filtered. The cake was collected, dried to give the titled compound (2.9 g, 40.5%). MS: M/e 221 (M+1).sup.+.
Step D: 8-((benzyloxy)methyl)-9-(2,2-difluoroethyl)-3-methyl-3,9-dihydro-1H-purine-2,6-dione
##STR00372##
[0599] A mixture of 5-amino-6-((2,2-difluoroethyl)amino)-1-methylpyrimidine-2,4 (1H,3H)-dione (2.9 g, 13.2 mmol) in 2-(benzyloxy)acetic acid (2.52 g, 14.5 mmol) was stirred at 240 C. for 1.5 hour. The mixture was allowed cool to room temperature and dissolved in aq.NaOH (2.0 M, 20 mL), then acidified to pH=56 with aq.HCl and filtered. The cake was collected, dried to give the titled compound (2.3 g, 50%). MS: M/e 351 (M+1).sup.+.
Step E: tert-butyl (2R,5S)-4-(8-((benzyloxy)methyl)-9-(2,2-difluoroethyl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00373##
[0600] To a stirred mixture of 8-((benzyloxy)methyl)-9-(2,2-difluoroethyl)-3-methyl-3,9-dihydro-1H-purine-2,6-dione (1.05 mg, 3 mmol) and tert-butyl (2R,5S)-2,5-dimethylpiperazine-1-carboxylate (1.28 g, 6 mmol) in CH.sub.3CN (15 mL) was added BOP (2.65 g, 6 mmol), followed by DBU (918 mg, 6 mmol). After the addition, the reaction mixture was concentrated to give the residue, which was dissolved in EtOAc (50 mL), washed with H.sub.2O (20 mL), brine, dried over Na.sub.2SO.sub.4, concentrated and purified by flash column chromatography to give the titled compound (1.1 g, 67%). MS: M/e 547 (M+1).sup.+.
Step F: tert-butyl (2R,5S)-4-(9-(2,2-difluoroethyl)-8-(hydroxymethyl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00374##
[0601] To a stirred solution of tert-butyl (2R,5S)-4-(8-((benzyloxy)methyl)-9-(2,2-difluoroethyl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (1.1 g, 2 mmol) in MeOH/AcOH (80 mL/2 mL) was added Pd/C (200 mg). After the addition, the reaction mixture was stirred over a weekend under H.sub.2 (1 atm). The reaction mixture was filtered, and filtrate was concentrated to give the titled compound (crude, 100%). MS: M/e 457 (M+1).sup.+.
Step G: tert-butyl (2R,5S)-4-(8-(cyanomethyl)-9-(2,2-difluoroethyl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00375##
[0602] To a stirred solution of tert-butyl (2R,5S)-4-(9-(2,2-difluoroethyl)-8-(hydroxymethyl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (crude, 2 mmol) in CH.sub.2Cl.sub.2 (10 mL) was added SOCl.sub.2 (0.26 g, 2.2 mmol). After stirred for 10 min, the reaction mixture was quenched with aq.NaHCO.sub.3, washed with brine, dried over Na.sub.2SO.sub.4, concentrated to give the intermediate, dissolved in CH.sub.2Cl.sub.2 (10 mL), TBAF (1.0 M, 4 mL, 4 mmol) was added followed by TMSCN (0.4 g, 4 mmol). After the addition, the reaction mixture was stirred for 3 hours. The reaction mixture was concentrated to give the residue, treated with H.sub.2O and extracted with EtOAc (15 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (250 mg, 27%). MS: M/e 466 (M+1).sup.+.
Step H: 2-(9-(2,2-difluoroethyl)-6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00376##
[0603] To a stirred solution of tert-butyl (2R,5S)-4-(8-(cyanomethyl)-9-(2,2-difluoroethyl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (250 mg, 0.54 mmol) in EtOAc (10 mL) was added dioxane/HCl (g) (4.0 M, 4 mL). After the addition, the reaction mixture was stirred for 4 hours. The reaction mixture was concentrated to give the residue, which was treated with CH.sub.3CN/aq.NaHCO.sub.3 (20 mL/3 mL), stirred for 10 min and filtered. The filtrate was concentrated, dried to give the titled compound (190 mg, 96%), which was directly used to the next step. MS: M/e 366 (M+1).sup.+.
Step I: 2-(9-(2,2-difluoroethyl)-6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo [b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00377##
[0604] A mixture of 2-(9-(2,2-difluoroethyl)-6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (190 mg, 0.52 mmol), 1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (324 mg, 1.56 mmol), (cyanomethyl)trimethylphosphonium iodide (505 mg, 2.08 mmol) and DIPEA (670 mg, 5.2 mmol) in CH.sub.3CN (6 mL) was stirred overnight at 100 C. in a sealed tube. The reaction mixture was diluted with EtOAc (20 mL), washed with H.sub.2O, brine, dried over Na.sub.2SO.sub.4, concentrated to give the residue, which was purified by Pre-TLC (CH.sub.2Cl.sub.2/MeOH=10:1) to give the desired compound, and further purified by Prep-HPLC (Method A) to give the titled compound (36 mg, 14%). .sup.1H NMR (400 MHz, CD.sub.3OD) 6.90-6.75 (m, 3H), 6.33 (t, J=54.0 Hz, 1H), 6.01-5.04 (m, 2H), 4.93-4.85 (m, 2H), 4.24-4.16 (m, 1H), 3.88 (d, J=3.2 Hz, 2H), 3.72 (s, 3H), 3.66-3.38 (m, 3H), 3.08-2.68 (m, 2.5H), 2.33 (d, J=11.2 Hz, 0.5H), 1.51-1.40 (m, 1.5H), 1.37-1.23 (m, 10.5H), 1.11-0.91 (m, 3H) ppm. MS: M/e 556 (M+1).sup.+.
Compound A88: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-9-propyl-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00378##
Step A: 1-methyl-6-(propylamino)pyrimidine-2,4 (1H,3H)-dione
##STR00379##
[0605] A solution of 6-chloro-1-methylpyrimidine-2,4 (1H,3H)-dione (5 g, 31.25 mmol) and propan-1-amine (3.7 g, 62.71 mmol) in EtOH (50 ml) was stirred at 60 C. for 4 h. The mixture was filtered to give the titled compound (4.6 g, 80%). MS: M/e 184 (M+1).sup.+.
Step B: 1-methyl-5-nitroso-6-(propylamino)pyrimidine-2,4 (1H,3H)-dione
##STR00380##
[0606] To a solution of 1-methyl-6-(propylamino)pyrimidine-2,4 (1H,3H)-dione (4.59 g, 25.08 mmol) in H.sub.2O (60 ml), NaNO.sub.2 (1.90 g, 27.54 mmol) was added at 05 C. Then AcOH (4.51 g, 75.17 mmol) was added dropwise to the above solution and stirred at 05 C. for 15 min. The reaction mixture was filtered to give the titled compound (5 g, 94%). MS: M/e 213 (M+1).sup.+.
Step C: 5-amino-1-methyl-6-(propylamino)pyrimidine-2,4 (1H,3H)-dione
##STR00381##
[0607] A solution of 1-methyl-5-nitroso-6-(propylamino)pyrimidine-2,4 (1H,3H)-dione (5 g, 23.58 mmol) and Na.sub.2S.sub.2O.sub.4 (12 g, 68.97 mmol) in H.sub.2O (50 ml) was stirred at RT for 30 min. The reaction mixture was filtered and dried to give the titled compound (3.8 g, 81%). MS: M/e 199 (M+1).sup.+.
Step D: 8-((benzyloxy)methyl)-3-methyl-9-propyl-3,9-dihydro-1H-purine-2,6-dione
##STR00382##
[0608] 5-amino-1-methyl-6-(propylamino)pyrimidine-2,4 (1H,3H)-dione (3.8 g, 19.19 mmol) and 2-(benzyloxy)acetic acid (3.5 g, 21.08 mmol) were stirred at 240 C. for 2 hrs. The mixture was cooled to RT, and then dissolved in aq. NaOH (2N, 20 ml). The solution was neutralized with aq. HCl (2N) to pH=7, which was then filtered. The resulting solid was dried to give the titled compound (3.7 g, 59%). MS: M/e 329 (M+1).sup.+.
Step E: tert-butyl (2R,5S)-4-(8-((benzyloxy)methyl)-3-methyl-2-oxo-9-propyl-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00383##
[0609] A solution of 8-((benzyloxy)methyl)-3-methyl-9-propyl-3,9-dihydro-1H-purine-2,6-dione (3.2 g, 9.76 mmol), tert-butyl (2R,5S)-2,5-dimethylpiperazine-1-carboxylate (2.3 g, 10.75 mmol), BOP (6.5 g, 14.71 mmol) and DBU (4.5 g, 29.41 mmol) in MeCN (35 ml) was stirred at RT overnight. The reaction solution was dissolved in EtOAc (50 ml) and washed with brine (20 ml2). The organic layer was dried over Na.sub.2SO.sub.4 and concentrated under reduced pressure. The residue was purified by flash column chromatography with 0-5% MeOH in DCM to give the titled compound (4 g, 78%). MS: M/e 525 (M+1).sup.+
Step F: 6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-8-(hydroxymethyl)-3-methyl-9-propyl-3,9-dihydro-2H-purin-2-one
##STR00384##
[0610] To a solution of tert-butyl (2R,5S)-4-(8-((benzyloxy)methyl)-3-methyl-2-oxo-9-propyl-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (4 g, 7.63 mmol) in DCM (50 ml) at RT, was added BBr.sub.3 (1M in DCM, 22.90 ml, 22.90 mmol) dropwise. The solution was stirred at RT for 6 hrs and then evaporated to dryness. The resulting residue was slurried in DCM & MeOH and then filtered. The filtrate was evaporated to dryness to give the titled compound (2.5 g, crude), which was used directly for the next step without further purification. MS: M/e 335 (M+1).sup.+.
Step G: tert-butyl (2R,5S)-4-(8-(hydroxymethyl)-3-methyl-2-oxo-9-propyl-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00385##
[0611] A solution of 6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-8-(hydroxymethyl)-3-methyl-9-propyl-3,9-dihydro-2H-purin-2-one (2.5 g, crude), (Boc).sub.2O (1.79 g, 8.21 mmol) and TEA (1.5 g, 14.85 mmol) in THE (30 ml) and H.sub.2O (2 ml) was stirred at RT overnight. The mixture was diluted with EtOAc (30 ml) and then washed with brine (20 ml2). The organic layer was dried over Na.sub.2SO.sub.4 and concentrated under reduced pressure. The residue was purified by flash column chromatography with 0-5% MeOH in DCM to give the titled compound (1.05 g). MS: M/e 435 (M+1).sup.+.
Step H: tert-butyl (2R,5S)-4-(8-(chloromethyl)-3-methyl-2-oxo-9-propyl-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00386##
[0612] A solution of tert-butyl (2R,5S)-4-(8-(hydroxymethyl)-3-methyl-2-oxo-9-propyl-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (500 mg, 1.15 mmol) and SOCl.sub.2 (411 mg, 3.45 mmol) in DCM (10 ml) was stirred at 0 C. for 15 min. The solution was washed with aq. NaHCO.sub.3 (10 ml), dried over Na.sub.2SO.sub.4 and evaporated to dryness to give the titled compound (520 mg, 100%), which was used directly for the next step.
Step I: tert-butyl (2R,5S)-4-(8-(cyanomethyl)-3-methyl-2-oxo-9-propyl-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00387##
[0613] A solution of tert-butyl (2R,5S)-4-(8-(chloromethyl)-3-methyl-2-oxo-9-propyl-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (520 mg, 1.15 mmol), trimethylsilanecarbonitrile (342 mg, 3.45 mmol) and Cs.sub.2CO.sub.3 (1.1 g, 3.37 mmol) in MeCN (10 ml) was stirred at 70 C. overnight. The mixture was diluted with EtOAc (20 ml) and washed with brine (15 ml). The organic layer was dried over Na.sub.2SO.sub.4 and concentrated. The resulting residue was purified by flash column chromatography with 0-5% MeOH in DCM to give the titled compound (210 mg, 41%). MS: M/e 444 (M+1).sup.+.
Step J: 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-9-propyl-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00388##
[0614] A solution of tert-butyl (2R,5S)-4-(8-(cyanomethyl)-3-methyl-2-oxo-9-propyl-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (210 mg, 0.47 mmol) and TFA (2 ml) in DCM (10 ml) was stirred at RT for 4 hrs. The solution was washed with aq. NaHCO.sub.3, dried over Na.sub.2SO.sub.4 and evaporated to dryness to give the titled compound (162.5 mg, 100%), which was used directly for the next step without further purification. MS: M/e 344 (M+1).sup.+.
Step K: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-9-propyl-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00389##
[0615] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-9-propyl-3,9-dihydro-2H-purin-8-yl)acetonitrile (160 mg, 0.47 mmol), 1-(3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (291 mg, 1.40 mmol), (cyanomethyl)trimethylphosphonium iodide (340 mg, 1.40 mmol) and DIPEA (602 mg, 4.67 mmol) in MeCN (4 ml) was stirred at 100 C. overnight. The reaction was diluted with EtOAc (10 ml) and washed with brine (10 ml). The organic layer was concentrated under reduced pressure. The resulting residue was purified by flash column chromatography with 0-5% MeOH in DCM and then Prep-HPLC (Method A) to give the titled compound (47.1 mg). .sup.1H NMR (400 MHz, DMSO-d6) 6.86-6.77 (m, 3H), 6.10-5.36 (m, 1H), 5.20-4.52 (m, 1H), 4.40 (s, 2H), 4.13 (s, 2H), 3.90 (d, J=4.5 Hz, 2H), 3.59 (s, 3H), 3.45 (q, J=6.1 Hz, 1.5H), 3.32 (d, J=6.4 Hz, 2H), 2.76 (d, J=36.8 Hz, 1H), 2.18 (d, J=10.8 Hz, 0.5H), 1.72 (q, J=7.4 Hz, 2H), 1.40-1.24 (m, 7.5H), 1.23-1.13 (m, 4.5H), 0.91 (t, J=7.0 Hz, 4.5H), 0.80 (d, J=6.2 Hz, 1.5H) ppm. MS. M/e 534 (M+1).sup.+.
Compound A91: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-methyl-4-oxo-4H-chromen-6-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00390##
Step A: 6-bromo-3-methyl-4H-chromen-4-one
##STR00391##
[0616] To a solution of 1-(5-bromo-2-hydroxyphenyl)propan-1-one (600 mg, 2.6 mmol) in DMF (10 mL) was added DMF-DMA (626 mg, 5.2 mol). The mixture was stirred at 120 C. for 3 hrs. The organic layer was diluted with EtOAc and washed with brine, dried over MgSO.sub.4, filtered and the solvent was removed under vacuum. The resulting residue was purified by flash column chromatography (PE:EA) to give the titled compound (510 mg, 88%). MS: M/e 221 (M+1-18).sup.+.
Step B: 6-acetyl-3-methyl-4H-chromen-4-one
##STR00392##
[0617] To a solution of 6-bromo-3-methyl-4H-chromen-4-one (400 mg, 1.7 mmol) in toluene (10 mL) was added 1-(vinyloxy) butane (1.7 g, 3.5 mmol), Pd(PPh.sub.3).sub.2Cl.sub.2 (120 mg, 0.17 mmol). The mixture was stirred at 100 C. under N.sub.2 for 16 hrs. The reaction was cooled to room temperature, treated with dioxane (4N HCl, 1 mL) dropwise, stirred at room temperature for one hour. The mixture was concentrated to give titled compound for the next step.
Step C: 6-(1-hydroxyethyl)-3-methyl-4H-chromen-4-one
##STR00393##
[0618] To a solution of 6-acetyl-3-methyl-4H-chromen-4-one (crude, 1.7 mmol) in MeOH (2 ml) at 0 C. was added NaBH.sub.4 (65 mg, 1.7 mmol) in some portions. After addition, the reaction was stirred at 0 C. for 5 min. The reaction was quenched by adding water and extracted with DCM, washed with brine, dried over Na.sub.2SO.sub.4, filtered, concentrated to give the titled compound (90 mg, two steps 27%). MS: M/e 187 (M+1-18).sup.+.
Step D: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-methyl-4-oxo-4H-chromen-6-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00394##
[0619] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (80 mg, 0.24 mol), 6-(1-hydroxyethyl)-3-methyl-4H-chromen-4-one (90 mg, 0.48 mol), (cyanomethyl)trimethylphosphonium iodide (175 mg, 0.72 mol) and DIPEA (310 mg, 2.4 mol) in CH.sub.3CN (2 ml). Then the mixture solution was stirred at 105 C. for 16 hours. The reaction mixture was concentrated under reduced pressure, then the crude was diluted with DCM and washed with brine, dried over MgSO.sub.4, filtered and the solvent was removed under vacuum. The resulting residue was purified by flash column chromatography (DCM:MeOH) to give the titled compound (42 mg). .sup.1H NMR (400 MHz, CD.sub.3OD) 8.17-8.07 (m, 2H), 7.87 (t, J=8.0 Hz, 1H), 7.56 (dd, J=12.5, 8.8 Hz, 1H), 6.15-4.91 (m, 2H), 4.42-4.31 (m, 2H), 3.79 (d, J=6.8 Hz, 1H), 3.76 (s, 3H), 3.70-3.40 (m, 2H), 3.28-3.15 (m, 1H), 2.99-2.80 (m, 2H), 2.79-2.10 (m, 1H), 2.02 (s, 3H), 1.50-1.43 (m, 4H), 1.41-1.22 (m, 5H), 1.10-0.87 (m, 3H) ppm. MS: M/e 516 (M+1).sup.+.
Compound A94: 2-(6-((2S,5R)-4-(1-(2,2-dimethylchroman-7-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00395##
Step A: 7-bromo-2,2-dimethylchroman-4-one
##STR00396##
[0620] To a solution of 1-(4-bromo-2-hydroxyphenyl)ethan-1-one (4 g, 0.019 mol) in Tol. (16 ml) was added pyrrolidine (4 ml) at 05 C. and stirred for 20 min. Acetone (1.62 g, 0.030 mmol) was added to the above solution and stirred at 80 C. overnight. The solution was diluted with EtOAc (20 ml), washed with brine (10 ml), dried and concentrated. The residue was purified by flash column chromatography with 20%-40% DCM in PE to give the titled compound (3.6 g, 75%). .sup.1H NMR (400 MHz, CDCl.sub.3) 7.71 (d, J=8.3 Hz, 1H), 7.14 (s, 1H), 7.11 (d, J=9.6 Hz, 1H), 2.71 (s, 2H), 1.45 (s, 6H) ppm.
Step B: 7-bromo-2,2-dimethylchroman-4-ol
##STR00397##
[0621] A solution of 7-bromo-2,2-dimethylchroman-4-one (3.1 g, 0.012 mol) and NaBH.sub.4 (0.46 g, 0.012 mol) in MeOH (30 ml) was stirred at RT for 15 min. The solution was quenched with water (10 ml) and then concentrated under reduced pressure. The residue was diluted with EtOAc (30 ml), washed with brine (10 ml), dried and evaporated to dryness to give the titled compound (3.1 g, 100%), which was used directly for the next step. MS: M/e 257 (M+1).sup.+.
Step C: 7-bromo-2,2-dimethylchromane
##STR00398##
[0622] A solution of 7-bromo-2,2-dimethylchroman-4-ol (3.1 g, 0.012 mol) in TFA (10 ml) and triethylsilane (10 ml) was stirred at RT for 30 min. The solution was diluted with EtOAc (30 ml), washed with brine (15 ml), died and then concentrated. The residue was purified by flash column chromatography with 0%-5% EtOAc in PE to give the titled compound (2.1 g, 72%). .sup.1H NMR (400 MHz, CDCl.sub.3) 6.97-6.87 (m, 3H), 2.71 (t, J=6.8 Hz, 2H), 1.79 (t, J=6.8 Hz, 2H), 1.32 (s, 6H) ppm.
Step D: 1-(2,2-dimethylchroman-7-yl)ethan-1-one
##STR00399##
[0623] A solution of 7-bromo-2,2-dimethylchromane (500 mg, 2.07 mmol), Pd(PPh.sub.3).sub.2Cl.sub.2 (73 mg, 0.10 mmol) and tributyl(1-ethoxyvinyl)stannane (1.1 g, 3.05 mmol) in Tol. (10 ml) was stirred at 100 C. overnight. The mixture was cooled to RT and HCl (4M in dioxane, 2 ml) was added, stirred for 30 min. The mixture was diluted with EtOAc (20 ml), washed with brine (10 ml), dried and concentrated. The residue was purified by flash column chromatography with 0%-10% EtOAc in PE to give the titled compound (250 mg, 59%). .sup.1H NMR (400 MHz, CDCl.sub.3) 7.43 (d, J=8.0 Hz, 1H), 7.36 (s, 1H), 7.14 (d, J=8.0 Hz, 1H), 2.82 (t, J=6.7 Hz, 2H), 2.54 (s, 3H), 1.84 (t, J=6.7 Hz, 2H), 1.35 (s, 6H) ppm.
Step E: 1-(2,2-dimethylchroman-7-yl)ethan-1-ol
##STR00400##
[0624] A solution of 1-(2,2-dimethylchroman-7-yl)ethan-1-one (250 mg, 1.23 mmol) and NaBH.sub.4 (47 mg, 1.24 mmol) in MeOH (4 ml) and THE (1 ml) was stirred at RT for 30 min. The solution was quenched with water (10 ml) and then concentrated under reduced pressure. The residue was diluted with EtOAc (15 ml), washed with brine (10 ml), dried and evaporated to dryness to give the titled compound (250 mg, 100%), which was used directly for the next step. MS: M/e 207 (M+1).sup.+.
Step F: 2-(6-((2S,5R)-4-(1-(2,2-dimethylchroman-7-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00401##
[0625] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (200 mg, 0.61 mmol), 1-(2,2-dimethylchroman-7-yl)ethan-1-ol (250 mg, 1.21 mmol), (cyanomethyl)trimethylphosphonium iodide (443 mg, 1.82 mmol) and DIPEA (784 mg, 6.08 mmol) in MeCN (3 ml) was stirred at 100 C. overnight. The reaction was diluted with EtOAc (10 ml) and washed with brine (10 ml). The organic layer was concentrated under reduced pressure. The resulting residue was purified by flash column chromatography with 0-5% MeOH in DCM to give the titled Compound A94 (70 mg), which was further separated into Compound A94a (21 mg) and Compound A94b (21 mg) by chiral Prep-HPLC. The chiral separation conditions are shown below.
TABLE-US-00018 Column CHIRALPAK IH Column Size 2 cm 25 cm, 5 um Mobile Phase A MtBE Mobile Phase B MeOH:DCM Flow Rate 20 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-HPLC Equipment Prep-HPLC-Gilson
[0626] Compound A94: .sup.1H NMR (400 MHz, DMSO-d6) 7.01 (t, J=7.6 Hz, 1H), 6.81 (d, J=7.6 Hz, 1H), 6.68 (s, 1H), 6.10-5.40 (m, 1H), 5.20-4.50 (m, 1H), 4.40 (s, 2H), 4.26 (d, J=6.4 Hz, 2H), 3.61 (s, 3H), 3.45 (s, 1H), 3.33 (s, 1.5H), 2.70 (q, J=5.3 Hz, 3H), 2.44 (s, 1H), 2.17 (d, J=12.3 Hz, 0.5H), 1.75 (q, J=4.2 Hz, 2H), 1.39-1.12 (m, 15H), 0.93 (s, 1.5H), 0.80 (d, J=6.0 Hz, 1.5H) ppm. MS: M/e 518 (M+1).sup.+.
[0627] Compound A94a (the earlier peak): .sup.1H NMR (400 MHz, DMSO-d6) 7.02 (d, J=7.3 Hz, 1H), 6.81 (d, J=7.5 Hz, 1H), 6.68 (s, 1H), 6.12-5.39 (m, 1H), 5.20-4.42 (m, 1H), 4.39 (s, 2H), 4.25 (s, 2H), 3.60 (s, 3H), 3.43 (s, 1H), 3.33 (s, 2H), 2.77-2.65 (m, 4H), 1.75 (t, J=7.8 Hz, 2H), 1.33 (s, 6H), 1.27 (s, 6H), 1.19 (d, J=5.5 Hz, 3H), 0.80 (d, J=5.1 Hz, 3H) ppm. MS: M/e 518 (M+1).sup.+.
[0628] Compound A94b (the later peak): .sup.1H NMR (400 MHz, DMSO-d6) 7.00 (d, J=7.1 Hz, 1H), 6.82 (d, J=7.3 Hz, 1H), 6.68 (s, 1H), 5.95-5.60 (m, 1H), 5.00-4.55 (m, 1H), 4.40 (s, 2H), 4.27 (s, 2H), 3.61 (s, 3H), 3.46 (s, 1H), 3.33 (s, 3H), 2.70 (t, J=4.0 Hz, 2H), 2.18 (d, J=7.8 Hz, 1H), 1.74 (t, J=7.8 Hz, 2H), 1.34 (t, J=6.8 Hz, 3H), 1.26 (d, J=4.3 Hz, 6H), 1.21 (d, J=5.8 Hz, 3H), 1.17 (s, 3H), 0.93 (s, 3H) ppm. MS: M/e 518 (M+1).sup.+.
Compound A95: 2-(9-ethyl-6-((2S,5R)-4-(1-(2-ethylbenzo[d]thiazol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00402##
Step A: 5-bromo-2-ethylbenzo[d]thiazole
##STR00403##
[0629] To a stirred solution of 5-bromo-2-methylbenzo[d]thiazole (1.3 g, 5.7 mmol) in THE (30 mL) was added dropwise LDA (2.0 M, 3.4 mL, 6.8 mmol) at 78 C. After stirred for 30 min, Mel (4.05 g, 28.5 mmol) was added dropwise at 78 C. The reaction mixture was quenched with aq.NH.sub.4Cl, extracted with EtOAc (15 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated and purified by flash column chromatography to give the titled compound (700 mg, 51%). MS: M/e 243 (M+1).sup.+.
Step B: 1-(2-ethylbenzo[d]thiazol-5-yl)ethan-1-one
##STR00404##
[0630] A mixture of 5-bromo-2-ethylbenzo[d]thiazole (400 mg, 1.65 mmol), tributyl(1-ethoxyvinyl)stannane (893 mg, 2.48 mmol) and Pd(PPh.sub.3).sub.2Cl.sub.2 (58 mg, 0.083 mmol) in toluene (10 mL) was stirred at 100 C. overnight under N.sub.2. The reaction mixture was treated with aq.HCl (2.0 M, 5 mL) and stirred for 10 min, then extracted with EtOAc (15 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated and purified by flash column chromatography to give the titled compound (250 mg, 74%). MS: M/e 206 (M+1).sup.+.
Step C: 1-(2-ethylbenzo[d]thiazol-5-yl)ethan-1-ol
##STR00405##
[0631] To a stirred solution of 1-(2-ethylbenzo[d]thiazol-5-yl)ethan-1-one (250 mg, 1.22 mmol) in MeOH (5 mL) was added NaBH.sub.4 (46.36 mg, 1.22 mmol). After then, the mixture was stirred for 10 min. The reaction mixture was poured into H.sub.2O (10 mL) and extracted with EtOAc (10 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated to give the titled compound (240 mg, 95%). MS: M/e 208 (M+1).sup.+.
Step D: 2-(9-ethyl-6-((2S,5R)-4-(1-(2-ethylbenzo[d]thiazol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00406##
[0632] A mixture of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (100 mg, 0.3 mmol), 1-(2-ethylbenzo[d]thiazol-5-yl)ethan-1-ol (94 mg, 0.45 mmol), (cyanomethyl)trimethylphosphonium iodide (218 mg, 0.9 mmol) and DIPEA (387 mg, 3 mmol) in CH.sub.3CN (4 mL) was stirred overnight at 100 C. in a sealed tube. The reaction mixture was diluted with EtOAc (20 mL), washed with H.sub.2O, brine, dried over Na.sub.2SO.sub.4, concentrated to give the residue, which was purified by Prep-HPLC (Method A) to give the titled Compound A95 (40 mg). which was further separated into Compound A95a (14 mg) and Compound A95b (15 mg) by chiral Prep-HPLC. The chiral separation conditions are shown below.
TABLE-US-00019 Column CHIRALPAK ID Column Size 2 cm 25 cm, 5 um Mobile Phase A MtBE Mobile Phase B MeOH:DCM Flow Rate 20 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-HPLC Equipment Prep-HPLC-Gilson
[0633] Compound A95: .sup.1H NMR (400 MHz, CD.sub.3OD) 7.97-7.87 (m, 2H), 7.50 (t, J=7.6 Hz, 1H), 6.25-4.94 (m, 2H), 4.43-4.31 (m, 2H), 4.27-3.80 (m, 2H), 3.76 (s, 3H), 3.73-3.64 (m, 1H), 3.34-3.31 (m, 2H), 3.21-3.12 (m, 2H), 3.00-2.91 (m, 1.5H), 2.81-2.68 (m, 0.5H), 1.53-1.23 (m, 12H), 1.12-0.92 (m, 3H) ppm. MS: M/e 519 (M+1).sup.+.
[0634] Compound A95a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.93 (s, 1H), 7.91 (d, J=8.4 Hz, 1H), 7.50 (d, J=7.6 Hz, 1H), 6.35-5.04 (m, 2H), 4.36 (dd, J=13.6, 6.4 Hz, 2H), 4.26-4.16 (m, 1H), 3.82-3.77 (m, 1H), 3.76 (s, 3H), 3.65-3.40 (m, 1H), 3.32-3.29 (m, 2H), 3.16 (q, J=7.6 Hz, 2H), 2.99-2.82 (m, 2H), 1.56-1.41 (m, 9H), 1.37 (d, J=6.4 Hz, 3H), 0.92 (d, J=6.4 Hz, 3H)) ppm. MS: M/e 519 (M+1).sup.+.
[0635] Compound A95b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.93 (s, 1H), 7.88 (d, J=8.4 Hz, 1H), 7.49 (d, J=8.4 Hz, 1H), 6.11-4.94 (m, 2H), 4.37 (q, J=7.2 Hz, 2H), 4.26-4.16 (m, 1H), 3.76 (s, 3H), 3.70-3.58 (m, 2H), 3.34-3.31 (m, 2H), 3.15 (q, J=7.6 Hz, 2H), 2.78-2.59 (m, 1H), 2.23 (d, J=12 Hz, 1H), 1.52-1.37 (m, 9H), 1.27 (d, J=6.4 Hz, 3H), 1.08 (d, J=6.4 Hz, 3H) ppm. MS: M/e 519 (M+1).sup.+.
Compound A96: 2-(9-ethyl-6-((2S,5R)-4-(1-(2-isopropylbenzo[d]thiazol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00407##
Step A: 5-bromo-2-isopropylbenzo[d]thiazole
##STR00408##
[0636] To a stirred solution of 5-bromo-2-methylbenzo[d]thiazole (2.28 g, 10 mmol) in THE (30 mL) was added dropwise LDA (2.0 M, 11 mL, 22 mmol) at 78 C. After stirred for 30 min, Mel (7.1 g, 50 mmol) was added dropwise at 78 C. The reaction mixture was quenched with aq.NH.sub.4Cl, extracted with EtOAc (15 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated and purified by flash column chromatography to give the titled compound (1.5 g, 59%). MS: M/e 257 (M+1).sup.+.
Step B: 1-(2-isopropylbenzo[d]thiazol-5-yl)ethan-1-one
##STR00409##
[0637] A mixture of 5-bromo-2-isopropylbenzo[d]thiazole (512 mg, 2 mmol), tributyl(1-ethoxyvinyl)stannane (1.08 g, 3 mmol) and Pd(PPh.sub.3).sub.2Cl.sub.2 (70 mg, 0.1 mmol) in toluene (10 mL) was stirred at 100 C. overnight under N.sub.2. The reaction mixture was treated with aq.HCl (2.0 M, 5 mL) and stirred for 10 min, then extracted with EtOAc (15 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated and purified by flash column chromatography to give the titled compound (385 mg, 88%). MS: M/e 220 (M+1).sup.+.
Step C: 1-(2-isopropylbenzo[d]thiazol-5-yl)ethan-1-ol
##STR00410##
[0638] To a stirred solution of 1-(2-isopropylbenzo[d]thiazol-5-yl)ethan-1-one (385 mg, 1.76 mmol) in MeOH (5 mL) was added NaBH.sub.4 (67 mg, 1.76 mmol). After then, the mixture was stirred for 10 min. The reaction mixture was poured into H.sub.2O (10 mL) and extracted with EtOAc (10 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated to give the titled compound (212 mg, 55%). MS: M/e 222 (M+1).sup.+.
Step D: 2-(9-ethyl-6-((2S,5R)-4-(1-(2-isopropylbenzo[d]thiazol-5-yl)ethyl)-2,5-dimethyl piperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00411##
[0639] A mixture of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (100 mg, 0.3 mmol), 1-(2-isopropylbenzo[d]thiazol-5-yl)ethan-1-ol (99.4 mg, 0.45 mmol), (cyanomethyl)trimethylphosphonium iodide (218 mg, 0.9 mmol) and DIPEA (387 mg, 3 mmol) in CH.sub.3CN (4 mL) was stirred overnight at 100 C. in a sealed tube. The reaction mixture was diluted with EtOAc (20 mL), washed with H.sub.2O, brine, dried over Na.sub.2SO.sub.4, concentrated to give the residue, which was purified by Prep-HPLC (Method A) to give the titled compound (55 mg). .sup.1H NMR (400 MHz, CD.sub.3OD) 8.00-7.87 (m, 2H), 7.54-7.45 (m, 1H), 6.34-4.97 (m, 2H), 4.46-4.31 (m, 2H), 4.29-3.82 (m, 2H), 3.76 (s, 3H), 3.73-3.63 (m, 1H), 3.48-3.37 (m, 1H), 3.04-2.64 (m, 4H), 1.54-1.24 (m, 15H), 1.03 (dd, J=56.4, 6.0 Hz, 3H) ppm. MS: M/e 533 (M+1).sup.+.
Compound A97: 2-(6-((2S,5R)-4-(1-(2,2-dimethyl-2,3-dihydrobenzofuran-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00412##
Step A: 1-(4-bromo-2-fluorophenyl)-2-methylpropan-2-ol
##STR00413##
[0640] To a solution of methyl 2-(4-bromo-2-fluorophenyl)acetate (1 g, 4.05 mmol) in THE (10 ml) at 70 C. under N.sub.2, was added MeMgBr (3M, 3.37 ml, 10.11 mmol) dropwise. The solution was warmed to RT and then stirred at RT for 2 hrs. The reaction was quenched with brine (10 ml) and extracted with EtOAc (10 ml2). The organic layer was concentrated under reduced pressure. The resulting residue was purified by flash column chromatography with 0-20% EtOAc in PE to give the titled compound (840 mg, 84%). .sup.1H NMR (400 MHz, CDCl.sub.3) 7.24 (d, J=8.4 Hz, 2H), 7.14 (t, J=8.0 Hz, 1H), 2.78 (s, 2H), 1.24 (s, 6H) ppm.
Step B: 6-bromo-2,2-dimethyl-2,3-dihydrobenzofuran
##STR00414##
[0641] A solution of 1-(4-bromo-2-fluorophenyl)-2-methylpropan-2-ol (740 mg, 3.00 mmol) and tBuOK (1M, 3.5 ml, 3.5 mmol) in THE (10 ml) was stirred at 60 C. overnight. The reaction was diluted with EtOAc (20 ml) and washed with brine (10 ml). The organic layer was concentrated under reduced pressure. The resulting residue was purified by flash column chromatography with 0-5% EtOAc in PE to give the titled compound (490 mg, 72%). .sup.1H NMR (400 MHz, CDCl.sub.3) 6.95 (dd, J=18.4, 7.9 Hz, 2H), 6.87 (s, 1H), 2.94 (s, 2H), 1.46 (s, 6H) ppm.
Step C: 1-(2,2-dimethyl-2,3-dihydrobenzofuran-6-yl)ethan-1-one
##STR00415##
[0642] A solution of 6-bromo-2,2-dimethyl-2,3-dihydrobenzofuran (570 mg, 2.51 mmol), ethyl tributylstannanecarboxylate (1.36 g, 3.77 mmol) and Pd(PPh.sub.3).sub.2Cl.sub.2 (88 mg, 0.13 mmol) in toluene (10 ml) under N.sub.2 was stirred at 100 C. overnight. The solution was cooled to RT. HCl/1,4-dioxane (4M, 2 ml) was added and stirred at RT for 10 min. The solution was diluted with EtOAc (15 ml), washed with aq. Na.sub.2CO.sub.3 (10 ml2), dried over Na.sub.2SO.sub.4 and concentrated to dryness. The resulting residue was purified by flash column chromatography with 0-10% EtOAc in PE to give the titled compound (0.2 g, 42%). .sup.1H NMR (400 MHz, CDCl.sub.3) 7.46 (d, J=7.8 Hz, 1H), 7.29 (s, 1H), 7.20 (d, J=7.6 Hz, 1H), 3.04 (s, 2H), 2.56 (s, 3H), 1.49 (s, 6H) ppm.
Step D: 1-(2,2-dimethyl-2,3-dihydrobenzofuran-6-yl)ethan-1-ol
##STR00416##
[0643] A solution of 1-(2,2-dimethyl-2,3-dihydrobenzofuran-6-yl)ethan-1-one (200 mg, 1.05 mmol) and NaBH.sub.4 (40 mg, 1.05 mmol) in MeOH (3 ml) was stirred at RT for 10 min. The solution was diluted with EtOAc (10 ml), washed with brine (5 ml2), dried over Na.sub.2SO.sub.4 and concentrated to dryness to give the titled compound (200 mg, 99%), which was used directly for the next step without further purification. MS: M/e 193 (M+1).sup.+.
Step E: 2-(6-((2S,5R)-4-(1-(2,2-dimethyl-2,3-dihydrobenzofuran-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00417##
[0644] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (115 mg, 0.35 mmol), 1-(2,2-dimethyl-2,3-dihydrobenzofuran-6-yl)ethan-1-ol (200 mg, 1.04 mmol), (cyanomethyl)trimethylphosphonium iodide (255 mg, 1.05 mmol) and DIPEA (451 mg, 3.50 mmol) in CH.sub.3CN (2 ml) was stirred at 100 C. overnight. The reaction was diluted with EtOAc (15 ml) and washed with brine (10 ml). The organic layer was concentrated under reduced pressure. The resulting residue was purified by flash column chromatography and then Prep-HPLC (Method A) to give the titled compound (73 mg). .sup.1H NMR (400 MHz, DMSO-d6) 7.10 (t, J=8.1 Hz, 1H), 6.80 (d, J=7.1 Hz, 1H), 6.70 (d, J=5.1 Hz, 1H), 6.10-5.40 (m, 1H), 5.20-4.47 (m, 1H), 4.41 (s, 2H), 4.25 (s, 2H), 3.61 (s, 3.5H), 3.52-3.20 (m, 2.5H), 2.97 (d, J=5.5 Hz, 2H), 2.79 (d, J=32.7 Hz, 1.5H), 2.19 (d, J=10.3 Hz, 0.5H), 1.46-1.29 (m, 10.5H), 1.26-1.14 (m, 4.5H), 0.94 (s, 1.5H), 0.80 (d, J=5.3 Hz, 1.5H) ppm. MS: M/e 504 (M+1).sup.+.
Compound A98: 2-(6-((2S,5R)-4-(1-(2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethyl piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00418##
[0645] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (100 mg, 0.32 mmol), 1-(2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethan-1-ol (62 mg, 0.31 mmol), (cyanomethyl)trimethylphosphonium iodide (230 mg, 0.95 mmol) and DIPEA (205 mg, 1.59 mmol) in MeCN (5 ml) was stirred at 100 C. overnight. After completed, the solution was concentrated under reduced pressure to dryness. The resulting residue was purified by Prep-HPLC (Method A) to give the Compound A98a (34 mg) and Compound A98b (32 mg).
[0646] Compound A98: .sup.1H NMR (400 MHz, CD.sub.3OD) 6.83 (s, 1H), 6.77 (t, J=7.0 Hz, 1H), 6.66 (t, J=8.4 Hz, 1H), 6.05-5.02 (m, 2H), 4.21 (s, 1H), 3.93 (s, 3H), 3.78 (s, 3H), 3.75-3.34 (m, 3H), 3.26-2.70 (m, 2H), 2.70-1.92 (m, 1H), 1.64 (d, J=5.1 Hz, 6H), 1.45-1.25 (m, 6H), 1.11-0.88 (m, 3H) ppm. MS: M/e 492 (M+1).sup.+
[0647] Compound A98a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.81 (s, 1H), 6.75 (d, J=7.1 Hz, 1H), 6.65 (d, J=6.9 Hz, 1H), 6.08-5.11 (m, 2H), 3.93 (s, 3H), 3.78 (s, 3H), 3.48 (s, 1H), 3.28-2.66 (m, 6H), 1.64 (d, J=2.2 Hz, 6H), 1.44 (s, 3H), 1.28 (s, 3H), 0.88 (s, 3H) ppm. MS: M/e 492 (M+1).sup.+
[0648] Compound A98b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.81 (s, 1H), 6.74 (d, J=8.1 Hz, 1H), 6.62 (d, J=7.7 Hz, 1H), 5.47 (d, J=324.8 Hz, 2H), 3.93 (s, 3H), 3.78 (s, 3H), 3.55 (s, 1H), 3.37 (s, 4H), 2.61 (s, 1H), 2.27 (d, J=12.2 Hz, 1H), 1.63 (s, 6H), 1.30 (d, J=5.5 Hz, 3H), 1.27 (s, 3H), 1.01 (d, J=6.2 Hz, 3H) ppm. MS: M/e 492 (M+1).sup.+
Compound A99: 2-(6-((2S,5R)-4-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethyl piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00419##
Step A: 1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)ethan-1-ol
##STR00420##
[0649] To a solution of 2,2-difluorobenzo[d][1,3]dioxole-5-carbaldehyde (200 mg, 1.07 mmol) in THE (20 mL) was added methylmagnesium bromide (1.3 mL, 1.29 mmol) at 78 C. and stirred for 1 h. The mixture was extracted with DCM (100 mL) and washed with water. The organic layer was dried, concentrated and purified by Prep-TLC (DCM:MeOH=20:1) to give the titled compound (180 mg, 83%). MS: M/e 203 (M+1).sup.+
Step B: 2-(6-((2S,5R)-4-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00421##
[0650] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.15 mmol) in CH.sub.3CN (3 mL) and was added 1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)ethan-1-ol (33 mg, 0.15 mmol), (cyanomethyl)trimethylphosphonium iodide (110 mg, 0.46 mmol) and DIPEA (98 mg, 0.76 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure to give the titled Compound A99 (crude), which was further purified and separated into Compound A99a (10 mg) and Compound A99b (14 mg) by Prep-HPLC (Method A).
[0651] Compound A99: .sup.1H NMR (400 MHz, CD.sub.3OD) 7.30 (s, 1H), 7.23-7.08 (m, 2H), 6.35-5.51 (m, 1H), 5.47-4.88 (m, 1H), 4.37 (q, J=7.2 Hz, 2H), 4.23 (s, 2H), 3.76 (s, 3H), 3.73-3.52 (m, 2H), 3.43-3.31 (m, 0.5H), 2.96-2.66 (m, 2H), 2.40-2.20 (m, 0.5H), 1.48-1.43 (m, 4H), 1.40-1.21 (m, 5H), 1.08-0.91 (m, 3H) ppm. MS: M/e 514 (M+1).sup.+
[0652] Compound A99a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.34-7.24 (m, 1H), 7.16 (dd, J=18.2, 8.2 Hz, 2H), 5.88-5.16 (m, 1H), 4.86-4.50 (m, 1H), 4.37 (d, J=6.3 Hz, 2H), 4.22 (s, 1H), 3.76 (s, 3H), 3.67 (s, 1H), 3.62-3.32 (m, 2H), 2.86 (s, 3H), 1.46 (s, 6H), 1.30 (d, J=5.7 Hz, 3H), 0.91 (d, J=5.4 Hz, 3H) ppm. MS: M/e 514 (M+1).sup.+
[0653] Compound A99b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.29 (s, 1H), 7.16 (d, J=8.0 Hz, 1H), 7.11 (d, J=8.4 Hz, 1H), 5.93-5.04 (m, 1H), 4.86-4.59 (m, 1H), 4.37 (d, J=7.2 Hz, 2H), 4.23 (s, 1H), 3.76 (s, 3H), 3.59 (s, 1H), 3.52 (s, 1H), 3.29-3.07 (m, 2H), 2.66 (s, 1H), 2.19 (d, J=12.1 Hz, 1H), 1.46 (t, J=7.1 Hz, 3H), 1.33 (d, J=5.7 Hz, 3H), 1.27 (s, 3H), 1.04 (d, J=5.1 Hz, 3H) ppm. MS: M/e 514 (M+1).sup.+
Compound A102: 2-(6-((2S,5R)-4-(1-(4,4-difluoro-3,4-dihydro-2H-pyrano[3,2-b]pyridin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00422##
[0654] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.15 mmol), 1-(4,4-difluoro-3,4-dihydro-2H-pyrano[3,2-b]pyridin-6-yl)ethan-1-ol (33 mg, 0.15 mmol), (cyanomethyl)trimethylphosphonium iodide (110 mg, 0.46 mmol) and DIPEA (98 mg, 0.76 mmol) in MeCN (3 ml) was stirred at 100 C. overnight. After completed, the solution was concentrated under reduced pressure. The resulting residue was purified by Prep-HPLC (Method A) to give the titled compound (24 mg, 30%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.67 (t, J=7.6 Hz, 1H), 7.43 (d, J=8.5 Hz, 1H), 6.36-4.80 (m, 2H), 4.47-4.30 (m, 4H), 4.30-3.98 (m, 2H), 3.98-3.80 (m, 1H), 3.77 (s, 3H), 3.74-3.48 (m, 1H), 3.44-3.31 (m, 1H), 3.06-2.84 (m, 2H), 2.63 (d, J=16.0 Hz, 2H), 1.57-1.39 (m, 6H), 1.38-1.28 (m, 3H), 1.19-0.96 (m, 3H) ppm. MS: M/e 527 (M+1).sup.+
Compound A103: 2-(6-((2S,5R)-4-(1-(4,4-difluoro-3,4-dihydro-2H-pyrano[3,2-b]pyridin-8-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00423##
[0655] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.15 mmol), 1-(4,4-difluoro-3,4-dihydro-2H-pyrano[3,2-b]pyridin-8-yl)ethan-1-ol (33 mg, 0.15 mmol), (cyanomethyl)trimethylphosphonium iodide (110 mg, 0.46 mmol) and DIPEA (98 mg, 0.76 mmol) in MeCN (3 ml) was stirred at 100 C. overnight. After completed, the solution was concentrated under reduced pressure. The resulting residue was purified and separated into Compound A103a (16 mg) and Compound A103b (20 mg) by Prep-HPLC (Method A).
[0656] Compound A103a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.24 (d, J=4.7 Hz, 1H), 7.80 (d, J=4.7 Hz, 1H), 6.00-4.50 (m, 2H), 4.49-4.40 (m, 2H), 4.37 (q, J=7.2 Hz, 2H), 4.24-3.97 (m, 2H), 3.76 (s, 3H), 3.57 (s, 1H), 3.30-2.36 (m, 5H), 2.16 (d, J=12.1 Hz, 1H), 1.46 (t, J=7.2 Hz, 3H), 1.42-1.20 (m, 6H), 1.05 (d, J=6.0 Hz, 3H) ppm. MS: M/e 527 (M+1).sup.+
[0657] Compound A103b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.26 (d, J=4.8 Hz, 1H), 7.73 (d, J=4.5 Hz, 1H), 6.23-4.50 (m, 2H), 4.45 (s, 2H), 4.37 (d, J=6.8 Hz, 2H), 4.24-4.17 (m, 2H), 3.76 (s, 3H), 3.30-2.74 (m, 5H), 2.69-2.58 (m, 2H), 1.46 (t, J=7.0 Hz, 6H), 1.27 (d, J=6.6 Hz, 3H), 0.94 (d, J=6.2 Hz, 3H) ppm. MS: M/e 527 (M+1).sup.+
Compound A108: 2-(9-ethyl-6-((2S,5R)-4-(1-(3-isopropylphenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00424##
Step A: 1-(3-isopropylphenyl)ethan-1-ol
##STR00425##
[0658] To a mixture of 1-bromo-3-isopropylbenzene (0.6 g, 3 mmol) in THE (10 mL) at 70 C. was added n-BuLi (2 mL, 1.5 mol/L in toluene). After 1 hour, a solution of acetaldehyde (264 mg, 6 mmol) in THE (2 mL) was added. The reaction was stirred for 1 hour. The reaction was quenched with saturated NH.sub.4Cl solution, extracted with EtOAc (60 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered and concentrated. The resulting residue was purified by flash column chromatography to give the titled compound (225 mg, 45%). .sup.1H NMR (400 MHz, DMSO-d6) 7.25-7.18 (m, 2H), 7.13 (d, J=7.2 Hz, 1H), 7.08 (d, J=7.6 Hz, 1H), 5.08 (brs, 1H), 4.74-4.62 (m, 1H), 2.91-2.80 (m, 1H), 1.30 (d, J=6.4 Hz, 3H), 1.19 (d, J=7.2 Hz, 6H) ppm.
Step B: 2-(9-ethyl-6-((2S,5R)-4-(1-(3-isopropylphenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00426##
[0659] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (100 mg, 0.3 mmol), 1-(3-isopropylphenyl)ethan-1-ol (75 mg, 0.45 mmol) and (cyanomethyl) trimethyl phosphonium iodide (218 mg, 0.9 mmol) in CH.sub.3CN (4 mL) was added DIPEA (387 mg, 3 mmol). The mixture was sealed in a bottle and heated at 100 C. for 16 hours. The mixture was cooled to room temperature, diluted with water, extracted with EtOAc (60 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by Prep-HPLC (Method A) to give the titled Compound A108 (60 mg, 41%), which was further separated into Compound A108a (12 mg) and Compound A108b (15 mg) by chiral-Prep-HPLC. The chiral separation conditions are shown below.
TABLE-US-00020 Column CHIRALPAK IH Column Size 2 cm 25 cm, 5 um Mobile Phase A Hex:DCM Mobile Phase B EtOH Flow Rate 20 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-HPLC Equipment Prep-HPLC-Gilson
[0660] Compound A108: .sup.1H NMR (400 MHz, CD.sub.3OD) 7.40-7.06 (m, 4H), 6.39-5.45 (m, 1H), 5.25-4.95 (m, 1H), 4.44-4.31 (m, 2H), 4.28-4.16 (m, 1.5H), 3.85-3.73 (m, 3.5H), 3.72-3.55 (m, 1.5H), 3.35-3.31 (m, 1H), 3.10-2.84 (m, 2.5H), 2.82-2.68 (m, 0.5H), 2.37-2.27 (m, 0.5H), 1.53-1.35 (m, 7H), 1.32-1.21 (m, 8H), 1.14-1.06 (m, 1.5H), 1.02-0.90 (m, 1.5H) ppm. MS: M/e 476 (M+1).sup.+.
[0661] Compound A108a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.27-7.06 (m, 4H), 6.39-5.41 (m, 1H), 5.25-4.90 (m, 1H), 4.42-4.30 (m, 2H), 3.76 (s, 3H), 3.65-3.50 (m, 1H), 3.34-3.25 (m 3H), 2.95-2.76 (m, 4H), 1.54-1.38 (m, 6H), 1.36-1.21 (m, 9H), 0.87 (d, J=6.0 Hz, 3H) ppm. MS: M/e 476 (M+1).sup.+.
[0662] Compound A108b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.30 (s, 1H), 7.20 (t, J=7.2 Hz, 1H), 7.13 (d, J=6.8 Hz, 1H), 7.09 (d, J=6.8 Hz, 1H), 6.15-5.65 (m, 1H), 5.27-4.95 (m, 1H), 4.45-4.30 (m, 2H), 3.76 (s, 3H), 3.66-3.54 (m, 1H), 3.50-3.39 (m, 1H), 3.34-3.25 (m 3H), 2.96-2.82 (m, 1H), 2.71-2.52 (m, 1H), 2.27-2.16 (m, 1H), 1.46 (t, J=7.2 Hz, 3H), 1.38-1.19 (m, 12H), 1.03 (d, J=5.6 Hz, 3H) ppm. MS: M/e 476 (M+1).sup.+.
Compound A110: 2-(6-((2S,5R)-4-(1-(3-(tert-butyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00427##
Step A: 1-(3-(tert-butyl)phenyl)ethan-1-ol
##STR00428##
[0663] To a mixture of 1-bromo-3-(tert-butyl)benzene (0.64 g, 3 mmol) in THE (10 mL) at 70 C. was added n-BuLi (2 mL, 1.5 mol/L in toluene). After 1 hour, a solution of acetaldehyde (264 mg, 6 mmol) in THE (2 mL) was added. The reaction was stirred for 1 hour. The reaction was quenched with saturated NH.sub.4Cl, extracted with EtOAc (60 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered and concentrated. The resulting residue was purified by flash column chromatography to give the titled compound (100 mg, 19%). .sup.1H NMR (400 MHz, DMSO-d6) 7.36 (s, 1H), 7.26-7.20 (m, 2H), 7.16-7.10 (m, 1H), 5.08 (d, J=4.0 Hz, 1H), 4.74-4.62 (m, 1H), 1.31 (d, J=6.4 Hz, 3H), 1.27 (s, 9H) ppm.
Step B: 2-(6-((2S,5R)-4-(1-(3-(tert-butyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00429##
[0664] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (100 mg, 0.3 mmol), 1-(3-(tert-butyl)phenyl)ethan-1-ol (100 mg, 0.56 mmol) and (cyanomethyl) trimethyl phosphonium iodide (218 mg, 0.9 mmol) in CH.sub.3CN (4 mL) was added DIPEA (387 mg, 3 mmol). The mixture was sealed in a bottle and heated at 100 C. for 16 hours. The mixture was cooled to room temperature, diluted with water, extracted with EtOAc (60 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by Prep-HPLC (Method A) to give the titled compound (65 mg, 44%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.49 (d, J=8.0 Hz, 1H), 7.36-7.19 (m, 2H), 7.15 (d, J=7.6 Hz, 1H), 6.20-5.60 (m, 1H), 5.25-4.95 (m, 1H), 4.37 (q, J=7.2 Hz 2H), 4.28-4.16 (m, 1.5H), 3.86-3.73 (m, 3.5H), 3.71-3.55 (m, 1.5H), 3.35-3.30 (m, 1H), 3.10-2.89 (m, 1.5H), 2.80-2.66 (m, 0.5H), 2.34-2.24 (m, 0.5H), 1.53-1.36 (m, 7.5H), 1.33 (s, 4.5H), 1.32 (s, 4.5H), 1.31-1.24 (m, 1.5H), 1.12-1.06 (m, 1.5H), 1.02-0.90 (m, 1.5H) ppm. MS: M/e 490 (M+1).sup.+.
Compound A112: 2-(6-((2S,5R)-4-(1-(3-(difluoromethyl)-4-fluorophenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00430##
Step A: 4-bromo-2-(difluoromethyl)-1-fluorobenzene
##STR00431##
[0665] A mixture of 5-bromo-2-fluorobenzaldehyde (1 g, 5 mmol), DAST (1.6 g, 10 mmol) in DCM (15 mL) was stirred at room temperature for 5 hours. The solution was diluted with DCM and washed with water. The organic layer was dried and purified by flash column chromatography to give the titled compound (1 g, 90%). MS: M/e 225 (M+1).sup.+
Step B: 1-(3-(difluoromethyl)-4-fluorophenyl)ethan-1-one
##STR00432##
[0666] A mixture of 4-bromo-2-(difluoromethyl)-1-fluorobenzene (1 g, 4.5 mmol), tributyl(1-ethoxyvinyl)stannane (2.4 g, 6.7 mmol) and Pd(PPh.sub.3).sub.2Cl.sub.2 (312 mg, 0.45 mmol) in toluene (15 mL) was stirred at 100 C. under N.sub.2 for 16 hours. The reaction mixture was diluted with HCl (4 mL, 3 M in ethyl acetate) in drops and the mixture was stirred at room temperature for 1 hour. The reaction was concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (600 mg, 71%). MS: M/e 189 (M+1).sup.+
Step C: 1-(3-(difluoromethyl)-4-fluorophenyl)ethan-1-ol
##STR00433##
[0667] To a solution of 1-(3-(difluoromethyl)-4-fluorophenyl)ethan-1-one (600 mg, 3.1 mmol) in MeOH (10 mL) was added NaBH.sub.4 (120 mg, 3.1 mmol) at room temperature and the resulting mixture was stirred at room temperature for 5 min. The reaction mixture was diluted with DCM and washed with water, dried over Na.sub.2SO.sub.4 and concentrated to give the titled compound (330 mg). MS: M/e 191 (M+1).sup.+.
Step D: 2-(6-((2S,5R)-4-(1-(3-(difluoromethyl)-4-fluorophenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00434##
[0668] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (100 mg, 0.3 mmol) in CH.sub.3CN (3 mL) and was added 1-(3-(difluoromethyl)-4-fluorophenyl)ethan-1-ol (116 mg, 0.6 mmol), (cyanomethyl)trimethylphosphonium iodide (219 mg, 0.9 mmol) and DIPEA (387 mg, 3 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure. The resulting residue was purified by flash column chromatography to give the titled compound (crude), which was further purified to give the titled compound (33 mg) by Prep-HPLC (Method A). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.73-7.52 (m, 2H), 7.25-7.14 (m, 1H), 7.13-6.82 (m, 1H), 6.28-4.98 (m, 2H), 4.37 (d, J=7.1 Hz, 2H), 4.23 (s, 2H), 3.76 (s, 3H), 3.74-3.36 (m, 2H), 3.11-2.75 (m, 2H), 2.75-2.06 (m, 1H), 1.49-1.42 (m, 4H), 1.39-1.21 (m, 5H), 1.11-0.85 (m, 3H) ppm. MS: M/e 502 (M+1).sup.+.
Compound A113: 2-(9-ethyl-6-((2S,5R)-4-(1-(4-fluoro-3-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00435##
Step A: 1-(4-fluoro-3-(trifluoromethyl)phenyl)ethan-1-ol
##STR00436##
[0669] To a solution of 1-(4-fluoro-3-(trifluoromethyl)phenyl)ethan-1-one (100 mg, 0.48 mmol) in MeOH (2 ml) at 0 C. was added NaBH.sub.4 (18 mg, 0.48 mmol) in some portions. After addition, the reaction was stirred at 0 C. for 5 min. The reaction was quenched by adding water and extracted with DCM, washed with brine, dried over Na.sub.2SO.sub.4, filtered, concentrated to give the titled compound (100 mg, crude). MS: M/e 191 (M+1-18).sup.+.
Step B: 2-(9-ethyl-6-((2S,5R)-4-(1-(4-fluoro-3-(trifluoromethyl)phenyl)ethyl)-2,5-dimethyl piperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00437##
[0670] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (100 mg, 0.3 mol), 1-(4-fluoro-3-(trifluoromethyl)phenyl)ethan-1-ol (94 mg, 0.45 mol), (cyanomethyl)trimethylphosphonium iodide (218 mg, 0.9 mol) and DIPEA (387 mg, 3 mol) in CH.sub.3CN (2 ml). Then the mixture solution was stirred at 105 C. for 16 hours. The reaction mixture was concentrated under reduced pressure and purified by flash column chromatography (DCM:MeOH) to give the titled compound (39.1 mg). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.80-7.61 (m, 2H), 7.35-7.22 (m, 1H), 6.50-4.99 (m, 2H), 4.37 (q, J=7.0 Hz, 2H), 4.22 (s, 1H), 3.76 (s, 3H), 3.74-3.65 (m, 1H), 3.60-3.48 (m, 1H), 3.30-3.21 (m, 1H), 2.99-2.73 (m, 2H), 2.70-2.03 (m, 1H), 1.46 (t, J=7.0 Hz, 4H), 1.37-1.21 (m, 5H), 1.25-0.78 (m, 3H) ppm. MS: M/e 520 (M+1).sup.+.
Compound A115: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(4-methyl-3-(trifluoromethyl)phenyl)ethyl) piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00438##
Step A: 1-(4-methyl-3-(trifluoromethyl)phenyl)ethan-1-one
##STR00439##
[0671] A solution of 4-bromo-1-methyl-2-(trifluoromethyl)benzene (500 mg, 2.09 mmol), Pd(PPh.sub.3).sub.2Cl.sub.2 (73 mg, 0.10 mmol) and tributyl(1-ethoxyvinyl)stannane (1.13 g, 3.13 mmol) in Tol. (10 ml) was stirred at 100 C. overnight. The mixture was cooled to RT and HCl (4M in dioxane, 2 ml) was added, stirred for 30 min. The mixture was diluted with EtOAc (20 ml), washed with brine (10 ml), dried and concentrated. The residue was purified by flash column chromatography with 0%-5% EtOAc in PE to give the titled compound (200 mg, 47%). .sup.1H NMR (400 MHz, CDCl.sub.3) 8.20 (s, 1H), 8.01 (d, J=7.6 Hz, 1H), 7.40 (d, J=7.8 Hz, 1H), 2.62 (s, 3H), 2.56 (s, 3H) ppm.
Step B: 1-(4-methyl-3-(trifluoromethyl)phenyl)ethan-1-ol
##STR00440##
[0672] A solution of 1-(4-methyl-3-(trifluoromethyl)phenyl)ethan-1-one (200 mg, 0.99 mmol) and NaBH.sub.4 (38 mg, 1.00 mmol) in MeOH (4 ml) was stirred at RT for 30 min. The solution was diluted with EtOAc (15 ml), washed with brine (10 ml), dried and evaporated to dryness to give the titled compound (200 mg, 100%), which was used directly for the next step. MS: M/e 205 (M+1).sup.+.
Step C: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(4-methyl-3-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00441##
[0673] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (108 mg, 0.33 mmol), 1-(4-methyl-3-(trifluoromethyl)phenyl)ethan-1-ol (200 mg, 0.98 mmol), (cyanomethyl)trimethylphosphonium iodide (239 mg, 0.98 mmol) and DIPEA (423 mg, 3.28 mmol) in MeCN (2 ml) was stirred at 100 C. overnight. The reaction was diluted with EtOAc (10 ml) and washed with brine (10 ml). The organic layer was concentrated under reduced pressure. The resulting residue was purified by flash column chromatography with 0-5% MeOH in DCM and then Prep-HPLC (Method A) to give the titled compound (44 mg). .sup.1H NMR (400 MHz, DMSO-d6) 7.68 (d, J=19.1 Hz, 1H), 7.56 (dd, J=22.6, 8.1 Hz, 1H), 7.44-7.36 (m, 1H), 6.15-5.35 (m, 1H), 5.20-4.55 (m, 1H), 4.41 (s, 2H), 4.26 (d, J=5.8 Hz, 2H), 3.68 (q, J=6.4 Hz, 1H), 3.61 (s, 3H), 3.52 (q, J=5.5 Hz, 1H), 3.32 (s, 1H), 2.75 (s, 1.5H), 2.43 (s, 3H), 2.05 (d, J=12.3 Hz, 0.5H), 1.34 (t, J=6.7 Hz, 4.5H), 1.25 (t, J=6.2 Hz, 3H), 1.16 (s, 1.5H), 0.96 (s, 1.5H), 0.83 (d, J=5.7 Hz, 1.5H) ppm. MS: M/e 516 (M+1).sup.+.
Compound A117: 2-(9-ethyl-6-((2S,5R)-4-(1-(6-isopropoxypyridin-2-yl)ethyl)-2,5-dimethyl piperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00442##
Step A: 1-(6-isopropoxyyridin-2-yl)ethan-1-ol
##STR00443##
[0674] To a solution of 1-(6-isopropoxypyridin-2-yl)ethan-1-one (150 mg, 0.84 mmol) in MeOH (15 mL) was added NaBH.sub.4 (32 mg, 0.84 mmol) at room temperature and the resulting mixture was stirred at room temperature for 15 mins. The reaction mixture was diluted with DCM (200 mL). The organic layer was washed with water, dried over Na.sub.2SO.sub.4 and concentrated to give the titled compound (100 mg, 66%). MS: M/e 182 (M+1).sup.+.
Step B: 2-(9-ethyl-6-((2S,5R)-4-(1-(6-isopropoxypyridin-2-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00444##
[0675] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.15 mmol) in CH.sub.3CN (3 mL) and was added 1-(6-isopropoxypyridin-2-yl)ethan-1-ol (28 mg, 0.15 mmol), (cyanomethyl)trimethylphosphonium iodide (110 mg, 0.46 mmol) and DIPEA (98 mg, 0.76 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure to dryness. The resulting residue was purified and separated into Compound A117a (26 mg) and Compound A117b (22 mg) by Prep-HPLC (Method A).
[0676] Compound A117a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.62 (t, J=7.9 Hz, 1H), 7.07 (d, J=7.4 Hz, 1H), 6.59 (d, J=8.2 Hz, 1H), 5.95 (s, 1H), 5.30-5.22 (m, 1H), 4.86 (s, 2H), 4.85-4.44 (m, 1H), 4.37 (q, J=7.2 Hz, 2H), 3.76 (s, 3H), 3.74-3.36 (m, 3H), 2.87 (s, 1H), 2.38 (d, J=11.8 Hz, 1H), 1.46 (t, J=7.2 Hz, 3H), 1.38 (d, J=6.5 Hz, 3H), 1.35-1.27 (m, 9H), 1.07 (d, J=6.1 Hz, 3H) ppm. MS: M/e 493 (M+1).sup.+
[0677] Compound A117b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.62 (t, J=7.7 Hz, 1H), 7.02 (d, J=7.3 Hz, 1H), 6.59 (d, J=8.2 Hz, 1H), 5.87-5.32 (m, 1H), 5.30-5.24 (m, 1H), 4.86 (s, 2H), 4.85-4.48 (m, 1H), 4.37 (q, J=7.0 Hz, 2H), 3.79 (s, 1H), 3.76 (s, 3H), 3.43 (s, 1H), 3.03 (s, 2H), 2.83 (d, J=10.9 Hz, 1H), 1.46 (t, J=7.2 Hz, 3H), 1.39 (dd, J=10.1, 6.9 Hz, 6H), 1.33 (d, J=6.0 Hz, 6H), 1.01 (d, J=6.4 Hz, 3H) ppm. MS: M/e 493 (M+1).sup.+
Compound A123: 2-(9-ethyl-6-((2S,5R)-4-(1-(6-isopropylpyridin-2-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00445##
Step A: 1-(6-isoproyloyridin-2-yl)ethan-1-ol
##STR00446##
[0678] To a solution of 1-(6-isopropylpyridin-2-yl)ethan-1-one (150 mg, 0.92 mmol) in MeOH (15 mL) was added NaBH.sub.4 (70 mg, 1.84 mmol) at room temperature and the resulting mixture was stirred at room temperature for 15 mins. The reaction mixture was diluted with DCM (200 mL). The organic layer was washed with water, dried over Na.sub.2SO.sub.4 and concentrated to give the titled compound (120 mg, 79%). MS: M/e 166 (M+1).sup.+.
Step B: 2-(9-ethyl-6-((2S,5R)-4-(1-(6-isopropylpyridin-2-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00447##
[0679] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.15 mmol) in CH.sub.3CN (3 mL) and was added 1-(6-isopropylpyridin-2-yl)ethan-1-ol (25 mg, 0.15 mmol), (cyanomethyl)trimethylphosphonium iodide (110 mg, 0.46 mmol) and DIPEA (98 mg, 0.76 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure to dryness. The resulting residue was purified by Prep-HPLC (Method A) to give the titled compound (28 mg, 38%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.77 (d, J=7.5 Hz, 1H), 7.41 (t, J=9.3 Hz, 1H), 7.31-7.20 (m, 1H), 6.45-4.89 (m, 2H), 4.37 (d, J=6.9 Hz, 2H), 4.25-4.02 (m, 2H), 3.83 (s, 1H), 3.77 (s, 3H), 3.62-3.31 (m, 1.5H), 3.16-2.99 (m, 3H), 2.50-2.20 (m, 0.5H), 1.51-1.44 (m, 6H), 1.43-1.28 (m, 9H), 1.25-1.03 (m, 3H) ppm. MS: M/e 477 (M+1).sup.+
Compound A124: 2-(9-ethyl-6-((2S,5R)-4-(1-(2-isopropoxypyridin-4-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00448##
Step A: 1-(2-isopropoxyyridin-4-yl)ethan-1-ol
##STR00449##
[0680] To a solution of 1-(2-isopropoxypyridin-4-yl)ethan-1-one (150 mg, 0.84 mmol) in MeOH (15 mL) was added NaBH.sub.4 (32 mg, 0.84 mmol) at room temperature and the resulting mixture was stirred at room temperature for 15 mins. The reaction mixture was diluted with DCM (200 mL). The organic layer was washed with water, dried over Na.sub.2SO.sub.4 and concentrated to give the titled compound (110 mg, 73%). MS: M/e 182 (M+1).sup.+.
Step B: 2-(9-ethyl-6-((2S,5R)-4-(1-(2-isopropoxypyridin-4-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00450##
[0681] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.15 mmol) in CH.sub.3CN (3 mL) and was added 1-(2-isopropoxypyridin-4-yl)ethan-1-ol (28 mg, 0.15 mmol), (cyanomethyl)trimethylphosphonium iodide (110 mg, 0.46 mmol) and DIPEA (98 mg, 0.76 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure to dryness. The resulting residue was further purified to give the titled compound (44 mg, 59%). .sup.1H NMR (400 MHz, CD.sub.3OD) 8.03 (dd, J=10.4, 5.3 Hz, 1H), 6.99 (d, J=4.8 Hz, 1H), 6.76 (s, 1H), 6.40-5.25 (m, 2H), 5.22-5.14 (m, 1H), 4.41-4.33 (m, 2H), 4.23 (s, 2H), 3.76 (s, 3H), 3.64-3.43 (m, 2H), 3.30-3.27 (m, 0.5H), 2.92-2.67 (m, 2H), 2.30-2.10 (m, 0.5H), 1.49-1.31 (m, 12H), 1.29 (d, J=6.2 Hz, 3H), 1.07-0.89 (m, 3H) ppm. MS: M/e 493 (M+1).sup.+
Compound A125: 2-(6-((2S,5R)-4-(1-(3-cyclopropylphenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00451##
Step A: 1-(3-cycloproylphenyl)ethan-1-ol
##STR00452##
[0682] To a stirred solution of 1-bromo-3-cyclopropylbenzene (394 mg, 2 mmol) in THE (8 mL) was added dropwise n-BuLi (1.6 M, 1.25 mL, 2 mmol) at 78 C. After stirred for 30 min, a solution of acetaldehyde (176 mg, 4 mmol) in THE (2 mL) was added dropwise. After the addition, the reaction mixture was stirred for an hour. The reaction was quenched with aq.NH.sub.4Cl, extracted with EtOAc (10 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated and purified by flash column chromatography to give the titled compound (280 mg, 86%). .sup.1H NMR (400 MHz, DMSO-d6) 7.16 (t, J=7.5 Hz, 1H), 7.10-7.03 (m, 2H), 6.89 (d, J=7.6 Hz, 1H), 5.07 (d, J=4.0 Hz, 1H), 4.71-4.60 (m, 1H), 1.94-1.81 (m, 1H), 1.29 (d, J=6.4 Hz, 3H), 0.98-0.89 (m, 2H), 0.69-0.60 (m, 2H) ppm.
Step B: 2-(6-((2S,5R)-4-(1-(3-cyclopropylphenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00453##
[0683] A mixture of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (100 mg, 0.3 mmol), 1-(3-cyclopropylphenyl)ethan-1-ol (84.1 mg, 0.45 mmol), (cyanomethyl)trimethylphosphonium iodide (218 mg, 0.9 mmol) and DIPEA (387 mg, 3 mmol) in CH.sub.3CN (4 mL) was stirred overnight at 100 C. in a sealed tube. The reaction mixture was diluted with EtOAc (20 mL), washed with H.sub.2O, brine, dried over Na.sub.2SO.sub.4, concentrated to give the residue, which was purified by Prep-HPLC (Method A) to give the titled compound (55 mg). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.28-7.12 (m, 3H), 6.99 (d, J=7.2 Hz, 1H), 6.32-4.96 (m, 2H), 4.44-4.32 (m, 2H), 4.23 (s, 1H), 3.86-3.79 (m, 0.5H), 3.76 (s, 3H), 3.73-3.20 (m, 3H), 3.12-2.65 (m, 2H), 2.34 (d, J=12.4 Hz, 0.5H), 1.96-1.85 (m, 1H), 1.50-0.62 (m, 16H) ppm. MS: M/e 474 (M+1).sup.+.
Compound A126: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-(1-methylcyclopropyl)phenyl)ethyl) piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00454##
Step A: 1-(3-(prop-1-en-2-yl)phenyl)ethan-1-one
##STR00455##
[0684] To a solution of 1-(3-bromophenyl)ethan-1-one (2 g, 10 mmol) and potassium isopropenyltrifluoroborate (1.5 g, 10 mmol) in IPA/H.sub.2O (2/1, 20 mL) was added TEA (1.5 g, 15 mmol) and Pd(dppf).sub.2Cl.sub.2 (292 mg, 0.4 mmol). The reaction mixture was protected by N.sub.2 atmosphere and stirred at 80 C. overnight. The mixture was cooled down to RT, diluted with water, extracted with EtOAc (80 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography to give the titled compound (1.3 g, 81%). .sup.1H NMR (400 MHz, CDCl.sub.3) 8.05 (s, 1H), 7.86 (d, J=7.2 Hz, 1H), 7.67 (d, J=7.2 Hz, 1H), 7.43 (t, J=7.6 Hz, 1H), 5.43 (s, 1H), 5.17 (s, 1H), 2.63 (s, 3H), 2.19 (s, 3H) ppm.
Step B: 1-(3-(1-methylcyclopropyl)phenyl)ethan-1-one
##STR00456##
[0685] To a solution of Et.sub.2Zn (4 mL, 1 mol/L in hexane) in DCM (4 mL) was added a solution of TFA (456 mg, 4 mmol) in DCM (4 mL) drop wise at ice-bath. After 20 min, a solution of CH.sub.2I.sub.2 (1.07 g, 4 mmol) in DCM (4 mL) was added drop wise. Then 20 min later, a solution of 1-(3-(prop-1-en-2-yl)phenyl)ethan-1-one (320 mg, 2 mmol) in DCM (4 mL) was added drop wise. Then the reaction mixture was warmed to room temperature overnight. The reaction mixture was diluted with H.sub.2O, extracted with DCM (60 mL), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography to give the titled compound (130 mg, 37%). .sup.1H NMR (400 MHz, CDCl.sub.3) 7.86 (s, 1H), 7.75 (d, J=7.6 Hz, 1H), 7.46 (d, J=7.2 Hz, 1H), 7.37 (t, J=7.2 Hz, 1H), 2.60 (s, 3H), 1.43 (s, 3H), 0.94-0.86 (m, 2H), 0.81-0.74 (m, 2H) ppm.
Step C: 1-(3-(1-methylcyclopropyl)phenyl)ethan-1-ol
##STR00457##
[0686] To a solution of 1-(3-(1-methylcyclopropyl)phenyl)ethan-1-one (130 mg, 0.75 mmol) in MeOH (5 mL) was added NaBH.sub.4 (28 mg, 0.75 mmol) slowly at 0 C. The resulting mixture was stirred at room temperature for 0.5 hour. The reaction mixture was quenched with H.sub.2O (10 mL) and extracted with EtOAc (80 mL), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography to give the product (125 mg, 96% yield). MS: M/e 177 (M+1).sup.+.
Step D: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-(1-methylcyclopropyl)phenyl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00458##
[0687] To a solution of 1-(3-(1-methylcyclopropyl)phenyl)ethan-1-ol (120 mg, 0.68 mmol), 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (100 mg, 0.3 mmol) and (cyanomethyl) trimethyl phosphonium iodide (218 mg, 0.9 mmol) in CH.sub.3CN (4 mL) was added DIPEA (387 mg, 3 mmol). The mixture was sealed in a bottle and heated at 100 C. for 16 hours. The mixture was cooled to room temperature, diluted with water, extracted with EtOAc (80 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by Prep-HPLC (Method A) to give the titled compound (62 mg, 42%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.36-7.05 (m, 4H), 6.19-5.65 (m, 1H), 5.31-4.95 (m, 1H), 4.43-4.30 (m, 2H), 4.28-4.16 (m, 1.5H), 3.76 (s, 3H), 3.70-3.40 (m, 2H), 3.35-3.31 (m, 1H), 2.97-2.85 (m, 1.5H), 2.74-2.51 (m, 0.5H), 2.30-2.16 (m, 0.5H), 1.54-1.15 (m, 12H), 1.12-0.51 (m, 7H) ppm. MS: M/e 488 (M+1).sup.+.
Compound A129: 2-(6-((2S,5R)-4-(1-(2-(tert-butyl)pyridin-4-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00459##
Step A: 1-(2-(tert-butyl)pyridin-4-yl)ethan-1-one
##STR00460##
[0688] A mixture of 4-bromo-2-(tert-butyl)pyridine (214 mg, 1 mmol), tributyl(1-ethoxyvinyl)stannane (541 mg, 1.5 mmol) and Pd(PPh.sub.3).sub.2Cl.sub.2 (70 mg, 0.1 mmol) in toluene (5 mL) was stirred at 100 C. under N.sub.2 overnight. To the resulting solution was added TFA (1 mL) in drops and the mixture was stirred at room temperature for 30 minutes. The mixture was diluted with EtOAc (20 mL), treated with saturated Na.sub.2CO.sub.3 aq. to pH 8, washed with brine (20 mL3), dried and concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (100 mg, 56%). MS: M/e 178 (M+1).sup.+.
Step B: 1-(2-(tert-butyl)pyridin-4-yl)ethan-1-ol
##STR00461##
[0689] To a solution of 1-(2-(tert-butyl)pyridin-4-yl)ethan-1-one (100 mg, 0.56 mmol) in EtOH (2 mL) was added NaBH.sub.4 (17 mg, 0.45 mmol) at room temperature and the mixture was stirred at room temperature for 1 hour. The mixture was treated with water, extracted with DCM. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated to dryness. The resulting residue (80 mg, crude) was used in the next step directly. MS: M/e 180 (M+1).sup.+.
Step C: 2-(6-((2S,5R)-4-(1-(2-(tert-butyl)pyridin-4-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00462##
[0690] A mixture of 1-(2-(tert-butyl)pyridin-4-yl)ethan-1-ol (80 mg, 0.45 mmol), 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (100 mg, 0.3 mmol), (cyanomethyl)trimethylphosphonium iodide (145 mg, 0.6 mmol) and DIPEA (155 mg, 1.2 mmol) in MeCN (3 mL) was stirred at 100 C. overnight. The reaction mixture was diluted with EtOAc, washed with brine, dried over Na.sub.2SO.sub.4 and concentrated to dryness. The resulting residue was purified by Prep-TLC (DCM/MeOH=13/1) to give the titled compound (19 mg, 13%). .sup.1H NMR (400 MHz, CD.sub.3OD) 8.40 (d, J=8.5 Hz, 1H), 7.57 (d, J=18.7 Hz, 1H), 7.28 (d, J=21.4 Hz, 1H), 5.93 (s, 2H), 4.37 (d, J=6.9 Hz, 2H), 4.23 (s, 2H), 3.89-3.48 (m, 5H), 3.05-2.60 (m, 2H), 2.13 (d, J=11.7 Hz, 1H), 1.58-1.21 (m, 18H), 1.11-0.86 (m, 3H) ppm. MS: M/e 491 (M+1).sup.+.
Compound A134: 2-(3-(1-((2R,5S)-4-(8-(cyanomethyl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazin-1-yl)ethyl)phenyl)-2-methylpropanenitrile
##STR00463##
Step A: 2-(3-acetylphenyl)-2-methylpropanenitrile
##STR00464##
[0691] To a solution of 2-(3-bromophenyl)-2-methylpropanenitrile (336 mg, 1.5 mmol) in toluene (6 mL) was added tributyl(1-ethoxyvinyl)stannane (812 mg, 2.25 mmol) and Pd(PPh.sub.3).sub.2Cl.sub.2 (53 mg, 0.075 mmol). The reaction mixture was protected by N.sub.2 atmosphere and stirred at 100 C. overnight. The mixture was cooled down to RT, added 2N HCl (4 ml) and stirred at room temperature for 30 mins. The mixture was concentrated in vacuo. The residue was added H.sub.2O, extracted with EtOAc (80 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography to give the titled compound (248 mg, 85%). .sup.1H NMR (400 MHz, CDCl.sub.3) 8.05 (s, 1H), 7.91 (d, J=8.0 Hz, 1H), 7.74 (d, J=7.6 Hz, 1H), 7.52 (t, J=8.0 Hz, 1H), 2.64 (s, 3H), 1.77 (s, 6H) ppm.
Step B: 2-(3-(1-hydroxyethyl)phenyl)-2-methylpropanenitrile
##STR00465##
[0692] To a solution of 2-(3-acetylphenyl)-2-methylpropanenitrile (93 mg, 0.5 mmol) in MeOH (5 mL) was added NaBH.sub.4 (19 mg, 0.5 mmol) slowly at 0 C. The resulting mixture was stirred at room temperature for 0.5 hour. The reaction mixture was quenched with H.sub.2O (10 mL) and extracted with EtOAc (80 mL), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography to give the titled compound (92 mg, 96% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.50 (s, 1H), 7.41-7.34 (m, 2H), 7.32-7.27 (m, 1H), 5.24 (s, 1H), 4.80-4.70 (m, 1H), 1.68 (s, 6H), 1.32 (d, J=6.4 Hz, 3H) ppm.
Step C: 2-(3-(1-((2R,5S)-4-(8-(cyanomethyl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazin-1-yl)ethyl)phenyl)-2-methylpropanenitrile
##STR00466##
[0693] To a solution of 2-(3-(1-hydroxyethyl)phenyl)-2-methylpropanenitrile (90 mg, 0.47 mmol), 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (100 mg, 0.3 mmol) and (cyanomethyl) trimethyl phosphonium iodide (218 mg, 0.9 mmol) in CH.sub.3CN (4 mL) was added DIPEA (387 mg, 3 mmol). The mixture was sealed in a bottle and heated at 100 C. for 16 hours. The mixture was cooled to room temperature, diluted with water, extracted with EtOAc (80 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by Prep-HPLC (Method A) to give the titled compound (70 mg, 46%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.67-7.60 (m, 1H), 7.46-7.30 (m, 3H), 6.25-5.62 (m, 1H), 5.31-4.95 (m, 1H), 4.43-4.30 (m, 2H), 4.28-4.16 (m, 2H), 3.85-3.50 (m, 5H), 3.35-3.31 (m, 1H), 3.00-2.85 (m, 1H), 2.75-2.60 (m, 0.5H), 2.26-2.16 (m, 0.5H), 1.74 (s, 6H), 1.54-1.23 (m, 9H), 1.07 (d, J=5.6 Hz, 1.5H), 0.93 (d, J=5.6 Hz, 1.5H) ppm. MS: M/e 501 (M+1).sup.+.
Compound A135, A136: 2-(9-ethyl-6-((2S,5R)-4-(1-(4-fluoro-3-(prop-1-en-2-yl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile & 2-(9-ethyl-6-((2S,5R)-4-(1-(4-fluoro-3-(1-methylcyclopropyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00467##
Step A: 1-(4-fluoro-3-(prop-1-en-2-yl)phenyl)ethan-1-one
##STR00468##
[0694] To a solution of 1-(3-bromo-4-fluorophenyl)ethan-1-one (2.17 g, 10 mmol) and K.sub.2CO.sub.3 (2.76 g, 20 mmol) in dioxane/H.sub.2O (10/1, 20 mL) was added 4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane (2 g, 12 mmol) and Pd(dppf).sub.2Cl.sub.2.Math.CH.sub.2Cl.sub.2 (818 mg, 1 mmol). The reaction mixture was protected by N.sub.2 atmosphere and stirred at 100 C. overnight. The mixture was cooled down to RT, diluted with water, extracted with EtOAc (80 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography to give the titled compound (1.6 g, 89%). .sup.1H NMR (400 MHz, CDCl.sub.3) 8.00-7.81 (m, 2H), 7.15-7.105 (m, 1H), 5.34-5.25 (m, 2H), 2.60 (s, 3H), 2.16 (s, 3H) ppm.
Step B: 1-(4-fluoro-3-(1-methylcyclopropyl)phenyl)ethan-1-one
##STR00469##
[0695] To a solution of Et.sub.2Zn (3 mL, 1 mol/L in hexane) in DCM (8 mL) was added a solution of TFA (684 mg, 6 mmol) in DCM (4 mL) drop wise at ice-bath. After 20 min, a solution of CH.sub.2I.sub.2 (1.57 g, 6 mmol) in DCM (4 mL) was added drop wise. Then 20 min later, a solution of 1-(4-fluoro-3-(prop-1-en-2-yl)phenyl)ethan-1-one (534 mg, 3 mmol) in DCM (4 mL) was added drop wise. Then the reaction mixture was warmed to room temperature overnight. The reaction mixture was diluted with H.sub.2O, extracted with DCM (60 mL), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography to give the titled compound (440 mg, a mixture of 1-(4-fluoro-3-(prop-1-en-2-yl)phenyl)ethan-1-one and 1-(4-fluoro-3-(1-methylcyclopropyl)phenyl)ethan-1-one, 3/1). .sup.1H NMR (400 MHz, CDCl.sub.3) 8.01-7.75 (m, 2H), 7.10-7.00 (m, 1H), 2.60 (s, 3H), 1.36 (s, 3H), 0.91-0.62 (m, 4H) ppm.
Step C: 1-(4-fluoro-3-(prop-1-en-2-yl)phenyl)ethan-1-ol and 1-(4-fluoro-3-(1-methylcyclopropyl) phenyl)ethan-1-ol
##STR00470##
[0696] To a solution of 1-(4-fluoro-3-(prop-1-en-2-yl)phenyl)ethan-1-one and 1-(4-fluoro-3-(1-methylcyclopropyl)phenyl)ethan-1-one (440 mg, 2.29 mmol) in MeOH (6 mL) was added NaBH.sub.4 (87 mg, 2.29 mmol) slowly at 0 C. The resulting mixture was stirred at room temperature for 0.5 hour. The reaction mixture was quenched with H.sub.2O (10 mL) and extracted with EtOAc (80 mL), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography to give the desired mixture (260 mg, 59% yield). MS: M/e 181 and 195 (M+1).sup.+.
Step D: 2-(9-ethyl-6-((2S,5R)-4-(1-(4-fluoro-3-(prop-1-en-2-yl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile and 2-(9-ethyl-6-((2S,5R)-4-(1-(4-fluoro-3-(1-methylcyclopropyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00471##
[0697] To a solution of 1-(4-fluoro-3-(prop-1-en-2-yl)phenyl)ethan-1-ol and 1-(4-fluoro-3-(1-methylcyclopropyl)phenyl)ethan-1-ol (260 mg, 1.36 mmol), 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (320 mg, 1 mmol) and (cyanomethyl) trimethyl phosphonium iodide (730 mg, 3 mmol) in CH.sub.3CN (5 mL) was added DIPEA (1.29 g, 10 mmol). The mixture was sealed in a bottle and heated at 100 C. for 16 hours. The mixture was cooled to room temperature, diluted with water, extracted with EtOAc (80 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified and separated into Compound A135 (40 mg) and A136 (80 mg) by Prep-HPLC (Method A).
[0698] Compound A135: .sup.1H NMR (400 MHz, CD.sub.3OD) 7.43-7.24 (m, 2H), 7.09-6.96 (m, 1H), 6.19-5.65 (m, 1H), 5.31-4.95 (m, 3H), 4.43-4.30 (m, 2H), 4.28-4.16 (m, 1.5H), 3.76 (s, 3H), 3.79-3.42 (m, 2H), 3.35-3.31 (m, 1H), 3.00-2.85 (m, 1.5H), 2.75-2.59 (m, 0.5H), 2.28-2.19 (m, 0.5H), 2.16-2.10 (m, 3H), 1.55-1.21 (m, 9H), 1.05 (d, J=6.0 Hz, 1.5H), 0.93 (d, J=6.4 Hz, 1.5H) ppm. MS: M/e 492 (M+1).sup.+.
[0699] Compound A136: .sup.1H NMR (400 MHz, CD.sub.3OD) 7.44-7.13 (m, 2H), 7.06-6.92 (m, 1H), 6.19-5.65 (m, 1H), 5.31-4.95 (m, 1H), 4.43-4.30 (m, 2H), 4.28-4.16 (m, 1.5H), 3.76 (s, 3H), 3.79-3.40 (m, 2H), 3.35-3.31 (m, 1H), 3.00-2.85 (m, 1.5H), 2.75-2.61 (m, 0.5H), 2.28-2.17 (m, 0.5H), 1.54-1.19 (m, 12H), 1.06 (d, J=6.8 Hz, 1.5H), 0.94 (d, J=6.4 Hz, 1.5H), 0.84-0.65 (m, 4H) ppm. MS: M/e 506 (M+1).sup.+.
Compound A137: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-methyl-4-(trifluoromethyl)phenyl) ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00472##
Step A: 1-(3-methyl-4-(trifluoromethyl)phenyl)ethan-1-ol
##STR00473##
[0700] To a 70 C. solution of 4-bromo-2-methyl-1-(trifluoromethyl)benzene (1.0 g, 4.2 mmol) in THE (10 mL) was added n-BuLi (1.6 M, 3.2 mL, 5.1 mmol) in drops under N.sub.2. After stirring for 20 minutes at 70 C., a solution of acetaldehyde (440 mg, 10 mmol) in THE (2 mL) was added. Then the mixture was allowed warm to room temperature and stirred for 16 hours. Aqueous solution of NH.sub.4Cl (20 mL) was added and the mixture was extracted with EtOAc (10 mL3), the organics were combined and washed with brine (10 mL2), dried over Na.sub.2SO.sub.4, filtered and concentrated to dryness. The resulting residue was purified by column chromatography to give the titled compound (678 mg, 79%). .sup.1H NMR (400 MHz, DMSO-d6) 7.60 (d, J=8.0 Hz, 1H), 7.39 (s, 1H), 7.34 (d, J=8.0 Hz, 1H), 5.32 (d, J=4.4 Hz, 1H), 4.85-4.64 (m, 1H), 2.43 (s, 3H), 1.32 (d, J=6.4 Hz, 3H) ppm.
Step B: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(3-methyl-4-(trifluoromethyl)phenyl) ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl) acetonitrile
##STR00474##
[0701] To a mixture of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.15 mmol), 1-(3-methyl-4-(trifluoromethyl)phenyl)ethan-1-ol (65 mg, 0.32 mmol) and (cyanomethyl)trimethylphosphonium iodide (120 mg, 0.50 mmol) in CH.sub.3CN (1 mL) was added DIPEA (155 mg, 1.2 mmol). The mixture was stirred at 100 C. in a sealed tube for 16 hours. The mixture was cooled and diluted with EtOAc (20 mL), washed with brine (10 mL3), dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography and Prep-HPLC (Method A) to give the titled compound (36 mg, 46%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.64-7.52 (m, 1H), 7.48-7.30 (m, 2H), 6.29-4.87 (m, 2H), 4.68-4.04 (m, 4H), 3.76 (s, 3H), 3.73-3.44 (m, 2H), 3.08-2.03 (m, 6H), 1.54-1.17 (m, 9H), 1.11-0.84 (m, 3H) ppm. MS: M/e 516 (M+1).sup.+.
Compound A144: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-phenylpyridin-4-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00475##
Step A: 4-bromo-2-phenylpyridine
##STR00476##
[0702] To a solution of 2,4-dibromopyridine (1.18 g, 5 mol) in MeCN (10 mL) was added 4,4,5,5-tetramethyl-2-phenyl-1,3,2-dioxaborolane (1 g, 5 mol), Pd(OAc).sub.2 (56 mg, 0.25 mmol), PPh.sub.3 (262 mg, 1 mmol) and KOH (560 mg, 10 mmol). The reaction mixture was protected by N.sub.2 atmosphere and stirred at 70 C. for 16 hrs. The mixture was cooled down to RT and added H.sub.2O (20 ml), The mixture was extracted with EtOAc (20 mL3), then concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (800 mg, 69%). MS: M/e 234 (M+1).sup.+.
Step B: 1-(2-phenylpyridin-4-yl)ethan-1-one
##STR00477##
[0703] To a solution of 4-bromo-2-phenylpyridine (232 mg, 1 mol) in toluene (10 mL) was added Pd(PPh.sub.3).sub.2Cl.sub.2 (70 mg, 0.1 mol) and tributyl(1-ethoxyvinyl)stannane (541 mg, 1.5 mmol). The reaction mixture was protected by N.sub.2 atmosphere and stirred at 90 C. for 16 hrs. The mixture was added 4N HCl in dioxane (0.5 ml) and stirred at RT for 15 mins. The mixture was added saturated NaHCO.sub.3 aqueous solution (10 ml) and extracted with EtOAc (20 mL3). The organic phase was dried with Na.sub.2SO.sub.4 and concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (40 mg, 20%). MS: M/e 198 (M+1).sup.+.
Step C: 1-(2-phenylpyridin-4-yl)ethan-1-ol
##STR00478##
[0704] To a solution of 1-(2-phenylpyridin-4-yl)ethan-1-one (40 mg, 0.2 mol) in MeOH was added NaBH.sub.4 (38 mg, 1 mmol). The reaction mixture was stirred at RT for 15 mins. The mixture was added H.sub.2O (20 ml) and extracted with DCM (20 mL3). The organic phase was dried with Na.sub.2SO.sub.4 and concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (40 mg, 100%). MS: M/e 200 (M+1).sup.+.
Step D: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-phenylpyridin-4-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00479##
[0705] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl) acetonitrile (30 mg, 0.1 mmol), 1-(2-phenylpyridin-4-yl)ethan-1-ol (40 mg, 0.2 mmol) and (cyanomethyl)trimethylphosphonium iodide (120 mg, 0.5 mmol) in CH.sub.3CN (10 mL) was added DIPEA (258 mg, 2 mmol). The reaction mixture was sealed in a bottle and heated at 105 C. for 16 hours, and then cooled to room temperature, diluted with water, extracted with EtOAc (20 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by Prep-HPLC (Method A) to give the titled compound (5 mg, 9%). .sup.1H NMR (400 MHz, CD.sub.3OD) 8.55 (dd, J=11.8, 5.1 Hz, 1H), 7.92 (d, J=8.0 Hz, 3H), 7.48 (ddd, J=21.5, 11.8, 6.2 Hz, 4H), 5.95 (m, 1H), 5.09 (m, 1H), 4.36 (m, 2H), 4.22 (s, 2H), 3.78 (m, 4H), 3.63 (d, J=6.2 Hz, 1H), 2.89 (m, 2H), 2.75 (s, 0.5H), 2.23 (d, J=12.2 Hz, 0.5H), 1.46 (t, J=7.2 Hz, 4H), 1.38 (m, 5H), 1.08 (d, J=6.4 Hz, 1.5H), 0.97 (d, J=6.4 Hz, 1.5H) ppm. MS: M/e 507 (M+1).sup.+.
Compound A149: 2-(6-((2S,5R)-2,5-diethyl-4-(1-(4-(trifluoromethyl)phenyl) ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00480##
Step A: tert-butyl (2R,5S)-2,5-diethyl-4-(1-methyl-6-(methylamino)-5-nitro-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxylate
##STR00481##
[0706] To a stirred mixture of 4-hydroxy-1-methyl-6-(methylamino)-5-nitropyrimidin-2 (1H)-one (600 mg, 3 mmol), tert-butyl (2R,5S)-2,5-diethylpiperazine-1-carboxylate (871.2 mg, 3.6 mmol) and BOP (2 g, 4.5 mmol) in DMF (10 mL) was added DBU (912 mg, 6 mmol). After the addition, the reaction mixture was stirred at 70 C. overnight. The reaction mixture was concentrated to give the residue, which was treated with H.sub.2O (50 mL) and extracted with EtOAc (60 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (1.0 g, 82%). MS: M/e 425 (M+1).sup.+.
Step B: tert-butyl (2R,5S)-4-(5-amino-1-methyl-6-(methylamino)-2-oxo-1,2-dihydropyrimidin-4-yl)-2,5-diethylpiperazine-1-carboxylate
##STR00482##
[0707] To a stirred solution of tert-butyl (2R,5S)-2,5-diethyl-4-(1-methyl-6-(methylamino)-5-nitro-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxylate (1.04 g, 2.45 mmol) in CH.sub.2Cl.sub.2/MeOH (8 mL/8 mL) was added NiCl.sub.2 (637 mg, 4.9 mmol), followed by NaBH.sub.4 (186.2 mg, 4.9 mmol) at 0 C. After stirred for 20 min, the reaction was quenched with aq.NH.sub.4Cl, extracted with EtOAc (20 mL4). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated to give the titled compound (960 mg, 100%). MS: M/e 395 (M+1).sup.+.
Step C: tert-butyl (2R,5S)-4-(5-(2-cyanoacetamido)-1-methyl-6-(methylamino)-2-oxo-1,2-dihydropyrimidin-4-yl)-2,5-diethylpiperazine-1-carboxylate
##STR00483##
[0708] A mixture of tert-butyl (2R,5S)-4-(5-amino-1-methyl-6-(methylamino)-2-oxo-1,2-dihydropyrimidin-4-yl)-2,5-diethylpiperazine-1-carboxylate (500 mg, 1.27 mmol), 2-cyanoacetic acid (129 mg, 1.52 mmol), HATU (580 mg, 1.52 mmol) and DIPEA (327 mg, 2.54 mmol) in CH.sub.2Cl.sub.2 (10 mL) was stirred overnight. The reaction mixture was diluted with CH.sub.2Cl.sub.2 (10 mL), washed with brine, dried over Na.sub.2SO.sub.4, concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (330 mg, 56%). MS: M/e 462 (M+1).sup.+.
Step D: 2-(6-((2S,5R)-2,5-diethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00484##
[0709] A solution of tert-butyl (2R,5S)-4-(5-(2-cyanoacetamido)-1-methyl-6-(methylamino)-2-oxo-1,2-dihydropyrimidin-4-yl)-2,5-diethylpiperazine-1-carboxylate (330 mg, 0.71 mmol) in AcOH (5 mL) was stirred at 100 C. overnight. The reaction mixture was concentrated to give the residue, which was dissolved in CH.sub.2Cl.sub.2 (10 mL) and EtOAc/HCl(g) (4.0 M, 5 mL) was added and stirred for 3 hours. The reaction mixture was concentrated, H.sub.2O (1 mL) was added, then basified to pH=910 with Na.sub.2CO.sub.3, concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (109 mg, 45%). MS: M/e 344 (M+1).sup.+.
Step E: 2-(6-((2S,5R)-2,5-diethyl-4-(1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00485##
[0710] A mixture of 2-(6-((2S,5R)-2,5-diethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (34.3 mg, 0.1 mmol), 1-(4-(trifluoromethyl)phenyl)ethan-1-ol (28.5 mg, 0.15 mmol), (cyanomethyl)trimethylphosphonium iodide (72.9 mg, 0.3 mmol) and DIPEA (64.5 mg, 0.5 mmol) in CH.sub.3CN (4 mL) was stirred overnight at 100 C. in a sealed tube. The reaction mixture was diluted with EtOAc (20 mL), washed with H.sub.2O, brine, dried over Na.sub.2SO.sub.4, concentrated to dryness. The resulting residue was purified and separated into Compound A149a (3 mg) and Compound A149b (5 mg) by Prep-HPLC (Method A).
[0711] Compound A149a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.67-7.54 (m, 4H), 6.02-5.08 (m, 2H), 3.92 (d, J=8.8 Hz, 3H), 3.85-3.79 (m, 1H), 3.78 (s, 3H), 3.48-3.36 (m, 1H), 3.05-2.65 (m, 3H), 2.42-1.91 (m, 4H), 1.53-1.26 (m, 2H), 1.33 (d, J=6.4 Hz, 3H), 0.93 (t, J=7.2 Hz, 3H), 0.75-0.63 (m, 3H) ppm. MS: M/e 516 (M+1).sup.+.
[0712] Compound A149b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.64-7.56 (m, 4H), 6.19-5.08 (m, 2H), 4.28-4.12 (m, 1H), 3.92 (s, 3H), 3.78 (s, 3H), 3.70-3.59 (m, 1.5H), 3.20-3.11 (m, 1.5H), 2.75-2.51 (m, 1H), 2.29-2.21 (m, 1H), 1.94-1.45 (m, 5H), 1.31 (d, J=6.4 Hz, 3H), 1.06-0.96 (m, 3H), 0.66 (t, J=7.2 Hz, 3H) ppm. MS: M/e 516 (M+1).sup.+.
Compound A150: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00486##
Step A: 6-chloro-1-methyl-5-nitropyrimidine-2,4 (1H,3H)-dione
##STR00487##
[0713] 6-chloro-1-methylpyrimidine-2,4 (1H,3H)-dione (4.8 g, 30 mmol) was dissolved in conc. H.sub.2SO.sub.4 (6 mL), then fluming HNO.sub.3 (6 mL) was added dropwise at 0 C. After the addition, the reaction mixture was stirred for 30 min. The reaction mixture was added to ice-H.sub.2O dropwise, then filtered. The cake was collected, dried to give the titled compound (3.8 g, 62%). MS: M/e 206 (M+1).sup.+.
Step B: 6-amino-1-methyl-5-nitropyrimidine-2,4 (1H,3H)-dione
##STR00488##
[0714] To a stirred suspension of 6-chloro-1-methyl-5-nitropyrimidine-2,4 (1H,3H)-dione (2.5 g, 12.2 mmol) in EtOH (20 mL) was added NH.sub.3.Math.H.sub.2O (2 mL, 24.2 mmol). Then the reaction mixture was stirred for 4 hours at RT. The reaction mixture was filtered and the cake was collected, which was suspended in H.sub.2O (30 mL) and acidified to pH=34 with aq.citric acid, the reaction mixture was filtered again and the cake was collected, dried to give the titled compound (1.8 g, 79%). MS: M/e 187 (M+1).sup.+.
Step C: tert-butyl (2R,5S)-4-(6-amino-1-methyl-5-nitro-2-oxo-1,2-dihydropyrimidin-4-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00489##
[0715] To a stirred mixture of 6-amino-1-methyl-5-nitropyrimidine-2,4 (1H,3H)-dione (617 mg, 3.32 mmol), tert-butyl (2R,5S)-2,5-dimethylpiperazine-1-carboxylate (852 mg, 4 mmol) and BOP (2.2 g, 4.98 mmol) in DMF (10 mL) was added DBU (1 g, 6.64 mmol). After the addition, the reaction mixture was stirred at 70 C. overnight. The reaction mixture was concentrated to give the residue, which was treated with H.sub.2O (50 mL) and extracted with EtOAc (60 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled crude compound (800 mg). MS: M/e 383 (M+1).sup.+.
Step D: 6-amino-4-((2S,5R)-2,5-dimethylpiperazin-1-yl)-1-methyl-5-nitropyrimidin-2 (1H)-one
##STR00490##
[0716] To a stirred solution of tert-butyl (2R,5S)-4-(6-amino-1-methyl-5-nitro-2-oxo-1,2-dihydropyrimidin-4-yl)-2,5-dimethylpiperazine-1-carboxylate (800 mg, 2.09 mmol) in CH.sub.2Cl.sub.2 (5 mL) was added EtOAc/HCl (g) (4.0 M, 10 mL). After then, the reaction mixture was stirred for 3 hours. The reaction mixture was filtered and the cake was collected, basified to pH=910 with aq.Na.sub.2CO.sub.3, then concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (260 mg, 28% over 2 steps). MS: M/e 283 (M+1).sup.+.
Step E: 6-amino-4-((2S,5R)-2,5-dimethyl-4-(1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-5-nitropyrimidin-2 (1H)-one
##STR00491##
[0717] A mixture of 6-amino-4-((2S,5R)-2,5-dimethylpiperazin-1-yl)-1-methyl-5-nitropyrimidin-2 (1H)-one (100 mg, 0.355 mmol), 1-(4-(trifluoromethyl)phenyl)ethan-1-ol (101 mg, 0.53 mmol), (cyanomethyl)trimethylphosphonium iodide (259 mg, 1.065 mmol) and DIPEA (228 mg, 1.775 mmol) in CH.sub.3CN (5 mL) was stirred overnight at 100 C. in a sealed tube. The reaction mixture was diluted with EtOAc (20 mL), washed with H.sub.2O, brine, dried over Na.sub.2SO.sub.4, concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (106 mg, 66%). MS: M/e 455 (M+1).sup.+.
Step F: 5,6-diamino-4-((2S,5R)-2,5-dimethyl-4-(1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methylpyrimidin-2 (1H)-one
##STR00492##
[0718] To a stirred solution of 6-amino-4-((2S,5R)-2,5-dimethyl-4-(1-(4-(trifluoromethyl)phenyl) ethyl)piperazin-1-yl)-1-methyl-5-nitropyrimidin-2 (1H)-one (30 mg, 0.066 mmol) in CH.sub.2Cl.sub.2/MeOH (4 mL/4 mL) was added NiCl.sub.2 (17.2 mg, 0.132 mmol), followed by NaBH.sub.4 (5 mg, 0.132 mmol) at 0 C. After stirred for 20 min, the reaction was quenched with aq.NH.sub.4Cl, extracted with EtOAc (10 mL2). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated to give the titled compound, which was directly used to the next step. MS: M/e 425 (M+1).sup.+.
Step G: N-(6-amino-4-((2S,5R)-2,5-dimethyl-4-(1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrimidin-5-yl)-2-cyanoacetamide
##STR00493##
[0719] A mixture of 5,6-diamino-4-((2S,5R)-2,5-dimethyl-4-(1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methylpyrimidin-2 (1H)-one (0.088 mmol), 2-cyanoacetic acid (7.48 mg, 0.088 mmol), HATU (40 mg, 0.105 mmol) and DIPEA (22.7 mg, 0.176 mmol) in CH.sub.2Cl.sub.2 (5 mL) was stirred overnight. The reaction mixture was diluted with CH.sub.2Cl.sub.2 (10 mL), washed with brine, dried over Na.sub.2SO.sub.4, concentrated and purified by Pre-TLC (CH.sub.2Cl.sub.2/MeOH=10:1) to give the titled compound (11 mg, 25%). MS: M/e 492 (M+1).sup.+.
Step H: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00494##
[0720] A solution of N-(6-amino-4-((2S,5R)-2,5-dimethyl-4-(1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrimidin-5-yl)-2-cyanoacetamide (11 mg, 0.022 mmol) in AcOH (3 mL) was stirred at 80 C. overnight. The reaction mixture was concentrated and purified by Prep-HPLC (Method A) to give the titled compound (4 mg). .sup.1H NMR (400 MHz, DMSO-d6) 7.75-7.58 (m, 4H), 5.92-4.31 (m, 2H), 4.14-4.02 (m, 2H), 3.76-3.50 (m, 2H), 3.37 (s, 3H), 2.88-2.57 (m, 2.5H), 2.06 (d, J=13.2 Hz, 0.5H), 1.46-1.17 (m, 6H), 1.03-0.81 (m, 3H) ppm. MS: M/e 474 (M+1).sup.+.
Compound A152: 8-(but-2-yn-1-yl)-6-((2S,5R)-2,5-diethyl-4-(1-(4-(trifluoromethyl)phenyl) ethyl)piperazin-1-yl)-3,9-dimethyl-3,9-dihydro-2H-purin-2-one
##STR00495##
Step A: tert-butyl (2R,5S)-2,5-diethyl-4-(1-methyl-6-(methylamino)-2-oxo-5-(pent-3-ynamido)-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxylate
##STR00496##
[0721] A mixture of tert-butyl (2R,5S)-4-(5-amino-1-methyl-6-(methylamino)-2-oxo-1,2-dihydropyrimidin-4-yl)-2,5-diethylpiperazine-1-carboxylate (460 mg, 1.17 mmol), pent-3-ynoic acid (137 mg, 1.4 mmol), HATU (535 mg, 1.4 mmol) and DIPEA (302 mg, 2.34 mmol) in CH.sub.2Cl.sub.2 (10 mL) was stirred overnight. The reaction mixture was diluted with CH.sub.2Cl.sub.2 (10 mL), washed with brine, dried over Na.sub.2SO.sub.4, concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (153 mg, 27.5%) as a crude product, which was directly used to the next step. MS: M/e 475 (M+1).sup.+.
Step B: tert-butyl (2R,5S)-4-(8-(but-2-yn-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-diethylpiperazine-1-carboxylate
##STR00497##
[0722] A solution of tert-butyl (2R,5S)-2,5-diethyl-4-(1-methyl-6-(methylamino)-2-oxo-5-(pent-3-ynamido)-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxylate (150 mg, 0.32 mmol) in AcOH (4 mL) was stirred at 80 C. for 6 hours. The reaction mixture was concentrated to give the residue, which was dissolved in CH.sub.2Cl.sub.2 (10 mL) and EtOAc/HCl(g) (4.0 M, 5 mL) was added and stirred for 3 hours. The reaction mixture was concentrated. The resulting residue was purified by Pre-TLC (CH.sub.2Cl.sub.2/MeOH=10:1) to give the titled compound (40 mg, 27%). MS: M/e 456 (M+1).sup.+.
Step C: 8-(but-2-yn-1-yl)-6-((2S,5R)-2,5-diethylpiperazin-1-yl)-3,9-dimethyl-3,9-dihydro-2H-purin-2-one
##STR00498##
[0723] To a stirred solution of tert-butyl (2R,5S)-4-(8-(but-2-yn-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-diethylpiperazine-1-carboxylate (40 mg, 0.088 mmol) in CH.sub.2Cl.sub.2 (4 mL) was added EtOAc/HCl(g) (4.0 M, 2 mL) and stirred for 3 hours. The reaction mixture was concentrated. The resulting residue was basified with MeOH/NH.sub.3 (g) and purified by Pre-TLC (CH.sub.2Cl.sub.2/MeOH(NH.sub.3)=10:1) to give the titled compound (20 mg, 64%). MS: M/e 357 (M+1).sup.+.
Step D: 8-(but-2-yn-1-yl)-6-((2S,5R)-2,5-diethyl-4-(1-(4-(trifluoromethyl)phenyl)ethyl) piperazin-1-yl)-3,9-dimethyl-3,9-dihydro-2H-purin-2-one
##STR00499##
[0724] A mixture of 8-(but-2-yn-1-yl)-6-((2S,5R)-2,5-diethylpiperazin-1-yl)-3,9-dimethyl-3,9-dihydro-2H-purin-2-one (20 mg, 0.056 mmol), 1-(4-(trifluoromethyl)phenyl)ethan-1-ol (21 mg, 0.112 mmol), (cyanomethyl)trimethylphosphonium iodide (40.8 mg, 0.168 mmol) and DIPEA (36 mg, 0.28 mmol) in CH.sub.3CN (3 mL) was stirred overnight at 100 C. in a sealed tube. The reaction mixture was diluted with EtOAc (20 mL), washed with H.sub.2O, brine, dried over Na.sub.2SO.sub.4, concentrated and purified by Pre-TLC (CH.sub.2Cl.sub.2/MeOH=10:1) to give crude product, which was further purified by Prep-HPLC (Method A) to give the titled compound (70% purity, 3 mg). .sup.1H NMR (500 MHz, CD.sub.3OD) 7.71-7.51 (m, 4H), 6.19-5.06 (m, 2H), 4.03-3.95 (m, 2H), 3.95-3.88 (m, 1H), 3.84-3.76 (m, 3H), 3.76-3.61 (m, 3H), 3.20-2.94 (m, 2H), 2.89-2.51 (m, 1H), 2.43-2.20 (m, 1H), 2.16-1.90 (m, 2H), 1.83-1.72 (m, 3H), 1.53-1.41 (m, 2H), 1.40-1.27 (m, 3H), 1.07-0.86 (m, 3H), 0.74-0.60 (m, 3H) ppm. MS: M/e 529 (M+1).sup.+.
Compound A153: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(quinoxalin-2-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00500##
Step A: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(quinoxalin-2-yl)ethyl)piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00501##
[0725] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.15 mmol) in CH.sub.3CN (5 mL) and was added 1-(quinoxalin-2-yl)ethan-1-ol (56 mg, 0.32 mmol), (cyanomethyl)trimethylphosphonium iodide (111 mg, 0.45 mmol) and DIPEA (205 mg, 1.57 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure. The resulting residue was purified by flash column chromatography to give the titled Compound A153 (crude), which was further purified and separated into Compound A153a (1 mg) and Compound A153b (2 mg) by Prep-HPLC (Method A).
[0726] Compound A153a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 9.17 (s, 1H), 8.08 (dd, J=12.9, 9.3 Hz, 2H), 7.85 (dd, J=9.5, 5.8 Hz, 2H), 6.18-4.88 (m, 2H), 4.10 (d, J=85.8 Hz, 2H), 3.93 (s, 3H), 3.78 (s, 3H), 3.73-3.33 (m, 3H), 2.84 (s, 1H), 2.11 (d, J=12.2 Hz, 1H), 1.54 (d, J=6.7 Hz, 3H), 1.28 (d, J=5.2 Hz, 3H), 1.12 (d, J=6.4 Hz, 3H) ppm. MS: M/e 472 (M+1).sup.+.
[0727] Compound A153b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 9.15 (s, 1H), 8.08 (dd, J=9.3, 6.7 Hz, 2H), 7.83 (p, J=7.8 Hz, 2H), 6.28-4.91 (m, 2H), 4.28-4.08 (m, 2H), 3.93 (s, 3H), 3.78 (s, 3H), 3.71-3.33 (m, 2H), 3.10-2.75 (m, 3H), 1.51 (d, J=6.8 Hz, 3H), 1.41 (d, J=5.6 Hz, 3H), 1.04 (d, J=6.5 Hz, 3H) ppm. MS: M/e 472 (M+1).sup.+.
Compound A156: 2-(9-ethyl-6-((2S,5R)-4-(1-(7-fluoro-2,2-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00502##
[0728] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (33 mg, 0.10 mmol), 1-(7-fluoro-2,2-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (45 mg, 0.20 mmol), (cyanomethyl)trimethylphosphonium iodide (73 mg, 0.30 mmol) and DIPEA (129 mg, 1.00 mmol) in CH.sub.3CN (2 ml) was stirred at 100 C. overnight. The reaction was diluted with EtOAc (10 ml) and washed with brine (10 ml). The organic layer was concentrated under reduced pressure. The resulting residue was purified by flash column chromatography to give the titled Compound A156, which was further separated into Compound A156a (5.99 mg) and Compound A156b (4.55 mg) by Prep-HPLC (Method A).
[0729] Compound A156a (the earlier peak): .sup.1H NMR (400 MHz, DMSO-d.sub.6): 6.96 (d, J=6.8 Hz, 1H), 6.72 (d, J=10.9 Hz, 1H), 5.95-5.55 (m, 1H), 4.95-4.60 (m, 1H), 4.40 (s, 2H), 4.26 (q, J=8.0 Hz, 2H), 3.94 (s, 2H), 3.67 (q, J=6.4 Hz, 2H), 3.61 (s, 3H), 3.44 (s, 2H), 2.17 (d, J=11.9 Hz, 1H), 1.34 (t, J=7.0 Hz, 3H), 1.27-1.21 (m, 9H), 1.17 (s, 3H), 0.92 (s, 3H) ppm. MS: M/e 538 (M+1).sup.+.
[0730] Compound A156b (the later peak): .sup.1H NMR (400 MHz, DMSO-d.sub.6): 6.90 (d, J=7.0 Hz, 1H), 6.75 (d, J=10.7 Hz, 1H), 6.05-5.37 (m, 1H), 5.12-4.54 (m, 1H), 4.39 (s, 2H), 4.25 (s, 2H), 3.95 (s, 2H), 3.83 (q, J=6.4 Hz, 1H), 3.61 (s, 3H), 3.27-3.00 (m, 1H), 2.89-2.64 (m, 3H), 1.37-1.25 (m, 12H), 1.22 (d, J=6.6 Hz, 3H), 0.83 (d, J=6.3 Hz, 3H) ppm. MS: M/e 538 (M+1).sup.+.
Compound A157: 2-(9-ethyl-6-((2S,5R)-4-(1-(7-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00503##
Step A: 7-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxine-6-carbaldehyde
##STR00504##
[0731] A solution of 7-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxine-6-carbonitrile (300 mg, 1.45 mmol) and Dibal-H (1.5 M, 1.93 ml, 2.90 mmol) in THE (10 ml) was stirred at 0 C. under N.sub.2 for 30 min and then RT for 2 hrs. The solution was extracted with EtOAc (15 ml), washed with brine (10 ml2), dried and evaporated. The residue was purified by flash column chromatography with 0-20% EtOAc in PE to give the titled compound (180 mg, 59%). MS: M/e 211 (M+1).sup.+.
Step B: 1-(7-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol
##STR00505##
[0732] A solution of 7-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxine-6-carbaldehyde (180 mg, 0.85 mmol) and MeMgBr (3M, 0.43 ml, 1.29 mmol) in THE (6 ml) was stirred at 0 C. under N.sub.2 for 30 min and then RT for 1 h. The solution was extracted with EtOAc (15 ml), washed with brine (10 ml2), dried and evaporated. The residue was purified by flash column chromatography with 0-25% EtOAc in PE to give the titled compound (170 mg, 88%). MS: M/e 227 (M+1).sup.+.
Step C: 2-(9-ethyl-6-((2S,5R)-4-(1-(7-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00506##
[0733] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (146 mg, 0.44 mmol), 1-(7-fluoro-3,3-dimethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-ol (100 mg, 0.44 mmol), (cyanomethyl)trimethylphosphonium iodide (324 mg, 1.33 mmol) and DIPEA (572 mg, 4.43 mmol) in MeCN (4 ml) was stirred at 100 C. overnight. The reaction was diluted with EtOAc (20 ml) and washed with brine (10 ml). The organic layer was concentrated under reduced pressure. The resulting residue was purified by flash column chromatography with 0-5% MeOH in DCM to give the titled Compound A157, which was further separated into Compound A157a (27 mg) and Compound A157b (27 mg) by chiral prep-HPLC. The chiral separation conditions are shown below.
TABLE-US-00021 Column CHIRALPAK IH Column Size 2 cm 25 cm, 5 um Mobile Phase A Hex:DCM Mobile Phase B IPA Flow Rate 20 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-HPLC Equipment Prep-HPLC-Gilson
[0734] Compound A157a (the earlier peak): .sup.1H NMR (500 MHz, DMSO-d6) 6.97 (d, J=6.9 Hz, 1H), 6.67 (d, J=10.9 Hz, 1H), 6.02-5.45 (m, 1H), 5.12-4.45 (m, 1H), 4.44-4.34 (m, 2H), 4.32-4.18 (m, 2H), 3.93 (s, 2H), 3.83 (q, J=6.6 Hz, 1H), 3.61 (s, 3H), 3.31-3.24 (m, 1H), 2.88-2.66 (m, 3H), 1.37-1.27 (m, 12H), 1.23 (d, J=6.4 Hz, 3H), 0.83 (d, J=5.9 Hz, 3H) ppm. MS: M/e 538 (M+1).sup.+.
[0735] Compound A157b (the later peak): .sup.1H NMR (500 MHz, DMSO-d6) 7.03 (d, J=6.9 Hz, 1H), 6.64 (d, J=11.0 Hz, 1H), 5.90-5.65 (m, 1H), 4.95-4.60 (m, 1H), 4.46-4.34 (m, 2H), 4.33-4.20 (m, 2H), 3.91 (s, 2H), 3.68 (q, J=6.0 Hz, 1H), 3.61 (s, 3H), 3.50-3.45 (m, 1H), 3.32-3.20 (m, 1H), 2.62-2.52 (m, 1H), 2.18 (d, J=11.7 Hz, 1H), 1.34 (t, J=7.1 Hz, 3H), 1.28 (s, 6H), 1.25 (d, J=6.4 Hz, 3H), 1.18 (s, 3H), 0.94 (s, 3H) ppm. MS: M/e 538 (M+1).sup.+.
Compound A158: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00507##
Step A: tert-butyl (2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-5-ethyl-2-methylpiperazine-1-carboxylate
##STR00508##
[0736] A mixture of tert-butyl (2S,5R)-5-ethyl-2-methylpiperazine-1-carboxylate (228 mg, 1 mmol), 1-(3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethan-1-ol (313.5 mg, 1.5 mmol), (cyanomethyl)trimethylphosphonium iodide (729 mg, 3 mmol) and DIPEA (1.29 g, 10 mmol) in CH.sub.3CN (5 mL) was stirred overnight at 100 C. in a sealed tube. The reaction mixture was concentrated to give the residue, which was purified by flash column chromatography to give the titled compound (240 mg, 57%). MS: M/e 420 (M+1).sup.+.
Step B: 6-(1-((2R,5S)-2-ethyl-5-methylpiperazin-1-yl)ethyl)-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridine
##STR00509##
[0737] To a stirred solution of tert-butyl (2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-5-ethyl-2-methylpiperazine-1-carboxylate (240 mg, 0.57 mmol) in CH.sub.2Cl.sub.2 (5 mL) was added TFA (2 mL). After the addition, the reaction mixture was stirred for 5 hours. The reaction mixture was concentrated, basified to pH=89 with aq.Na.sub.2CO.sub.3, and extracted with CH.sub.2Cl.sub.2 (10 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated to give the titled compound (180 mg, 98%) as colorless oil. MS: M/e 320 (M+1).sup.+.
Step C: 9-ethyl-6-hydroxy-8-(hydroxymethyl)-3-methyl-3,9-dihydro-2H-purin-2-one
##STR00510##
[0738] A mixture of 8-((benzyloxy)methyl)-9-ethyl-6-hydroxy-3-methyl-3,9-dihydro-2H-purin-2-one (4.4 g, 14 mmol) in MeOH/AcOH (300 mL/8 mL) was stirred at 70 C. for an hour to become a solution. Then Pd/C (0.5 g) was added, and the mixture was stirred at 40 C. overnight under H.sub.2 (1 atm). The reaction mixture was filtered, and the filtrate was concentrated to give the titled compound (3 g, 95%). MS: M/e 225 (M+1).sup.+.
Step D: 8-(chloromethyl)-9-ethyl-6-hydroxy-3-methyl-3,9-dihydro-2H-purin-2-one
##STR00511##
[0739] To a stirred mixture of 9-ethyl-6-hydroxy-8-(hydroxymethyl)-3-methyl-3,9-dihydro-2H-purin-2-one (1 g, 4.46 mmol) in CH.sub.2Cl.sub.2 (20 mL) was added SOCl.sub.2 (3.18 g, 26.78 mmol) at 40 C. overnight. The reaction mixture was filtered, and the cake was collected, dried to give the titled compound (1.15 g, 100%). MS: M/e 243 (M+1).sup.+.
Step E: 2-(9-ethyl-6-hydroxy-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00512##
[0740] To a stirred mixture of 8-(chloromethyl)-9-ethyl-6-hydroxy-3-methyl-3,9-dihydro-2H-purin-2-one (1.15 g, 4.74 mmol) in CH.sub.2Cl.sub.2 (20 mL) was added TMSCN (0.94 g, 9.48 mmol), followed by TBAF (1.0M, 9.48 mL, 9.48 mmol). After the addition, the reaction was stirred overnight. The reaction mixture was filtered, and the cake was collected to give part 1 (338 mg), and the filtrate was concentrated and purified by flash column chromatography to give part 2 (222 mg), then combined to give the titled compound (560 mg, 51%). MS: M/e 234 (M+1).sup.+.
Step F: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00513##
[0741] To a stirred mixture of 2-(9-ethyl-6-hydroxy-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (70 mg, 0.3 mmol) in C.sub.2H.sub.4Cl.sub.2 (4 mL) was added DIPEA (77.4 mg, 0.6 mmol), followed by POCl.sub.3 (92.1 mg, 0.6 mmol). After then, the reaction mixture was stirred at 60 C. for 2 hours. The reaction mixture was concentrated to give the residue, which was dissolved in C.sub.2H.sub.4Cl.sub.2 (10 mL) again, DIPEA (77.4 mg, 0.6 mmol) and 6-(1-((2R,5S)-2-ethyl-5-methylpiperazin-1-yl)ethyl)-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridine (95 mg, 0.3 mmol) were added and the mixture was stirred at 100 C. for 3 days. The reaction mixture was washed with H.sub.2O, brine, extracted with CH.sub.2Cl.sub.2 (5 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated. The resulting residue was purified by Prep-HPLC (Method A) to give the titled Compound A158 (17 mg), which was further separated into Compound A158a (7 mg) and Compound A158b (7 mg) by chiral Prep-HPLC. The chiral separation conditions are shown below.
TABLE-US-00022 Column CHIRALPAK IF Column Size 2 cm 25 cm, 5 um Mobile Phase A MtBE Mobile Phase B MeOH:DCM Flow Rate 20 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-HPLC Equipment Prep-HPLC-Gilson
[0742] Compound A158: .sup.1H NMR (400 MHz, CD.sub.3OD) 7.31 (t, J=7.2 Hz, 1H), 7.12 (dd, J=22.8, 8.0 Hz, 1H), 6.20-4.98 (m, 2H), 4.43-4.31 (m, 2H), 4.22 (s, 0.5H), 4.00-3.91 (m, 2H), 3.76 (s, 3H), 3.73-3.43 (m, 1.5H), 3.30-3.20 (m, 2H), 3.19-2.73 (m, 2H), 2.50-2.16 (m, 1H), 1.56-1.22 (m, 17H), 1.07-0.72 (m, 3H) ppm. MS: M/e 535 (M+1).sup.+.
[0743] Compound A158a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.32 (d, J=8.0 Hz, 1H), 7.09 (d, J=8.0 Hz, 1H), 6.14-5.06 (m, 2H), 4.45-4.26 (m, 2H), 4.21 (s, 0.5H), 3.97 (s, 2H), 3.76 (s, 3H), 3.74-3.67 (m, 1H), 3.52-3.38 (m, 0.5H), 3.30-3.20 (m, 2H), 2.96-2.72 (m, 2H), 2.50-2.38 (m, 1H), 1.61-1.26 (m, 17H), 0.76 (t, J=6.6 Hz, 3H) ppm. MS: M/e 535 (M+1).sup.+.
[0744] Compound A158b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.30 (d, J=8.0 Hz, 1H), 7.15 (d, J=8.0 Hz, 1H), 6.18-4.99 (m, 2H), 4.43-4.32 (m, 2H), 3.95 (s, 2H), 3.76 (s, 3H), 3.72-3.49 (m, 2H), 3.30-3.20 (m, 2H), 3.18-3.04 (m, 1H), 2.85-2.60 (m, 1H), 2.20 (d, J=11.2 Hz, 1H), 1.55-1.22 (m, 17H), 1.01 (t, J=6.4 Hz, 3H) ppm. MS: M/e 535 (M+1).sup.+.
Compound A159: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00514##
Step A: tert-butyl (2R,5S)-4-(8-((benzyloxy)methyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2-ethyl-5-methylpiperazine-1-carboxylate
##STR00515##
[0745] To a stirred mixture of 8-((benzyloxy)methyl)-6-hydroxy-3,9-dimethyl-3,9-dihydro-2H-purin-2-one (1.5 g, 5 mmol) and tert-butyl (2R,5S)-2-ethyl-5-methylpiperazine-1-carboxylate (1.37 g, 6 mmol) in DMF (10 mL) was added BOP (3.3 g, 7.5 mmol), followed by DBU (1.52 g, 10 mmol). After the addition, the reaction mixture was stirred overnight. The reaction mixture was concentrated to give the residue, which was treated with H.sub.2O (20 mL), extracted with EtOAc (30 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated and purified by flash column chromatography to give the titled compound (2.1 g, 82%). MS: M/e 511 (M+1).sup.+.
Step B: tert-butyl (2R,5S)-2-ethyl-4-(8-(hydroxymethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-5-methylpiperazine-1-carboxylate
##STR00516##
[0746] To a stirred solution of tert-butyl (2R,5S)-4-(8-((benzyloxy)methyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2-ethyl-5-methylpiperazine-1-carboxylate (2.1 g, 4.11 mmol) in MeOH/AcOH (150 mL/5 mL) was added Pd/C (300 mg). After the addition, the reaction mixture was stirred at 35 C. over a weekend under H.sub.2 (1 atm). The reaction mixture was filtered, and the filtrate was concentrated to give the titled compound (1.7 g, 98%). MS: M/e 421 (M+1).sup.+.
Step C: tert-butyl (2R,5S)-4-(8-(cyanomethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2-ethyl-5-methylpiperazine-1-carboxylate
##STR00517##
[0747] To a stirred solution of tert-butyl (2R,5S)-2-ethyl-4-(8-(hydroxymethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-5-methylpiperazine-1-carboxylate (1.7 g, 4 mmol) in CH.sub.2Cl.sub.2 (15 mL) was added SOCl.sub.2 (0.96 g, 8 mmol). After the addition, the reaction mixture was stirred for 30 min. The reaction mixture was concentrated to give the residue, which was dissolved in CH.sub.2Cl.sub.2 (20 mL) again. TMSCN (0.8 g, 8 mmol) was added followed by TBAF (1.0 M, 8 mL, 8 mmol). After then, the reaction was stirred for 4 hours. The reaction mixture was washed with H.sub.2O, brine, dried over Na.sub.2SO.sub.4, concentrated and purified by flash column chromatography to give the titled compound (400 mg, 23%). MS: M/e 430 (M+1).sup.+.
Step D: 2-(6-((2S,5R)-5-ethyl-2-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00518##
[0748] To a stirred solution of tert-butyl (2R,5S)-4-(8-(cyanomethyl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2-ethyl-5-methylpiperazine-1-carboxylate (400 mg, 0.93 mmol) in CH.sub.2Cl.sub.2 (5 mL) was added EtOAc/HCl(g) (1.0 M, 10 mL). After then, the reaction mixture was stirred for 4 hours. The reaction mixture was concentrated to give the residue, which was treated with H.sub.2O (2 mL), then Na.sub.2CO.sub.3 (0.5 g) was added, then washed with CH.sub.3CN (15 mL), stirred for 10 min and filtered. The filtrate was dried over Na.sub.2SO.sub.4, concentrated to give the titled compound (300 mg, 98%). MS: M/e 330 (M+1).sup.+.
Step E: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00519##
[0749] A mixture of 2-(6-((2S,5R)-5-ethyl-2-methylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (100 mg, 0.304 mmol), 1-(3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethan-1-ol (190 mg, 0.9 mmol), (cyanomethyl)trimethylphosphonium iodide (221 mg, 0.9 mmol) and DIPEA (196 mg, 1.52 mmol) in CH.sub.3CN (4 mL) was stirred overnight at 100 C. in a sealed tube. The reaction mixture was diluted with EtOAc (20 mL), washed with H.sub.2O, brine, dried over Na.sub.2SO.sub.4, concentrated to dryness. The resulting residue was purified by Prep-HPLC (Method A) to give the titled Compound A159 (28 mg), which was further separated into Compound A159a (5 mg) and Compound A159b (6 mg) by chiral Prep-HPLC. The chiral separation conditions are shown below.
TABLE-US-00023 Column CHIRALPAK ID Column Size 2 cm 25 cm, 5 um Mobile Phase A MtBE Mobile Phase B MeOH:DCM Flow Rate 20 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-HPLC Equipment Prep-HPLC-Gilson
[0750] Compound A159: .sup.1H NMR (400 MHz, CD.sub.3OD) 7.32 (t, J=6.8 Hz, 1H), 7.11 (t, J=8.4 Hz, 1H), 6.24-5.00 (m, 2H), 4.22 (s, 1H), 3.97 (d, J=2.4 Hz, 2H), 3.93 (s, 3H), 3.88-3.82 (m, 1H), 3.79 (s, 3H), 3.75-3.50 (m, 1H), 3.27-2.36 (m, 4H), 1.64-1.23 (m, 14H), 1.11-0.75 (m, 3H) ppm. MS: M/e 521 (M+1).sup.+.
[0751] Compound A159a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.32 (d, J=8.0 Hz, 1H), 7.09 (d, J=8.0 Hz, 1H), 6.15-5.04 (m, 2H), 3.96 (s, 2H), 3.92 (s, 3H), 3.78 (s, 3H), 3.76-3.63 (m, 1H), 3.50-3.38 (m, 1H), 3.30-3.20 (m, 1H), 3.14-2.71 (m, 3H), 2.48-2.36 (m, 1H), 1.57-1.26 (m, 14H), 0.74 (t, J=6.8 Hz, 3H) ppm. MS: M/e 521 (M+1).sup.+.
[0752] Compound A159b (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.30 (d, J=8.0 Hz, 1H), 7.15 (d, J=8.0 Hz, 1H), 6.17-4.95 (m, 2H), 3.95 (s, 2H), 3.93 (s, 3H), 3.78 (s, 3H), 3.71-3.43 (m, 2H), 3.30-3.20 (m, 2H), 3.15-3.03 (m, 1H), 2.79-2.60 (m, 1H), 2.19 (d, J=11.2 Hz, 1H), 1.52-1.42 (m, 2H), 1.40 (s, 6H), 1.35-1.22 (m, 6H), 1.00 (t, J=6.4 Hz, 3H) ppm. MS: M/e 521 (M+1).sup.+.
Compound A161: 2-(9-ethyl-6-((2S,5R)-4-(1-(7-fluoro-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00520##
Step A: 2-chloro-5-fluoro-6-(hydroxymethyl)pyridin-3-ol
##STR00521##
[0753] A mixture of 2-chloro-5-fluoropyridin-3-ol (2 g, 13.5 mmol) in water (10 mL) was added NaHCO.sub.3 (1.7 g, 20.3 mmol) at room temperature and the resulting mixture was stirred at room temperature for 15 mins. Then was added HCHO (5.5 mL in water, 37%) at room temperature and the resulting mixture was stirred at 90 C. under N.sub.2 for 5 hours. Then the reaction mixture was treated with HCl (1M) to pH4, extracted with EtOAc dried over Na.sub.2SO.sub.4 and concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (1.8 g, 75%). MS: M/e 178 (M+1).sup.+.
Step B: 1-((2-chloro-5-fluoro-6-(hydroxymethyl)pyridin-3-yl)oxy)propan-2-one
##STR00522##
[0754] To a solution of 2-chloro-5-fluoro-6-(hydroxymethyl)pyridin-3-ol (1.8 g, 10.1 mmol) in CH.sub.3CN (20 mL) was added K.sub.2CO.sub.3 (2.8 g, 20.3 mmol) and 1-chloropropan-2-one at (1.58 g, 16.9 mmol) room temperature and the resulting mixture was stirred at 60 C. for 15 hours. The reaction mixture was concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (1.5 g, 63%). MS: M/e 234 (M+1).sup.+.
Step C: 6-chloro-3-fluoro-5-(2-oxopropoxy)picolinaldehyde
##STR00523##
[0755] To a solution of 1-((2-chloro-5-fluoro-6-(hydroxymethyl)pyridin-3-yl)oxy)propan-2-one (1.5 g, 6.4 mmol) in DCM (50 mL) was added Dess Martin (4.1 g, 9.6 mmol) at room temperature and the resulting mixture was stirred at room temperature for 1.5 hours. The reaction mixture was quenched with saturated NaHCO.sub.3 aq., extracted with EtOAc combined, washed brine, dried and concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (800 mg, 53%). MS: M/e 232 (M+1).sup.+.
Step D: 1-((2-chloro-5-fluoro-6-(1-hydroxyethyl)pyridin-3-yl)oxy)-2-methylpropan-2-ol
##STR00524##
[0756] To a solution of 6-chloro-3-fluoro-5-(2-oxopropoxy)picolinaldehyde (800 mg, 3.5 mmol) in THE (50 mL) was added CH.sub.3MgBr (3M, 5.7 mL, 17.3 mmol) at room temperature and the resulting mixture was stirred at room temperature for 1 hour. The reaction mixture was quenched with saturated NH.sub.4Cl aq. (50 mL), extracted with EtOAc washed brine, dried and concentrated to give the titled compound (820 mg, 89%). MS: M/e 264 (M+1).sup.+.
Step E: 1-(7-fluoro-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethan-1-ol
##STR00525##
[0757] A mixture of 1-((2-chloro-5-fluoro-6-(1-hydroxyethyl)pyridin-3-yl)oxy)-2-methylpropan-2-ol (800 mg, 3 mmol), BINAP (379 mg, 0.6 mmol), Cs.sub.2CO.sub.3 (1.9 g, 6 mmol) and Pd(AcO).sub.2 (68 mg, 0.3 mmol) in toluene (50 mL) was stirred at 100 C. under N.sub.2 for 15 hours. The reaction mixture was concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (370 mg, 53%). MS: M/e 228 (M+1).sup.+.
Step F: 2-(9-ethyl-6-((2S,5R)-4-(1-(7-fluoro-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00526##
[0758] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (300 mg, 0.9 mmol) in CH.sub.3CN (8 mL) and was added 1-(7-fluoro-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethan-1-ol (310 mg, 1.35 mmol), (cyanomethyl)trimethylphosphonium iodide (875 mg, 3.6 mmol) and DIPEA (1.16 g, 9 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure and purified by flash column chromatography (DCM/MeOH) to give the titled Compound A161, which was further separated into Compound A161a (113 mg) and Compound A161b (115 mg) by chiral Prep-HPLC. The chiral separation conditions are shown below.
TABLE-US-00024 Column CHIRALPAK IA Column Size 2 cm 25 cm, 5 um Mobile Phase A MtBE Mobile Phase B MeOH:DCM Flow Rate 20 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-HPLC Equipment Prep-HPLC-Gilson
[0759] Compound A161: .sup.1H NMR (400 MHz, DMSO-d6) 7.22-7.14 (m, 1H), 6.15-4.94 (m, 2H), 4.36 (d, J=7.4 Hz, 2H), 4.23 (s, 1H), 4.07 (s, 1H), 3.99 (s, 2H), 3.76 (s, 3H), 3.73-3.36 (m, 2H), 3.10-2.67 (m, 2H), 2.49-2.10 (m, 1H), 1.54-1.22 (m, 15H), 1.10-0.90 (m, 3H) ppm. MS: M/e 539 (M+1).sup.+.
[0760] Compound A161a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.17 (d, J=9.6 Hz, 1H), 6.17-4.92 (m, 2H), 4.36 (d, J=7.0 Hz, 2H), 4.03 (d, J=6.6 Hz, 1H), 3.98 (s, 2H), 3.76 (s, 3H), 3.60-3.34 (m, 3H), 3.10-2.62 (m, 3H), 1.54-1.27 (m, 15H), 0.97 (d, J=6.2 Hz, 3H) ppm. MS: M/e 539 (M+1).sup.+.
[0761] Compound A161b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.14 (d, J=9.4 Hz, 1H), 6.11-4.88 (m, 2H), 4.36 (d, J=7.3 Hz, 2H), 3.98 (s, 2H), 3.89 (s, 1H), 3.75 (s, 3H), 3.73-3.35 (m, 4H), 2.80 (s, 1H), 2.20 (d, J=11.4 Hz, 1H), 1.46 (t, J=7.0 Hz, 3H), 1.38 (d, J=7.9 Hz, 9H), 1.24 (s, 3H), 1.00 (d, J=5.6 Hz, 3H) ppm. MS: M/e 539 (M+1).sup.+.
Compound A162: 2-(6-((2S,5R)-4-(1-(7-fluoro-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00527##
[0762] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.16 mmol) in CH.sub.3CN (8 mL) and was added 1-(7-fluoro-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethan-1-ol (54 mg, 0.24 mmol), (cyanomethyl)trimethylphosphonium iodide (115 mg, 0.48 mmol) and DIPEA (204 mg, 9 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure and purified by flash column chromatography (DCM/MeOH) to give the titled compound (16 mg). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.21-7.11 (m, 1H), 6.01-4.91 (m, 2H), 4.21 (s, 1H), 4.04 (s, 1H), 3.98 (s, 2H), 3.93 (s, 3H), 3.78 (s, 3H), 3.70-3.36 (m, 2H), 3.05-2.09 (m, 3H), 1.43-1.22 (m, 12H), 1.04-0.85 (m, 3H) ppm. MS: M/e 525 (M+1).sup.+.
Compound A163: 2-(9-ethyl-6-((2S,5R)-4-(1-(8-fluoro-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00528##
Step A: 2-chloro-4-fluoro-6-(hydroxymethyl)pyridin-3-ol
##STR00529##
[0763] A mixture of 2-chloro-4-fluoropyridin-3-ol (0.6 g, 4.05 mmol) in water (3 mL) was added NaHCO.sub.3 (0.51 g, 6.08 mmol) at room temperature and the resulting mixture was stirred at room temperature for 15 mins. Then was added HCHO (8.2 mL, in water, 37%) at room temperature and the resulting mixture was stirred at 110 C. under N.sub.2 for 3 days. Then the reaction mixture was treated with HCl (1M) to pH4, extracted with EtOAc dried over Na.sub.2SO.sub.4 and concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (0.42 g, 58%). MS: M/e 178 (M+1).sup.+.
Step B: 1-((2-chloro-4-fluoro-6-(hydroxymethyl)pyridin-3-yl)oxy)propan-2-one
##STR00530##
[0764] To a solution of 2-chloro-4-fluoro-6-(hydroxymethyl)pyridin-3-ol (0.7 g, 3.9 mmol) in CH.sub.3CN (20 mL) was added K.sub.2CO.sub.3 (1.1 g, 7.9 mmol) and 1-chloropropan-2-one at (0.55 g, 5.9 mmol) room temperature and the resulting mixture was stirred at 60 C. for 15 hours. The reaction mixture was concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (0.8 g, 86%). MS: M/e 234 (M+1).sup.+.
Step C: 6-chloro-4-fluoro-5-(2-oxopropoxy)picolinaldehyde
##STR00531##
[0765] To a solution of 1-((2-chloro-4-fluoro-6-(hydroxymethyl)pyridin-3-yl)oxy)propan-2-one (0.8 g, 3.4 mmol) in DCM (50 mL) was added Dess-Martin periodinane (2.2 g, 51 mmol) at room temperature and the resulting mixture was stirred at room temperature for 1.5 hours. The reaction mixture was quenched with saturated NaHCO.sub.3 aq., extracted with EtOAc combined, washed brine, dried and concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (320 mg, 40%). MS: M/e 232 (M+1).sup.+.
Step D: 1-((2-chloro-4-fluoro-6-(1-hydroxyethyl)pyridin-3-yl)oxy)-2-methylpropan-2-ol
##STR00532##
[0766] To a solution of 6-chloro-4-fluoro-5-(2-oxopropoxy)picolinaldehyde (320 mg, 1.38 mmol) in THE (40 mL) was added CH.sub.3MgBr (3M, 2.3 mL, 6.9 mmol) at room temperature and the resulting mixture was stirred at room temperature for 1 hour. The reaction mixture was quenched with saturated NH.sub.4Cl aq. (50 mL), extracted with EtOAc washed brine, dried and concentrated to give the titled compound (350 mg, 96%). MS: M/e 264 (M+1).sup.+.
Step E: 1-(8-fluoro-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethan-1-ol
##STR00533##
[0767] A mixture of 1-((2-chloro-4-fluoro-6-(1-hydroxyethyl)pyridin-3-yl)oxy)-2-methylpropan-2-ol (350 mg, 1.3 mmol), BINAP (165 mg, 0.25 mmol), Cs.sub.2CO.sub.3 (0.86 g, 2.6 mmol) and Pd(OAc).sub.2 (30 mg, 0.13 mmol) in toluene (10 mL) was stirred at 100 C. under N.sub.2 for 15 hours. The reaction mixture was concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (170 mg, 56%). MS: M/e 228 (M+1).sup.+.
Step F: 2-(9-ethyl-6-((2S,5R)-4-(1-(8-fluoro-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00534##
[0768] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (120 mg, 0.36 mmol) in CH.sub.3CN (8 mL) and was added 1-(8-fluoro-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethan-1-ol (124 mg, 0.54 mmol), (cyanomethyl)trimethylphosphonium iodide (353 mg, 1.4 mmol) and DIPEA (464 mg, 3.6 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure and purified by by flash column chromatography (DCM/MeOH) to give the titled Compound A163, which was further separated into Compound A163a (44 mg) and Compound A163b (42 mg) by chiral Prep-HPLC. The chiral separation conditions are shown below.
TABLE-US-00025 Column CHIRALPAK IG Column Size 2 cm 25 cm, 5 um Mobile Phase A MtBE Mobile Phase B EtOH Flow Rate 20 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-HPLC Equipment Prep-HPLC-Gilson
[0769] Compound A163: .sup.1H NMR (400 MHz, CD.sub.3OD) 7.05 (t, J=9.5 Hz, 1H), 6.12-4.99 (m, 2H), 4.37 (d, J=6.9 Hz, 2H), 4.22 (s, 1H), 4.01 (s, 2H), 3.76 (s, 3H), 3.69-3.33 (m, 3H), 3.07-2.10 (m, 3H), 1.51-1.39 (m, 10H), 1.37-1.23 (m, 5H), 1.07-0.89 (m, 3H) ppm. MS: M/e 539 (M+1).sup.+.
[0770] Compound A163a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.04 (d, J=10.1 Hz, 1H), 6.28-4.87 (m, 2H), 4.52-4.15 (m, 3H), 4.01 (s, 2H), 3.76 (s, 3H), 3.69-3.32 (m, 3H), 2.89 (s, 2H), 2.78 (d, J=10.9 Hz, 1H), 1.54-1.37 (m, 12H), 1.26 (d, J=6.1 Hz, 3H), 0.94 (d, J=5.9 Hz, 3H) ppm. MS: M/e 539 (M+1).sup.+.
[0771] Compound A163b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.06 (d, J=10.6 Hz, 1H), 6.14-4.91 (m, 2H), 4.58-4.09 (m, 3H), 4.01 (s, 2H), 3.76 (s, 3H), 3.74-3.64 (m, 1H), 3.46 (d, J=52.3 Hz, 3H), 2.72 (s, 1H), 2.17 (d, J=11.9 Hz, 1H), 1.54-1.25 (m, 15H), 1.01 (d, J=4.5 Hz, 3H) ppm. MS: M/e 539 (M+1).sup.+.
Compound A164: 2-(6-((2S,5R)-4-(1-(8-fluoro-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00535##
[0772] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.16 mmol) in CH.sub.3CN (8 mL) and was added 1-(8-fluoro-3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethan-1-ol (54 mg, 0.24 mmol), (cyanomethyl)trimethylphosphonium iodide (115 mg, 0.48 mmol) and DIPEA (204 mg, 9 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure and purified by flash column chromatography (DCM/MeOH) to give the titled compound (12 mg). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.05 (t, J=9.1 Hz, 1H), 6.18-4.95 (m, 2H), 4.21 (s, 1H), 4.01 (s, 2H), 3.93 (s, 3H), 3.79 (s, 3H), 3.73-3.33 (m, 3H), 3.10-2.03 (m, 3H), 1.42 (s, 6H), 1.36-1.24 (m, 6H), 1.05-0.90 (m, 3H) ppm. MS: M/e 525 (M+1).sup.+.
Compound A165: 2-(9-(2,2-difluoroethyl)-6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00536##
Step A: tert-butyl (2R,5S)-4-(8-((benzyloxy)methyl)-9-(2,2-difluoroethyl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00537##
[0773] To a stirred mixture of 8-((benzyloxy)methyl)-9-(2,2-difluoroethyl)-6-hydroxy-3-methyl-3,9-dihydro-2H-purin-2-one (1.5 g, 4.28 mmol) and tert-butyl (2R,5S)-2,5-dimethylpiperazine-1-carboxylate (1.83 g, 8.57 mmol) in CH.sub.3CN (15 mL) was added BOP (4.60 g, 8.57 mmol), followed by DBU (1.3 g, 8.57 mmol). After the addition, the reaction mixture was concentrated to give the residue, which was dissolved in EtOAc (50 mL), washed with H.sub.2O (20 mL), brine, dried over Na.sub.2SO.sub.4, concentrated and purified by flash column chromatography to give the titled compound (1.8 g, 77%). MS: M/e 547 (M+1).sup.+.
Step B: tert-butyl (2R,5S)-4-(9-(2,2-difluoroethyl)-8-(hydroxymethyl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00538##
[0774] To a stirred solution of tert-butyl (2R,5S)-4-(8-((benzyloxy)methyl)-9-(2,2-difluoroethyl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (1.8 g, 3.3 mmol) in MeOH/AcOH (80 mL/2 mL) was added Pd/C (200 mg). After the addition, the reaction mixture was stirred over a weekend under H.sub.2 (4 atm). The reaction mixture was filtered, and filtrate was concentrated to give the titled compound (1.22 g, 81%). MS: M/e 457 (M+1).sup.+.
Step C: tert-butyl (2R,5S)-4-(8-(cyanomethyl)-9-(2,2-difluoroethyl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00539##
[0775] To a stirred solution of tert-butyl (2R,5S)-4-(9-(2,2-difluoroethyl)-8-(hydroxymethyl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (1.22 g, 2.68 mmol) in CH.sub.2Cl.sub.2 (50 mL) was added SOCl.sub.2 (319 mg, 2.68 mmol). After stirred for 10 min, the reaction mixture was quenched with aq.NaHCO.sub.3, washed with brine, dried over Na.sub.2SO.sub.4, concentrated to give the intermediate, dissolved in CH.sub.2Cl.sub.2 (10 mL), TBAF (1.0 M, 3.2 mL, 3.2 mmol) was added followed by TMSCN (0.32 g, 3.2 mmol). After the addition, the reaction mixture was stirred for 3 hours. The reaction mixture was concentrated to give the residue, treated with H.sub.2O and extracted with EtOAc (15 mL3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, concentrated and purified by flash column chromatography to give the titled compound (900 mg, 72%). MS: M/e 466 (M+1).sup.+.
Step D: 2-(9-(2,2-difluoroethyl)-6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00540##
[0776] To a stirred solution of tert-butyl (2R,5S)-4-(8-(cyanomethyl)-9-(2,2-difluoroethyl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-6-yl)-2,5-dimethylpiperazine-1-carboxylate (900 mg, 1.93 mmol) in EtOAc (10 mL) was added dioxane/HCl (g) (4.0 M, 4 mL). After the addition, the reaction mixture was stirred for 4 hours. The reaction mixture was concentrated to give the residue, which was treated with CH.sub.3CN/aq.NaHCO.sub.3 (20 mL/3 mL), stirred for 10 min and filtered. The filtrate was concentrated, dried to give the titled compound (560 mg, 79%), which was directly used to the next step. MS: M/e 366 (M+1).sup.+.
Step E: 2-(9-(2,2-difluoroethyl)-6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00541##
[0777] To a solution of 2-(9-(2,2-difluoroethyl)-6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.14 mmol) in CH.sub.3CN (3 mL) and was added 1-(3,3-dimethyl-2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)ethan-1-ol (29 mg, 0.14 mmol), (cyanomethyl)trimethylphosphonium iodide (100 mg, 0.41 mmol) and DIPEA (88 mg, 0.68 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure to dryness. The resulting residue was purified by Prep-HPLC (Method A) to give the titled compound (15 mg, 20%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.31 (d, J=7.6 Hz, 1H), 7.12 (d, J=8.1 Hz, 1H), 6.33 (t, J=54.0 Hz, 1H), 5.93-5.06 (m, 2H), 4.93 (s, 2H), 4.21 (s, 1H), 3.96 (s, 2H), 3.72 (s, 3H), 3.70-3.32 (m, 3H), 3.05-2.26 (m, 3H), 1.40 (s, 6H), 1.35 (dd, J=16.3, 5.8 Hz, 6H), 1.13-0.96 (m, 3H) ppm. MS: M/e 557 (M+1).sup.+.
Compound A166: 2-(9-(2,2-difluoroethyl)-6-((2S,5R)-4-(1-(2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00542##
[0778] To a solution of 2-(9-(2,2-difluoroethyl)-6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.14 mmol) in CH.sub.3CN (3 mL) and was added 1-(2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethan-1-ol (27 mg, 0.14 mmol), (cyanomethyl)trimethylphosphonium iodide (100 mg, 0.41 mmol) and DIPEA (88 mg, 0.68 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure to dryness. The resulting residue was purified by Prep-HPLC (Method A) to give the titled compound (12 mg, 16%). .sup.1H NMR (400 MHz, CD.sub.3OD) 6.83 (s, 1H), 6.76 (s, 1H), 6.65 (t, J=8.9 Hz, 1H), 6.33 (t, J=53.8 Hz, 1H), 6.14-4.98 (m, 2H), 4.93 (s, 2H), 4.86-4.79 (m, 1H), 4.21 (s, 1H), 3.72 (s, 3H), 3.64-3.41 (m, 2H), 3.05-2.29 (m, 3H), 1.64 (s, 6H), 1.47-1.28 (m, 6H), 1.09-0.90 (m, 3H) ppm. MS: M/e 542 (M+1).sup.+.
Compound A167: 2-(6-((2S,5R)-4-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethyl piperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00543##
[0779] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3,9-dimethyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (100 mg, 0.32 mmol), 1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)ethan-1-ol (64 mg, 0.32 mmol), (cyanomethyl)trimethylphosphonium iodide (230 mg, 0.95 mmol) and DIPEA (205 mg, 1.59 mmol) in MeCN (5 ml) was stirred at 100 C. overnight. After completed, the solution was concentrated under reduced pressure to dryness. The resulting residue containing Compound A167 was purified by Prep-HPLC (Method A) to give the titled Compound A167 (46 mg, 29%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.29 (s, 1H), 7.21-7.08 (m, 2H), 6.18-4.89 (m, 2H), 4.21 (s, 1H), 3.93 (s, 3H), 3.78 (s, 3H), 3.68-3.48 (m, 2H), 3.29-2.52 (m, 4H), 1.59-1.32 (m, 3H), 1.31-1.21 (m, 3H), 1.05-0.88 (m, 3H) ppm. MS: M/e 500 (M+1).sup.+
Compound A168: 2-(9-ethyl-6-((2S,5R)-4-(1-(2-ethyl-2-methylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00544##
Step A: 5-bromo-2-ethyl-2-methylbenzo[d][1,3]dioxole
##STR00545##
[0780] A solution of 4-bromobenzene-1,2-diol (0.5 g, 2.65 mmol), butan-2-one (381 mg, 5.29 mmol) and PBr.sub.3 (287 mg, 1.06 mmol) in Tol. (6 ml) was stirred at 100 C. for 1 h. The solution was diluted with EtOAc (20 ml), washed with brine (10 ml), dried over Na.sub.2SO.sub.4 and evaporated to dryness to give the titled compound (0.64 g, 100%). .sup.1H NMR (400 MHz, CDCl.sub.3) 6.92-6.83 (m, 2H), 6.59 (d, J=8.3 Hz, 1H), 1.94 (q, J=7.1 Hz, 2H), 1.60 (s, 3H), 1.00 (t, J=7.4 Hz, 3H) ppm.
Step B: 1-(2-ethyl-2-methylbenzo[d][1,3]dioxol-5-yl)ethan-1-one
##STR00546##
[0781] A solution of 5-bromo-2-ethyl-2-methylbenzo[d][1,3]dioxole (640 mg, 2.63 mmol), Pd(PPh.sub.3).sub.2Cl.sub.2 (92 mg, 0.13 mmol) and tributyl(1-ethoxyvinyl)stannane (1.43 g, 3.96 mmol) in Tol. (10 ml) was stirred at 100 C. overnight. The mixture was cooled to RT and HCl (4M in dioxane, 2 ml) was added, stirred for 30 min. The mixture was diluted with EtOAc (20 ml), washed with brine (10 ml), dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by flash column chromatography with 0%-10% EtOAc in PE to give the titled compound (160 mg, 29%).
[0782] .sup.1H NMR (400 MHz, CDCl.sub.3) 7.51 (d, J=8.8 Hz, 1H), 7.35 (s, 1H), 6.75 (d, J=7.7 Hz, 1H), 2.53 (s, 3H), 1.97 (q, J=7.3 Hz, 2H), 1.64 (s, 3H), 1.01 (t, J=7.3 Hz, 3H) ppm.
Step C: 1-(2-ethyl-2-methylbenzo[d][1,3]dioxol-5-yl)ethan-1-ol
##STR00547##
[0783] A solution of 1-(2-ethyl-2-methylbenzo[d][1,3]dioxol-5-yl)ethan-1-one (160 mg, 0.78 mmol) and NaBH.sub.4 (29.5 mg, 0.78 mmol) in MeOH (3 ml) and THE (1 ml) was stirred at RT for 10 min. The solution was quenched with water (10 ml), diluted with EtOAc (15 ml), washed with brine (10 ml), dried and evaporated to dryness to give the titled compound (160 mg, 100%), which was used directly for the next step. MS: M/e 209 (M+1).sup.+
Step D: 2-(9-ethyl-6-((2S,5R)-4-(1-(2-ethyl-2-methylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00548##
[0784] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (100 mg, 0.30 mmol), 1-(2-ethyl-2-methylbenzo[d][1,3]dioxol-5-yl)ethan-1-ol (160 mg, 0.77 mmol), (cyanomethyl)trimethylphosphonium iodide (222 mg, 0.91 mmol) and DIPEA (392 mg, 3.04 mmol) in MeCN (3 ml) was stirred at 100 C. overnight. The reaction was diluted with EtOAc (10 ml) and washed with brine (10 ml). The organic layer was concentrated under reduced pressure. The resulting residue was purified by flash column chromatography with 0-5% MeOH in DCM and then prep-HPLC (Method A) to give the titled compound (53 mg). .sup.1H NMR (400 MHz, CD.sub.3OD) 6.97-6.79 (m, 2H), 6.78-6.68 (m, 1H), 6.34-5.65 (m, 1H), 5.49-5.01 (m, 1H), 4.86-4.85 (m, 3H), 4.37 (q, J=6.8 Hz, 2H), 4.24 (s, 1.5H), 3.77 (s, 4H), 3.48 (s, 1H), 3.11-2.90 (m, 0.5H), 1.96 (s, 2H), 1.60 (s, 3H), 1.56-1.40 (m, 7H), 1.35-0.96 (m, 8H) ppm. MS: M/e 520 (M+1).sup.+.
Compound A169: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(spiro[benzo[d][1,3]dioxole-2,1-cyclobutan]-5-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00549##
Step A: 5-bromospiro[benzo[d][1,3]dioxole-2,1-cyclobutane]
##STR00550##
[0785] To a solution of 4-bromobenzene-1,2-diol (1.89 g, 10 mol) in toluene (8 mL) was added cyclobutanone (1.4 g, 20 mol) and PBr.sub.3 (1.62 g, 6 mmol). The reaction mixture was sealed and stirred at 80 C. for overnight. The mixture was cooled down to RT and added H.sub.2O. The mixture was extracted with EtOAc and then concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (1.7 g, 71%). MS: M/e 241 (M+1).sup.+.
Step B: 1-(spiro[benzo[d][1,3]dioxole-2,1-cyclobutan]-5-yl)ethan-1-one
##STR00551##
[0786] To a solution of 5-bromospiro[benzo[d][1,3]dioxole-2,1-cyclobutane](570 mg, 2.4 mol) in toluene (10 mL) was added Pd(PPh.sub.3).sub.2Cl.sub.2 (210 mg, 0.3 mol) and tributyl(1-ethoxyvinyl)stannane (1.3 g, 3.6 mmol). The reaction mixture was protected by N.sub.2 atmosphere and stirred at 100 C. for overnight. The mixture was added H.sub.2O extracted with EtOAc. The organic phase was dried with Na.sub.2SO.sub.4 and concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (500 mg, 100%). MS: M/e 205 (M+1).sup.+.
Step C: 1-(spiro[benzo[d][1,3]dioxole-2,1-cyclobutan]-5-yl)ethan-1-ol
##STR00552##
[0787] To a solution of 1-(spiro[benzo[d][1,3]dioxole-2,1-cyclobutan]-5-yl)ethan-1-one (500 mg, 2.5 mol) in MeOH was added NaBH.sub.4 (38 mg, 1 mmol). The reaction mixture was stirred at RT for 15 mins. The mixture was added H.sub.2O and extracted with DCM. The organic phase was dried with Na.sub.2SO.sub.4 and concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (370 mg, 74%). MS: M/e 207 (M+1).sup.+.
Step D: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(spiro[benzo[d][1,3]dioxole-2,1-cyclobutan]-5-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00553##
[0788] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (480 mg, 1.5 mmol), 1-(spiro[benzo[d][1,3]dioxole-2,1-cyclobutan]-5-yl)ethan-1-ol (450 mg, 2.2 mmol) and (cyanomethyl)trimethylphosphonium iodide (723 mg, 3 mmol) in CH.sub.3CN (10 mL) was added DIPEA (774 mg, 6 mmol). The reaction mixture was sealed in a bottle and heated at 105 C. for 16 hours, and then cooled to room temperature, diluted with water, extracted with EtOAc. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by Prep-HPLC (Method A) to give the titled Compound A169 (200 mg, 26%), which was further separated into Compound A169a (65 mg) and Compound A169b (60 mg) by chiral Prep-HPLC. The chiral separation conditions are shown below.
TABLE-US-00026 Column CHIRALPAK IH Column Size 2 cm 25 cm, 5 um Mobile Phase A MtBE Mobile Phase B EtOH Flow Rate 20 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-HPLC Equipment Prep-HPLC-Gilson
[0789] Compound A169: .sup.1H NMR (400 MHz, DMSO-d6) 6.89 (d, J=5.7 Hz, 1H), 6.79 (d, J=4.1 Hz, 2H), 6.25-5.30 (m, 1H), 5.30-4.50 (m, 1H), 4.40 (s, 2H), 4.27-4.23 (m, 2H), 3.61 (s, 3H), 3.50-3.45 (m, 1H), 3.30-3.28 (m, 1H), 2.70 (d, J=26.8 Hz, 2H), 2.60-2.55 (m, 4H), 2.33-2.18 (m, 1H), 1.79 (d, J=7.5 Hz, 2H), 1.34 (s, 4H), 1.26-1.13 (m, 5H), 0.93 (s, 1.5H), 0.81 (s, 1.5H) ppm. MS: M/e 518 (M+1).sup.+.
[0790] Compound A169a (the earlier peak): .sup.1HNMR (400 MHz, CD.sub.3OD) 6.85 (s, 1H), 6.77 (d, J=7.3 Hz, 1H), 6.68 (d, J=7.8 Hz, 1H), 6.1-5.64 (m, 1H), 5.21-4.60 (m, 1H), 4.36 (d, J=6.6 Hz, 2H), 3.76 (s, 3H), 3.60-3.40 (m, 2H), 3.30-3.13 (m, 2H), 3.00-2.65 (m, 3H), 2.62-2.58 (m, 4H), 1.90-1.80 (m, 2H), 1.46 (t, J=6.7 Hz, 6H), 1.27 (s, 3H), 0.88 (s, 3H) ppm. MS: M/e 518 (M+1).sup.+.
[0791] Compound A169b (the later peak): .sup.1HNMR (400 MHz, CD.sub.3OD) 6.85 (s, 1H), 6.76 (d, J=7.6 Hz, 1H), 6.66 (d, J=7.4 Hz, 1H), 6.10-5.60 (m, 1H), 5.25-4.50 (m, 1H), 4.37 (q, J=6.9 Hz, 2H), 3.76 (s, 4H), 3.56-3.37 (m, 2H), 3.20-3.00 (m, 1H), 2.59 (d, J=4.3 Hz, 6H), 2.26 (d, J=10.3 Hz, 1H), 1.90-1.80 (m, 2H), 1.46 (t, J=7.1 Hz, 3H), 1.29 (s, 6H), 1.02 (s, 3H) ppm. MS: M/e 518 (M+1).sup.+.
Compound A172: 2-(6-((2S,5R)-4-1-(4-chloro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00554##
Step A: 5-bromo-4-chloro-2,2-dimethylbenzo[d][1,3]dioxole
##STR00555##
[0792] To a solution of 5-bromo-2,2-dimethylbenzo[d][1,3]dioxole (0.92 g, 4 mmol) in THE (10 mL) was added a solution of LDA in THE (2.6 ml, 5.2 mmol) drop wise at 70 C. After 1 h, a solution of 1,1,2-trichloro-1,2,2-trifluoroethane (1.12 g, 6 mmol) in THE (1 mL) was added. The reaction mixture was slowly warmed to room temperature over 16 h. The mixture was diluted with water, extracted with EtOAc (60 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography to give the titled compound (1 g, 95%). .sup.1H NMR (400 MHz, CDCl.sub.3) 7.07 (d, J=6.4 Hz, 1H), 6.56 (d, J=6.8 Hz, 1H), 1.74 (s, 6H) ppm.
Step B: 1-(4-chloro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethan-1-one
##STR00556##
[0793] To a solution of 5-bromo-4-chloro-2,2-dimethylbenzo[d][1,3]dioxole (2.3 g, 8.77 mmol) in toluene (20 mL) was added tributyl(1-ethoxyvinyl)stannane (4.15 g, 13.2 mmol) and Pd(PPh.sub.3).sub.2Cl.sub.2 (310 mg, 0.44 mmol). The reaction mixture was protected by N.sub.2 atmosphere and stirred at 100 C. overnight. The mixture was cooled down to RT, diluted with H.sub.2O, extracted with EtOAc (80 mL3), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography to give the titled compound (1.2 g, 60%). MS: M/e 227 (M+1).sup.+.
Step C: 1-(4-chloro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethan-1-ol
##STR00557##
[0794] To a solution of 1-(4-chloro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethan-1-one (1.2 g, 5.3 mmol) in MeOH (15 mL) was added NaBH.sub.4 (160 mg, 4.24 mmol) slowly at 0 C. The resulting mixture was stirred at room temperature for 0.5 hour. The reaction mixture was quenched with H.sub.2O (10 mL) and extracted with EtOAc (100 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography to give the titled compound (700 mg, 58%). .sup.1H NMR (400 MHz, DMSO-d6) 7.01 (d, J=8.0 Hz, 1H), 6.81 (d, J=8.4 Hz, 1H), 5.21 (s, 1H), 4.95-4.85 (m, 1H), 1.68 (s, 3H), 1.66 (s, 3H), 1.27 (d, J=6.0 Hz, 3H) ppm.
Step D: 2-(6-((2S,5R)-4-1-(4-chloro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00558##
[0795] To a solution of 1-(4-chloro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethan-1-ol (340 mg, 1.5 mmol), 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (450 mg, 1.5 mmol) and (cyanomethyl) trimethyl phosphonium iodide (1.09 g, 4.5 mmol) in CH.sub.3CN (10 mL) was added DIPEA (1.9 g, 15 mmol). The mixture was sealed in a bottle and heated at 100 C. for 16 hours. The mixture was cooled to room temperature, diluted with water, extracted with EtOAc (80 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by combi flash with 0-5% MeOH in DCM to give the titled Compound A172 (260 mg, 23%), which was further separated into Compound A172a (80 mg) and Compound A172b (73 mg) by chiral-Prep-HPLC. The chiral separation conditions are shown below.
TABLE-US-00027 Column CHIRALPAK IA Column Size 2 cm 25 cm, 5 um Mobile Phase A MtBE Mobile Phase B MeOH:DCM Flow Rate 20 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-HPLC Equipment Prep-HPLC-Gilson
[0796] Compound A172a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.09 (d, J=8.0 Hz, 1H), 6.71 (d, J=8.0 Hz, 1H), 6.25-6.05 (m, 0.5H), 5.75-5.60 (m, 0.5H), 5.28-5.14 (m, 0.5H), 4.70-4.55 (m, 0.5H), 4.45-4.31 (m, 2H), 4.26-3.99 (m, 2H), 3.76 (s, 3H), 3.61-3.45 (m, 0.5H), 3.34-3.25 (m 1H), 3.27-3.14 (m, 0.5H), 3.04-2.72 (m 3H), 1.70 (s, 6H), 1.56-1.32 (m, 6H), 1.23 (d, J=6.4 Hz, 3H), 0.91 (d, J=4.8 Hz, 3H) ppm. MS: M/e 540 (M+1).sup.+.
[0797] Compound A172a (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.16 (d, J=8.4 Hz, 1H), 6.70 (d, J=8.0 Hz, 1H), 6.11-5.70 (m, 1H), 5.20-4.98 (m, 1H), 4.43-4.32 (m, 2H), 4.27-4.18 (m, 1H), 4.00-3.91 (m, 1H), 3.76 (m, 3H), 3.65-3.50 (m, 1H), 3.34-3.25 (m 2H), 2.79-2.60 (m, 1H), 2.31-2.21 (m 1H), 1.69 (s, 6H), 1.46 (t, J=6.8 Hz, 3H), 1.29 (d, J=6.4 Hz, 6H), 1.05 (d, J=5.6 Hz, 3H) ppm. MS: M/e 540 (M+1).sup.+.
Compound A173: 2-(9-ethyl-6-((2S,5R)-4-(1-(7-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00559##
Step A: 6-bromo-4-fluoro-2,2-dimethylbenzo[d][1,3]dioxole
##STR00560##
[0798] To a solution of 5-bromo-3-fluorobenzene-1,2-diol (1.65 g, 8 mmol) in toluene (10 mL) was added acetone (0.93 g, 16 mmol) and POBr.sub.3 (2.28 g, 8 mmol). The reaction mixture was stirred at 80 C. in a sealed bottle overnight. The mixture was cooled down to RT. Weight 6 eq of Na.sub.2CO.sub.3, treated with ice and EtOAc. The reaction solution was added drop wise to keep the temperature below 10 C. (pH>8). extracted with EtOAc (80 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography to give the titled compound (1.2 g, 61%). .sup.1H NMR (400 MHz, CDCl.sub.3) 6.81 (d, J=9.6 Hz, 1H), 6.71 (s, 1H), 1.71 (s, 6H) ppm.
Step B: 7-fluoro-2,2-dimethylbenzo[d][1,3]dioxole-5-carbaldehyde
##STR00561##
[0799] To a solution of 6-bromo-4-fluoro-2,2-dimethylbenzo[d][1,3]dioxole (1.2 g, 4.85 mmol) and 3-oxobenzo[d]isothiazole-2 (3H)-carbaldehyde 1,1-dioxide (1.54 g, 7.28 mmol) in DMF (20 mL) was added Pd(OAc).sub.2 (107 mg, 0.48 mmol), 1,4-Bis(diphenylphosphino)butane (413 mg, 0.97 mmol), Na.sub.2CO.sub.3 (1.02 g, 9.7 mmol) and Et.sub.3SiH (675 mg, 5.82 mmol). The reaction was heated at 85 C. under N.sub.2 overnight. Then the reaction mixture was cooled to room temperature, diluted with H.sub.2O, extracted with EtOAc (60 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography to give the titled compound (700 mg, crude). .sup.1H NMR (400 MHz, CDCl.sub.3) 9.76 (s, 1H), 7.22 (d, J=8.8 Hz, 1H), 7.11 (s, 1H), 1.76 (s, 6H) ppm.
Step C: 1-(7-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethan-1-ol
##STR00562##
[0800] To a solution of 7-fluoro-2,2-dimethylbenzo[d][1,3]dioxole-5-carbaldehyde (670 mg, 3.4 mmol) in THE (7 mL) was added CH.sub.3MgBr (1.2 mL, 3.6 mmol, 3 mol/L in hexane) slowly at 0 C. The resulting mixture was stirred at room temperature for 0.5 hour. The reaction mixture was quenched with H.sub.2O (10 mL) and extracted with EtOAc (80 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The residue was purified by flash column chromatography to give the titled compound (530 mg, 51% yield for two steps). .sup.1H NMR (400 MHz, DMSO-d6) 6.72 (d, J=11.6 Hz, 1H), 6.69 (s, 1H), 5.17 (s, 1H), 4.66-4.54 (m, 1H), 1.67 (s, 6H), 1.26 (d, J=6.0 Hz, 3H) ppm.
Step D: 2-(9-ethyl-6-((2S,5R)-4-(1-(7-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00563##
[0801] To a solution of 1-(7-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethan-1-ol (530 mg, 2.5 mmol), 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (800 mg, 2.5 mmol) and (cyanomethyl) trimethyl phosphonium iodide (1.82 g, 7.5 mmol) in CH.sub.3CN (10 mL) was added DIPEA (3.2 g, 25 mmol). The mixture was sealed in a bottle and heated at 100 C. for 16 hours. The mixture was cooled to room temperature, diluted with water, extracted with EtOAc (100 mL2), washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by combi flash (DCM/MeOH=20/1) to give the titled Compound A173 (300 mg, 23%), which was further separated into Compound A173a (110 mg) and Compound A173b (125 mg) by chiral-Prep-HPLC. The chiral separation conditions are shown below.
TABLE-US-00028 Column CHIRALPAK IH Column Size 2 cm 25 cm, 5 um Mobile Phase A MtBE Mobile Phase B EtOH Flow Rate 20 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-HPLC Equipment Prep-HPLC-Gilson
[0802] Compound A173: .sup.1H NMR (400 MHz, CD.sub.3OD) 6.74-6.62 (m, 2H), 6.19-5.65 (m, 1H), 5.31-4.95 (m, 1H), 4.43-4.31 (m, 2H), 4.26-4.18 (m, 1H), 3.76 (s, 3H), 3.60-3.44 (m, 2H), 3.35-3.31 (m, 1H), 3.01-2.57 (m, 2.5H), 2.30-2.16 (m, 0.5H), 1.68 (s, 6H), 1.53-1.39 (m, 4.5H), 1.33-1.22 (m, 4.5H), 1.21 (d, J=6.0 Hz, 1.5H), 0.90 (d, J=6.4 Hz, 1.5H) ppm. MS: M/e 524 (M+1).sup.+.
[0803] Compound A173a (the earlier peak): .sup.1H NMR (400 MHz, DMSO-d6) 6.76 (d, J=11.3 Hz, 1H), 6.73 (s, 1H), 6.02-4.41 (m, 2H), 4.40-4.31 (m, 2H), 4.30-4.12 (m, 2H), 3.58 (s, 3H), 3.52-3.38 (m, 1H), 3.28-2.96 (m, 1H), 2.85-2.63 (m, 3H), 1.68-1.64 (m, 6H), 1.37-1.25 (m, 6H), 1.16 (d, J=6.4 Hz, 3H), 0.80 (d, J=6.0 Hz, 3H) ppm. MS: M/e 524 (M+1).sup.+.
[0804] Compound A173b (the later peak): .sup.1H NMR (400 MHz, DMSO-d6) 6.75 (d, J=11.3 Hz, 1H), 6.70 (s, 1H), 5.85-4.45 (m, 2H), 4.38 (s, 2H), 4.28-4.18 (m, 2H), 3.58 (s, 3H), 3.48-3.34 (m, 2H), 3.29-3.20 (m, 1H), 2.60-2.53 (m, 1H), 2.20-2.07 (m, 1H), 1.65 (s, 6H), 1.31 (t, J=7.1 Hz, 3H), 1.24-1.07 (m, 6H), 0.95-0.85 (m, 3H) ppm. MS: M/e 524 (M+1).sup.+.
Compound A174: 2-(9-ethyl-6-((2S,5R)-4-(1-(6-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00564##
Step A: 4-bromo-5-fluorobenzene-1,2-diol
##STR00565##
[0805] To a solution of 1-bromo-2-fluoro-4,5-dimethoxybenzene (235 mg, 1 mol) in DCM (15 mL) was added BBr.sub.3 (1M, 4 mmol, 4 mL). The reaction mixture was stirred at RT for 2 hrs. The mixture was added H.sub.2O and extracted with EtOAc and then concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (200 mg, 98%). MS: M/e 206 (M+1).sup.+.
Step B: 5-bromo-6-fluoro-2,2-dimethylbenzo[d][1,3]dioxole
##STR00566##
[0806] To a solution of 4-bromo-5-fluorobenzene-1,2-diol (501 mg, 2.43 mol) in toluene (5 mL) was added acetone (278 mg, 4.8 mol) and PCl.sub.3 (137 mg, 0.97 mmol). The reaction mixture was sealed and stirred at 80 C. for overnight. The mixture was cooled down to RT and added NaHCO.sub.3 aqueous solution. The mixture was extracted with EtOAc and then concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (248 mg, 42%). MS: M/e 247 (M+1).sup.+.
Step C: 1-(6-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethan-1-one
##STR00567##
[0807] To a solution of 5-bromo-6-fluoro-2,2-dimethylbenzo[d][1,3]dioxole (120 mg, 0.5 mol) in toluene (10 mL) was added Pd(PPh.sub.3).sub.2Cl.sub.2 (7 mg, 0.01 mol) and tributyl(1-ethoxyvinyl)stannane (252 mg, 0.7 mmol). The reaction mixture was protected by N.sub.2 atmosphere and stirred at 100 C. for overnight. The mixture was added 4N Hcl in dionxane and stirred at RT for 5 mins. The mixture was added saturated NaHCO.sub.3 aqueous solution and extracted with EtOAc. The organic phase was dried with Na.sub.2SO.sub.4 and concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (30 mg, 29%). MS: M/e 211 (M+1).sup.+.
Step D: 1-(6-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethan-1-ol
##STR00568##
[0808] To a solution of 1-(6-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethan-1-one (30 mg, 0.14 mol) in MeOH was added NaBH.sub.4 (38 mg, 1 mmol). The reaction mixture was stirred at RT for 15 mins. The mixture was added H.sub.2O and extracted with DCM. The organic phase was dried with Na.sub.2SO.sub.4 and concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (10 mg, 33%). MS: M/e 213 (M+1).sup.+.
Step D: 2-(9-ethyl-6-((2S,5R)-4-(1-(6-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00569##
[0809] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (30 mg, 0.14 mmol), 1-(6-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethan-1-ol (10 mg, 0.05 mmol) and (cyanomethyl)trimethylphosphonium iodide (120 mg, 0.5 mmol) in CH.sub.3CN (5 mL) was added DIPEA (258 mg, 2 mmol). The reaction mixture was sealed in a bottle and heated at 105 C. for 16 hours, and then cooled to room temperature, diluted with water, extracted with EtOAc. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified and separated into Compound A174a (0.7 mg) and Compound A174b (0.6 mg) by Prep-HPLC (Method B).
[0810] Compound A174a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.93 (d, J=5.2 Hz, 1H), 6.50 (d, J=9.7 Hz, 1H), 6.10-5.70 (m, 1H), 5.20-4.50 (m, 1H), 4.37 (d, J=7.1 Hz, 2H), 4.23 (s, 1H), 3.80-3.76 (m, 1H), 3.76 (s, 3H), 3.60-3.54 (m, 1H), 3.30-3.13 (m, 1H), 2.90-2.50 (m, 1H), 2.65 (s, 1H), 2.31 (d, J=12.4 Hz, 1H), 1.64 (d, J=6.4 Hz, 6H), 1.46 (t, J=6.8 Hz, 3H), 1.31 (d, J=6.4 Hz, 6H), 1.04 (d, J=5.2 Hz, 3H) ppm. MS: M/e 524 (M+1).sup.+.
[0811] Compound A174b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.87 (d, J=5.4 Hz, 1H), 6.52 (d, J=9.5 Hz, 1H), 6.30-5.50 (m, 1H), 5.50-4.50 (m, 1H), 4.37 (d, J=7.0 Hz, 2H), 4.22 (s, 1H), 3.98 (d, J=5.8 Hz, 1H), 3.76 (s, 3H), 3.55-3.10 (m, 2H), 2.87 (d, J=36.8 Hz, 3H), 1.66 (d, J=8.7 Hz, 6H), 1.49-1.40 (m, 6H), 1.28 (d, J=6.2 Hz, 3H), 0.92 (d, J=5.5 Hz, 3H) ppm. MS: M/e 524 (M+1).sup.+.
Compound A175: 2-(9-ethyl-6-((2S,5R)-4-(1-(4-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00570##
Step A: 4-bromo-3-fluorobenzene-1,2-diol
##STR00571##
[0812] To a solution of 4-bromo-3-fluoro-2-methoxyphenol (442 mg, 2 mol) in DCM (15 mL) was added BBr.sub.3 (1M, 4 mmol, 4 mL). The reaction mixture was stirred at RT for 2 hrs. The mixture was added H.sub.2O and extracted with EtOAc and then concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (350 mg, 85%). MS: M/e 206 (M+1).sup.+.
Step B: 5-bromo-4-fluoro-2,2-dimethylbenzo[d][1,3]dioxole
##STR00572##
[0813] To a solution of 4-bromo-3-fluorobenzene-1,2-diol (280 mg, 1.35 mol) in toluene (5 mL) was added acetone (156 mg, 2.7 mol) and PCl.sub.3 (95 mg, 0.72 mmol). The reaction mixture was sealed and stirred at 80 C. for overnight. The mixture was cooled down to RT and added NaHCO.sub.3 aqueous solution. The mixture was extracted with EtOAc and then concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (180 mg, 54%). MS: M/e 247 (M+1).sup.+.
Step C: 1-(4-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethan-1-one
##STR00573##
[0814] To a solution of 5-bromo-4-fluoro-2,2-dimethylbenzo[d][1,3]dioxole (180 mg, 0.7 mol) in toluene (10 mL) was added Pd(PPh.sub.3).sub.2Cl.sub.2 (49 mg, 0.07 mol) and tributyl(1-ethoxyvinyl)stannane (360 mg, 1 mmol). The reaction mixture was protected by N.sub.2 atmosphere and stirred at 100 C. for overnight. The mixture was added 4N Hcl in dionxane and stirred at rt for 5 mins. The mixture was added saturated NaHCO.sub.3 aqueous solution and extracted with EtOAc. The organic phase was dried with Na.sub.2SO.sub.4 and concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (120 mg, 82%). MS: M/e 211 (M+1).sup.+.
Step D: 1-(4-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethan-1-ol
##STR00574##
[0815] To a solution of 1-(4-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethan-1-one (120 mg, 0.57 mol) in MeOH was added NaBH.sub.4 (38 mg, 1 mmol). The reaction mixture was stirred at rt for 15 mins. The mixture was added H.sub.2O and extracted with DCM. The organic phase was dried with Na.sub.2SO.sub.4 and concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (80 mg, 67%). MS: M/e 213 (M+1).sup.+.
Step D: 2-(9-ethyl-6-((2S,5R)-4-(1-(4-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00575##
[0816] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.15 mmol), 1-(4-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethan-1-ol (80 mg, 0.37 mmol) and (cyanomethyl)trimethylphosphonium iodide (120 mg, 0.5 mmol) in CH.sub.3CN (5 mL) was added DIPEA (258 mg, 2 mmol). The reaction mixture was sealed in a bottle and heated at 105 C. for 16 hours, and then cooled to room temperature, diluted with water, extracted with EtOAc. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified and separated into Compound A175a (7 mg) and Compound A175b (5 mg) by Prep-HPLC (Method B).
[0817] Compound A175: .sup.1H NMR (400 MHz, CD.sub.3OD) 6.89 (d, J=5.7 Hz, 1H), 6.79 (d, J=4.1 Hz, 1H), 5.98-5.47 (m, 1H), 5.06-4.64 (m, 1H), 4.40 (s, 2H), 4.27-4.24 (m, 2H), 3.94-3.84 (m, 1H) 3.61 (s, 3H), 3.48-3.46 (m, 1H), 3.30-3.28 (m, 1H), 2.80-2.60 (m, J=26.8 Hz, 2H), 2.23-2.18 (m, 1H), 1.79 (d, J=7.5 Hz, 6H), 1.34 (s, 4H), 1.20 (m, 5H), 0.93 (s, 1.5H), 0.81 (s, 1.5H) ppm. MS: M/e 524 (M+1).sup.+.
[0818] Compound A175a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.99 (t, J=7.2 Hz, 1H), 6.57 (d, J=8.2 Hz, 1H), 6.00-5.50 (m, 1H), 5.50-5.00 (m, 1H), 4.37 (q, J=6.7 Hz, 2H), 4.23 (s, 2H), 3.84-3.82 (m, 1H), 3.76-3.74 (m, 4H), 3.58-3.56 (m, 1H), 2.70-2.68 (m, 1H), 2.33 (d, J=11.8 Hz, 1H), 1.69 (s, 6H), 1.46 (t, J=7.0 Hz, 3H), 1.35 (d, J=6.1 Hz, 3H), 1.28 (s, 3H), 1.05 (d, J=5.6 Hz, 3H) ppm. MS: M/e 524 (M+1).sup.+.
[0819] Compound A175b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.93 (t, J=7.1 Hz, 1H), 6.59 (d, J=7.8 Hz, 1H), 6.30-5.50 (m, 1H), 5.50-4.50 (m, 1H), 4.36 (d, J=6.6 Hz, 2H), 4.22 (s, 2H), 3.94 (d, J=5.2 Hz, 1H), 3.76 (s, 3H), 3.52-3.46 (m, 1H), 2.91 (d, J=22.0 Hz, 2H), 2.78 (d, J=11.5 Hz, 1H), 1.69 (s, 6H), 1.44 (dd, J=17.1, 10.0 Hz, 6H), 1.32 (d, J=6.0 Hz, 3H), 0.92 (d, J=5.8 Hz, 3H) ppm. MS: M/e 524 (M+1).sup.+.
Compound A176: 2-(6-((2S,5R)-4-(1-(4,6-difluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00576##
Step A: 3,5-difluorobenzene-1,2-diol
##STR00577##
[0820] To a solution of 3,5-difluoro-2-hydroxybenzaldehyde (500 mg, 3.2 mmol) in THE (50 mL) was added dropwise aq. NaOH solution (152 mg, 3.8 mmol, 2 mL) at 0 C., and then 30% H.sub.2O.sub.2 solution (1.4 mL). The mixture was stirred for 2 hours at RT and followed by the addition of a second portion of 30% H.sub.2O.sub.2 (1.4 mL). After stirred for 4 hours, it was cooled to 0 C. and aq. NaOH solution (2 N) was added until pH=1011 was reached, and then the mixture was stirred for 0.5 h and quenched with cone. HCl at 0 C. to pH=23. It was extracted with dichloromethane (50 mL3) and washed with brine (30 mL2), dried over Na.sub.2SO.sub.4, and concentrated to give residue. The resulting residue was purified by flash column chromatography (MeOH:DCM=0-10%) to give 3,5-difluorobenzene-1,2-diol (200 mg, 43%). MS: M/e 147 (M+1).sup.+.
Step B: 4,6-difluoro-2,2-dimethylbenzo[d][1,3]dioxole
##STR00578##
[0821] A solution of 3,5-difluorobenzene-1,2-diol (200 mg, 1.37 mmol), POBr.sub.3 (392 mg, 1.37 mmol), acetone (159 mg, 2.74 mmol) in Tol (5 mL) was stirred at 85 C. overnight. The mixture was quenched with water (10 mL), extracted with dichloromethane (50 mL3) and washed with brine (30 mL2), dried over Na.sub.2SO.sub.4, and concentrated to give residue. The resulting residue was purified by flash column chromatography (MeOH:DCM=0-10%) to give 4,6-difluoro-2,2-dimethylbenzo[d][1,3]dioxole (50 mg, 20%). MS: M/e 187 (M+1).sup.+.
Step C: 5-bromo-4,6-difluoro-2,2-dimethylbenzo[d][1,3]dioxole
##STR00579##
[0822] A solution of 4,6-difluoro-2,2-dimethylbenzo[d][1,3]dioxole (50 mg, 0.27 mmol), NBS (57 mg, 0.32 mmol) in DCM (5 mL) was stirred at RT overnight. The mixture was quenched with water (10 mL), extracted with dichloromethane (50 mL3) and washed with brine (30 mL2), dried over Na.sub.2SO.sub.4, and concentrated to give residue. The resulting residue was purified by flash column chromatography (MeOH:DCM=0-10%) to give 5-bromo-4,6-difluoro-2,2-dimethylbenzo[d][1,3]dioxole (70 mg, 98%). MS: M/e 265 (M+1).sup.+.
Step D: 1-(4,6-difluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethan-1-ol
##STR00580##
[0823] To a solution of 5-bromo-4,6-difluoro-2,2-dimethylbenzo[d][1,3]dioxole (70 mg, 0.26 mmol) in THE (20 mL) was added n-Butyllithium (0.2 mL, 0.32 mmol) at 78 C. and stirred for 1 h. Then acetaldehyde was added and stirred at 78 C. for 1 h. the mixture was quenched by water, extracted with dichloromethane (50 mL3) and washed with brine (30 mL2), dried over Na.sub.2SO.sub.4, and concentrated to give residue. The resulting residue was purified by flash column chromatography (MeOH:DCM=0-10%) to give 1-(4,6-difluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethan-1-ol (50 mg, 82%). MS: M/e 231 (M+1).sup.+.
Step E: 2-(6-((2S,5R)-4-(1-(4,6-difluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00581##
[0824] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.15 mmol) in CH.sub.3CN (3 mL) and was added 1-(4,6-difluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethan-1-ol (35 mg, 0.15 mmol), (cyanomethyl) trimethyl phosphonium iodide (111 mg, 0.46 mmol) and DIPEA (98 mg, 0.76 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure to dryness. The resulting residue was purified by Prep-TLC (DCM:MeOH=20:1) and further separated into Compound A176a (20 mg) and Compound A176b (14 mg) by Prep-HPLC (Method A).
[0825] Compound A176a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.44 (d, J=9.6 Hz, 1H), 6.14-4.89 (m, 2H), 4.37 (d, J=7.4 Hz, 2H), 4.23 (s, 1H), 3.99 (s, 1H), 3.76 (s, 3H), 3.53 (s, 1H), 3.30-3.13 (m, 2H), 2.76 (s, 1H), 2.27 (d, J=11.7 Hz, 1H), 1.69 (s, 6H), 1.47 (d, J=7.1 Hz, 6H), 1.25 (s, 3H), 0.99 (s, 3H) ppm. MS: M/e 542 (M+1).sup.+.
[0826] Compound A176b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.47 (d, J=9.7 Hz, 1H), 5.93-5.04 (m, 2H), 4.35 (s, 2H), 4.23 (s, 1H), 4.10 (s, 1H), 3.76 (s, 3H), 3.47 (s, 2H), 2.89 (s, 2H), 2.75 (s, 1H), 1.70 (s, 6H), 1.45 (d, J=7.0 Hz, 6H), 1.37 (s, 3H), 0.95 (s, 3H) ppm. MS: M/e 542 (M+1).sup.+.
Compound A178: 2-(9-ethyl-6-((2S,5R)-4-(1-(2-isopropyl-1,3-dioxoisoindolin-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00582##
Step A: 5-(1-hydroxyethyl)-2-isopropylisoindoline-1,3-dione
##STR00583##
[0827] To a solution of 5-acetyl-2-isopropylisoindoline-1,3-dione (150 mg, 0.65 mmol) in MeOH (15 mL) was added NaBH.sub.4 (25 mg, 0.65 mmol) at room temperature and the resulting mixture was stirred at room temperature for 15 mins. The reaction mixture was diluted with DCM (200 mL). The organic layer was washed with water, dried over Na.sub.2SO.sub.4 and concentrated to give the titled compound (130 mg, 86%). MS: M/e 234 (M+1).sup.+.
Step B: 2-(9-ethyl-6-((2S,5R)-4-(1-(2-isopropyl-1,3-dioxoisoindolin-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00584##
[0828] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.15 mmol) in CH.sub.3CN (3 mL) and was added 5-(1-hydroxyethyl)-2-isopropylisoindoline-1,3-dione (35 mg, 0.15 mmol), (cyanomethyl)trimethylphosphonium iodide (110 mg, 0.46 mmol) and DIPEA (98 mg, 0.76 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure to dryness. The resulting residue containing compound was purified by Prep-HPLC (Method A) to give the titled compound (32 mg, 39%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.98 (s, 1H), 7.89 (s, 2H), 6.29-5.06 (m, 2H), 4.51 (s, 1H), 4.38 (d, J=7.1 Hz, 2H), 4.26 (s, 2H), 4.11-3.80 (m, 1H), 3.78 (s, 3H), 3.70-3.48 (m, 1H), 3.29-2.68 (m, 2H), 2.67-2.05 (m, 1H), 1.49 (s, 3H), 1.47 (d, J=6.6 Hz, 9H), 1.32 (s, 6H) ppm. MS: M/e 545 (M+1).sup.+
Compound A179: 2-(6-((2S,5R)-4-(1-(2,2-dimethyl-3-oxo-2,3-dihydrobenzofuran-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00585##
Step A: 1-(4-bromo-2-fluorophenyl)-2-methylpropan-2-ol
##STR00586##
[0829] To a solution of methyl 2-(4-bromo-2-fluorophenyl) acetate (2 g, 8.10 mmol) in THE (20 ml) at 60 C. under N.sub.2 was added MeMgBr (3M, 6.75 ml, 20.25 mmol). The solution was warmed to rt naturally and stirred for 1 h. The solution was diluted with EtOAc (30 ml), washed with brine (10 ml), dried and concentrated. The residue was purified by flash column chromatography with 0%-20% EtOAc in PE to give the titled compound (1.7 g, 85%). MS: M/e 247 (M+1).sup.+.
Step B: 6-bromo-2,2-dimethyl-2,3-dihydrobenzofuran
##STR00587##
[0830] A solution of 1-(4-bromo-2-fluorophenyl)-2-methylpropan-2-ol (1.7 g, 6.91 mmol) and tBuOK (1M in THF, 8.2 ml, 8.2 mmol) in THE (20 ml) was stirred at 60 C. for 1 h. The solution was diluted with EtOAc (30 ml), washed with brine (10 ml), dried and evaporated to dryness to give the titled compound (1.4 g, 90%), which was used directly for the next step.
Step C: 6-bromo-2,2-dimethylbenzofuran-3 (2H)-one
##STR00588##
[0831] A solution of 6-bromo-2,2-dimethyl-2,3-dihydrobenzofuran (1.2 g, 5.29 mmol), K.sub.2S.sub.2O.sub.8 (4.3 g, 15.93 mmol) and CuSO.sub.4 (0.85 g, 5.31 mmol) in MeCN (12 ml) and H.sub.2O (12 ml) was stirred at 100 C. for 1 h. The solution was diluted with EtOAc (30 ml), washed with brine (15 ml2), died and then concentrated. The residue was purified by flash column chromatography with 0%-5% EtOAc in PE to give the titled compound (1 g, 78%). MS: M/e 241,243 (M+1).sup.+.
Step D: 6-acetyl-2,2-dimethylbenzofuran-3 (21H)-one
##STR00589##
[0832] A solution of 6-bromo-2,2-dimethylbenzofuran-3 (2H)-one (500 mg, 2.07 mmol), Pd(PPh.sub.3).sub.2Cl.sub.2 (73 mg, 0.10 mmol) and tributyl(1-ethoxyvinyl)stannane (1.1 g, 3.05 mmol) in Tol. (8 ml) was stirred at 100 C. overnight. The mixture was cooled to RT and HCl (4M in dioxane, 2 ml) was added, stirred for 30 min. The mixture was diluted with EtOAc (20 ml), washed with brine (10 ml), dried and concentrated. The residue was purified by flash column chromatography with 0%-15% MEA in PE to give the titled compound (400 mg, 95%). .sup.1H NMR (400 MHz, CDCl.sub.3) 7.75 (d, J=7.8 Hz, 1H), 7.63 (d, J=8.4 Hz, 2H), 2.65 (s, 3H), 1.49 (s, 6H) ppm.
Step E: 6-(1-hydroxyethyl)-2,2-dimethylbenzofuran-3 (2H)-one
##STR00590##
[0833] A solution of 6-acetyl-2,2-dimethylbenzofuran-3 (2H)-one (100 mg, 0.49 mmol) and NaBH.sub.4 (9.3 mg, 0.24 mmol) in MeOH (2 ml) was stirred at RT for 10 min. The solution was quenched with water (10 ml), diluted with EtOAc (15 ml), washed with brine (10 ml), dried and evaporated to dryness to give the titled compound (100 mg, 100%), which was used directly for the next step. .sup.1H NMR (400 MHz, CDCl.sub.3) 7.64 (d, J=7.3 Hz, 1H), 7.12 (s, 1H), 7.06 (d, J=7.8 Hz, 1H), 4.96 (q, J=6.4 Hz, 1H), 1.52 (d, J=6.1 Hz, 3H), 1.47 (s, 6H) ppm.
Step F: 2-(6-((2S,5R)-4-(1-(2,2-dimethyl-3-oxo-2,3-dihydrobenzofuran-6-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00591##
[0834] A solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (100 mg, 0.30 mmol), 6-(1-hydroxyethyl)-2,2-dimethylbenzofuran-3 (2H)-one (100 mg, 0.48 mmol), (cyanomethyl)trimethylphosphonium iodide (222 mg, 0.91 mmol) and DIPEA (392 mg, 3.04 mmol) in MeCN (3 ml) was stirred at 100 C. overnight. The reaction was diluted with EtOAc (10 ml) and washed with brine (10 ml). The organic layer was concentrated under reduced pressure. The resulting residue was purified by flash column chromatography with 0-5% MeOH in DCM and then Prep-HPLC (Method A) to give the titled compound (14 mg). .sup.1H NMR (400 MHz, DMSO-d6) 7.62 (t, J=8.2 Hz, 1H), 7.23 (d, J=11.1 Hz, 2H), 6.13-5.46 (m, 1H), 5.20-4.47 (m, 1H), 4.43 (d, J=18.5 Hz, 2H), 4.26 (d, J=7.3 Hz, 2H), 3.71 (d, J=5.5 Hz, 0.5H), 3.61 (s, 3H), 3.59-3.47 (m, 1H), 3.31 (s, 1.5H), 2.80 (s, 1.5H), 2.13 (d, J=11.3 Hz, 0.5H), 1.40 (s, 6H), 1.34 (s, 4.5H), 1.26 (dd, J=13.7, 5.9 Hz, 3H), 1.20 (s, 1.5H), 0.97 (s, 1.5H), 0.86 (s, 1.5H) ppm. MS: M/e 518 (M+1).sup.+.
Compound A181: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrofuro[3,2-b]pyridin-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00592##
Step A: 5-hydroxy-6-iodopicolinic acid
##STR00593##
[0835] To a stirred solution of 5-hydroxypyridine-2-carboxylic acid (1 g, 6.8 mmol, 95%) in NH.sub.3.Math.H.sub.2O (30 mL) were added a solution of KI (5.97 g, 34 mmol) and 12 (1.82 g, 6.8 mmol) in H.sub.2O (40 mL) dropwise at room temperature. The resulting mixture was stirred for overnight at room temperature. The mixture was acidified to pH=3 with HCl (1M). The resulting mixture was extracted with EtOAc (3150 mL). The combined water layers were concentrated under reduced pressure to give the titled compound (1 g, 55%). MS: M/e 266 (M+1).sup.+.
Step B: methyl 5-hydroxy-6-iodopicolinate
##STR00594##
[0836] To a stirred solution of 5-hydroxy-6-iodopyridine-2-carboxylic acid (1 g, 3.7 mmol) in MeOH was added SOCl.sub.2 (1.3 mL, 18 mmol) dropwise at 0 C. The resulting mixture was stirred for 2 hours at 70 C. The mixture was allowed to cool down to room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flash chromatography to give the titled compound (600 mg, 53%). MS: M/e 280 (M+1).sup.+.
Step C: methyl 6-iodo-5-((2-methylallyl)oxy)picolinate
##STR00595##
[0837] A solution of methyl 5-hydroxy-6-iodopyridine-2-carboxylate (500 mg, 1.6 mmol), 3-bromo-2-methylprop-1-ene (250 mg, 1.8 mmol) and K.sub.2CO.sub.3 (609 mg, 4.1 mmol) in acetone was stirred at 60 C. overnight under nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography to give the titled compound (500 mg, 89%). MS: M/e 334 (M+1).sup.+.
Step D: methyl 3,3-dimethyl-2,3-dihydrofuro[3,2-b]pyridine-5-carboxylate
##STR00596##
[0838] A solution of methyl 6-iodo-5-[(2-methylprop-2-en-1-yl)oxy]pyridine-2-carboxylate (500 mg, 1.5 mmol), n-Bu.sub.3SnH (689 mg, 2.25 mmol) and AIBN (26 mg, 0.15 mmol) in benzene was stirred at 80 C. overnight under nitrogen atmosphere. The mixture was allowed to cool down to room temperature. The resulting mixture was added KF (10%, 5 mL) at room temperature. The resulting mixture was stirred for 3.5 hours at room temperature. The resulting mixture was extracted with EtOAc (350 mL). The combined organic layers were dried over anhydrous Na.sub.2SO.sub.4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography to give the titled compound (240 mg, 77%). MS: M/e 208 (M+1).sup.+.
Step E: 3,3-dimethyl-2,3-dihydrofuro[3,2-b]pyridine-5-carboxylic acid
##STR00597##
[0839] To a solution of methyl 3,3-dimethyl-2,3-dihydrofuro[3,2-b]pyridine-5-carboxylate (0.2 g, 0.97 mmol) in MeOH (4 mL)/water (1 mL) was added LiOH.Math.H.sub.2O (58 mg, 1.5 mmol). The mixture solution was stirred at room temperature for 5 hours. The reaction was quenched with water and acidified to pH=56 with citric acid, extracted with DCM (20 mL2). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated to give the titled compound (0.18 g, 96%). MS: M/e 194 (M+1).sup.+.
Step F: N-methoxy-N, 3,3-trimethyl-2,3-dihydrofuro[3,2-b]pyridine-5-carboxamide
##STR00598##
[0840] To a solution of 3,3-dimethyl-2,3-dihydrofuro[3,2-b]pyridine-5-carboxylic acid (0.14 g, 0.72 mmol) in DMF (2 mL) were added HATU (548 mg, 1.4 mmol), DIPEA (465 mg, 3.6 mol) and N,N-dimethyl-hydroxylamine hydrochloride salt (210 mg, 2.2 mol). The mixture was stirred at room temperature for overnight. The reaction was diluted with DCM and washed with water. The organic layer was separated, dried by Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (120 mg, 70%). MS: M/e 237 (M+1).sup.+.
Step G: 1-(3,3-dimethyl-2,3-dihydrofuro[3,2-b]pyridin-5-yl)ethan-1-one
##STR00599##
[0841] A mixture of N-methoxy-N, 3,3-trimethyl-2,3-dihydrofuro[3,2-b]pyridine-5-carboxamide (120 mg, 0.51 mmol) in THE (5 mL) at 0 C. was added methyl magnesium bromide (0.33 mL, 3M, 1.02 mol). The reaction mixture was stirred at room temperature for 30 mins. The reaction was quenched by adding aqueous NH.sub.4Cl. The resulting mixture was extracted with EtOAc and then concentrated by using a rotary evaporator, to give the titled compound (110 mg). MS: M/e 192 (M+1).sup.+.
Step H: 1-(3,3-dimethyl-2,3-dihydrofuro[3,2-b]pyridin-5-yl)ethan-1-ol
##STR00600##
[0842] To a solution of 1-(3,3-dimethyl-2,3-dihydrofuro[3,2-b]pyridin-5-yl)ethan-1-one (110 mg, 0.57 mmol) in MeOH (5 mL) was added NaBH.sub.4 (21 mg, 0.57 mmol) at room temperature and the resulting mixture was stirred at room temperature for 5 min. The reaction mixture was diluted with DCM and washed with water, dried over Na.sub.2SO.sub.4 and concentrated to give the titled compound (100 mg). MS: M/e 194 (M+1).sup.+.
Step I: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrofuro[3,2-b]pyridin-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00601##
[0843] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.15 mmol) in CH.sub.3CN (2 mL) and was added 1-(3,3-dimethyl-2,3-dihydrofuro[3,2-b]pyridin-5-yl)ethan-1-ol (44 mg, 0.23 mmol), (cyanomethyl)trimethylphosphonium iodide (110 mg, 0.45 mmol) and DIPEA (194 mg, 1.5 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure. The resulting residue was purified to give the titled compound (7 mg) by Prep-HPLC (Method A). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.33 (d, J=8.4 Hz, 1H), 7.14 (dd, J=8.3, 3.5 Hz, 1H), 6.21-4.89 (m, 2H), 4.39 (d, J=7.0 Hz, 1H), 4.35 (s, 3H), 4.22 (s, 1H), 3.94-3.81 (m, 1H), 3.77 (s, 3H), 3.73-3.32 (m, 2H), 3.08-2.10 (m, 3H), 1.51-1.24 (m, 15H), 1.21-0.89 (m, 3H) ppm. MS: M/e 505 (M+1).sup.+.
Compound A183: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzofuran-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00602##
Step A: 1-bromo-2-((2-methylallyl)oxy)benzene
##STR00603##
[0844] To a solution of 2-bromophenol (2.457 g, 14.2 mol) in actone (75 mL) was added 3-bromo-2-methylprop-1-ene (2 ml, 14.9 mol) and K.sub.2CO.sub.3 (4.9 g, 35 mmol). The reaction mixture was protected by N.sub.2 atmosphere and stirred at 70 C. for overnight. The mixture was cooled down to RT and added H.sub.2O. The mixture was extracted with EtOAc then concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (2.6 g, 81%). MS: M/e 227 (M+1).sup.+.
Step B: 3,3-dimethyl-2,3-dihydrobenzofuran
##STR00604##
[0845] To a solution of 1-bromo-2-((2-methylallyl)oxy)benzene (1.1 g, 4.8 mol) in toluene (15 mL) was added Bu.sub.3SnH (2.2 g, 7.6 mmol) and AIBN (80 mg, 0.48 mmol). The reaction mixture was protected by N.sub.2 atmosphere and stirred at 100 C. for overnight. The mixture was cooled down to RT and added H.sub.2O. The mixture was extracted with EtOAc then concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (0.7 g, 99%). MS: M/e 149 (M+1).sup.+
Step C: 5-bromo-3,3-dimethyl-2,3-dihydrobenzofuran
##STR00605##
[0846] To a solution of 3,3-dimethyl-2,3-dihydrobenzofuran (0.7 g, 4.69 mol) in DCM (15 mL) was added NBS (1.7 g, 7 mmol). The reaction mixture was stirred at RT for 2 h. The mixture was added H.sub.2O and extracted with EtOAc then concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (0.6 g, 60%). MS: M/e 227 (M+1).sup.+
Step D: 1-(3,3-dimethyl-2,3-dihydrobenzofuran-5-yl)ethan-1-one
##STR00606##
[0847] To a solution of 5-bromo-3,3-dimethyl-2,3-dihydrobenzofuran (200 mg, 0.88 mol) in toluene (10 mL) was added Pd(PPh.sub.3).sub.2Cl.sub.2 (70 mg, 0.1 mol) and tributyl(1-ethoxyvinyl)stannane (476 mg, 1.32 mmol). The reaction mixture was protected by N.sub.2 atmosphere and stirred at 90 C. for overnight. The mixture was added 4N Hcl in dionxane and stirred at RT for 15 mins. The mixture was added saturated NaHCO.sub.3 aqueous solution and extracted with EtOAc. The organic phase was dried with Na.sub.2SO.sub.4 and concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (100 mg, 59.8%). MS: M/e 191 (M+1).sup.+.
Step E: 1-(3,3-dimethyl-2,3-dihydrobenzofuran-5-yl)ethan-1-ol
##STR00607##
[0848] To a solution of 1-(3,3-dimethyl-2,3-dihydrobenzofuran-5-yl)ethan-1-one (100 mg, 0.5 mol) in MeOH was added NaBH.sub.4 (38 mg, 1 mmol). The reaction mixture was stirred at RT for 15 mins. The mixture was added H.sub.2O and extracted with DCM. The organic phase was dried with Na.sub.2SO.sub.4 and concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (50 mg, 50%). MS: M/e 193 (M+1).sup.+.
Step F: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydrobenzofuran-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00608##
[0849] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.14 mmol), 1-(3,3-dimethyl-2,3-dihydrobenzofuran-5-yl)ethan-1-ol (50 mg, 0.26 mmol) and (cyanomethyl)trimethylphosphonium iodide (120 mg, 0.5 mmol) in CH.sub.3CN (10 mL) was added DIPEA (258 mg, 2 mmol). The reaction mixture was sealed in a bottle and heated at 105 C. for 16 hours, and then cooled to room temperature, diluted with water, extracted with EtOAc. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (16 mg, 24%). .sup.1H NMR (400 MHz, CD.sub.3OD) 7.21 (d, J=13.7 Hz, 1H), 7.11 (dd, J=29.3, 8.1 Hz, 1H), 6.69 (dd, J=19.4, 8.1 Hz, 1H), 5.92 (s, 1H), 4.89 (s, 1H), 4.37 (q, J=7.1 Hz, 2H), 4.22 (d, J=4.6 Hz, 3H), 3.76 (s, 3H), 3.65 (s, 1H), 3.48 (s, 1H), 3.13-2.89 (m, 2H), 2.69 (s, 1H), 2.31 (s, 1H), 1.46 (t, J=7.1 Hz, 4H), 1.31 (m, 11H), 1.07 (d, J=6.3 Hz, 1.5H), 0.95 (d, J=5.4 Hz, 1.5H) ppm. MS: M/e 507 (M+1).sup.+.
Compound A184: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydro-1H-inden-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00609##
Step A: 1-(3,3-dimethyl-2,3-dihydro-1H-inden-5-yl)ethan-1-one
##STR00610##
[0850] To a solution of 6-bromo-1,1-dimethyl-2,3-dihydro-1H-indene (220 mg, 1 mol) in toluene (10 mL) was added Pd(PPh.sub.3).sub.2Cl.sub.2 (70 mg, 0.1 mol) and tributyl(1-ethoxyvinyl)stannane (541 mg, 1.5 mmol). The reaction mixture was protected by N.sub.2 atmosphere and stirred at 100 C. for overnight. The mixture was added 4N HCl in dionxane and stirred at RT for 15 mins. The mixture was added saturated NaHCO.sub.3 aqueous solution and extracted with EtOAc. The organic phase was dried with Na.sub.2SO.sub.4 and concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (150 mg, 79%). MS: M/e 189 (M+1).sup.+.
Step B: 1-(3,3-dimethyl-2,3-dihydro-1H-inden-5-yl)ethan-1-ol
##STR00611##
[0851] To a solution of 1-(3,3-dimethyl-2,3-dihydro-1H-inden-5-yl)ethan-1-one (150 mg, 0.79 mol) in MeOH was added NaBH.sub.4 (38 mg, 1 mmol). The reaction mixture was stirred at RT for 15 mins. The mixture was added H.sub.2O and extracted with DCM. The organic phase was dried with Na.sub.2SO.sub.4 and concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (70 mg, 47%). MS: M/e 191 (M+1).sup.+.
Step C: 2-(6-((2S,5R)-4-(1-(3,3-dimethyl-2,3-dihydro-1H-inden-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00612##
[0852] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.15 mmol), 1-(3,3-dimethyl-2,3-dihydro-1H-inden-5-yl)ethan-1-ol (100 mg, 0.5 mmol) and (cyanomethyl)trimethylphosphonium iodide (160 mg, 0.66 mmol) in CH.sub.3CN (10 mL) was added DIPEA (258 mg, 2 mmol). The reaction mixture was sealed in a bottle and heated at 105 C. for 16 hours, and then cooled to room temperature, diluted with water, extracted with EtOAc. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (6 mg, 8%). .sup.1H NMR (400 MHz, DMSO-d6) 7.20 (d, J=21.1 Hz, 1H), 7.15 (s, 1H), 7.09 (s, 1H), 5.93 (s, 1H), 5.07 (s, 1H), 4.37 (q, J=7.1 Hz, 2H), 4.21 (s, 1H), 3.78 (d, J=14.2 Hz, 4H), 3.63 (s, 1H), 3.48 (s, 2H), 2.87 (dd, J=13.2, 6.8 Hz, 3H), 2.65-2.27 (m, 1H), 1.93 (dd, J=10.9, 6.9 Hz, 2H), 1.46 (t, J=7.1 Hz, 4H), 1.35 (s, 3H), 1.25 (d, J=7.8 Hz, 8H), 1.06 (d, J=5.4 Hz, 1.5H), 0.92 (s, 1.5H) ppm. MS: M/e 502 (M+1).sup.+.
Compound A185: 2-(9-ethyl-6-((2S,5R)-4-(1-(7-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-4-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00613##
Step A: 3-bromo-6-fluoro-2-hydroxybenzaldehyde
##STR00614##
[0853] A mixture of 2-bromo-5-fluorophenol (19.1 g, 100 mmol), MgCl.sub.2 (19.0 g, 200 mmol), Et.sub.3N (20.2 g, 200 mmol) and paraformaldehyde (9.0 g, 300 mmol) in THF (150 mL) was stirred at 85 C. for 24 hours. An aqueous solution of HCl (2M) was added to pH3. The mixture was extracted with EtOAc (200 mL3). The combined extracts was washed with brine (200 mL3), dried over Na.sub.2SO.sub.4, filtered and concentrated to dryness. The resulted residue was purified by flash column chromatography to give a light yellow suspension. PE (30 mL) was added and stirred for 10 minutes. The suspension was filtered and the filter cake was washed with PE (20 mL), dried under high vacuum to give the titled compound (5.2 g, 24%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) 11.87 (s, 1H), 10.17 (s, 1H), 7.95 (t, J=7.2 Hz, 1H), 6.90 (t, J=9.6 Hz, 1H) ppm.
Step B: 3-bromo-6-fluorobenzene-1,2-diol
##STR00615##
[0854] To a suspension of 3-bromo-6-fluoro-2-hydroxybenzaldehyde (5.2 g, 23.7 mmol) in H.sub.2O (40 mL) was added H.sub.2O.sub.2 (3.5 g, 30%, 30.8 mmol) at 0 C. and followed by aqueous solution of NaOH (3M, 10.3 mL, 30.8 mmol) in drops. The resulted mixture was stirred at 0 C. for 30 min and room temperature for 30 min. The mixture was acidified with HCl (2.5 M) to Ph-3 and extracted with EtOAc (50 mL2). The extracts were combined and washed with brine (50 mL2), dried over Na.sub.2SO.sub.4, filtered and concentrated to dryness. The resulted residue was purified by flash column chromatography to give the titled compound (4.6 g, 94%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) 9.83 (s, 1H), 9.66 (s, 1H), 6.94 (t, J=6.8 Hz, 1H), 6.65 (t, J=9.6 Hz, 1H) ppm.
Step C: 4-bromo-7-fluoro-2,2-dimethylbenzo[d][1,3]dioxole
##STR00616##
[0855] To a mixture of 3-bromo-6-fluorobenzene-1,2-diol (1.04 g, 5.0 mmol) and acetone (580 mg, 10.0 mmol) in toluene (8 mL) was added POBr.sub.3 (1.43 g, 5.0 mmol) and the resulted mixture was stirred at 80 C. for 20 hours. The mixture was cooled and diluted with EtOAc (20 mL), basified with aqueous solution of NaHCO.sub.3 to Ph-8, washed with brine (20 mL2), dried over Na.sub.2SO.sub.4, filtered and concentrated to dryness. The resulted residue was purified by column chromatography to give the titled compound (618 mg, 50%). .sup.1H NMR (400 MHz, CDCl.sub.3) 6.93-6.80 (m, 1H), 6.56 (t, J=9.2 Hz, 1H), 1.77 (s, 6H) ppm.
Step D: 1-(7-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-4-yl)ethan-1-ol
##STR00617##
[0856] To a 70 C. solution of 4-bromo-7-fluoro-2,2-dimethylbenzo[d][1,3]dioxole (100 mg, 0.40 mmol) in THF (2 mL) was added n-BuLi (1.6 M, 0.35 mL, 1.56 mmol) in drops under N.sub.2. After stirring for 10 minutes at 70 C., a solution of acetaldehyde (52 mg, 1.2 mmol) in THF (0.5 mL) was added. Then the mixture was allowed warm to room temperature and stirred for 2 hours. Aqueous solution of NH.sub.4Cl (10 mL) was added and the mixture was extracted with EtOAc (5 mL3), the organics were combined and washed with brine (10 mL2), dried over Na.sub.2SO.sub.4, filtered and concentrated to dryness. The resulted residue was purified by column chromatography to give the titled compound (76 mg, 89%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) 6.97-6.90 (m, 1H), 6.80 (t, J=9.6 Hz, 1H), 5.25 (s, 1H), 4.85-4.75 (m, 1H), 1.74 (s, 6H), 1.34 (d, J=6.0 Hz, 3H) ppm.
Step E: 2-(9-ethyl-6-((2S,5R)-4-(1-(7-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-4-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00618##
[0857] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (60 mg, 0.18 mmol), 1-(7-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-4-yl)ethan-1-ol (60 mg, 0.28 mmol) and (cyanomethyl)trimethylphosphonium iodide (130 mg, 0.55 mmol) in CH.sub.3CN (1 mL) was added DIPEA (93 mg, 0.72 mmol). The mixture was stirred at 100 C. in a sealed tube for 16 hours. The mixture was cooled and diluted with EtOAc (5 mL), washed with brine (5 mL2), dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to dryness. The resulting residue was purified by flash column chromatography and further separated into to give Compound 185a (3.5 mg) and Compound A185b (4.0 mg) by Prep-HPLC (Method A).
[0858] Compound A185a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.95-6.88 (m, 1H), 6.63 (t, J=9.4 Hz, 1H), 6.20-5.70 (m, 1H), 5.20-4.50 (m, 1H), 4.37 (q, J=6.8 Hz, 2H), 4.23 (s, 1H), 3.90-3.36 (m, 6H), 3.29-3.07 (m, 1H), 2.80-2.55 (m, 1H), 2.31 (d, J=11.6 Hz, 1H), 1.69 (s, 6H), 1.46 (t, J=6.8 Hz, 3H), 1.37-1.22 (m, 6H), 1.02 (d, J=5.2 Hz, 3H) ppm. MS: M/e 524 (M+1).sup.+.
[0859] Compound 185b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.96-6.79 (m, 1H), 6.66 (t, J=9.6 Hz, 1H), 6.30-4.95 (m, 2H), 4.37 (d, J=7.6 Hz, 2H), 4.26-4.15 (m, 1H), 3.86-3.70 (m, 4H), 3.64-3.32 (m, 2H), 3.00-2.84 (m, 2H), 2.81-2.71 (m, 1H), 1.71 (s, 3H), 1.68 (s, 3H), 1.46 (t, J=6.8 Hz, 3H), 1.40 (d, J=5.6 Hz, 3H), 1.32 (d, J=6.0 Hz, 3H), 0.93 (d, J=6.0 Hz, 3H) ppm. MS: M/e 524 (M+1).sup.+.
Compound A188: 2-(7-((2S,5R)-4-(1-(2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-3-ethyl-6-fluoro-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00619##
[0860] To a solution of 2-(7-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3-ethyl-6-fluoro-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (50 mg, 0.14 mmol) in CH.sub.3CN (2 mL) and was added 1-(2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethan-1-ol (54 mg, 0.28 mmol), (cyanomethyl)trimethylphosphonium iodide (110 mg, 0.45 mmol) and DIPEA (194 mg, 1.5 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure. The resulting residue was purified to give the titled compound (2 mg) by Prep-HPLC (Method A). .sup.1H NMR (400 MHz, CD.sub.3OD) 6.85-6.72 (m, 2H), 6.69-6.61 (m, 1H), 4.75-4.55 (m, 1H), 4.45-4.37 (m, 2H), 4.08-3.89 (m, 1H), 3.88-3.86 (m, 3H), 3.70-3.65 (m, 2H), 3.55-3.34 (m, 2H), 3.17-2.76 (m, 2H), 2.75-2.20 (m, 1H), 1.66-1.59 (m, 6H), 1.48-1.43 (m, 3H), 1.40-1.25 (m, 6H), 1.21-0.99 (m, 3H) ppm. MS: M/e 523 (M+1).sup.+.
Compound A189: 2-(3-ethyl-6-fluoro-7-((2S,5R)-4-(1-(7-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00620##
[0861] To a solution of 2-(7-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3-ethyl-6-fluoro-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (50 mg, 0.14 mmol) in CH.sub.3CN (2 mL) and was added 1-(7-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethan-1-ol (61 mg, 0.28 mmol), (cyanomethyl)trimethylphosphonium iodide (105 mg, 0.43 mmol) and DIPEA (185 mg, 1.5 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure. The resulting residue was purified by flash column chromatography to give the titled compound (crude), which was further purified to give the titled compound (4 mg) by Prep-HPLC (Method A). .sup.1H NMR (400 MHz, CD.sub.3OD) 6.75-6.63 (m, 2H), 4.79-4.55 (m, 1H), 4.45-4.37 (m, 2H), 4.26-4.20 (m, 1H), 4.10-3.89 (m, 1H), 3.87 (s, 3H), 3.79-3.40 (m, 3H), 3.10-2.73 (m, 2H), 2.65-2.17 (m, 1H), 1.73-1.65 (m, 6H), 1.50-1.37 (m, 4H), 1.33-1.23 (m, 5H), 1.20-0.99 (m, 3H) ppm. MS: M/e 541 (M+1).sup.+.
Compound A190: 2-(3-ethyl-6-fluoro-7-((2S,5R)-4-(1-(4-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile
##STR00621##
[0862] To a solution of 2-(7-((2S,5R)-2,5-dimethylpiperazin-1-yl)-3-ethyl-6-fluoro-4-methyl-5-oxo-4,5-dihydro-3H-imidazo[4,5-b]pyridin-2-yl)acetonitrile (50 mg, 0.14 mmol) in CH.sub.3CN (2 mL) and was added 1-(4-fluoro-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)ethan-1-ol (61 mg, 0.28 mmol), (cyanomethyl)trimethylphosphonium iodide (110 mg, 0.45 mmol) and DIPEA (194 mg, 1.5 mmol). The resulting mixture was stirred at 105 C. overnight. The reaction solvent was removed under reduced pressure and purified by flash column chromatography (DCM:MeOH) to give the titled Compound A190, which was further separated into Compound A190a (22 mg) and Compound A190b (11 mg) by Prep-HPLC (Method A) and Prep-chiral HPLC. The chiral separation conditions are shown below.
TABLE-US-00029 Column IE Column Size 20.0 mm 250 mm, 5 um Mobile Phase A Hex Mobile Phase B EtOH (0.2% 2M NH.sub.3 MeOH) Flow Rate 18 mL/min Gradient Mobile Phase A:Mobile Phase B = 80:20(v/v) Wave Length UV 200 nm and 290 nm Temperature RT Prep-HPLC Equipment Prep-HPLC-Gilson
[0863] Compound A190: .sup.1H NMR (400 MHz, CD.sub.3OD) 7.01-6.91 (m, 1H), 6.65-6.51 (m, 1H), 4.74-4.53 (m, 1H), 4.45-4.38 (m, 2H), 3.98-4.10 (m, 1H), 3.91 (d, J=11.6 Hz, 1H), 3.87 (d, J=1.7 Hz, 3H), 3.81-3.74 (m, 1H), 3.49-3.40 (m, 2H), 3.18-2.78 (m, 2H), 2.69-2.21 (m, 1H), 1.74-1.65 (m, 6H), 1.49-1.25 (m, 9H), 1.22-1.01 (m, 3H) ppm. MS: M/e 541 (M+1).sup.+.
[0864] Compound A190a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 6.97-6.88 (m, 1H), 6.58 (d, J=8.2 Hz, 1H), 4.74-4.65 (m, 1H), 4.42 (q, J=7.2 Hz, 2H), 4.05 (q, J=6.5 Hz, 1H), 3.87 (s, 3H), 3.75-3.67 (m, 2H), 3.37-3.33 (m, 2H), 3.10-3.01 (m, 1H), 2.88-2.79 (m, 1H), 2.67-2.57 (m, 1H), 1.69 (s, 6H), 1.45 (t, J=7.2 Hz, 3H), 1.35 (d, J=6.5 Hz, 6H), 1.04 (d, J=6.5 Hz, 3H) ppm. MS: M/e 541 (M+1).sup.+.
[0865] Compound A190b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 7.03-6.93 (m, 1H), 6.56 (t, J=6.8 Hz, 1H), 4.67-4.57 (m, 1H), 4.41 (q, J=7.2 Hz, 2H), 4.03 (d, J=12.9 Hz, 1H), 3.951-3.88 (m, 2H), 3.87 (s, 3H), 3.47-3.33 (m, 3H), 2.85 (d, J=11.8 Hz, 1H), 2.24 (d, J=11.9 Hz, 1H), 1.68 (s, 6H), 1.45 (t, J=7.2 Hz, 3H), 1.33 (t, J=7.0 Hz, 3H), 1.28 (d, J=6.6 Hz, 3H), 1.14 (d, J=6.5 Hz, 3H) ppm. MS: M/e 541 (M+1).sup.+.
Compound A191: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-methyl-4-oxo-4H-chromen-7-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00622##
Step A: 1-(4-bromo-2-hydroxyphenyl)butane-1,3-dione
##STR00623##
[0866] To a solution of 1-(4-bromo-2-hydroxyphenyl)ethan-1-one (2.15 g, 10 mol) in THE (15 mL) was added ethyl acetate (2.2 g, 25 mol) and NaH (60% in oil, 2 g, 50 mmol). The reaction mixture was protected by N.sub.2 atmosphere and stirred at 70 C. for overnight. The mixture was cooled down to RT and added H.sub.2O, The mixture was adjusted PH 2-3 with 1 N HCl solution and extracted with EtOAc then concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (1.8 g, 70%). MS: M/e 257 (M+1).sup.+.
Step B: 7-bromo-2-methyl-4H-chromen-4-one
##STR00624##
[0867] To a solution of 1-(4-bromo-2-hydroxyphenyl)butane-1,3-dione (1.8 g, 7 mol) in MeOH (15 mL) was added concentrated H.sub.2SO.sub.4 (1 mL). The reaction mixture was stirred at 50 C. for overnight. The mixture was cooled down to RT and added H.sub.2O, The mixture was added H.sub.2O and extracted with EtOAc then concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (1.5 g, 90%). MS: M/e 239 (M+1).sup.+
Step C: 7-acetyl-2-methyl-4H-chromen-4-one
##STR00625##
[0868] To a solution of 7-bromo-2-methyl-4H-chromen-4-one (240 mg, 1 mol) in toluene (10 mL) was added Pd(PPh.sub.3).sub.2Cl.sub.2 (70 mg, 0.1 mol) and tributyl(1-ethoxyvinyl)stannane (720 mg, 2 mmol). The reaction mixture was protected by N.sub.2 atmosphere and stirred at 90 C. for overnight. The mixture was added 4N HCl in dionxane and stirred at RT for 15 mins. The mixture was added saturated NaHCO.sub.3 aqueous solution and extracted with EtOAc. The organic phase was dried with Na.sub.2SO.sub.4 and concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (180 mg, 89%). MS: M/e 203 (M+1).sup.+.
Step D: 7-(1-hydroxyethyl)-2-methyl-4H-chromen-4-one
##STR00626##
[0869] To a solution of 7-acetyl-2-methyl-4H-chromen-4-one (180 mg, 0.89 mol) in MeOH was added NaBH.sub.4 (38 mg, 1 mmol). The reaction mixture was stirred at RT for 15 mins. The mixture was added H.sub.2O and extracted with DCM. The organic phase was dried with Na.sub.2SO.sub.4 and concentrated by using a rotary evaporator to give a residue. The resulting residue was purified by flash column chromatography to give the titled compound (120 mg, 66%). MS: M/e 205 (M+1).sup.+.
Step E: 2-(6-((2S,5R)-2,5-dimethyl-4-(1-(2-methyl-4-oxo-4H-chromen-7-yl)ethyl)piperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile
##STR00627##
[0870] To a solution of 2-(6-((2S,5R)-2,5-dimethylpiperazin-1-yl)-9-ethyl-3-methyl-2-oxo-3,9-dihydro-2H-purin-8-yl)acetonitrile (50 mg, 0.14 mmol), 7-(1-hydroxyethyl)-2-methyl-4H-chromen-4-one (60 mg, 0.29 mmol) and (cyanomethyl)trimethylphosphonium iodide (120 mg, 0.5 mmol) in CH.sub.3CN (10 mL) was added DIPEA (258 mg, 2 mmol). The reaction mixture was sealed in a bottle and heated at 105 C. for 16 hours, and then cooled to room temperature, diluted with water, extracted with EtOAc. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by flash column chromatography to give the titled compound (16 mg, 24%). .sup.1H NMR (400 MHz, CD.sub.3OD)) 8.07 (t, J=9.5 Hz, 1H), 7.61 (s, 1H), 7.56 (s, 1H), 6.24 (s, 1H), 6.10-5.50 (m, 1H), 5.50-5.00 (m, 1H), 4.37 (d, J=6.7 Hz, 2H), 4.23 (s, 2H), 3.79 (d, J=22.7 Hz, 4H), 3.68 (d, J=7.2 Hz, 1H), 2.90 (s, 2H), 2.73 (s, 0.5H), 2.45 (s, 3H), 2.19 (d, J=12.5 Hz, 0.5H), 1.47 (d, J=7.1 Hz, 4.5H), 1.38 (dd, J=12.1, 6.4 Hz, 3H), 1.30 (s, 1.5H), 1.08 (d, J=5.5 Hz, 1.5H), 0.94 (d, J=5.4 Hz, 1.5H) ppm. MS: M/e 507 (M+1).sup.+.
Compound A194: 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(quinoxalin-6-yl)ethyl)piperazin-1-yl)-4-methyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridin-2-yl)acetonitrile
##STR00628##
Step A: methyl 5-(methylamino)thiazole-4-carboxylate
##STR00629##
[0871] Methyl 2-isocyanoacetate (50 g, 0.5 mol) and isothiocyanatomethane (37 g, 0.5 mol) were dissolved in THF (800 ml) followed by addition of t-BuOK (1M in THF, 650 ml, 0.65 mol) dropwise. The mixture was stirred at rt overnight. The solvent was removed under reduced pressure and water was added to quench the reaction. The reaction was adjusted pH to 7 with saturated citric acid solution and extracted with EtOAc. The organic phase was washed with brine, dried (Na.sub.2SO.sub.4), and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography (EtOAc:PE=0-70% in 50 minutes) to give the titled compound (43 g, 50%). MS: M/e 173 (M+1).sup.+.
Step B: methyl 2-bromo-5-(methylamino)thiazole-4-carboxylate
##STR00630##
[0872] NBS (45 g, 0.25 mol) was added to the mixture of methyl 5-(methylamino)thiazole-4-carboxylate (43 g, 0.25 mol) in MeCN (300 ml). The reaction was stirred at rt for overnight. The reaction was purified by c.c (EtOAc:PE=0-50% in 20 minutes) to give the titled compound (48 g, 77%). .sup.1H NMR (400 MHz, CDCl.sub.3) 7.38 (s, 1H), 3.94-3.81 (m, 3H), 3.03 (s, 3H) ppm. MS: M/e 251 (M+1).sup.+.
Step C: methyl 2-bromo-5-(N-methylacetamido)thiazole-4-carboxylate
##STR00631##
[0873] Methyl 2-bromo-5-(methylamino)thiazole-4-carboxylate (25 g, 0.1 mol) was dissolved in DCE (100 ml) followed by addition of acetic anhydride (51 g, 0.5 mol). The reaction was stirred at 100 C. for 3 days in a sealed tube. The reaction was quenched with NaHCO.sub.3 (aq.) The aqueous phase was extracted with DCM. The combined organic layers were washed with brine, dried over MgSO.sub.4, and concentrated to dryness. The resulting residue was slurried from PE to give the titled compound (27 g, 93%). .sup.1H NMR (400 MHz, CDCl.sub.3) 3.94 (s, 3H), 3.33 (d, J=72.1 Hz, 3H), 2.37-1.94 (m, 3H) ppm. MS: M/e 293 (M+1).sup.+.
Step D: 2-bromo-7-hydroxy-4-methylthiazolo[5,4-b]pyridin-5 (4H)-one
##STR00632##
[0874] LiHMDS (1M in THF, 25 ml) was added to the mixture of methyl 2-bromo-5-(N-methylacetamido)thiazole-4-carboxylate (5 g, 17 mmol) in THF (200 ml) at 65 C. dropwise. The reaction was stirred at 60 C. to rt for 2 hours. Water was added to quench the reaction and extracted with EtOAc. The aqueous phase was adjusted pH to 3-4 with saturated citric acid solution and extracted with DCM. The organic phase was then washed with brine, dried (Na.sub.2SO.sub.4), and concentrated under reduced pressure to give the titled compound (1.5 g, crude). MS: M/e 261 (M+1).sup.+.
Step E: 2-bromo-4-methyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridin-7-yl trifluoromethanesulfonate
##STR00633##
[0875] The 2-bromo-7-hydroxy-4-methylthiazolo[5,4-b]pyridin-5 (4H)-one (2.6 g, crude) was dissolved in the mixture of DMF/THF (20 ml/20 ml) and K.sub.2CO.sub.3 (2.7 g, 20 mmol) and N,N-bis(trifluoromethylsulfonyl) aniline (7.1 g, 20 mmol) was added to the solution. The reaction was stirred at rt overnight. Water was added to quench the reaction and extracted with EtOAc. The organic phase was then washed with brine, dried (Na.sub.2SO.sub.4), and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (petroleum ether/ethyl acetate, 0-80% in 20 minutes) to give the titled compound (600 mg, 9% over 2 steps). MS: M/e 393 (M+1).sup.+.
Step F: tert-butyl (2R,5S)-4-(2-bromo-4-methyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00634##
[0876] A sealed tube was charged with 2-bromo-4-methyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridin-7-yl trifluoromethanesulfonate (700 mg, 1.8 mmol), tert-butyl (2R,5S)-2,5-dimethylpiperazine-1-carboxylate (458 mg, 2.1 mmol), DIPEA (464 mg, 3.6 mmol) and acetonitrile (10 ml). The mixture was stirred at 90 C. for overnight, cooled to rt. The reaction was purified by flash column chromatography (EtOAc:PE=0-80% in 20 minutes) to give the titled compound (650 mg, 80%). MS: M/e 457 (M+1).sup.+.
Step G: methyl 7-((2S,5R)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin-1-yl)-4-methyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridine-2-carboxylate
##STR00635##
[0877] To a mixture of tert-butyl (2R,5S)-4-(2-bromo-4-methyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (650 mg, 1.4 mmol) was dissolved in MeOH (10 ml) added Pd(dppf)Cl.sub.2 (208 mg, 0.28 mmol), Et.sub.3N (283 mg, 2.8 mmol). The reaction was stirred at 90 C. for overnight under carbon monoxide (20 atm) atmosphere. The reaction was quenched with water and extracted with EtOAc. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, and concentrated. The residue was purified by flash column chromatography (EtOAc:PE=0-100% in 20 minutes) to give the titled compound (470 mg, 77%). MS: M/e 437 (M+1).sup.+.
Step H: tert-butyl (2R,5S)-4-(2-(hydroxymethyl)-4-methyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00636##
[0878] To a mixture of methyl 7-((2S,5R)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin-1-yl)-4-methyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridine-2-carboxylate (470 mg, 1.1 mmol) in THF (10 mL) was added NaBH.sub.4 (164 mg, 4.3 mmol). The resulting mixture was stirred at 70 C. for overnight. The reaction was diluted with water, extracted with EtOAc, washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated. The residue was purified by flash column chromatography (MeOH:DCM=0-10% in 20 minutes) to give the titled compound (390 mg, 88%). .sup.1H NMR (400 MHz, CD.sub.3OD) 5.74 (s, 1H), 5.49 (s, 1H), 4.89 (s, 1H), 4.83 (s, 2H), 4.45-4.10 (m, 2H), 3.76 (d, J=13.4 Hz, 1H), 3.63 (s, 3H), 3.57-3.39 (m, 2H), 1.49 (s, 9H), 1.26 (d, J=6.7 Hz, 3H), 1.18 (d, J=6.1 Hz, 3H) ppm. MS: M/e 409 (M+1).sup.+.
Step I: tert-butyl (2R,5S)-4-(2-(chloromethyl)-4-methyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00637##
[0879] To a mixture of tert-butyl (2R,5S)-4-(2-(hydroxymethyl)-4-methyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (370 mg, 0.9 mmol) in DCM (5 mL) was added SOCl.sub.2 (322 mg, 2.7 mmol). The resulting mixture was stirred at rt for 30 minutes. The reaction was diluted with NaHCO.sub.3 (aq.), extracted with DCM, washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated. The resulting residue was used in the next step (400 mg, crude). MS: M/e 427 (M+1).sup.+.
Step G: tert-butyl (2R,5S)-4-(2-(cyanomethyl)-4-methyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate
##STR00638##
[0880] To a mixture of tert-butyl (2R,5S)-4-(2-(chloromethyl)-4-methyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (400 mg, crude) in DCM (4 mL) was added TMSCN (278 mg, 2.81 mmol), TBAF (1M in THF, 2.8 ml, 2.8 mmol). The resulting mixture was stirred at rt for 2 hours. The reaction was diluted with water, extracted with DCM, washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated. The residue was purified by Combi-flash (EtOAc:PE=0-100% in 20 minutes) to give the titled compound (130 mg, 33%). MS: M/e 418 (M+1).sup.+.
Step K: 2-(7-((2S,5R)-2,5-dimethylpiperazin-1-yl)-4-methyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridin-2-yl)acetonitrile
##STR00639##
[0881] To tert-butyl (2R,5S)-4-(2-(cyanomethyl)-4-methyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridin-7-yl)-2,5-dimethylpiperazine-1-carboxylate (130 mg) in DCM (4 mL) was added TFA (1.5 ml). The mixture was stirred at rt for 4 hours. The mixture was concentrated to dryness and diluted with water. The mixture was extracted with EtOAc. The aqueous phase was adjusted pH to 12-13 with saturated sodium carbonate solution and extracted with DCM. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, and concentrated. The residue 80 mg (yield=81%) was used in the next step. MS: M/e 318 (M+1).sup.+.
Step L: 2-(7-((2S,5R)-2,5-dimethyl-4-(1-(quinoxalin-6-yl)ethyl)piperazin-1-yl)-4-methyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridin-2-yl)acetonitrile
##STR00640##
[0882] To the 2-(7-((2S,5R)-2,5-dimethylpiperazin-1-yl)-4-methyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridin-2-yl)acetonitrile (80 mg, 0.25 mmol) in MeCN (3 mL) was added 1-(quinoxalin-6-yl)ethan-1-ol (66 mg, 0.38 mmol), (cyanomethyl)trimethylphosphonium iodide (121 mg, 0.5 mmol) and DIPEA (129 mg, 1 mmol). The resulted mixture was stirred at 90 C. for overnight. The mixture was concentrated to dryness. The reaction was quenched with water, extracted with EtOAc, washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated. The resulting residue was purified by flash column chromatography (MeOH:DCM=0-10% in 20 minutes) to give the titled compound, which was further separated into Compound A194a (12 mg) and Compound A194b (11 mg) by chiral prep-HPLC. The chiral separation conditions are shown below.
TABLE-US-00030 Column CHIRALPAK IH Column Size 2 cm 25 cm, 5 um Mobile Phase A MtBE (0.5% 2 mM NH3MeOH) Mobile Phase B MeOH Flow Rate 20 mL/min Wave Length UV 220 nm Temperature 25 C. Prep-HPLC Equipment Prep-HPLC-Gilson
[0883] Compound A194a (the earlier peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.87 (d, J=3.9 Hz, 2H), 8.14-8.02 (m, 3H), 5.71 (s, 1H), 4.39 (s, 1H), 3.98 (d, J=6.6 Hz, 1H), 3.60 (s, 3H), 3.49 (d, J=3.4 Hz, 1H), 3.33 (s, 1H), 3.14-3.05 (m, 1H), 2.93 (d, J=12.5 Hz, 4H), 1.45 (dd, J=11.7, 6.5 Hz, 6H), 1.09 (d, J=6.5 Hz, 3H) ppm. MS: M/e 474 (M+1).sup.+.
[0884] Compound A194b (the later peak): .sup.1H NMR (400 MHz, CD.sub.3OD) 8.87 (d, J=4.2 Hz, 2H), 8.06 (q, J=8.8 Hz, 3H), 5.71 (s, 1H), 4.68-4.35 (m, 2H), 3.83 (d, J=6.1 Hz, 1H), 3.69 (d, J=10.4 Hz, 2H), 3.60 (s, 3H), 3.32 (s, 1H), 2.87 (d, J=8.5 Hz, 2H), 2.20 (d, J=11.7 Hz, 1H), 1.47 (d, J=6.5 Hz, 3H), 1.26 (t, J=9.4 Hz, 6H) ppm. MS: M/e 474 (M+1).sup.+.
Compound A195: 2-(7-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-4-methyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridin-2-yl)acetonitrile
##STR00641##
Step 1: tert-butyl (2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazine-1-carboxylate
##STR00642##
[0885] A sealed tube was charged with 1-(4-fluoro-2-(trifluoromethyl)phenyl)ethan-1-ol (1.56 g, 7.5 mmol), tert-butyl (2S,5R)-2,5-dimethylpiperazine-1-carboxylate (1.07 g, 5 mmol), (cyanomethyl)trimethylphosphonium iodide (2.4 g, 10 mmol), DIPEA (2.58 g, 20 mmol) and acetonitrile (15 ml). The mixture was stirred at 100 C. overnight, cooled to rt. The reaction was purified by flash column chromatography (EtOAc:PE=0-60% in 25 minutes) to give the titled compound (2.5 g, crude). MS: M/e 405 (M+1).sup.+.
Step 2: (2R,5S)-1-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazine
##STR00643##
[0886] To tert-butyl (2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazine-1-carboxylate (2.5 g) in DCM (20 mL) was added TFA (7 ml). The mixture was stirred at rt for 4 hours. The mixture was concentrated to dryness and diluted with water. The mixture was extracted with EtOAc. The aqueous phase was adjusted pH to 12-13 with saturated sodium carbonate solution and extracted with DCM. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, and concentrated to give the titled compound (1.1 g, 59%). MS: M/e 305 (M+1).sup.+.
Step 3: 2-bromo-7-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-4-methylthiazolo[5,4-b]pyridin-5 (4H)-one
##STR00644##
[0887] To tert-butyl (2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazine-1-carboxylate (85 mg, crude) in MeCN (3 mL) was added 2-bromo-4-methyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridin-7-yl trifluoromethanesulfonate (100 mg, 0.25 mmol) and DIPEA (129 mg, 1 mmol). The resulted mixture was stirred at 90 C. for overnight. The mixture was concentrated to dryness. The reaction was quenched with water, extracted with EtOAc, washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated. The resulting residue was purified by flash column chromatography (EtOAc:PE=0-60% in 20 minutes) to give the titled compound (100 mg, 72%). MS: M/e 547 (M+1).sup.+.
Step 4: methyl 7-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-4-methyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridine-2-carboxylate
##STR00645##
[0888] To a mixture of 2-bromo-7-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-4-methylthiazolo[5,4-b]pyridin-5 (4H)-one (100 mg, 0.18 mmol) was dissolved in MeOH (5 ml) added Pd(dppf)Cl.sub.2 (30 mg, 0.01 mmol), Et.sub.3N (36 mg, 0.36 mmol). The reaction was stirred at 90 C. for overnight under carbon monoxide (20 atm) atmosphere. The reaction was quenched with water and extracted with EtOAc. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4, and concentrated. The residue was purified by flash column chromatography (EtOAc:PE=0-100% in 20 minutes) to give the titled compound (70 mg, 73%). MS: M/e 527 (M+1).sup.+.
Step 5: 7-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-2-(hydroxymethyl)-4-methylthiazolo[5,4-b]pyridin-5 (4H)-one
##STR00646##
[0889] To a mixture of methyl 7-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-4-methyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridine-2-carboxylate (70 mg, 0.13 mmol) in THF (4 mL) was added NaBH.sub.4 (20 mg, 0.53 mmol). The resulting mixture was stirred at 70 C. for overnight. The reaction was diluted with water, extracted with EtOAc washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was used in the next step (80 mg, crude). MS: M/e 499 (M+1).sup.+.
Step 6: 2-(chloromethyl)-7-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-4-methylthiazolo[5,4-b]pyridin-5 (4H)-one
##STR00647##
[0890] To a mixture of 7-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-2-(hydroxymethyl)-4-methylthiazolo[5,4-b]pyridin-5 (4H)-one (80 mg, crude) in DCM (4 mL) was added SOCl.sub.2 (57 mg, 0.48 mmol). The resulting mixture was stirred at 0 C. for 30 minutes. The reaction was diluted with NaHCO.sub.3 (aq.), extracted with DCM, washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was used in the next step (60 mg, crude). MS: M/e 517 (M+1).sup.+.
Step 7: 2-(7-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-4-methyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridin-2-yl)acetonitrile
##STR00648##
[0891] To a mixture of 2-(chloromethyl)-7-((2S,5R)-4-(1-(4-fluoro-2-(trifluoromethyl)phenyl)ethyl)-2,5-dimethylpiperazin-1-yl)-4-methylthiazolo[5,4-b]pyridin-5 (4H)-one (60 mg, crude) in DCM (4 mL) was added TMSCN (23 mg, 0.23 mmol), TBAF (1M in THF, 0.23 ml, 0.23 mmol). The resulting mixture was stirred at rt for 2 hours. The reaction was diluted with water, extracted with DCM, washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated to dryness. The resulting residue was purified by Prep-TLC (MeOH:DCM=1:13) to give the titled compound (13 mg). .sup.1H NMR (400 MHz, CD.sub.3OD) 8.14-8.01 (m, 1H), 7.40 (t, J=9.0 Hz, 2H), 5.70 (s, 1H), 4.40 (s, 1H), 4.01 (d, J=57.5 Hz, 1H), 3.65 (s, 1H), 3.61 (d, J=7.5 Hz, 3H), 3.48-3.33 (m, 1H), 2.99 (dd, J=42.5, 9.4 Hz, 1H), 2.82 (d, J=8.4 Hz, 2H), 2.00 (d, J=12.1 Hz, 1H), 1.48-1.30 (m, 4H), 1.23 (dd, J=28.7, 6.5 Hz, 5H), 1.04 (d, J=6.4 Hz, 1H) ppm. MS: M/e 508 (M+1).sup.+.
[0892] The following compounds were prepared according to the similar procedures under appropriate conditions:
TABLE-US-00031 Procedures similar to or Compound Name Structure Yield following Characterization A2 2-(7-((2S,5R)-4-(1-(4-fluoro-2- (trifluoromethyl)phenyl)ethyl)- 2,5-dimethylpiperazin-1-yl)-3,4- dimethyl-5-oxo-4,5-dihydro-3H- imidazo[4,5-b]pyridin-2- yl)acetonitrile
Assay
Biochemical DGK IC50 Assays
[0893] Enzymatic reactions of DGK, DGK and DGK were performed using ADP-Glo assay with lipid micelle substrate. Full length DGK (in-house protein M1-V929 with SEQ ID No: 2) was expressed in baculovirus expression system. Full length DGK (D21-10BG, SignalChem) and full length DGK (D23-10G, SignalChem) were purchased. Lipid micelle was prepared by dissolving DAG (Sigma, 317505-10MG) and PS (Sigma, P7769-100MG) with chloroform which was furtherly removed by rotary evaporation. The obtained residue was resuspended in buffer containing 25 mM HEPES pH 7.0, 0.5 mM EDTA and 160 mM Octyl -D glucopyranoside by vigorous mixing and ultrasonic (IID, Scientz).
TABLE-US-00032 Final concentration in reaction Enzyme Enzyme (nM) ATP (M) DAG (M) DGK 7.5 140 80 DGK 10 140 80 DGK 0.5 140 80
[0894] The inhibition activities testing for the compound disclosed herein were carried out at room temperature in assay buffer containing 50 mM HEPES, 10 mM MgCl.sub.2, 0.01% BSA, 0.1 mM Na.sub.3VO.sub.4, 0.005% Tween-20 and 0.01 mM CaCl.sub.2. Compounds in DMSO were dispensed into wells of a black 384 well plate (Corning 4514) using D300e digital dispenser (Tecan). The ranges of compounds final concentration were 1.5510000 nM or 23.3150000 nM. 3 L 2 enzyme solution was added to wells. After incubation for 1 hour, 3 L 2 substates solution containing 160 M DAG and 280 M ATP was added to the wells to initiate reaction. After 1 hour reaction, 5 L ADP-Glo reagent (Promga V9101) was added and incubated for 40 minutes. 10 L Kinase Detection reagent was added and incubated for 30 minutes. Luminescence was measured on a microplate reader (PHERAstar FSX, BMG labtech). The IC.sub.50s are calculated based on inhibition of enzyme activity in the presence of increasing concentrations of compounds. Selected compounds had no inhibitory activity on DGK. IC.sub.50s of the compounds disclosed herein for DGK and DGK are shown in Table 1.
Baculovirus Expression of Human DGK
[0895] Human His-TEV-DGK-zeta-pFastBacl and human baculovirus samples was generated using the Bac-to-Bac baculovirus expression system (Invitrogen) according to the manufacturer's protocol. The DNA used for expression of DGK-zeta, have SEQ ID Nos: 1. Baculovirus amplification was achieved using infected SF9 cells at 1:2000 virus/cells ratios, and grown for 96 hours at 27 C. post-transfected.
[0896] The expression scaled up for each protein was carried out in the flask 3 L from CORNING. 4 L of 310.sup.6 cells/mL Sf9 cells (Expression System, Invitrogen) grown in SF900 II SFM insect medium (Expression System) was infected with virus stock at 1:200 virus/cells ratio, and grown for 48 hours at 27 C. post-transfection. The infected cell culture was harvested by centrifugation at 6000 rpm for 15 min 4 C. in a SORVALL LYNX6000 centrifuge. The cell pellets were stored at 80 C.
Purification of Human DGK-zeta
[0897] Full length human DGK produced as described above, was purified from Sf9 baculovirus-infected insect cell paste. The cells were lysed using sonication method, and the lysates were clarified by centrifugation. The clarified lysates were purified to 90% homogeneity, using two successive column chromatography steps on an AKTA Purifier system. The two steps column chromatography included nickel affinity resin capture (i.e. Ni-NTA Agarose, Qiagen), followed by size exclusion chromatography (i.e. Hiload16/60 Superdex200 prep grade, GE Healthcare. The protein was delivered and stored at 80 C. The formulation buffers were identical for the protein: 25 mM Tris, 150 mmol/L NaCl, 2 mM DTT, pH8.0.
TABLE-US-00033 SEQIDNo:1isNucleotidesequenceencodingHis-TEV-hDGK-(M1-V929): ATGCATCACCATCACCATCACCATCACGAGAACCTGTACTTCCAGGGATCCATGGA ACCCCGTGATGGTAGCCCCGAAGCTCGTAGCTCCGATTCCGAGTCCGCCAGCGCTTCCTCCT CCGGTAGCGAACGTGACGCTGGTCCCGAGCCCGACAAAGCTCCCCGTCGTCTGAATAAGCG CCGTTTTCCCGGTCTCCGTCTGTTCGGCCACCGCAAGGCCATCACTAAGTCCGGTCTCCAGCA TCTGGCTCCTCCTCCTCCTACCCCCGGTGCTCCTTGCTCCGAATCCGAGCGCCAGATTCGCTC CACTGTGGATTGGTCCGAAAGCGCCACCTATGGTGAGCATATCTGGTTCGAGACCAACGTCT CCGGCGACTTCTGTTATGTCGGTGAGCAATACTGTGTGGCTCGTATGCTGCAGAAGTCCGTG TCCCGCCGTAAATGCGCCGCTTGCAAAATCGTGGTCCATACCCCTTGCATCGAGCAACTGGA GAAAATCAACTTCCGCTGCAAGCCCAGCTTTCGTGAGTCCGGTTCCCGCAACGTGCGCGAAC CTACTTTCGTGCGCCACCACTGGGTGCATCGTCGTCGCCAAGACGGCAAATGCCGCCACTGC GGCAAAGGTTTTCAGCAGAAATTCACCTTCCACAGCAAGGAGATCGTCGCCATCAGCTGCAG CTGGTGCAAACAAGCTTACCATTCCAAAGTGAGCTGCTTCATGCTCCAGCAGATCGAAGAGC CTTGCTCTCTGGGTGTGCATGCTGCTGTCGTGATTCCCCCTACTTGGATTCTGCGTGCTCGCC GTCCCCAGAACACTCTGAAGGCCTCCAAAAAGAAGAAGCGCGCCAGCTTCAAGCGTAAGAG CTCCAAAAAGGGTCCCGAAGAGGGCCGTTGGCGTCCCTTCATCATCCGCCCTACTCCTTCCC CTCTGATGAAGCCTCTGCTGGTCTTCGTCAACCCTAAGAGCGGCGGCAACCAAGGTGCTAAA ATCATCCAGTCCTTCCTCTGGTATCTGAACCCTCGTCAAGTGTTCGACCTCAGCCAAGGCGGT CCTAAGGAGGCTCTGGAGATGTACCGCAAGGTCCACAATCTGCGCATCCTCGCTTGTGGTGG CGATGGCACCGTGGGCTGGATTCTGTCCACTCTGGACCAACTGCGTCTGAAACCTCCCCCCC CCGTGGCTATTCTGCCTCTCGGTACCGGCAACGATCTGGCTCGTACTCTGAATTGGGGTGGT GGCTACACCGATGAGCCCGTGTCCAAGATTCTGTCCCACGTCGAAGAAGGCAATGTCGTCCA ACTGGACCGTTGGGACCTCCACGCCGAACCCAACCCCGAGGCTGGCCCCGAGGACCGTGAC GAGGGCGCTACTGACCGTCTGCCCCTCGACGTCTTCAATAATTACTTCTCTCTGGGCTTTGAC GCTCACGTGACTCTGGAATTTCATGAAAGCCGCGAGGCCAACCCCGAGAAGTTCAATTCCCG TTTCCGCAACAAGATGTTCTACGCTGGCACCGCCTTCAGCGACTTCCTCATGGGCTCCAGCA AGGACCTCGCTAAGCATATCCGCGTGGTGTGCGATGGCATGGATCTGACCCCTAAGATCCAA GATCTGAAGCCCCAATGTGTCGTGTTTCTGAACATCCCCCGCTACTGCGCTGGTACTATGCCT TGGGGCCATCCCGGTGAACACCATGACTTCGAACCTCAGCGTCATGATGACGGCTATCTGGA GGTGATCGGTTTCACCATGACCTCCCTCGCTGCTCTGCAAGTGGGTGGCCACGGCGAACGTC TGACTCAATGCCGCGAGGTGGTGCTGACCACCAGCAAAGCCATCCCCGTCCAAGTGGATGGT GAGCCTTGCAAGCTGGCCGCCTCCCGTATCCGTATCGCTCTCCGCAATCAAGCTACCATGGT CCAGAAGGCCAAACGCCGCAGCGCTGCTCCTCTCCACAGCGACCAACAACCCGTCCCCGAA CAGCTGCGCATCCAAGTGTCCCGTGTCAGCATGCATGACTACGAGGCTCTGCACTACGACAA GGAACAGCTGAAGGAAGCCAGCGTGCCTCTGGGTACTGTGGTCGTGCCCGGTGACAGCGAT CTGGAGCTCTGCCGTGCCCACATCGAGCGTCTGCAGCAAGAGCCCGACGGTGCTGGTGCCAA GAGCCCTACTTGCCAAAAACTCTCCCCCAAGTGGTGTTTCCTCGACGCTACCACCGCCAGCC GCTTCTACCGCATTGATCGCGCCCAAGAGCATCTGAACTATGTCACCGAGATCGCTCAAGAC GAGATCTACATCCTCGACCCCGAACTCCTCGGTGCTAGCGCCCGTCCCGACCTCCCCACTCCT ACCTCCCCTCTGCCCACTTCCCCTTGTTCCCCCACCCCTCGTAGCCTCCAAGGTGATGCTGCC CCTCCTCAAGGTGAGGAGCTCATTGAGGCCGCTAAGCGTAACGATTTCTGCAAGCTCCAAGA GCTGCATCGTGCTGGTGGCGACCTCATGCACCGCGATGAGCAGAGCCGCACTCTGCTGCACC ACGCTGTGTCCACTGGTAGCAAGGACGTGGTGCGCTATCTGCTGGACCACGCTCCTCCCGAG ATCCTCGACGCTGTGGAAGAAAACGGCGAGACTTGCCTCCACCAAGCTGCTGCTCTGGGTCA ACGTACCATCTGCCACTACATCGTCGAAGCTGGTGCTTCTCTGATGAAGACCGACCAGCAAG GTGATACTCCCCGTCAGCGCGCCGAGAAAGCCCAAGACACCGAACTGGCTGCCTATCTGGA GAACCGTCAGCACTACCAGATGATTCAGCGTGAAGACCAAGAGACCGCCGTGTAA SEQIDNo:2isAminoacidsequenceofHis-TEV-hDGK-(M1-V929): MEPRDGSPEARSSDSESASASSSGSERDAGPEPDKAPRRLNKRRFPGLRLFGHRKAITKS GLQHLAPPPPTPGAPCSESERQIRSTVDWSESATYGEHIWFETNVSGDFCYVGEQYCVARMLQKS VSRRKCAACKIVVHTPCIEQLEKINFRCKPSFRESGSRNVREPTFVRHHWVHRRRQDGKCRHCG KGFQQKFTFHSKEIVAISCSWCKQAYHSKVSCFMLQQIEEPCSLGVHAAVVIPPTWILRARRPQN TLKASKKKKRASFKRKSSKKGPEEGRWRPFIIRPTPSPLMKPLLVFVNPKSGGNQGAKIIQSFLWY LNPRQVFDLSQGGPKEALEMYRKVHNLRILACGGDGTVGWILSTLDQLRLKPPPPVAILPLGTGN DLARTLNWGGGYTDEPVSKILSHVEEGNVVQLDRWDLHAEPNPEAGPEDRDEGATDRLPLDVF NNYFSLGFDAHVTLEFHESREANPEKFNSRFRNKMFYAGTAFSDFLMGSSKDLAKHIRVVCDGM DLTPKIQDLKPQCVVFLNIPRYCAGTMPWGHPGEHHDFEPQRHDDGYLEVIGFTMTSLAALQVG GHGERLTQCREVVLTTSKAIPVQVDGEPCKLAASRIRIALRNQATMVQKAKRRSAAPLHSDQQP VPEQLRIQVSRVSMHDYEALHYDKEQLKEASVPLGTVVVPGDSDLELCRAHIERLQQEPDGAGA KSPTCQKLSPKWCFLDATTASRFYRIDRAQEHLNYVTEIAQDEIYILDPELLGASARPDLPTPTSPL PTSPCSPTPRSLQGDAAPPQGEELIEAAKRNDFCKLQELHRAGGDLMHRDEQSRTLLHHAVSTGS KDVVRYLLDHAPPEILDAVEENGETCLHQAAALGQRTICHYIVEAGASLMKTDQQGDTPRQRAE KAQDTELAAYLENRQHYQMIQREDQETAV*
TABLE-US-00034 TABLE 1 Compound DGK ADP-Glo DGK ADP-Glo No. assay IC50(nM) assay IC50(nM) A1 19 20,200 A1a 9 14,900 A1b 2,350 >10,000 A2 92 >150,000 A3 398 822 A4a 566 >10,000 A4b 64 >10,000 A5a 878 5,770 A5b 88 616 A6a 528 >10,000 A6b 13 >10,000 A7a 3,110 >10,000 A7b 35 97,600 A8a 1,480 >10,000 A8b 97 >150,000 A9 262 >10,000 A9a 8,590 >10,000 A9b 169 >10,000 A10 25 >10,000 A10a 405 >10,000 A10b 14 >10,000 A11a 764 57,800 A11b 14 11,900 A12a 2.7 18,800 A12b 57 27,400 A13a 5,060 >150,000 A13b 54 >150,000 14a 1,420 >10,000 A14b 180 >10,000 A15a 1,410 >10,000 A15b 291 >10,000 A16 24 58,200 A17a 133 1,460 A17b 409 1,580 A18a 19 23,600 A18b 327 6,380 A19a 2.8 27,500 A19b 82 6,940 A20 5 93,000 A21 61 >150,000 A22a 26 >10,000 A23a 8 63,200 A23b 94 16,200 A24a 975 >10,000 A24b 42 131,000 A25 15 36,900 A26 74 4,240 A27a 569 >10,000 A27b 15 99,500 A28 3 96,900 A29a 437 >150,000 A29b 4 136,000 A30 14 44,300 A31 4 110,000 A32a 3,740 >10,000 A32b 138 5,590 A33a 804 >10,000 A33b 3 >10,000 A34a 392 >10,000 A34b 9 >10,000 A35a 2,760 >150,000 A35b 24 >150,000 A36 17 104,000 A37 18 128,000 A38 165 143,000 A39 211 >150,000 A40 398 25,900 A41 16 9,700 A42 42 61,900 A42a 24 78,700 A42b 285 118,000 A43 79 101,000 A44 34 51,200 A45 410 79,300 A46 301 >150,000 A47 518 >150,000 A48 29 44,600 A49 35 123,000 A49a 6 77,900 A49b 400 >150,000 A50a 4 24,700 A50b 332 13,600 A51a 3 14,000 A51b 265 22,300 A52a 32 >150,000 A52b 1,030 >150,000 A53a 24 60,300 A53b 662 >150,000 A54 37 >150,000 A55 31 68,700 A56a 810 108,000 A56b 23 50,900 A57 3,360 >150,000 A58 40 >10,000 A58a 20 13,300 A58b 1,660 90,800 A59a 238 40,300 A59b 2.3 14,200 A60a 3.3 72,500 A60b 206 >150,000 A61a 2 10,100 A61b 420 37,900 A62 279 >150,000 A63a 2 10,300 A63b 22 7,400 A64a 0.88 5,280 A64b 14 4,750 A65 2.3 24,200 A66a 217 15,800 A66b 1 6,860 A67 10 118,000 A68 7 43,800 A68a 2 20,300 A68b 199 72,800 A69 6 3,070 A69a 2 1,190 A69b 147 6,090 A70 8 9,690 A71 48 124,000 A72 64 >150,000 A73 639 >150,000 A74 45 >150,000 A74a 46 >150,000 A74b 4,870 >150,000 A75 253 >150,000 A76 419 >150,000 A77 6 19,700 A78 7 102,000 A79 16 16,400 A79a 38 7,280 A79b 529 14,800 A80 31 95,000 A80a 7 65,600 A80b 664 >150000 A81 7 >150,000 A82 201 9,520 A83 40 >150,000 A84 232 >150,000 A85 25 103,000 A85a 1,250 148,247 A85b 14 88,617 A86 67 >150,000 A87 2 21,900 A88 4 111,000 A89 334 >150,000 A90 213 >150,000 A91 20 >150,000 A92 123 >150,000 A93 414 >150,000 A94 14 23,200 A94a 6 16,600 A94b 532 >150,000 A95 42 50,500 A95a 11 24,700 A95b 555 108,000 A96 63 49,600 A97 11 51,100 A98 5 11,600 A98a 5 14,300 A98b 262 19,700 A99 128 55,600 A99a 71 34,500 A99b 536 46,500 A100 4,110 >150,000 A101 4,230 >150,000 A102 37 >150,000 A103a 524 >150,000 A103b 52 >150,000 A104 839 >150,000 A105 767 >150,000 A106 >10,000 >150,000 A107 >10,000 >150,000 A108 32 95,100 A108a 7 65,200 A108b 822 95,800 A109 161 52,400 A110 7 >150,000 A111 771 >150,000 A112 35 >150,000 A113 18 >150,000 A114 246 135,000 A115 19 >150,000 A116 350 >150,000 A116a 7,760 >150,000 A116b 228 >150,000 A117a 3,300 >150,000 A117b 75 145,000 A118a 4,820 >150,000 A118b 103 65,600 A119 172 >150,000 A120 1,280 >150,000 A121 212 >150,000 A122 834 >150,000 A123 49 128,000 A124 48 111,000 A125 42 >150,000 A126 24 124,000 A127 428 >150,000 A128 203 94,000 A129 4 >150,000 A130 3,900 >150,000 A131 5,850 >150,000 A132 1,580 >150,000 A133 111 >150,000 A134 22 113,000 A135 14 >150,000 A136 2 >150,000 A137 64 48,700 A138 949 >150,000 A139 2,190 120,000 A140 594 >150,000 A141 1,290 >150,000 A142 6,300 >150,000 A143 >10,000 >150,000 A144 60 74,100 A145 480 5,620 A146a 4,190 9,150 A146b 290 610 A147 1,100 2,580 A148a 1,100 3,630 A148b 176 666 A149a 37 1,620 A149b 15 308 A150 789 1,180 A151a 187 113 A151b 1,950 529 A152 707 2,690 A153a 1,420 1,490 A153b 21 1,270 A154 178 2,430 A155 1,210 101,000 A156a 496 68,600 A156b 6 14,100 A157a 28 11,600 A157b 346 55,700 A158 33 122,000 A158a 8 61,900 A158b 407 >150,000 A159 11 31,500 A159a 25 27,400 A159b 691 63,400 A160 331 >150,000 A161 81 >150,000 A16la 24 71,800 A161b 2,630 >150,000 A162 33 60,500 A163 48 >150,000 A163a 13 >150,000 A163b 2,070 >150,000 A164 31 125,000 A165 69 >150,000 A166 14 26,100 A167 90 19,000 A168 26 64,300 A169 31 28,500 A169a 10 31,700 A169b 455 78,200 A170 1,180 >150,000 A171 107 65,800 A172a 170 132,000 A172b 8 116,000 A173 44 143,000 A173a 12 51,000 A173b 412 83,300 A174a 1,030 51,200 A174b 69 30,600 A175 5 22,200 A175a 291 37,200 A175b 4 21,400 A176a 40 13,100 A176b 7 7,190 A177 168 72,600 A178 15 92,400 A179 6 69,000 A180 4,440 >150,000 A181 17 >150,000 A182 499 >150,000 A183 4 >150,000 A184 6 88,500 A185a 806 121,000 A185b 20 82,500 A186 156 143,000 A187 407 115,000 A188 74 >150,000 A189 34 >150,000 A190 13 106,000 A190a 3 >150,000 A190b 41 38,200 A191 78 40,900 A192 119 >150,000 A193 >10,000 >150,000 A194 7 4,080 A194a 7 3,950 A194b 873 7,020 A195 25 >150,000 A196 749 >10,000
Cell Culture and DGK/Knockout Jurkat Cell Line Construction
[0898] Jurkat cells and human PBMC were maintained in RPMI 1640 medium (Gibco) supplemented with 10% fetal bovine serum (FBS, Thermo Scientific), 100 units/mL penicillin and 0.1 mg/mL streptomycin (Gibco) in a humidified 37 C. environment with 500 COz. HepG2-OS8 cells, which express the single chain variable fragment (scFv) of an anti-human CD.sub.3 mAb OKT3 fused to the C-terminal domain (113-220) of mouse CD.sub.8a which includes hinge, transmembrane and cytoplasmic domains, were maintained in MEM medium (Gibco) supplemented with 10% fetal bovine serum (FBS, Thermo Scientific), 100 units/mL penicillin and 0.1 mg/mL streptomycin (Gibco) in a humidified 37 C. environment with 5% CO.sub.2.
[0899] Jurkat cells were infected with the lentivirus expressing spCas9 and sgRNA targeting human DGK or DGK. Cell clones that stably knockout with DGK/ were established and maintained in the RPMI 1640 complete medium. Knockout efficiency of eSPCas9-Lenticrispr DGK or DGK sgRNA in single cell clone was determined using genomic sequencing and immunoblotting method. Selected compounds did not induce DGK or DGK independent IL-2 production in DGK/ KO jurkat cells.
Non-Stimulated Phosphorylated ERK Detection Assay
[0900] Cellular non-stimulated phospho-ERK were measured using a AlphaLISA-based method (Beaudet, Lucille, et al. Nature Methods. 2008, 5.12: an8-an9). Jurkat cells were subcultured in T75 flasks. The next day, growth medium was replaced to serum free RPMI 1640 for 4 hours or overnight. The cells were then seeded into 96-well plates and treated with compounds. After 2 h compound treatment, lysis buffer (PerkinElmer) was added to each well. Plates were then incubated at room temperature with shaking for 30 minutes. A total of 10 L of cell lysate from each well of a 96-well plate was transferred to a 384-well white assay plate. Phosphor-ERK was quantitated using the AlphaLISA kit (Cat #ALSU-PERK-A10K) as described by the manufacturer manual (PerkinElmer). AlphaLISA signals were measured using a PHERAstar FSX reader (BMG Labtech). Selected compounds did not elevate ERK phosphorylation in Jurkat cells without TCR activation.
IL-2 Production Assay in Human PBMC
[0901] Frozen human PBMC were thawed in RPMI 1640 medium and incubated at 37 C. overnight. OS8 overexpressing HepG2 cells were seeded into 384-well plates overnight. The next day, PBMC were added into the 384-well plates and then treated with compounds. PBMC and HepG2-OS8 cells were co-cultured for 48 hours at 37 C. Culture supernatant was collected for subsequent measurement of IL-2 concentration by a TR-FRET-based method (Degorce, Frangois, et al. Current chemical genomics. 2009, 3: 22) as described by the manufacturer manual (Cisbio). FRET signals were measured using a PHERAstar FSX reader (BMG Labtech). Selected compounds showed good potency in Human PBMC assay.
Explorative Acute Toxicity Study in BALB/c Mice
[0902] The test compound was dissolved in vehicle formulation (DMA: 30% solutol HS-15 (w/v): saline=20:20:60) and injected through tail vein into BALB/c mice at the doses of 2 and/or 10 mg/kg. Continuous clinical observation within 2 hours post injection was performed. Selected compounds were well tolerant at the doses of 2 and/or 10 mg/kg.
[0903] It is to be understood that, if any prior art publication is referred to herein, such reference does not constitute an admission that the publication forms a part of the common general knowledge in the art in any country.
[0904] In the claims which follow and in the preceding description of the invention, except where the context requires otherwise due to express language or necessary implication, the word comprise or variations such as comprises or comprising is used in an inclusive sense, i.e., to specify the presence of the stated features but not to preclude the presence or addition of further features in various embodiments of the invention.
[0905] The disclosures of all publications, patents, patent applications and published patent applications referred to herein by an identifying citation are hereby incorporated herein by reference in their entirety.