Heparanase inhibitors and use thereof
11787783 · 2023-10-17
Assignee
Inventors
- Keats Nelms (Canberra, AU)
- Brett Schwartz (Canberra, AU)
- Colin Jackson (Canberra, AU)
- Martin Banwell (Canberra, AU)
- Edward Hammond (Canberra, AU)
Cpc classification
C07D239/95
CHEMISTRY; METALLURGY
C07D403/06
CHEMISTRY; METALLURGY
C07D403/12
CHEMISTRY; METALLURGY
C07D413/12
CHEMISTRY; METALLURGY
International classification
A61K31/498
HUMAN NECESSITIES
A61P35/00
HUMAN NECESSITIES
C07D239/95
CHEMISTRY; METALLURGY
C07D403/04
CHEMISTRY; METALLURGY
C07D403/06
CHEMISTRY; METALLURGY
C07D403/12
CHEMISTRY; METALLURGY
C07D405/06
CHEMISTRY; METALLURGY
C07D413/06
CHEMISTRY; METALLURGY
C07D413/12
CHEMISTRY; METALLURGY
Abstract
The invention relates to functionalized dihydro- and tetrahydro-quinazoline compounds, pharmaceutical compositions comprising such compounds, and the use of such compounds as heparanase inhibitors for the treatment of diseases or conditions related to heparanase.
Claims
1. A compound of formula ##STR00161## or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: X is S or O; R.sup.1 is selected from H, hydroxyl, halo, C.sub.1-6alkyl, C.sub.1-4alkoxy, O-CH.sub.2phenyl, O-phenyl; R.sup.2 is selected from H, hydroxyl, halo, C.sub.1-6alkyl, C.sub.1-4alkoxy, O-CH.sub.2phenyl, O-phenyl; R.sup.3 is selected from H, hydroxyl, halo, C.sub.1-6alkyl, C.sub.1-4alkoxy, O-CH.sub.2phenyl, O-phenyl; R.sup.4 is selected from H, hydroxyl, halo, C.sub.1-6alkyl, C.sub.1-4alkoxy, O-CH.sub.2phenyl, O-phenyl; or R.sup.1 and R.sup.2, or R.sup.2 and R.sup.3, or R.sup.3 and R.sup.4 together form C.sub.1-3alkylenedioxy; R.sup.5 is selected from H and C.sub.1-6 alkyl; L is selected from C.sub.1-6 alkylene-indolyl, C.sub.1-6alkylene-NHC(O)—, azetidinyl-C(O)—, C.sub.1-6alkylene-NHC(O)-indolyl, and C.sub.1-6alkylene-NHSO.sub.2—; R.sup.6 is selected from H, halo, hydroxyl, C.sub.1-6alkyl, C.sub.2-6alkenyl, C.sub.2-6alkynyl, C.sub.6-10aryl optionally substituted with 1 or 2 R.sup.X groups, C.sub.1-9heteroaryl optionally substituted with 1 or 2 R.sup.X groups, C.sub.2-5heterocycloalkyl optionally substituted with 1 or 2 R.sup.X groups, or is absent; each R.sup.X is independently selected from hydroxyl, halo, nitro, NR′R″; C.sub.1-4alkyl, C.sub.3-6cycloalkyl, haloC.sub.1-4alkyl, C.sub.1-4alkoxy, C.sub.6-10aryl optionally substituted with 1 or 2 R.sup.Y groups, C.sub.1-9heteroaryl, C.sub.1-4alkyl-(C.sub.1-9heteroaryl), C(O)OC.sub.1-4alkyl, C(O)NHR.sup.Y, C.sub.2-5heterocycloalkyl optionally substituted with 1 or 2 C.sub.1-4alkyl groups, C(O)—(C.sub.2-5heterocycloalkyl) optionally substituted with 1 or 2 C.sub.1-4alkyl groups, or two adjacent R.sup.X groups together form C.sub.1-3alkylenedioxy; R.sup.Y is selected from H, hydroxyl, halo, C.sub.1-4 alkyl, haloC.sub.1-4 alkyl, C.sub.1-4 alkoxy, C.sub.1-4 alkylheterocycloalkyl, C(O)—(C.sub.1-4 alkylC.sub.2-5heterocycloalkyl), C.sub.1-4alkylNR′R″; R′ and R″ are independently selected from H, C.sub.1-4alkyl, C.sub.1-4 alkylC.sub.2-5heterocycloalkyl; R.sup.7 is selected from H, C.sub.1-4alkyl, C.sub.1-6alkylC.sub.1-9heteroaryl, wherein the term heterocycloalkyl refers to a cycloalkyl ring in which one or more carbon atoms has been replaced by one or more heteroatoms.
2. A compound of formula ##STR00162## or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: X is S or O; R.sup.1 is selected from H, hydroxyl, halo, C.sub.1-6alkyl, C.sub.1-4alkoxy, O-CH.sub.2phenyl, O-phenyl; R.sup.2 is selected from H, hydroxyl, halo, C.sub.1-6alkyl, C.sub.1-4alkoxy, O-CH.sub.2phenyl, O-phenyl; R.sup.3 is selected from H, hydroxyl, halo, C.sub.1-6alkyl, C.sub.1-4alkoxy, O-CH.sub.2phenyl, O-phenyl; R.sup.4 is selected from H, hydroxyl, halo, C.sub.1-6alkyl, C.sub.1-4alkoxy, O-CH.sub.2phenyl, O-phenyl; or R.sup.1 and R.sup.2, or R.sup.2 and R.sup.3, or R.sup.3 and R.sup.4 together form C.sub.1-3alkylenedioxy; R.sup.5′ is selected from C.sub.1-6 alkyl, C.sub.1-3alkylC(O)OC.sub.1-4alkyl and C.sub.1-3alkylC.sub.6-10aryl optionally substituted with 1 or 2 groups independently selected from haloC.sub.1-3alkyl and haloC.sub.1-3alkoxy; L is selected from C.sub.1-6 alkylene-indolyl, C.sub.1-6alkylene-NHC(O)—, azetidinyl-C(O)—, C.sub.1-6alkylene-NHC(O)-indolyl, and C.sub.1-6alkylene-NHSO.sub.2—; R.sup.6 is selected from H, halo, hydroxyl, C.sub.2-6alkyl, C.sub.2-6alkenyl, C.sub.2-6alkynyl, C.sub.6-10aryl optionally substituted with 1 or 2 R.sup.X groups, C.sub.1-9heteroaryl optionally substituted with 1 or 2 R.sup.X groups, C.sub.2-5heterocycloalkyl optionally substituted with 1 or 2 R.sup.X groups, or is absent; each R.sup.X is independently selected from hydroxyl, halo, nitro, NR′R″; C.sub.1-4alkyl, C.sub.3-6cycloalkyl, C.sub.1-4alkoxy, C.sub.6-10aryl optionally substituted with 1 or 2 R.sup.Y groups, C.sub.1-9heteroaryl, C.sub.1-4alkyl-(C.sub.1-9heteroaryl), C(O)OC.sub.1-4alkyl, C(O)NHR.sup.Y, C.sub.2-5heterocycloalkyl optionally substituted with 1 or 2 C.sub.1-4alkyl groups, C(O)—(C.sub.2-5heterocycloalkyl) optionally substituted with 1 or 2 C.sub.1-4alkyl groups, or two adjacent R.sup.X groups together form C.sub.1-3alkylenedioxy; R.sup.Y is selected from H, hydroxyl, halo, C.sub.1-4 alkyl, haloC.sub.1-4 alkyl, C.sub.1-4 alkoxy, C.sub.1-4 alkylheterocycloalkyl, C(O)—(C.sub.1-4 alkylC.sub.2-5heterocycloalkyl), C.sub.1-4alkylNR′R″; R′ and R″ are independently selected from H, C.sub.1-4alkyl, C.sub.1-4 alkylC.sub.2-5heterocycloalkyl; R.sup.7 is selected from H, C.sub.1-6alkylC.sub.1-9heteroaryl, wherein the term heterocycloalkyl refers to a cycloalkyl ring in which one or more carbon atoms has been replaced by one or more heteroatoms.
3. The compound according to claim 1, wherein each heteroaryl and each heterocycloalkyl group has at least one nitrogen heteroatom.
4. The compound according to claim 1, wherein X is S.
5. The compound according to claim 1, wherein R.sup.1 and R.sup.4 are hydrogen.
6. The compound according to claim 1, wherein R.sup.2 and R.sup.3 are independently selected from hydroxyl, halo, and C.sub.1-3 alkoxy.
7. The compound according to claim 1, wherein R.sup.2 and R.sup.3 are both methoxy, or R.sup.2 and R.sup.3 together are methylenedioxy.
8. The compound according to claim 2, wherein R.sup.5′ is a benzyl group optionally substituted with 1 or 2 groups selected from CF.sub.3 and OCF.sub.3.
9. The compound according to claim 1, wherein each heteroaryl is independently selected from indolyl, pyridyl, pyrazolyl, pyrrolyl, oxazolyl, ozadiazolyl, benzoxadiazolyl and triazolyl, each of which is optionally substituted with 1 or 2 R.sup.X groups.
10. The compound according to claim 1, wherein each heterocycloalkyl is independently selected from aziridinyl, morpholinyl, piperidinyl, piperazinyl, each of which is optionally substituted with 1 or 2 R.sup.X groups.
11. The compound according to claim 1, wherein R.sup.6 is a group selected from indolyl, phenyl, pyridyl, N-morpholinyl, N-piperidinyl, N-piperazinyl, triazolyl, pyrrolyl, pyrazolyl, oxazolyl, ozadiazolyl, benzodiazolyl and pyrrolopyridinyl, wherein each group is optionally substituted with 1 or 2 R.sup.X groups.
12. The compound according to claim 1, wherein L is C.sub.1-2alkylene-NHC(O)— and R.sup.6 is 2-indolyl, pyridyl, oxazolyl or oxadiazolyl, each of which is optionally substituted with 1 or 2 groups.
13. The compound according to claim 1, wherein L is C.sub.1-2alkylene-NHC(O)-indolyl and R.sup.6 is selected from phenyl optionally substituted with 1 or 2 R.sup.X groups, C.sub.3-8heteroaryl optionally substituted with 1 or 2 R.sup.X groups, C(O)(C.sub.3-8heteroaryl) optionally substituted with 1 or 2 R.sup.X groups, C(O)(C.sub.2-5heterocycloalkyl) optionally substituted with 1 or 2 R.sup.X groups, C(O)NH.sup.Y.
14. The compound according to claim 1, wherein each R.sup.X is independently selected from hydroxyl, halo, C.sub.1-3alkyl, C.sub.1-4alkoxy, C(O)OC.sub.1-4alkyl, phenyl, NR′R″ wherein R′ and R″ are independently selected from H, C.sub.1-3alkyl, morpholinyl optionally substituted with 1 or 2 C.sub.1-3alkyl groups, piperazinyl optionally substituted with 1 or 2 C.sub.1-3alkyl groups, C(O)morpholinyl optionally substituted with 1 or 2 C.sub.1-3alkyl groups, C(O)piperazinyl optionally substituted with 1 or 2 C.sub.1-3alkyl groups, C.sub.6-10aryl optionally substituted with 1 or 2 C(O)—(C.sub.1-4alkylC.sub.2-5heterocycloalkyl), or two adjacent R.sup.X groups together form methylenedioxy, pyridyl.
15. The compound according to claim 1 having formula (IA), formula (IB), formula (IC), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof: ##STR00163## wherein: R.sup.2 is H, halo, C.sub.1-3alkoxy, O-CH.sub.2phenyl, O-phenyl; R.sup.3 is H, halo, C.sub.1-3alkoxy, O-CH.sub.2phenyl, O-phenyl; R.sup.7 is H; A is C═O or SO.sub.2; X is C or N; R.sup.E is H, C.sub.1-3alkyl, C(O)—C.sub.2-5heterocycloalkyl (e.g., C(O)—(N-morpholinyl)); R.sup.A, R.sup.B, R.sup.C and R.sup.D are independently selected from H, OH, C.sub.1-3 alkyl, OC.sub.1-3 alkyl, C(O)—(N-heterocycloalkyl) optionally substituted with 1 or 2 C.sub.1-3alkyl groups; N-heteroaryl optionally substituted with 1 or 2 groups selected from OH, halo, C.sub.1-3alkyl, C.sub.1-3alkoxy; phenyl optionally substituted with 1 or 2 groups selected from OH, halo, C.sub.1-3alkyl, C.sub.1-3alkoxy, C(O)NHC.sub.1-3alkyl-[N(C.sub.1-3alkyl).sub.2], C(O)—C.sub.2-5heterocycloalkyl optionally substituted with 1 or 2 C.sub.1-3alkyl groups.
16. The compound according to claim 1 selected from: ##STR00164## ##STR00165## ##STR00166## ##STR00167## ##STR00168## ##STR00169## ##STR00170## ##STR00171## ##STR00172## or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.
17. The compound according to claim 1 selected from ##STR00173## ##STR00174## or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.
18. A pharmaceutical composition comprising a compound of claim 1, or a pharmaceutically acceptable salt thereof, together with one or more pharmaceutically acceptable carriers, diluents or excipients.
19. A pharmaceutical composition comprising a compound of claim 2, or a pharmaceutically acceptable salt thereof, together with one or more pharmaceutically acceptable carriers, diluents or excipients.
20. The compound according to claim 2 having formula (IIA) or formula (IIB), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof: ##STR00175## wherein: R.sup.2 is H, halo, C.sub.1-3alkoxy, O-CH.sub.2phenyl, O-phenyl; R.sup.3 is H, halo, C.sub.1-3alkoxy, O-CH.sub.2phenyl, O-phenyl; R.sup.5′ is selected from H, C.sub.1-6 alkyl, C.sub.1-3alkylC(O)OC.sub.1-4alkyl and C.sub.1-3alkylC.sub.6-10aryl optionally substituted with 1 or 2 groups independently selected from haloC.sub.1-3alkyl and haloC.sub.1-3alkoxy; R.sup.7 is H; A is C═O or SO.sub.2; X is C or N; R.sup.A, R.sup.B, R.sup.C and R.sup.D are independently selected from H, OH, C.sub.1-3 alkyl, OC.sub.1-3 alkyl, C(O)—(N-heterocycloalkyl) optionally substituted with 1 or 2 C.sub.1-3alkyl groups; N-heteroaryl optionally substituted with 1 or 2 groups selected from OH, halo, C.sub.1-3alkyl, C.sub.1-3alkoxy; phenyl optionally substituted with 1 or 2 groups selected from OH, halo, C.sub.1-3alkyl, C.sub.1-3alkoxy, C(O)NHC.sub.1-3alkyl-[N(C.sub.1-3alkyl).sub.2], C(O)—C.sub.2-5heterocycloalkyl optionally substituted with 1 or 2 C.sub.1-3alkyl groups.
21. The compound according to claim 2 selected from: ##STR00176## ##STR00177## ##STR00178## or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.
22. The compound according to claim 2 selected from ##STR00179## ##STR00180## ##STR00181## or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
(1)
(2)
(3)
(4)
(5)
(6)
DETAILED DESCRIPTION OF EMBODIMENTS OF THE INVENTION
(7) The present invention relates to functionalized dihydro- and tetrahydro-quinazoline compounds as defined herein, and to the use of such compounds in the treatment of diseases or conditions associated with heparanase activity. In preferred embodiments, the compounds of the invention are heparanase inhibitors.
(8) In one aspect the present invention relates to compounds of general formula (I) or general formula (II):
(9) ##STR00004##
(10) or a salt, hydrate, solvate, tautomer or stereoisomer thereof,
(11) wherein:
(12) X is S or O; R.sup.1 is selected from H, hydroxyl, halo, C.sub.1-6 alkyl, C.sub.1-4 alkoxy, O—CH.sub.2phenyl, O-phenyl; R.sup.2 is selected from H, hydroxyl, halo, C.sub.1-6 alkyl, C.sub.1-4 alkoxy, O—CH.sub.2phenyl, O-phenyl; R.sup.3 is selected from H, hydroxyl, halo, C.sub.1-6 alkyl, C.sub.1-4 alkoxy, O—CH.sub.2phenyl, O-phenyl; R.sup.4 is selected from H, hydroxyl, halo, C.sub.1-6 alkyl, C.sub.1-4 alkoxy, O—CH.sub.2phenyl, O-phenyl; or R.sup.1 and R.sup.2, or R.sup.2 and R.sup.3, or R.sup.3 and R.sup.4 together form C.sub.1-3 alkylenedioxy; R.sup.5 and R.sup.5′ are independently selected from H, C.sub.1-6 alkyl, C.sub.1-3alkylC(O)OC.sub.1-4alkyl and C.sub.1-3alkylC.sub.6-10aryl optionally substituted with 1 or 2 groups independently selected from haloC.sub.1-3alkyl and haloC.sub.1-3alkoxy; L is selected from C.sub.1-6 alkyl, azetidinyl, C.sub.1-6 alkyl-indolyl, NH, C.sub.1-6alkyl-NHC(O)O, azetidinyl-C(O)—, C.sub.1-6 alkyl-NHC(O)-indolyl, C.sub.1-6 alkyl-NHSO.sub.2—, or is absent; R.sup.6 is selected from H, halo, hydroxyl, C.sub.1-6alkyl, C.sub.1-6alkenyl, C.sub.1-6alkynyl, C.sub.6-10aryl optionally substituted with 1 or 2 R.sup.X groups, C.sub.1-9heteroaryl optionally substituted with 1 or 2 R.sup.X groups, C.sub.2-5heterocycloalkyl optionally substituted with 1 or 2 R.sup.X groups, C(O)(C.sub.1-9heteroaryl) optionally substituted with 1 or 2 R.sup.X groups, C(O)(C.sub.2-5heterocycloalkyl) optionally substituted with 1 or 2 R.sup.X groups, C(O)NHR.sup.Y, or is absent; each R.sup.X is independently selected from hydroxyl, halo, nitro, NR′R″, C.sub.1-4 alkyl, C.sub.3-6 cycloalkyl, haloC.sub.1-4 alkyl, C.sub.1-4alkoxy, C.sub.6-10 aryl optionally substituted with 1 or 2 R.sup.Y groups, C.sub.1-9 heteroaryl, C.sub.1-4 alkyl-(C.sub.1-9heteroaryl), C(O)OC.sub.1-4 alkyl, C(O)NHR.sup.Y, C.sub.2-5heterocycloalkyl optionally substituted with 1 or 2 C.sub.1-4 alkyl groups, C(O)-(heterocycloalkyl) optionally substituted with 1 or 2 C.sub.1-4 alkyl groups, or two adjacent R.sup.X groups together form C.sub.1-3alkylenedioxy; R.sup.Y is selected from H, hydroxyl, halo, C.sub.1-4 alkyl, haloC.sub.1-4 alkyl, C.sub.1-4 alkoxy, C.sub.1-4 alkylheterocycloalkyl, C(O)—(C.sub.1-4 alkylheterocycloalkyl), C.sub.1-4 alkylNR′R″; R′ and R″ are independently selected from H, C.sub.1-4alkyl, and C.sub.1-4 alkylheterocycloalkyl; R.sup.7 is selected from H, C.sub.1-4 alkyl, C.sub.1-6 alkylC.sub.1-9 heteroaryl.
(13) In preferred embodiments each heteroaryl and each heterocycloalkyl group comprises at least one nitrogen heteroatom.
(14) In preferred embodiments X is S.
(15) In preferred embodiments the salt is a pharmaceutically acceptable salt.
(16) In one or more embodiments, each heteroaryl is independently selected from indolyl (e.g., N-indolyl, 2-indolyl, 3-indolyl), pyridyl (e.g., 2-pyridyl, 3-pyridyl, 4-pyridyl), pyrazolyl, pyrrolyl, benzoxadiazolyl, triazolyl, oxazolyl, oxadiazolyl, each of which may be optionally substituted with 1 or 2 R.sup.X groups.
(17) In one or more embodiments, each heterocycloalkyl is independently selected from aziridinyl, morpholinyl, piperidinyl, piperazinyl, each of which may be optionally substituted with 1 or 2 R.sup.X groups.
(18) In one or more embodiments R.sup.1 and R.sup.4 are H. In one or more embodiments R.sup.2 and R.sup.3 are independently H, halo, or C.sub.1-4alkoxy. In one or more embodiments R.sup.2 and R.sup.3 are not both H. In one or more embodiments R.sup.2 and R.sup.3 are both C.sub.1-4alkoxy e.g., methoxy, ethoxy. In one or more embodiments R.sup.2 and R.sup.3 together are methylenedioxy.
(19) In one or more embodiments R.sup.5 or R.sup.5′ is independently H. In one or more embodiments R.sup.5 or R.sup.5′ is independently a C.sub.1-6 alkyl group, e.g., methyl, ethyl. In one or more embodiments R.sup.5 or R.sup.5′ is independently C.sub.1-3alkylC(O)OC.sub.1-3alkyl. In one or more embodiments R.sup.5 or R.sup.5′ is independently a benzyl group optionally substituted with 1 or 2 groups selected from CF.sub.3 and OCF.sub.3.
(20) In one or more embodiments L is C.sub.1-3 alkyl, C.sub.1-3 alkylNHC(O)—, or azetidinyl. In one or more embodiments L is C.sub.1-6 alkyl-NHC(O)-indolyl.
(21) In or more embodiments R.sup.6 is a group selected from indolyl (e.g., 2-indolyl or 3-indolyl), phenyl, pyridyl (e.g., 2-pyridyl, 3-pyridyl or 4-pyridyl), N-morpholinyl, N-piperidinyl, N-piperazinyl, pyrrolyl, diazolyl, triazolyl (e.g., 4-triazolyl), pyrazolyl (e.g., N(1)-pyrazolyl); oxazolyl, ozadiazolyl, benzodiazolyl and pyrrolopyridinyl (e.g., pyrrolo[2,3-b]pyridinyl), wherein each group is optionally substituted with 1 or 2 R.sup.X groups.
(22) In one or more embodiments R.sup.7 is H or C.sub.1-4 alkyl. In more or more embodiments, R.sup.7 is C.sub.1-6 alkylC.sub.1-9 heteroaryl. In one or more embodiments, R.sup.7 is C.sub.1-2 alkyl-(2-indolyl) or C.sub.1-2 alkyl-(3-indolyl).
(23) In one or more embodiments L is absent and R.sup.6 is selected from C.sub.1-6alkyl, C.sub.1-6alkenyl. C.sub.1-6 alkynyl, and C.sub.2-5heterocycloalkyl (e.g., piperidinyl, piperazinyl or morpholinyl) optionally substituted with 1 or 2 R.sup.X groups.
(24) In one or more embodiments L is NH and R.sup.6 is H or phenyl.
(25) In one or more embodiments L is C.sub.1-2alkyl and R.sup.6 is indolyl (e.g., 2-indolyl or 3-indolyl) optionally substituted with 1 or 2 R.sup.X groups. In one or more embodiments L is C.sub.1-2 alkyl-NHC(O)— and R.sup.6 is C.sub.1-9 heteroaryl (e.g., indolyl, pyrrolyl, pyridinyl, oxazolyl, oxadiazolyl) optionally substituted with 1 or 2 R.sup.X groups, R.sup.5 or R.sup.5′ is independently H and R.sup.7 is H.
(26) In one or more embodiments L is C.sub.1-2 alkyl-NHC(O)— and R.sup.6 is C.sub.1-9 heteroaryl (e.g., indolyl, pyrrolyl, pyridinyl, oxazolyl, oxadiazolyl) optionally substituted with 1 or 2 R.sup.X groups. In one or more embodiments, L is C.sub.1-3 alkylNHC(O)— and R.sup.6 is indolyl (e.g., 2-indolyl or 3-indolyl) optionally substituted with 1 or 2 R.sup.X groups or 3-indolyl optionally substituted with 1 or 2 R.sup.X groups. In one or more embodiments L is C.sub.1-6 alkyl-NHC(O)— and R.sup.6 is C.sub.1-9 heteroaryl (e.g., pyrrolyl, pyridinyl, oxazolyl, oxadiazolyl) optionally substituted with 1 or 2 R.sup.X groups. In one or more embodiments L is C.sub.1-3 alkylNHC(O)— and R.sup.6 is indolyl (e.g., 2-indolyl or 3-indolyl) optionally substituted with a pyridinyl group (e.g., 2-, 3- or 4-pyridinyl). In one or more embodiments L is C.sub.1-6 alkyl-NHC(O)— and R.sup.6 is C.sub.1-9 heteroaryl (e.g., pyrrolyl, pyridinyl, oxazolyl, oxadiazolyl) optionally substituted with 1 or 2 R.sup.X groups, R.sup.5 or R.sup.5′ is independently selected from C.sub.1-3 alkyl, C.sub.1-3alkylC(O)OC.sub.1-4alkyl and C.sub.1-3alkylC.sub.6-10 aryl optionally substituted with 1 or 2 groups independently selected from haloC.sub.1-3alkyl (e.g., CF.sub.3) and haloC.sub.1-3alkoxy (e.g., OCF.sub.3), and R.sup.7 is H.
(27) In one or more embodiments L is C.sub.1-3 alkyl-NHC(O)-indolyl and R.sup.6 is selected from phenyl optionally substituted with 1 or 2 R.sup.X groups, C.sub.3-8 heteroaryl optionally substituted with 1 or 2 R.sup.X groups, C(O)(C.sub.3-8 heteroaryl) optionally substituted with 1 or 2 R.sup.X groups, C(O)(C.sub.2-5heterocycloalkyl) optionally substituted with 1 or 2 R.sup.X groups, C(O)NHR.sup.Y, or is absent. In one or more embodiments L is C.sub.1-3 alkyl-NHC(O)-indolyl and R.sup.6 is selected from phenyl optionally substituted with C(O)NHR.sup.Y, C(O)-(heterocycloalkyl) optionally substituted with 1 or 2 C.sub.1-4 alkyl groups (e.g., methyl, ethyl).
(28) In one or more embodiments L is C.sub.1-2 alkyl-NHC(O)-indolyl, R.sup.6 is pyridyl, and R.sup.5 or R.sup.5′ is independently selected from H, C.sub.1-6 alkyl, C.sub.1-3alkylC(O)OC.sub.1-4alkyl and C.sub.1-3alkylC.sub.6-10aryl optionally substituted with 1 or 2 groups independently selected from haloC.sub.1-3alkyl or haloC.sub.1-3alkoxy.
(29) In one or more embodiments L is azetidinylC(O)— and R.sup.6 is indolyl (e.g., 2-indolyl or 3-indolyl) optionally substituted with 1 or 2 R.sup.X groups. In one or more embodiments L is azetidinylC(O)— and R.sup.6 is 2-indolyl optionally substituted with a C.sub.1-9 heteroaryl group. In one or more embodiments L is azetidinylC(O)— and R.sup.6 is 2-indolyl optionally substituted with a pyridinyl group (e.g., 2-, 3- or 4-pyridinyl). In one or more embodiments L is azetidinylC(O)— and R.sup.6 is 2-indolyl optionally substituted with a C.sub.1-9 heteroaryl group, R.sup.5 or R.sup.5′ is independently H and R.sup.7 is H. In one or more embodiments L is azetidinylC(O)— and R.sup.6 is 2-indolyl optionally substituted with a C.sub.1-9 heteroaryl group, R.sup.5 or R.sup.5′ is independently selected from C.sub.1-3 alkyl, C.sub.1-3alkylC(O)OC.sub.1-4alkyl and C.sub.1-3alkylC.sub.6-10aryl optionally substituted with 1 or 2 groups independently selected from haloC.sub.1-3alkyl and haloC.sub.1-3alkoxy, and R.sup.7 is H.
(30) In one or more embodiments, R.sup.5 or R.sup.5′ is independently H and R.sup.7 is H.
(31) In one or more embodiments, R.sup.5 or R.sup.5′ is independently H and R.sup.7 is C.sub.1-2alkyl-(3-indolyl).
(32) In one or more embodiments each R.sup.X is independently selected from hydroxyl, halo, C.sub.1-3 alkyl, C.sub.1-4alkoxy, C(O)OC.sub.1-4 alkyl, phenyl, NR′R″ wherein R′ and R″ are independently selected from H and C.sub.1-3 alkyl, morpholinyl optionally substituted with 1 or 2 C.sub.1-3 alkyl groups, piperazinyl optionally substituted with 1 or 2 C.sub.1-3 alkyl groups, C(O)morpholinyl optionally substituted with 1 or 2 C.sub.1-3alkyl groups, C.sub.6-10aryl optionally substituted with 1 or 2 C(O)—(C.sub.1-4alkylheterocycloalkyl) groups, C(O)piperazinyl optionally substituted with 1 or 2 C.sub.1-3 alkyl groups, or two adjacent R.sup.X groups together form methylenedioxy or 1,2-ethylenedioxy. In one or more embodiments R.sup.X is pyridyl (e.g., 2-, 3- or 4-pyridyl).
(33) In one or more embodiments the invention relates to compounds of formula (IA) or formula (IB), formula (IC), formula (IIA), or formula (IIB) or a salt, hydrate, solvate, tautomer or stereoisomer thereof:
(34) ##STR00005##
wherein: R.sup.1 and R.sup.4 are both H; R.sup.2 is H, halo, C.sub.1-3alkoxy, O—CH.sub.2phenyl, O-phenyl; R.sup.3 is H, halo, C.sub.1-3alkoxy, O—CH.sub.2phenyl, O-phenyl; R.sup.7 is H or C.sub.1-3 alkyl; A is C═O or SO.sub.2; X is C or N; R.sup.E is H, C.sub.1-3alkyl, or C(O)-heterocycloalkyl (e.g., C(O)—(N-morpholino)); R.sup.5′ is H, C.sub.1-6 alkyl, C.sub.1-3alkylC(O)OC.sub.1-3alkyl or C.sub.1-3 alkylC.sub.6-10 aryl optionally substituted with 1 or 2 groups independently selected from haloC.sub.1-3alkyl or haloC.sub.1-3 alkoxy; R.sup.A, R.sup.B, R.sup.C and R.sup.D are independently selected from H, OH, C.sub.1-3 alkyl, OC.sub.1-3 alkyl, C(O)—(N-heterocycloalkyl) (e.g., C(O)morpholinyl), C(O)piperazinyl) optionally substituted with 1 or 2 C.sub.1-3 alkyl groups; N-heteroaryl (e.g., 3-pyridyl, 4-pyridyl, 2,1,3-benzoxadiazolyl, pyrazolyl) optionally substituted with 1 or 2 groups selected from OH, halo, C.sub.1-3 alkyl, OC.sub.1-3 alkyl; phenyl optionally substituted with 1 or 2 groups selected from OH, halo, C.sub.1-3 alkyl, OC.sub.1-3 alkyl, C(O)NHC.sub.1-3 alkyl-[N(C.sub.1-3 alkyl).sub.2], C(O)(heterocycloalkyl) (e.g., morpholinyl, piperazinyl, piperidinyl) optionally substituted with 1 or 2 C.sub.1-3 alkyl groups.
(35) It will be apparent to those skilled in the art that general formulae (IA)-(IC) are subsets of general formula (I), and formulae (IIA) and (IIB) are a subsets of general formula (II).
(36) In one or more embodiments R.sup.A and R.sup.D are H. In one or more embodiments, R.sup.A, R.sup.C and R.sup.D are H. In one or more embodiments, R.sup.A, R.sup.B, R.sup.C and R.sup.D are H.
(37) In on or more embodiments R.sup.E is H. In one or more embodiments R.sup.E is C.sub.1-3alkyl.
(38) In one or more embodiments A is C═O.
(39) Those skilled in the art will recognise that when the linker group L comprises an amide or cyclic moiety (e.g., a heterocycloalkyl, cycloalkyl moiety), rotation of the groups attached to the linker will be restricted. In one or more embodiments of the invention, such compounds may have increased activity.
(40) Another aspect of the invention relates to compounds of general formula (III):
(41) ##STR00006##
or a salt, hydrate, solvate, tautomer or stereoisomer thereof,
wherein groups R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.A, R.sup.B, R.sup.C, R.sup.D, and R.sup.E are as defined for formulae (I) and (II), formulae (IA)-(IC) and formulae (IIA)-(IIB), including the preferred embodiments.
(42) Those skilled in the art will recognise that general formula (III) is based on a tautomeric form of the imine group of general formula (I), wherein the amine group has been alkylated.
(43) For the avoidance of doubt, throughout this specification a general reference to ‘a compound of the invention’ refers to compounds of general formula (I), general formulae (IA)-(IC), general formula (II), general formulae (IIA)-(IIB), general formula (III) and salts, hydrates, solvates, tautomers or stereoisomers thereof unless expressly stated otherwise. For convenience, a reference to ‘a compound of formula (I)’ or ‘a compound of general formula (I)’ includes compounds of general formulae (IA)-(IC) and salts, hydrates, solvates, tautomers or stereoisomers thereof unless expressly stated otherwise. Similarly, a reference to ‘a compound of formula (II)’ or ‘a compound of general formula (II)’ includes compounds of general formulae (IIA)-(IIB) and salts, hydrates, solvates, tautomers or stereoisomers thereof unless expressly stated otherwise.
(44) Compounds of the invention, or salts, hydrates, solvates, tautomers or stereoisomers thereof may be prepared using methods known to those skilled in the art, the illustrative reaction schemes and General Procedures disclosed herein, the specific methods described in the Examples section, or by routine modifications thereof. The present invention also encompasses any one or more of these processes for preparing the compounds of the invention, in addition to any novel intermediates used therein.
(45) Suitable reagents and reaction conditions for performing reactions will be known to the skilled person and are described in the literature and text books, including for example March, J. Advanced Organic Chemistry, 4.sup.th Ed (John Wiley & Sons, New York, 1992) and Vogel's Textbook of Practical Organic Chemistry, 5.sup.th Ed (John Wiley & Sons, New York, 1989).
(46) The reaction schemes presented below are illustrative of general methods that may be employed to prepare the compounds of the invention. Alternative methods, including routine modifications of the methods disclosed herein, will be apparent to those skilled in the art. Groups R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.5′, R.sup.6, R.sup.A, R.sup.B, R.sup.C, R.sup.D are as previously defined for general formula (I), general formula (II) and general formula (III).
(47) Compounds of general formula (I) may be prepared via the condensation reaction described by Pazdera, P., et al, Chem. Papers, 1989, 43, 771 which involves the condensation of a 2-isothiocyanatobenzonitrile 1 (X=S) or 2-isocyanatobenzonitrile 1 (X=O) with a primary amine 2 as illustrated in Scheme 1:
(48) ##STR00007##
(49) The reaction scheme depicted in Scheme 1 is versatile and allows ready access to the 4-imino-3,4-dihydroquinazoline-2(1H)-thione (X=S) or 4-imino-3,4-dihydroquinazoline-2(1H)-one (X=O) scaffold, with substituent groups R.sup.1-R.sup.4 in formula (I) derived from the benzonitrile reactant 1. Advantageously, such reactions may be performed in one-pot.
(50) Typical solvents include but are not limited to alcohols (e.g., ethanol, propanol), tetrahydrofuran, petroleum spirit/dichloromethane mixtures.
(51) The reaction is typically performed at a temperature in the range of about 20° C. to about 100° C., e.g., about 20° C. to about 80° C., or about 25° C. to about 80° C.
(52) An exemplary reaction scheme for preparing compounds of general formula (I) is shown in Scheme 1A in which linker L is an alkylene and R.sup.6 is an optionally substituted 3-indole.
(53) ##STR00008##
Scheme 1A
(54) Suitable acids for removal of the t-butoxycarbonyl (Boc) protecting group from the amine in step 2 of Scheme 1A include but are not limited to an inorganic acid such as hydrochloric acid or trifluoracetic acid, and suitable solvents include alcohols such as methanol or ethanol, dioxane, tetrahydrofuran or acetonitrile. Suitable bases for the condensation reaction shown in the third step include but are not limited to triethylamine, diisopropylethylamine, 2,6-dimethylpyridine, and suitable solvents include alcohols e.g., methanol, ethanol, propanol, tetrahydrofuran, and petroleum spirit/dichloromethane mixtures. The reactions are typically carried out at temperatures below 60° C., typically at room temperature.
(55) Another exemplary reaction scheme for preparing compounds of general formula (I) is depicted in Scheme 1B:
(56) ##STR00009##
(57) The Suzuki coupling step to introduce functionality onto the indole ring is well known to those skilled in the art and can be performed, for example, by reacting aryl bromide with a boronic acid or ester in base, e.g, potassium carbonate, sodium carbonate in a degassed mixture of dimethoxyethane, water and ethanol, followed by addition of bis(triphenylphosphine)palladium(II) dichloride. The reaction mixture is typically heated to a temperature above 100° C. The deprotection step to remove the Boc protecting group from the amine is also well known in the art and suitable conditions include trifluoroacetic acid/CH.sub.2Cl.sub.2, or hydrochloric acid/dioxane. The final step typically may use a base such as triethylamine, diisopropylethylamine, 2,6-dimethylpyridine, among others, and suitable solvents include alcohols e.g., methanol, ethanol, propanol, tetrahydrofuran, and petroleum spirit/dichloromethane mixtures. The reactions are typically carried out at temperatures below 60° C., typically at room temperature.
(58) Another exemplary reaction scheme useful in the preparation of compounds of general formula (I) is depicted in Scheme 1C below:
(59) ##STR00010##
(60) Scheme 1C illustrates methods for introducing amide functionality at the 5-position of the indole ring. In various embodiments the groups R.sup.p and R.sup.q together with the nitrogen atom to which they are attached may form a morpholinyl, piperazinyl or piperidinyl ring. The first step shown in Scheme 1C involves standard methodology to protect the carboxylic acid as an ester. Suitable alcohols (R′OH) include methanol, ethanol, butanol (e.g., n-butanol), etc. A suitable base for use in the fifth step to convert the ester into a carboxylic acid without removing the Boc protecting group includes but is not limited to lithium hydroxide. A suitable exemplary solvent for the sixth step which involves formation of the amide bond, is N,N-dimethylformamide. The amide product shown in Scheme 1C may then be deprotected to remove the Boc protecting group and reacted with a 2-isothiocyanatobenzonitrile (X=S) or 2-isocyanatobenzonitrile (X=O) following the general procedures known in the art, including those described above for Scheme 1, Scheme 1A and Scheme 1B.
(61) Compounds of general formula (II) may be prepared from compounds of general formula (I) (when R.sup.5═H). For example, as illustrated in general Scheme 2, a compound of formula (I) represented by quinazoline compound 3 is first treated with base to form an intermediate thioamide enolate (X=S) or an amide enolate (X=O), followed by a nucleophilic substitution reaction with a halide compound 4 (R.sup.5′—X′) (where X′=Cl, Br, I) to afford the compound of general formula (II).
(62) ##STR00011##
(63) An exemplary reaction scheme for preparing compounds of general formula (II) is shown in Scheme 2A in which linker L is an alkyl amide, R.sup.6 is an optionally substituted 2-indole and R.sup.5′—X′ is methyl iodide.
(64) ##STR00012##
(65) As illustrated in Scheme 2A, a quinazoline compound 5 of general formula (I) is reacted with a base (e.g., potassium carbonate) in a suitable solvent such as acetone. The reaction mixture is stirred, typically at room temperature, until substantially complete. The reaction will typically require at least 2-4 hours, more typically about 12-48 hours, although longer periods may be required for large scale reactions. If necessary, routine methods of purification such as flash column chromatography can be used to isolate compounds of general formula (II).
(66) Compounds of general formula (III) may be prepared by performing a nucleophilic reaction at the 4-position of the dihydroquinazoline 6 (X=S or O) according to the method described in Eur J Med Chem 2012 47 283 and Lee et al. J Med Chem. 1995, 38 (18), pp 3547-3557, as illustrated in general Scheme 3:
(67) ##STR00013##
(68) Suitable solvents for step i include, for example, DMF, THF. Typically the reaction mixture is heated at reflux until substantially complete. Step ii is typically performed by heating the reaction mixture at temperatures above 60° C., typically about 75° C. until the reaction is judged to be substantially complete. Typical reaction times are about 2-24 hours, e.g., about 12 hours or 15 hours, depending on the scale of the reaction.
(69) In an alternative procedure, compounds of general formula (III) may be prepared by thionation of the corresponding 2-chloroquinazoline 9 as illustrated in Scheme 4:
(70) ##STR00014##
(71) As illustrated in Scheme 4, a 2-chloro-4-amino substituted dihydro-quinazoline 6 is reacted with a salt of thioacetate (e.g., potassium thioacetate) in a suitable solvent such as dioxane or THF. The reaction mixture is typically heated to a temperature above 60° C., typically above 100° C., more typically about 120° C. and the reaction is allowed to proceed for a period of time sufficient for the reaction to proceed substantially to completion. Those skilled in the art know how to monitor the progress of a chemical reaction using standard techniques, such as Thin Layer Chromatography (TLC), .sup.1H NMR, etc. The reaction will typically require at least 1 hour, more typically about 3 hours, although longer periods may be required for large scale reactions. When the reaction is judged to be sufficiently complete, the reaction mixture is typically cooled, and the reaction solvent removed (e.g., under vacuum), after which the residue is treated with ammonia in a protic solvent such as methanol. The compound of general formula (III) may then be isolated using routine methods such as flash column chromatography.
(72) Where appropriate or necessary, protecting groups may be employed at any stage in the synthesis of compounds of the invention. Similarly, those skilled in the art will also appreciate that compounds of the invention can also be prepared with certain protecting groups that are useful for purification or storage that can be removed before administration to a patient. Thus, this invention also encompasses compounds of the invention containing protective groups. Suitable protecting groups and their use are well known to those skilled in the art and include, for example, protecting groups described in Peter G. M. Wuts, Theodora W. Greene, “Greene's Protective Groups in Organic Synthesis”, 4.sup.th Edition. (John Wiley & Sons, Inc., 2007). The protection and deprotection of functional groups is also described in “Protective Groups in Organic Chemistry”, edited by J. W. F. McOmie, Plenum Press (1973).
(73) Those skilled in the art will recognise the versatility of the reactions illustrated in the above Schemes, which can provide access to a wide range of substituted dihydro and tetrahydro quinazoline compounds of general formulae (I), (II) and (III). The methods described above are merely representative and routine modifications and variations that would be apparent to persons skilled in the art fall within the broad scope and ambit of the invention disclosed herein.
(74) Compounds of general formula (I), general formula (II) or general formula (III) may be isolated or purified using standard techniques known to those skilled in the art. Such techniques include precipitation, crystallisation, recrystallization, column chromatography (including flash column chromatography), HPLC, formation of salts, lyophilisation, among others. Suitable solvents for use in these techniques will be known or can be readily ascertained by those skilled in the art using routine practices.
(75) Salts, including pharmaceutically acceptable salts of compounds of the invention may be prepared by methods known to those skilled in the art, including for example: (i) by reacting the compound of the invention with the desired acid or base; (ii) by removing an acid- or base-labile protecting group from a suitable precursor of the compound of the invention or by ring-opening a suitable cyclic precursor, for example, a lactone or lactam, using the desired acid or base; or (iii) by converting one salt of the compound of the invention into another salt by reaction with an appropriate acid or base, or by means of a suitable ion exchange column.
(76) The above reactions are typically carried out in solution. Suitable solvent systems (including mixed solvent systems) are well known to those skilled in the art and those skilled in the art can readily select or determine a suitable solvent system using routine methods taking into consideration the nature of the compound, the particular salt being formed, and the amount of the compound. Exemplary solvent systems include methanol, ethanol, water, acetone, tetrahydrofuran, dichloromethane, pentane, hexane, diethyl ether, ethyl acetate, and any mixture of two or more such solvents. The resulting salt may precipitate out and be collected by filtration or may be recovered by evaporation of the solvent. The degree of ionisation in the resulting salt may vary from completely ionised to almost non-ionised.
(77) In Vitro Screening Assays
(78) The present disclosure also includes in vitro screening assays for assessing the heparanase inhibitor activity of test compounds.
(79) Suitable assays for determining heparanase inhibitory activity are known in the art, see, for example, the in vitro assays described in Rivara et al. (2016) Future Med Chem, 8(6): 647-680. For example, the method may include contacting a preparation comprising heparanase and a heparanase substrate (e.g. heparan sulfate or fondaparinux) with a test compound and detecting the amount of the intact substrate in comparison to a reference level of intact substrate in the absence of the test compound, or detecting the modulation of the activity of a downstream target of the intact heparanase substrate. Detecting the amount of intact substrate or modulation may be achieved using techniques including, but not limited to, ELISA, cell-based ELISA, inhibition ELISA, western blots, RIA, immunoprecipitation, immunostaining, a solid-phase labeled substrate assay such as a solid phase radio- or fluorescently-labeled or biotinylated substrate, an ultrafiltration assay, proximity assays such as HTRF and scintillation proximity assays, fluorescent assays using e.g. fluorescent substrate-heparanase substrate conjugates such as fluorescein or rhodamine, colorimetric assays and fluorescent immunoassays, all of which are well known to those skilled in the art.
(80) In some embodiments, test compounds may be screened using commercially available assays, illustrative examples of which include Cisbio heparanse assay toolbox (Biotin-Heparan sulfate-Eu cryptate; Catalogue No. 61BHSKAA; Cisbio Bioassays, Codolet France), Amsbio heparanase assay kit (Catalogue No. Ra001-BE-K; AMS Biotechnology Ltd, Abington UK) and InSight heparanase activity kit (Catalogue No. INS-26-4-0000-10; InSight Biopharmaceuticals, Rehovot, Israel).
(81) Heparanase inhibitor compounds according to the present invention may elicit their heparanase inhibitory activity through one or more modes of action. For example, heparanase inhibitor compounds may decrease, inhibit or impair any one or more of heparanase catalytic activity, heparanase protein binding, heparanase-mediated modulation of gene transcription, heparanase-mediated initiation of cell signaling and/or angiogenesis.
(82) Pharmaceutical Use
(83) Compounds of general formula (I), general formula (II) and general formula (III) according to the present invention are heparanase inhibitors and the invention also relates to a method of treating a condition or disease related to heparanase activity. Thus, an embodiment of the present invention relates to a method of treatment of a disease or condition associated with heparanase activity in a subject, the method comprising administering to the subject an effective amount of a compound of general formula (I), general formula (II) or general formula (III), or a pharmaceutical composition thereof.
(84) A further embodiment of the invention relates to the use of a compound of formula (I) or a salt, hydrate, solvate, tautomer or stereoisomer thereof or a pharmaceutical composition thereof in the manufacture of a medicament for treating a disease or condition associated with heparanase activity.
(85) In another aspect the present invention relates to the use of a compound of formula (I) or a salt, hydrate, solvate, tautomer or stereoisomer thereof or a pharmaceutical composition thereof for the treatment of a disease or condition associated with heparanase activity in a subject.
(86) The expression “a disease or condition associated with heparanase activity” means that the heparanase enzyme plays a role in the disease or condition. However, other enzymes, pathways and mechanisms may also be implicated in the disease of condition. The compounds of the present invention may be an inhibitor of one or more activities of heparanase, including, but not limited to, heparanase catalytic activity. Furthermore, compounds of the present invention may partially or substantially inhibit any one or more heparanase activities.
(87) In preferred embodiments the subject is a human.
(88) In various embodiments the disease or condition is selected from Type 1 diabetes, Type 2 diabetes, nephritis, glomerulonephritis, cell-mediated autoimmune inflammation, diabetic nephropathy, gestational diabetes, diabetic ketoacidosis, hyperglycemia, hyperosmolar state, hypoglycemia, diabetic coma, diabetic cardiomyopathy, diabetic neuropathy, diabetic foot, diabetic retinopathy, diabetic myonecrosis, and diabetic encephalopathy. In other embodiments the disease or condition related to heparanase activity is selected from cancer, allergies, dermatitis, psoriasis, an ocular inflammatory disorder, such as, macular degeneration, retinitis pigmentosa and pancreatitis.
(89) In preferred embodiments the disease or condition is selected from Type 1 diabetes, Type 2 diabetes, diabetic nephropathy, nephritis, glomerulonephritis, and cell-mediated autoimmune inflammation indications involving heparanase. In particularly preferred embodiments the disease is Type 1 diabetes or Type 2 diabetes.
(90) In another aspect the invention relates to a method of controlling blood glucose levels in a subject, the method comprising administering to the subject an effective amount of a compound of general formula (I), general formula (II), or general formula (III), or a pharmaceutical composition thereof.
(91) In a further aspect the invention relates to a method of treating or preventing rejection of a pancreatic islet transplant in a subject, the method comprising administering to the subject an effective amount of a compound of general formula (I), general formula (II), or general formula (III) or a pharmaceutical composition thereof.
(92) In another aspect the invention relates to a method of preserving beta-cell function in a subject, the method comprising administering to the subject an effective amount of a compound of general formula (I), general formula (II), or general formula (III), or a pharmaceutical composition thereof.
(93) Ocular inflammatory disorders such as diabetic retinopathy, age-related macular degeneration (AMD), retinitis pigmentosa, uveitis and viral corneal inflammation, occur wholly or in part from progressive inflammation in the eye. In a diseased eye, excessive macrophage activation and accumulation in particular tissues, such as the activation and accumulation of microglia in the sub-retinal space in degenerative disorders, can disrupt the immune privilege of the eye (Li et al. (2015) Experimental Eye Research, 136: 116-130). Heparanase has been shown to be important for ocular inflammation associated with viral infection of the cornea (Agelidis et al (2017) Cell, 20:439-450) as well as for the activation of macrophages (Gutter-Kapon et al. (2016) PNAS, 113(48): E7808-E781). Activated macrophages, including retinal microglia, produce different kinds of products including complement proteins, pro-inflammatory cytokines, reactive oxygen species, growth factors and other products, which can result in a chronic local inflammation and can typically lead to further damage (Li et al. (2015) Experimental Eye Research, 136: 116-130). For example, in the pathogenesis of AMD, microglia activated by cell death migrate to the damaged area to phagocytose cellular debris but also secrete molecules that kill neighboring photoreceptors around the area of primary degeneration (Li et al. (2015) Experimental Eye Research, 136: 116-130). Ocular macrophage, including microglial, activation is therefore an important target for the treatment and prevention of ocular inflammatory disorders. Accordingly, drugs that target heparanase, such as heparanase inhibitors, may prevent or reduce macrophage activation and may be useful for treating ocular inflammatory disorders, such as diabetic retinopathy and AMD among others.
(94) As described in the applicant's co-pending PCT application filed 13 Dec. 2017, titled “Heparanase inhibitors and use thereof” which claims priority from U.S. provisional application Ser. No. 62/433,652, and co-pending PCT application filed 13 Dec. 2017, titled “Methods of treating ocular disorders” which claims priority from Australian Provisional Patent Application No 2017902346 filed 20 Jun. 2017, compounds possessing heparanase inhibitory activity are useful for treating ocular inflammatory disease such as age-related macular degeneration.
(95) Thus, in another aspect the invention relates to a method for treating, or inhibiting the progression or development of, an ocular inflammatory disorder in a subject, the method comprising administering to the subject a compound of general formula (I) or a pharmaceutical composition thereof. The ocular inflammatory disorder may be any disorder of the eye which has an inflammatory component. Exemplary ocular inflammatory disorders include, but are not limited to, age-related macular degeneration (AMD) including the exudative or ‘wet’ AMD, dry AMD, diabetic retinopathy, retinitis pigmentosa, retinal vein occlusion, retinoblastoma, uveitis, macular edema, dry eye, viral infection and/or keratoconus; especially AMD, diabetic retinopathy and retinitis pigmentosa. In preferred embodiments, the ocular inflammatory disorder is AMD or diabetic retinopathy, preferably AMD. In preferred embodiments, the ocular inflammatory disorder is dry AMD. In other preferred embodiments the ocular inflammatory disorder is wet AMD. Although acute ocular inflammatory disorders are contemplated by the invention, in particularly preferred embodiments, the ocular inflammatory disorder is a chronic disorder.
(96) Pharmaceutical Formulations
(97) The compounds of the invention described herein may be administered as a formulation comprising a pharmaceutically effective amount of the compound, in association with one or more pharmaceutically acceptable excipients including carriers, vehicles and diluents. The term “excipient” herein means any substance, not itself a therapeutic agent, used as a diluent, adjuvant, or vehicle for delivery of a therapeutic agent to a subject or added to a pharmaceutical composition to improve its handling or storage properties or to permit or facilitate formation of a solid dosage form such as a tablet, capsule, or a solution or suspension suitable for oral, parenteral, intradermal, subcutaneous, or topical application. Excipients can include, by way of illustration and not limitation, diluents, disintegrants, binding agents, adhesives, wetting agents, polymers, lubricants, glidants, stabilizers, and substances added to mask or counteract a disagreeable taste or odor, flavors, dyes, fragrances, and substances added to improve appearance of the composition. Acceptable excipients include (but are not limited to) stearic acid, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric and sulfuric acids, magnesium carbonate, talc, gelatin, acacia gum, sodium alginate, pectin, dextrin, mannitol, sorbitol, lactose, sucrose, starches, gelatin, cellulosic materials, such as cellulose esters of alkanoic acids and cellulose alkyl esters, low melting wax, cocoa butter or powder, polymers such as polyvinyl-pyrrolidone, polyvinyl alcohol, and polyethylene glycols, and other pharmaceutically acceptable materials. Examples of excipients and their use is described in Remington's Pharmaceutical Sciences, 20th Edition (Lippincott Williams & Wilkins, 2000). The choice of excipient will to a large extent depend on factors such as the particular mode of administration, the effect of the excipient on solubility and stability, and the nature of the dosage form.
(98) The compounds and pharmaceutical compositions of the invention may be formulated for oral, injectable, rectal, parenteral, subcutaneous, intravenous, topical, intravitreal or intramuscular delivery. Non-limiting examples of particular formulation types include tablets, capsules, caplets, powders, granules, injectables, ampoules, vials, ready-to-use solutions or suspensions, lyophilized materials, creams, lotions, ointments, drops, suppositories and implants. Solid formulations such as the tablets or capsules may contain any number of suitable pharmaceutically acceptable excipients or carriers described above. The compounds of the invention may also be formulated for sustained delivery.
(99) Tablets and capsules for oral administration may be in unit dose presentation form, and may contain conventional excipients such as binding agents, for example, acacia, gelatin, sorbitol, tragacanth, or polyvinylpyrollidone; fillers, for example lactose, sugar, maize-starch, calcium phosphate, sorbitol or glycine; tabletting lubricants, for example, magnesium stearate, talc, polyethylene glycol or silica; disintegrants, for example, potato starch; or acceptable wetting agents such as sodium lauryl sulphate. The tablets may be coated according to methods well known in normal pharmaceutical practice.
(100) Oral liquid preparations may be in the form of, for example, aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, or may be presented as a dry product for reconstitution with water or other suitable vehicle before use. Such liquid preparations may contain conventional additives, such as suspending agents, for example, sorbitol, methyl cellulose, glucose syrup, gelatin, hydroxyethyl cellulose, carboxymethyl cellulose, aluminium stearate gel or hydrogenated edible fats, emulsifying agents, for example, lecithin, sorbitan monooleate, or acacia; non-aqueous vehicles (which may include edible oils), for example, almond oil, oily esters such as glycerin, propylene glycol, or ethyl alcohol; preservatives, for example, methyl or propyl p-hydroxybenzoate or sorbic acid; and, if desired, conventional flavouring or colouring agents.
(101) For parenteral administration, including intravenous, intramuscular, subcutaneous, intravitreal, or intraperitoneal administration, fluid unit dosage forms may be prepared by combining the compound and a sterile vehicle, typically a sterile aqueous solution which is preferably isotonic with the blood of the recipient. Depending on the vehicle and concentration used, the compound may be either suspended or dissolved in the vehicle or other suitable solvent. In preparing solutions, the compound may be dissolved in water for injection and filter sterilized before filling into a suitable vial or ampoule and sealing. Advantageously, agents such as a local anaesthetic, preservative and buffering agents can be dissolved in the vehicle. To enhance the stability, the composition may be frozen after filling into the vial and the water removed under vacuum. The dry lyophilized powder may then be sealed in the vial and an accompanying vial of water for injection or other suitable liquid may be supplied to reconstitute the liquid prior to use. Parenteral suspensions are prepared in substantially the same manner except that the compound is suspended in the vehicle instead of being dissolved and sterilization cannot be accomplished by filtration. The compound can be sterilized by exposure to ethylene oxide before suspending in the sterile vehicle. A surfactant or wetting agent may be included in the composition to facilitate uniform distribution of the compound.
(102) In one or more preferred embodiments the compounds of the invention are formulated as an injectable solution, suspension or emulsion. In preferred embodiments, the compounds of the invention are formulated for intravitreal injection into the eye of a subject. Such formulations may be particularly preferred for treatment of ocular inflammatory disorders, such as age-related macular degeneration (AMD) including the exudative or ‘wet’ and ‘dry’ form of AMD, diabetic retinopathy, retinitis pigmentosa, retinal vein occlusion, retinoblastoma, uveitis, macular edema, dry eye, viral infection and/or keratoconus; especially AMD, diabetic retinopathy and retinitis pigmentosa.
(103) In preferred embodiments, ophthalmic formulations, including intravitreal formulations and other ophthalmic formulations, such as eye drops, typically may comprise one or more co-solvent(s), such as one or more organic co-solvents; one or more tonicity agent(s); a buffering system comprising one or more buffering agents; a stabilizing agent; pH between about 3-8. In preferred embodiments, the organic co-solvent may be polysorbate, for example, polysorbate 20 or polysorbate 80, polyethylene glycol (PEG), for example, PEG 3350, or propylene glycol, or a combination thereof; the tonicity agent may be, for example, sodium chloride or potassium chloride; the stabilizing agent may be sucrose, sorbitol, glycerol, trehalose, or mannitol; and the buffering agent may be, for example, phosphate buffer, such as a sodium phosphate buffer.
(104) Intravitreal formulations are preferably sterile, isotonic and preferably have a pH within the range pH 3-8, preferably pH 5-7 or pH 3-5. Such formulations may contain one or more buffers as part of a buffer system, however, the concentration of buffers is preferably kept as low as possible. Buffer stressing studies may be carried out to select the minimal buffer amount needed to safely maintain the desired pH range. Exemplary intravitreal formulations are described in Shikari H, Samant P M. Intravitreal injections: A review of pharmacological agents and techniques. J Clin Ophthalmol Res 2016; 4:51-9.
(105) In preferred embodiments, ophthalmically acceptable formulations may be lyophilizable. Lyophilizable formulations can be reconstituted into solutions, suspensions, emulsions, or any other suitable form for administration or use. Lyophilizable formulations are typically first prepared as liquids, then frozen and lyophilized. The total liquid volume before lyophilization can be less, equal to, or more than, the final reconstituted volume of the lyophilized formulation. The lyophilization process is well known to those of ordinary skill In the art, and typically Includes sublimation of water from a frozen formulation under controlled conditions. Lyophilized formulations typically can be stored at a wide range of temperatures. For example, lyophilized formulations may be stored below 25° C., for example, refrigerated at 2-8° C., or at room temperature (e.g., approximately 25° C.). Preferably, lyophilized formulations are stored below about 25° C., more preferably, at about 4-20° C.; below about 4° C.; or below about 0° C.
(106) Lyophilized formulations are preferably reconstituted with a solution consisting primarily of water (e.g., USP WFI, or water for injection) or bacteriostatic water (e.g., USP WFI with 0.9% benzyl alcohol). Alternatively, solutions comprising buffers and/or excipients and/or one or more pharmaceutically acceptable carriers may be used. The liquid that is to undergo freeze-drying or lyophilization preferably comprises all components desired in a final reconstituted liquid formulation.
(107) The pharmaceutical compositions of the invention may be administered locally to an eye using a variety of routes including, but not limited to, topical, through an ocular implant or direct injection into the eye. In particular embodiments, the pharmaceutical composition of the invention is administered locally to the eye using intravitreal injection, subconjunctival injection, sub-tenon injection, retrobulbar injection, suprachoroidal injection, intrascleral injection, intracorneal injection, subretinal injection or intracameral injection; especially intravitreal injection. In some embodiments, the composition is administered using a microneedle, for example, through intrascleral or intracorneal injection.
(108) In some embodiments, the composition is administered using an ocular implant, for example, a biodegradable implant such as those made from, for example, polylactic acid (PLA), polyglycolic acid, poly(lactide-co-glycolide) (PLGA), cross-linked gelatin derivatives, hypromellose, polyesters and/or polycaprolactones; or a non-biodegradable implant such as those made from, for example, polyvinyl alcohol, ethylene vinyl acetate, silicon and/or polysulfone capillary fiber.
(109) In some embodiments, the composition of the invention is formulated in a sustained release formulation or depot. Exemplary sustained release formulations or depots include a microsphere; matrix; emulsion; lipid-based; polymer-based; nanomicelle; micelle; nanovesicle such as a liposome, noisome, transfersome, discome, pharmacosome, emulsome or spanlastic, especially a liposome; microparticle; nanoparticle such as a nanocapsule or nanosphere composed of e.g. lipids, proteins, natural or synthetic polymers such as albumin, sodium alginate, chitosan, PLGA, PLA and/or polycaprolactone; or in situ gel such as an in situ hydrogel drug delivery system.
(110) In some embodiments, the composition of the invention is formulated for topical administration to the eye. Thus, the composition may be in the form of an eye drop, gel or ointment; especially an eye drop. The composition may be in a single unit dose or multiple unit dose form.
(111) The amount of therapeutically effective compound that is administered and the dosage regimen for treating a disease condition with the compounds and/or pharmaceutical compositions of the present invention depends on a variety of factors, including the age, weight, sex, and medical condition of the subject, the severity of the disease, the route and frequency of administration, the particular compound employed, as well as the pharmacokinetic properties (e.g., adsorption, distribution, metabolism, excretion) of the individual treated, and thus may vary widely. Such treatments may be administered as often as necessary and for the period of time judged necessary by the treating physician. One of skill in the art will appreciate that the dosage regime or therapeutically effective amount of the compound to be administrated may need to be optimized for each individual. The pharmaceutical compositions may contain active ingredient in the range of about 0.1 mg to 2000 mg, typically in the range of about 0.5 mg to 500 mg and more typically between about 1 mg and 200 mg. A daily dose of about 0.01 mg/kg to 100 mg/kg body weight, typically between about 0.1 mg/kg and about 50 mg/kg body weight, may be appropriate, depending on the route and frequency of administration. The daily dose will typically be administered in one or multiple, e.g., two, three or four, doses per day.
(112) The compounds of the present invention may be administered along with a pharmaceutical carrier, diluent or excipient as described above. Alternatively, or in addition, the compounds may be administered in combination with other agents, for example, other antidiabetic therapeutic agents, or VEGF inhibitor drugs.
(113) The terms “combination therapy” or “adjunct therapy” in defining use of a compound of the present invention and one or more other pharmaceutical agents, are intended to embrace administration of each agent in a sequential manner in a regimen that will provide beneficial effects of the drug combination, and is intended as well to embrace co-administration of these agents in a substantially simultaneous manner, such as in a single formulation having a fixed ratio of these active agents, or in multiple, separate formulations of each agent.
(114) In accordance with various embodiments of the present invention one or more compounds of formula (I), formula (II), or formula (III) may be formulated or administered in combination with one or more other therapeutic agents. Thus, in accordance with various embodiments of the present invention, one or more compounds of formula (I), formula (II), or formula (III) may be included in combination treatment regimens with surgery and/or other known treatments or therapeutic agents, and/or adjuvant or prophylactic agents.
(115) A number of agents are available in commercial use, in clinical evaluation and in pre-clinical development, which could be selected for treatment of Type 1 diabetes, Type 2 diabetes, diabetic nephropathy, nephritis, glomerulonephritis, and other cell-mediated autoimmune inflammation indications, cancer, psoriasis, dermatitis, allergy, macular degeneration, retinitis pigmentosa and pancreatitis as part of combination drug therapy. Suitable agents which may be used in combination therapy will be recognized by those of skill in the art. Suitable agents are listed, for example, in the Merck Index, An Encyclopaedia of Chemicals, Drugs and Biologicals, 12.sup.th Ed., 1996, and subsequent editions, the entire contents of which are incorporated herein by reference.
(116) For example, when used in the treatment of Type 1 diabetes or Type 2 diabetes, compounds of the present invention may be administered with an additional anti-diabetic agent, or combinations thereof, such as: biguanides (e.g., metformin), insulin secretagogues (e.g., sulfonylureas and glinides), glitazones, non-glitazone PPARy agonists, PPARp agonists, inhibitors of DPPIV, inhibitors of PDE5, inhibitors of GSK-3, glucagon antagonists, inhibitors of f-1,615 BPase (Metabasis/Sankyo), GLP-1/analogs (AC 2993, also known as exendin-4), insulin and insulin mimetics (Merck natural products). Other examples include PKC-p inhibitors, AGE breakers, SGLT2 inhibitors, T cell inhibitors (including anti-CD3 monoclonal antibodies), B cell inhibitors (including anti-CD20 monoclonal antibodies such as Rituximab), CTLA-4-Ig (Abatacept/Bristol-Myers Squibb) and inflammatory cytokine inhibitors including blocking monoclonal antibodies, among others.
(117) In other embodiments, when used for the treatment of ocular inflammatory disorders, such as, for example, age-related macular degeneration (AMD), the compound of formula (I) may be administered in combination with a growth factor inhibitor. Suitable growth factor inhibitors include, but are not limited to, a vascular endothelial growth factor (VEGF) inhibitor, such as ranibizumab, aflibercept, bevacizumab, pegaptanib, conbercept, abicipar pegol (MP0112) and MP0250; a platelet derived growth factor (PDGF) inhibitor, such as E10030 (anti-PDGF PEGylated aptamer), trapidil and pegpleranib; and pharmaceutically acceptable salts and combinations thereof. In some embodiments, the one or more pharmaceutically active agent is a VEGF inhibitor selected from the group consisting of ranibizumab, aflibercept, bevacizumab, pegaptanib, conbercept and pharmaceutically acceptable salts and combinations thereof.
(118) Combination regimens may involve the active agents being administered together, sequentially, or spaced apart as appropriate in each case. Combinations of active agents including compounds of the invention may be synergistic.
(119) The co-administration of compounds of the invention may be effected by the compounds being in the same unit dose as another active agent, or the compounds and one or more other active agent(s) may be present in individual and discrete unit doses administered at the same, or at a similar time, or at different times according to a dosing regimen or schedule. Sequential administration may be in any order as required, and may require an ongoing physiological effect of the first or initial compound to be current when the second or later compound is administered, especially where a cumulative or synergistic effect is desired.
(120) Embodiments of the invention will now be discussed in more detail with reference to the examples which are provided for exemplification only and which should not be considered limiting on the scope of the invention in any way.
EXAMPLES
Abbreviations
(121) BOC refers to a t-butoxycarbonyl group T3P® refers to propylphosphonic anhydride solution (>50 wt. % in ethyl acetate) DIPEA refers to diisopropylethylamine DEA refers to diethylamine DCM refers to dichloromethane THF refers to tetrahydrofuran DMF refers to N,N-dimethyformamide DMSO refers to dimethyl sulfoxide EtOAc refers to ethyl acetate rt refers to room temperature
General Procedures
(122) Unless otherwise specified, proton (.sup.1H) and carbon (.sup.13C) NMR spectra were recorded at room temperature in base-filtered CDCl.sub.3 with a Bruker spectrometer operating at 400 MHz for proton and 100 MHz for carbon nuclei. For .sup.1H NMR spectra, signals arising from the residual protio forms of the solvent were used as the internal standards. .sup.1H NMR spectroscopic data are recorded as follows: chemical shift (δ) [multiplicity, coupling constant(s) J (Hz), relative integral] where multiplicity is defined as: s=singlet; d=doublet; t=triplet; q=quartet; m=multiplet; br=broad or combinations of thereof. The signal due to residual CHCl.sub.3 appearing at δH=7.26 ppm and the central resonance of the CDCl.sub.3 “triplet” appearing at δC=77.0 ppm were used to reference .sup.1H and .sup.13C NMR spectra, respectively. The quintet due to residual DMSO-d.sub.5 appearing at δH=2.50 ppm and the central resonance of the DMSO-d.sub.6 “multiplet” appearing at δC=39.52 ppm were used to reference 1H and .sup.13C NMR spectra, respectively. Infrared spectra (IR: max) were recorded with a Perkin-Elmer 1800 series FTIR spectrometer or a Perkin-Elmer UATR Spectrum Two FTIR spectrometer. Samples were analyzed as thin films on KBr plates or compressed and flattened on a diamond window. Low-resolution ESI mass spectra were recorded on a single quadrupole liquid chromatograph-mass spectrometer, while high-resolution measurements were conducted on a time-of-flight instrument. Low- and high-resolution EI mass spectra were recorded with a magnetic-sector machine. Melting points were measured with an Optimelt automated melting point system and are uncorrected. Analytical thin layer chromatography (TLC) was performed on aluminium-backed 0.2 mm thick silica gel 60 F254 plates. Eluted plates were visualized with a 254 nm UV lamp and/or by treatment with a suitable dip followed by heating. These dips included phosphomolybdic acid/ceric sulfate/sulfuric acid (concd.)/water (37.5 g:7.5 g:37.5 g:720 mL) or potassium permanganate/potassium carbonate/5% sodium hydroxide aqueous solution/water (3 g:20 g:5 mL:300 mL). Flash chromatographic separations were carried out according to protocols defined by Still et al. J. Org. Chem. 1978, 43, 2923 with silica gel 60 (40-63 μm) as the stationary phase and with the AR- or HPLC-grade solvents indicated.
(123) Microwave reactions were conducted with a CEM Explorer microwave reactor. Microwave vessels were sealed with a snap-cap and irradiated for the time and at the temperatures specified, typically with a ramp time of 1 minute to the specified temperature at a maximum power of 200 W.
(124) Starting materials and reagents were generally available from the Sigma-Aldrich, Merck, TCI, Strem, AK Scientific or Lancaster chemical companies and were used as supplied. Drying agents and other inorganic salts were purchased from the AJAX, BDH or Unilab chemical companies. Tetrahydrofuran (THF), diethyl ether, methanol and dichloromethane were dried by using a Glass Contour solvent purification system that is based upon a technology originally described by Grubbs et al. Organometallics 1996, 15, 1518. Where necessary, reactions were performed under nitrogen.
I. SYNTHESIS EXAMPLES
General Experimental Procedures
General Procedure A
Preparation of 2-isothiocyanatobenzonitriles
(125) ##STR00015##
(126) Following the procedure of Calestani, G et al. (Tetrahedron, 2001, 57 (33), 7221) a magnetically stirred mixture of thiophosgene (1.35 mL, 17.5 mmol) in DCM (2 mL) and water (4 mL) was treated dropwise with a solution of the substituted amino-benzonitrile (14.3 mmol) in DCM (15 mL). The mixture was stirred for two hours and was then transferred to a separatory funnel and the aqueous layer extracted with DCM (20 mL). The organic layers were combined, washed with water (2×20 mL) then dried over Na.sub.2SO.sub.4. The DCM was removed with a gentle stream of nitrogen with heating at 40° C. and the residue obtained was then held under high vacuum (1 mmHg) for 4 h to afford the desired 2-isothiocyanatobenzonitrile which may be used without further purification.
General Procedure B
Reaction of Primary Amines with 2-isothiocyanatobenzonitriles
(127) ##STR00016##
(128) Following a modified procedure analogous to that reported by Pazdera, P (Chem. Papers 1989, 43 (3), 465), a magnetically stirred solution of substituted 2-isothiocyanatobenzonitrile (1.36 mmol) in a mixture of DCM and petroleum Spirit (2.5 mL: 2.5 mL) was treated with a solution of primary amine in DCM (7 mL). The mixture was stirred for 0.5 h and then pet. spirit (15 mL) was added. The solid was collected by vacuum filtration and then magnetically stirred in a solution of ethanol (30 mL) and heated at 70° C. for 2 h. The mixture was cooled to 0° C. and the solid was collected by vacuum filtration, washed with ethanol (10 mL) and dried at the pump (1 mmHg).
General Procedure C
Reaction of Primary Amines with 2-isothiocyanatobenzonitriles—Alternative Procedure
(129) ##STR00017##
(130) Crude amine hydrochloride (0.27 mmol) suspended in ethanol (6 mL), was treated with triethylamine (96 μL, 0.68 mmol) and then a solution of 2-isothiocyanato-4,5-dimethoxybenzonitrile (50 mg, 0.27 mmol) in DCM (3 mL) was added. The mixture was stirred at rt for 1 h then concentrated by a gentle stream of nitrogen and re-suspended in ethanol (5 mL). The mixture was heated at reflux for 4 h and the precipitate that formed was collected and washed with ethanol (15 mL) then ether (10 mL) and dried at the pump.
General Procedure D
Suzuki Cross-Coupling Procedure
(131) ##STR00018##
(132) A 10 mL snap-cap microwave vessel, fitted with a magnetic stirring bar, was charged with a mixture of boronic acid or ester (77.2 μmol), aryl bromide or triflate (52.3 μmol) and potassium carbonate (38 mg, 274 μmol) then treated with a degassed mixture of dimethoxyethane, water and ethanol (7:3:2, 1 mL). Bis(triphenylphosphine)palladium(II) dichloride (1.8 mg, 5 mol %) was added and the mixture was sparged with nitrogen for 0.05 hr, sealed then subjected to microwave irradiation (120° C./0.33 h, ramp time 1 minute, maximum power 200 W). The mixture was treated with water (1 mL) and extracted with EtOAc (3×2 mL) and the combined organic layers washed with brine and concentrated under a gentle stream of nitrogen. The residue obtained was subjected to flash column chromatography [silica, 1:10 v/v MeOH/DCM elution (unless otherwise specified) to give, after concentration of the appropriate fractions the desired compound.
General Procedure E
Boc Deprotection with Hydrogen Chloride in Dioxane
(133) ##STR00019##
(134) A solution of hydrogen chloride (2 mL, 4M in dioxane) at 4° C. was added to the Boc-protected compound and magnetically stirred for 1 h at rt. Diethyl ether (5 mL) was added and the mixture vigorously stirred for 0.05 h then magnetic stirring was ceased and the precipitate or gum allowed to settle. The solvent was decanted off the precipitate and this procedure was repeated a further 3 times and the resulting compound placed under high vacuum for 3 hours and used without further purification.
General Procedure F
Boc Deprotection with Trifluoroacetic Acid in Dichloromethane
(135) ##STR00020##
(136) A magnetically stirred suspension of Boc-protected compound (1.80 mmol) in DCM (4 mL) maintained at 0° C. was treated with trifluoroacetic acid (1 mL) and magnetically stirred for 2 h. The cold bath was removed and the mixture was then stirred for a further 1 h at rt. The solvent was removed with a gentle stream of nitrogen and the remaining gum was triturated with diethyl ether (3×10 mL) then placed under high vacuum for 1 h to afford the amine trifluoroacetate salt as a powder and used directly without further purification.
General Procedure G
Preparation of Amides with T3P®
(137) ##STR00021##
(138) A magnetically stirred mixture of carboxylic acid (100 mg, 0.29 mmol) in DMF (1 mL) was sequentially treated with, triethylamine (320 μL, 2.30 mmol), propylphosphonic anhydride (T3P®) (366 μL, 0.58 mmol, 50% wt in EtOAc) and amine (0.29 mmol). The reaction was stirred for 24 h at 18° C. and then the mixture was concentrated in vacuo and purified by flash chromatography.
General Procedure H
Acid Chloride Formation and Amine Addition
(139) ##STR00022##
(140) A magnetically stirred suspension of carboxylic acid (6.25 mmol) in DCM (30 mL) at 0° C. was treated with oxalyl chloride (563 μL, 6.57 mmol) dropwise, followed by DMF (1 drop). The mixture was stirred at 0° C. for 1 h and then the cold-bath was removed and stirring was continued at rt for 2.5 hr. The clear solution was then concentrated with a gentle stream of nitrogen with heating, in a water bath at 40° C. The resulting cream coloured powder was then placed under high vacuum (1 mm Hg) for 1 hr and redissolved in THF (20 mL) and added transferred dropwise by syringe to a magnetically stirred, ice-cold solution of amine (6.25 mmol) and N,N-diisopropylethylamine (1.75 mL, 12.5 mmol) in DCM (20 mL). After 1 hr at 0° C. the cold bath was removed and stirred a further one hr at 18° C. then a solution of sodium hydrogen carbonate (50 mL, half saturated) was added and the DCM and THF were removed in vacuo. The precipitate was collected by vacuum filtration and washed with water (200 mL) and was dried under high vacuum for 18 hours.
General Procedure I
Thionation of 2-chloroquinazolines
(141) The general procedure is illustrated with respect to the synthesis of the compound BT2004:
(142) ##STR00023##
(143) A 10 mL sealed-tube was charged with 2-chloro-4-amino substituted quinazoline (100 mg, 0.31 mmol), potassium thioacetate (106 mg, 0.93 mmol) and dioxane (3 mL). The magnetically stirred mixture was heated at 120° C. for three hours and then the solvent was removed in vacuo. An ammoniacal solution of methanol (1 mL, saturated) was added and then the solution was subjected to flash column chromatography [silica, 1:10 v/v MeOH/DCM elution] to give, after concentration of the appropriate fractions the compound BT2004 (47 mg, 47%) as a tan solid. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.37 (s, 1H), 10.83 (s, 1H), 8.83 (t, J=5.7 Hz, 1H), 8.08 (d, J=8.2 Hz, 1H), 7.74 (d, J=7.9 Hz, 1H), 7.65 (t, J=7.9 Hz, 1H), 7.38 (d, J=8.3 Hz, 1H), 7.34 (d, J=8.2 Hz, 1H), 7.27 (t, J=7.6 Hz, 1H), 7.20 (s, 1H), 7.07 (t, J=7.6 Hz, 1H), 6.98 (t, J=7.4 Hz, 1H), 3.86-3.78 (m, 2H), 3.07 (app. t, J=7.5 Hz, 2H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) δ 180.9, 156.5, 141.2, 136.7, 134.4, 127.7, 123.8, 123.7, 123.1, 121.4, 119.2, 118.8, 116.0, 112.1, 111.8, 110.3, 42.1, 24.9; (+)-LRESIMS m/z (rel. int.) 343 (100) [M+Na].sup.+, 321 (60) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.18H.sub.16N.sub.4S [M+H].sup.+ 321.1168, found 321.1176; v.sub.max 1621, 1604, 1569, 1538, 1372, 1356, 1333, 1278, 1218, 1193, 1124, 954 cm.sup.−1.
General Procedure J
(144) The general procedure is illustrated with respect to the synthesis of the compound BT2152:
(145) ##STR00024##
(146) A mixture of BT2070 (50 mg, 0.10 mmol), potassium carbonate (28 mg, 0.20 mmol) in acetone (1.5 mL) at rt was treated with 1-(bromomethyl)-4-(trifluoromethyl)benzene (24 μL, 0.10 mmol) in one portion. The mixture was stirred for 24 h and then the mixture was purified by flash column chromatography [silica, 1:20 v/v ammoniacal MeOH/DCM elution] to give, after concentration of the appropriate fractions, BT2152 as a white powder (11 mg, 17%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.73 (s, 1H), 8.91 (d, J=2.0 Hz, 1H), 8.82 (app. t, J=5.9 Hz, 1H), 8.52 (dd, J=4.7, 1.6 Hz, 1H), 8.48 (br s, 1H), 8.07 (ddd, J=8.0, 2.4, 1.6 Hz, 1H), 7.98 (s, 1H), 7.65 (s, 1H), 7.58 (dd, J=8.8, 1.7 Hz, 1H), 7.54 (d, J=8.8 Hz, 1H), 7.47 (dd, J=8.0, 4.7 Hz, 1H), 7.39 (d, J=8.0 Hz, 2H), 7.28 (d, J=8.0 Hz, 2H), 7.12 (d, J=2.0 Hz, 1H), 6.86 (s, 1H), 4.42 (s, 2H), 4.34-4.27 (s, 2H), 3.86 (s, 3H), 3.85 (s, 3H), 3.71-3.64 (m, 2H). (+)-LRESIMS m/z (rel. int.) 659 (100) [M+H].sup.+; v.sub.max 1623, 1601, 1547, 1510, 1328, 1233, 1106, 1018, 799 cm.sup.−1.
Experimental Procedures and Product Characterization
Preparation (i). 2-Isothiocyanato-4,5-dimethoxybenzonitrile
(147) ##STR00025##
(148) Prepared according to General Procedure A, from reaction of 2-amino-4,5-dimethoxybenzonitrile (2.50 g, 14.3 mmol) and thiophosgene (1.35 mL, 17.5 mmol) to afford 2-Isothiocyanato-4,5-dimethoxybenzonitrile as a pale-orange powder (2.60 g, 84%). .sup.1H NMR (CDCl.sub.3, 400 MHz) δ 6.96 (s, 1H), 6.75 (s, 1H), 3.93 (s, 3H), 3.90 (s, 3H); .sup.13C NMR (CDCl.sub.3, 100 MHz) δ 153.4, 148.1, 139.8, 128.9, 115.8, 113.6, 109.4, 101.0, 56.5, 56.4; LRMS (EI, 70 eV) m/z (rel. int.) 220 (100, M+), 205 (65), 177 (39), 119 (28); HREIMS calcd. for C.sub.10H.sub.8N.sub.2O.sub.2S [M+.Math. 220.0301, found 220.0312; v.sub.max 3311, 2221, 1713, 1598, 1527, 1406, 1239, 1213, 1191, 1118, 1000, 688 cm.sup.−1.
Preparation (ii). 5-Methoxy-2-isothiocyanatobenzonitrile
(149) ##STR00026##
(150) Prepared according to General Procedure A, from reaction of 2-amino-5-methoxybenzonitrile (492 mg, 3.32 mmol) and thiophosgene (318 μL, 4.15 mmol) to afford 5-methoxy-2-isothiocyanatobenzonitrile (402 mg, 64%) as a tan powder and used without further purification. .sup.1H NMR (CDCl.sub.3, 400 MHz) δ 7.26 (d, J=8.7 Hz, 1H), 7.12-7.07 (m, 2H), 3.84 (s, 2H); .sup.13C NMR (CDCl.sub.3, 100 MHz) δ 157.8, 139.7, 128.2, 127.1, 120.5, 117.3, 115.3, 110.3, 56.0; (+)-LRESIMS m/z (rel. int.) 191 (100) [M+H].sup.+; v.sub.max 2235, 2093, 1603, 1567, 1487, 1299, 1229, 1159, 1112, 1027, 939, 868, 824 cm.sup.−1.
Preparation (iii). 5-Iodo-2-isothiocyanatobenzonitrile
(151) ##STR00027##
(152) Prepared according to General Procedure A from reaction of 2-amino-5-iodobenzonitrile (2.44 g, 10.0 mmol) and thiophosgene (960 μL, 12.5 mmol) to afford 5-iodo-2-isothiocyanatobenzonitrile (1.88 g, 64%) as an orange powder and used without further purification. .sup.1H NMR (CDCl.sub.3, 400 MHz) δ 7.93 (d, J=2.0 Hz, 1H), 7.88 (dd, J=8.5, 2.0 Hz, 1H), 7.07 (d, J=8.5 Hz, 1H); .sup.13C NMR (CDCl.sub.3, 100 MHz) δ 143.0, 142.1, 141.6, 134.5, 128.2, 113.9, 111.3, 90.2; LRMS (EI, 70 eV) m/z (rel. int.) 286 (100, M+), 159 (54); HREIMS calcd. for C.sub.8H.sub.3IN.sub.2S [M+] 285.9056, found 285.9062; v.sub.max 2233, 2063, 1468, 1072, 935, 819 cm.sup.−1.
Preparation (iv). 5-Bromo-2-isothiocyanatobenzonitrile
(153) ##STR00028##
(154) Prepared according to General Procedure A, from reaction of 2-amino-5-bromobenzonitrile (1.97 g, 10.0 mmol) and thiophosgene (960 μL, 12.5 mmol) to afford 5-bromo-2-isothiocyanatobenzonitrile (1.52 g, 64%) as a yellow powder and used without further purification. .sup.1H NMR (CDCl.sub.3, 400 MHz) δ 7.75 (d, J=2.2 Hz, 1H), 7.70 (dd, J=8.6, 2.2 Hz, 1H), 7.22 (d, J=8.6 Hz, 1H); .sup.13C NMR (CDCl.sub.3, 100 MHz) δ 142.1, 137.2, 135.8, 134.0, 128.2, 120.0, 114.1, 111.1; LRMS (EI, 70 eV) m/z (rel. int.) 240 (100, M+.Math.), 238 (97, M+.Math.), 159 (47); HREIMS calcd. for C.sub.8H.sub.3.sup.79BrN.sub.2S [M+.Math.] 237.9195, found 237.9203; v.sub.max 2230, 2034, 1476, 1462, 888, 830 cm.sup.−1.
Preparation (v): Phenyl (2-cyano-4,5-dimethoxyphenyl)carbamate
(155) ##STR00029##
(156) Following a procedure analogous to that used by Vovk, M. B. (Russ. J. Org. Chem. 2007, 43 (2), 312): A magnetically stirred solution of 2-amino-4,5-dimethoxybenzonitrile (500 mg, 2.81 mmol) in toluene (3 mL) at 0° C. was treated with a solution of phenyl chloroformate (388 μL, 3.09 mmol) in toluene (5 mL) dropwise. The mixture was then refluxed for 3 h and cooled to rt over 18 h. The precipitate was collected by vacuum filtration and washed with ether (30 mL) to afford phenyl (2-cyano-4,5-dimethoxyphenyl)carbamate (347 mg, 41-61%) as a beige powder. .sup.1H NMR (CDCl.sub.3, 400 MHz) δ 7.90 (s, 1H), 7.48-7.37 (m, 2H), 7.39 (s, 1H), 7.33-7.25 (m, 1H), 7.25-7.17 (m, 2H), 6.97 (s, 1H), 3.92 (s, 3H), 3.88 (s, 3H); .sup.13C NMR (CDCl.sub.3, 100 MHz) δ 153.9, 151.5, 150.1, 145.3, 136.1, 129.6 (2C), 126.2, 121.5 (2C), 116.7, 112.9, 103.2, 91.8 56.3, 56.3; v.sub.max 3312, 2221, 1712, 1598, 1527, 1511, 1492, 1406, 1357, 1238, 1213, 1190, 1118, 1026, 999, 874, 793, 725, 688 cm.sup.−1.
Preparation (vi): tert-Butyl (2-(5-bromo-1H-indole-2-carboxamido)ethyl)carbamate
(157) ##STR00030##
(158) Following General Procedure H, 5-bromo-1H-indole-2-carboxylic acid (1.50 g, 6.25 mmol) was converted to 5-bromo-1H-indole-2-carbonyl chloride and reacted with tert-butyl (2-aminoethyl)carbamate (1.00 g, 6.25 mmol) to afford tert-butyl (2-(5-bromo-1H-indole-2-carboxamido)ethyl)carbamate (2.17 g, 91%) as a white powder. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.78 (s br, 1H), 8.57 (t, J=5.7 Hz, 1H), 7.84 (d, J=2.0 Hz, 1H), 7.39 (d, J=8.7 Hz, 1H), 7.28 (dd, J=8.7, 2.0 Hz, 1H), 7.08 (s, 1H), 6.92 (t, J=5.7 Hz, 1H), 3.32 (app. q, J=6.3 Hz, 2H), 3.13 (app. q, J=6.3 Hz, 2H), 1.37 (s, 9H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) 2 signals obscured by DMSO-d.sub.6 δ 160.9, 155.7, 135.0, 133.1, 128.9, 125.8, 123.6, 114.3, 112.1, 101.9, 77.7, 28.2 (3C); (+)-LRESIMS m/z (rel. int.) 404 (100) [M+H], 406 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.16H.sub.20N.sub.3O.sub.3Na.sup.81Br [M+Na].sup.+ 406.0560, found 406.0561; v.sub.max 3357, 3269, 1684, 1629, 1546, 1531, 1367, 1275, 1252, 1167, 880, 797, 768 cm.sup.−1.
Preparation (vii): tert-Butyl (2-(6-bromo-1H-indole-2-carboxamido)ethyl)carbamate
(159) ##STR00031##
(160) Following a procedure analogous to General Procedure H, 6-bromo-1H-indole-2-carboxylic acid (1.00 g, 4.17 mmol) was converted to 6-bromo-1H-indole-2-carbonyl chloride and reacted with tert-butyl (2-aminoethyl)carbamate (0.73 g, 4.56 mmol) to afford tert-butyl (2-(6-bromo-1H-indole-2-carboxamido)ethyl)carbamate (1.20 g, 76%) as a white solid. 1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.71 (s, 1H), 8.55 (t, J=5.3 Hz, 1H), 7.59 (d, J=8.1 Hz, 1H), 7.58 (s, 1H), 7.15 (dd, J=8.1, 1.9 Hz, 1H), 7.10 (s, 1H), 6.92 (t, J=5.3 Hz, 1H), 3.29 (app. q, J=6.2 Hz, 2H), 3.11 (app. q, J=6.2 Hz, 2H), 1.37 (s, 9H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) δ 160.9, 155.7, 137.1, 132.6, 126.1, 123.4, 122.7, 115.9, 114.7, 102.5, 77.7, 39.7, 39.2, 28.2 (3C); (+)-LRESIMS m/z (rel. int.) 404 (100), 406 (100) [M+Na].sup.+; (+)-HRESIMS calcd. for C.sub.16H.sub.20.sup.79BrN.sub.3NaO.sub.3 [M+Na].sup.+404.0586, found 404.0589; v.sub.max 1696, 1647, 1549, 1365, 1289, 1247, 1152 cm.sup.−1.
Preparation (viii): tert-Butyl (3-(6-bromo-1H-indole-2-carboxamido)propyl)carbamate
(161) ##STR00032##
(162) Following a procedure analogous to General Procedure H, 6-bromo-1H-indole-2-carboxylic acid (1.00 g, 4.17 mmol) was converted to 6-bromo-1H-indole-2-carbonyl chloride and reacted with N-(3-aminopropyl)-6-bromo-1H-indole-2-carboxamide (0.73 g, 4.17 mmol) to afford tert-butyl (3-(6-bromo-1H-indole-2-carboxamido)propyl)carbamate (0.88 g, 53%) as a white solid. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.71 (s, 1H), 8.52 (t, J=5.7 Hz, 1H), 7.58 (d, J=8.4 Hz, 1H), 7.58 (d, J=1.2 Hz, 1H), 7.15 (dd, J=8.4, 1.7 Hz, 1H), 7.11 (d, J=1.2 Hz, 1H), 6.83 (t, J=5.3 Hz, 1H), 3.28 (app. q, J=6.8 Hz, 2H), 2.99 (app. q, J=6.5 Hz, 2H), 1.68-1.61 (m, J=6.8 Hz, 2H), 1.37 (s, 9H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) δ 160.7, 155.6, 137.1, 132.7, 126.1, 123.3, 122.7, 115.9, 114.7, 102.3, 77.5, 37.7, 36.6, 29.7, 28.3. (3C); (+)-LRESIMS m/z (rel. int.) 396 (100), 398 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.17H.sub.23.sup.79BrN.sub.3O.sub.3[M+H].sup.+ 396.0917, found 396.0917; v.sub.max 3381, 3269, 2979, 1686, 1615, 1549, 1523, 1365, 1267, 1168, 906, 861, 814, 788, 732 cm.sup.−1.
Preparation (ix): tert-Butyl (1-(5-bromo-1H-indole-2-carbonyl)azetidin-3-yl)carbamate
(163) ##STR00033##
(164) Following a procedure analogous to General Procedure H, 5-bromo-1H-indole-2-carboxylic acid (0.74 g, 3.08 mmol) was converted to 5-bromo-1H-indole-2-carbonyl chloride and reacted with 3-N-Boc-amino-azetidine hydrochloride (0.64 g, 3.08 mmol) to afford tert-butyl (1-(5-bromo-1H-indole-2-carbonyl)azetidin-3-yl)carbamate (1.12 g, 92%) as a white solid. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.82 (s, 1H), 7.82 (s, 1H), 7.65 (d, J=6.7 Hz, 1H), 7.39 (d, J=8.8 Hz, 1H), 7.33-7.29 (m, 1H), 6.79 (s, 1H), 4.75-4.69 (m, 1H), 4.48-4.37 (m, 1H), 4.36-4.23 (m, 2H), 3.99-3.89 (m, 1H), 1.40 (s, 9H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) δ 161.0, 154.8, 134.6, 130.5, 129.2, 126.2, 123.8, 114.3, 112.1, 103.8, 78.3, 59.6, 55.6, 40.7, 28.2 (3C). (+)-LRESIMS m/z (rel. int.) 394 (100), 396 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.17H.sub.21.sup.79BrN.sub.3O.sub.3[M+H].sup.+ 394.0761, found 394.0765; v.sub.max 3360, 3225, 1680, 1603, 1532, 1455, 1410, 1272, 1169, 800, 767, 639 cm.sup.−1.
Preparation (x): 5-(Methoxycarbonyl)-1H-indole-2-carboxylic acid
(165) ##STR00034##
(166) The title compound was prepared following a procedure analogous to that used by Boger et. al. (Bioorg. Med. Chem. Lett. 2010, 20, 2722). A mixture of methyl 4-amino-3-iodobenzoate (2.92 g, 10.5 mmol), DMF (30 mL), DABCO (3.55 g, 31.5 mmol) and pyruvic acid (2.18 mL, 31.5 mmol) was degassed by sparging with a gentle stream of nitrogen for 10 minutes and then palladium(II) acetate (12 mg, 52.5 μmol) was added. The mixture was sparged with nitrogen for a further minute and then heated at 105° C. under an atmosphere of nitrogen for 6 hours. The reaction was cooled to rt and the mixture was acidified to pH 2-3 with an aqueous solution of HCl (1M) and water added (100 mL). The resulting precipitate was collected to afford 5-(methoxycarbonyl)-1H-indole-2-carboxylic acid (1.37 g, 59%) as a tan solid and used without further purification. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 13.15 (s br, 1H), 12.14 (s, 1H), 8.38 (s, 1H), 7.84 (d, J=8.5 Hz, 1H), 7.51 (d, J=8.5 Hz, 1H), 7.26 (s, 1H), 3.85 (s, 3H); .sup.13C NMR (CDCl.sub.3, 100 MHz) δ 166.9, 162.5, 139.6, 130.3, 126.5, 125.0, 124.8, 121.6, 112.6, 108.9, 51.9. v.sub.max 3324, 1695, 1661, 1433, 1340, 1301, 1256, 1202, 1191, 923, 761 cm.sup.−1.
Preparation (xi): [2-((2-((tert-butoxycarbonyl)amino)ethyl)carbamoyl)-1H-indole-5-carboxylic acid
(167) ##STR00035##
(168) Following a procedure analogous to General Procedure H, 5-(methoxycarbonyl)-1H-indole-2-carboxylic acid (1.18 g, 4.25 mmol) was converted to its acid chloride and reacted with tert-butyl (2-aminoethyl)carbamate. The resulting amide (1.49 g, 4.12 mmol) was then suspended in a solution of methanol/THF/water (3:3:9, 30 mL) and treated with lithium hydroxide monohydrate (3.47 g, 82.7 mmol) and stirred at 18° C. for 1 hour. The mixture was then heated at 55° C. for two hours and the methanol and THF removed in vacuo. Water (100 mL) was added and the mixture transferred to a separatory funnel and washed with EtOAc (2×30 mL) and the aqueous layer brought to pH 3 with an aqueous solution of HCl (2M). The precipitated solid was collected and washed with water (100 mL) to afford the desired carboxylic acid product (1.33 g, 93%) as a beige coloured powder which was used without further purification. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.51 (s, 1H), 11.92 (s, 1H), 8.59 (t, J=5.7 Hz, 1H), 8.30 (s, 1H), 7.78 (dd, J=8.6, 1.6 Hz, 1H), 7.47 (d, J=8.6 Hz, 1H), 7.23 (s, 1H), 6.92 (t, J=5.7 Hz, 1H), 3.35-3.29 (m, 2H), 3.15-3.09 (m, 2H), 1.37 (s, 9H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) δ 168.1, 160.9, 155.7, 138.7, 133.3, 126.6, 124.4, 124.2, 122.3, 112.0, 103.7, 77.7, 39.7, 39.3, 28.2 (3C); (+)-LRESIMS m/z (rel. int.) 370 (100) [M+Na].sup.+; (+)-HRESIMS calcd. for C.sub.17H.sub.21N.sub.3NaO.sub.5 [M+Na].sup.+ 370.1373, found 370.1376; v.sub.max 3277, 1686, 1640, 1560, 1338, 1259, 1169, 760 cm.sup.−1.
Preparation (xii): Morpholine-4-carbonyl cyanide
(169) ##STR00036##
(170) Following a method analogous to that reported by Schwartz et al. (Eur. J. Org. Chem 2017, 34, 5110), in a fully operational and well maintained fume cupboard, behind a safety shield, a 10 mL sealed tube vessel fitted with a magnetic stirring-bar was charged with a mixture of trimethylsilyl cyanide (452 μL, 3.62 mmol, CAUTION!) and (1H-imidazol-1-yl)(morpholino)methanone (624 mg, 3.44 mmol) prepared by the method described by Grzyb, J. A.; et al. Tetrahedron 2005, 61, 7153. The vessel was sealed under an atmosphere of nitrogen and heated with magnetic stirring at 100° C. in an oil bath for 18 h. The sealed tube was cooled and the contents dissolved in DCM (5 mL) and poured into a saturated aqueous solution of sodium bicarbonate (5 mL) and extracted into DCM (2×15 mL). The combined organic layers were washed with brine (5 mL), dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The resulting residue was subjected to flash column chromatography [silica, 1:1 v/v EtOAc/pet. spirit elution] to give, after concentration of the appropriate fractions morpholine-4-carbonyl cyanide as a white solid (433 mg, 90%). Mp. 59-61° C.; .sup.1H NMR (CDCl.sub.3, 500 MHz) 3.77-3.74 (m, 4H), 3.70-3.68 (m, 2H), 3.64-3.62 (m, 2H); .sup.13C NMR (CDCl.sub.3, 125 MHz) 143.3, 109.9, 66.5, 65.9, 47.0, 42.6; LRMS (EI, 70 eV) m/z (rel. int.) 140 (87, M+.Math.), 113 (30), 110 (59), 86 (35), 56 (100), 54 (93); HREIMS calcd. for C.sub.6H.sub.8N.sub.2O.sub.2 [M+.Math.] 140.0580, found 140.0586; v.sub.max 2871, 2233, 1669, 1448, 1434, 1273, 1246, 1113, 1036, 958, 854, 721, 628 cm.sup.−1.
Preparation (xiii): tert-butyl (2-(6-bromopicolinamido)ethyl)carbamate
(171) ##STR00037##
(172) Following a procedure analogous to General Procedure H, 6-bromopicolinic acid (2.00 g, 9.90 mmol) was converted to 6-bromopicolinoyl chloride with oxalyl chloride (0.88 mL, 10.40 mmol) and reacted with tert-butyl (2-aminoethyl)carbamate (1.59 g, 9.90 mmol) to afford tert-butyl (2-(6-bromopicolinamido)ethyl)carbamate (2.41 g, 70%) as a white powder. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 8.66 (t, J=5.0 Hz, 1H), 8.03 (d, J=7.8 Hz, 1H), 7.93 (app. t, J=7.8 Hz, 1H), 7.85 (d, J=7.8 Hz, 1H), 6.92 (t, J=4.7 Hz, 1H), 3.34 (app. q, J=6.0 Hz 2H), 3.12 (app. q, J=5.8 Hz, 2H), 1.36 (s, 9H). (+)-LRESIMS m/z (rel. int.) 366 (100) C.sub.13H.sub.18.sup.79BrN.sub.3NaO.sub.3 [M+Na].sup.+, 368 (98) C.sub.13H.sub.18.sup.81BrN.sub.3NaO.sub.3 [M+Na].sup.+; v.sub.max 3331, 1695, 1657, 1526, 1428, 1271, 1168, 1120, 677 cm.sup.−1.
Preparation (xiv): tert-butyl (1-(6-bromo-1H-indole-2-carbonyl)azetidin-3-yl)carbamate
(173) ##STR00038##
(174) Following a procedure analogous to General Procedure H, 6-bromo-1H-indole-2-carboxylic acid (1.00 g, 4.17 mmol) was converted to 6-bromo-1H-indole-2-carbonyl chloride and reacted with 3-N-Boc-amino-azetidine hydrochloride (0.96 g, 4.58 mmol) to afford tert-butyl (1-(6-bromo-1H-indole-2-carbonyl)azetidin-3-yl)carbamate (1.08 g, 66%) as a tan solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.75 (s, 1H), 7.65 (d, J=7.3 Hz, 1H), 7.62-7.56 (m, 2H), 7.17 (dd, J=8.6, 1.8 Hz, 1H), 6.83 (d, J=1.5 Hz, 1H), 4.73 (app. t, J=7.6 Hz, 1H), 4.49-4.36 (m, 1H), 4.34-4.32 (m, 2H), 3.93 (dd, J=10.3, 5.3 Hz, 1H), 1.40 (s, 9H); .sup.13C NMR (100 MHz, DMSO-d.sub.6) δ 161.1, 154.8, 136.7, 130.1, 126.4, 123.6, 122.8, 116.4, 114.7, 104.6, 78.3, 59.6, 55.6, 40.7, 28.2 (3C). (+)-LRESIMS m/z (rel. int.) 416 (95) C.sub.17H.sub.20.sup.79BrN.sub.3NaO.sub.3 [M+Na].sup.+, 418 (100) C.sub.17H.sub.20.sup.81BrN.sub.3NaO.sub.3 [M+Na].sup.+; v.sub.max 3355, 3237, 1606, 1680, 1531, 1456, 1342, 1167, 815, 645 cm.sup.−1.
Synthesis Example 1—BT1057
(175) ##STR00039##
(176) Prepared according to General Procedure I from reaction of N-(2-(1H-indol-3-yl)ethyl)-2-chloro-6,7-dimethoxyquinazolin-4-amine (70 mg, 0.18 mmol) with potassium thioacetate (60 mg, 0.53 mmol) to afford a residue that was subjected to flash column chromatography [silica, 1:20-1:10 v/v MeOH/DCM elution] to give, after concentration of the appropriate fractions the compound BT1057 as a white powder (51 mg, 76%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.18 (s, 1H), 10.84 (s, 1H), 8.54 (t, J=5.4 Hz, 1H), 7.74 (d, J=7.8 Hz, 1H), 7.55 (s, 1H), 7.34 (d, J=8.1 Hz, 1H), 7.20 (d, J=2.0 Hz, 1H), 7.07 (t, J=7.5 Hz, 1H), 6.98 (t, J=7.4 Hz, 1H), 6.94 (s, 1H), 3.82 (s, 3H), 3.82 (s, 3H), 3.80-3.78 (m, 2H), 3.08-3.02 (m, 2H); (+)-LRESIMS m/z (rel. int.) 403 (100) [M+Na].sup.+; (+)-HRESIMS calcd. for C.sub.20H.sub.21N.sub.4O.sub.2S [M+H].sup.+ 381.1380, found 381.1385; v.sub.max 3391, 3303, 1625, 1609, 1577, 1539, 1511, 1345, 1266, 1205, 1155, 739, 608 cm.sup.−1.
Synthesis Example 2—BT2011
(177) ##STR00040##
(178) Prepared according to General Procedure C from reaction of N-(2-(1H-indol-3-yl)ethyl)-7-bromo-2-chloroquinazolin-4-amine (25 mg, 62 μmol) with potassium thioacetate (20 mg, 124 μmol) to afford a residue that was subjected to flash column chromatography [silica, 1:20 v/v ammoniacal MeOH/DCM elution] to give, after concentration of the appropriate fractions the compound BT2011 as a white powder (18 mg, 73%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.37 (s, 1H), 10.83 (s, 1H), 8.92 (t, J=5.6 Hz, 1H), 8.01 (d, J=8.7 Hz, 1H), 7.72 (d, J=8.0 Hz, 1H), 7.55 (d, J=2.0 Hz, 1H), 7.46 (dd, J=8.7, 2.0 Hz, 1H), 7.34 (d, J=8.1 Hz, 1H), 7.20 (d, J=2.3 Hz, 1H), 7.07 (t, J=7.5 Hz, 1H), 6.97 (t, J=7.5 Hz, 1H), 3.84-3.74 (m, 2H), 3.05 (t, J=7.5 Hz, 2H); (+)-LRESIMS m/z (rel. int.) 399 (20), 401 (20) [M+H].sup.+, 421 (100), 423 (100) [M+Na].sup.+; (+)-HRESIMS calcd. for C.sub.18H.sub.16.sup.79BrN.sub.4S [M+H].sup.+ 399.0274, found 399.0280. v.sub.max3399, 1604, 1540, 1482, 1333, 1216, 1188, 1144, 754, 740 cm.sup.−1.
Synthesis Example 3—BT2012
(179) ##STR00041##
(180) Prepared according to General Procedure B, from reaction of 2-isothiocyanato-4,5-dimethoxybenzonitrile (300 mg, 1.36 mmol) and tryptamine (218 mg, 1.36 mmol) to afford BT2012 as a cream powder (395 mg, 76%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.92 (s br, 1H), 10.83 (s, 1H), 9.18 (s, 1H), 7.94 (d, J=8.0 Hz, 1H), 7.64 (s, 1H), 7.35 (d, J=8.0 Hz, 1H), 7.19 (s, 1H), 7.12-7.04 (m, 1H), 7.02-6.95 (m, 1H), 6.86 (s, 1H), 4.92-4.78 (m, 2H), 3.84 (s, 3H), 3.80 (s, 3H), 3.18-3.09 (m, 2H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) δ 172.8, 153.8, 153.3, 146.6, 136.3, 131.9, 127.4, 122.9, 121.0, 119.0, 118.2, 111.4, 111.2, 107.1, 106.5, 98.4, 56.3, 55.8, 47.8, 21.6; (+)-LRESIMS m/z (rel. int.) 380 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.20H.sub.21N.sub.4O.sub.2S [M+H].sup.+ 381.1380, found 381.1386. v.sub.max 3181, 3122, 1632, 1583, 1551, 1515, 1456, 1441, 1401, 1350, 1285, 1265, 1243, 1163, 1126, 1011 cm.sup.−1.
Synthesis Example 4—BT2014
(181) ##STR00042##
(182) Prepared according to General Procedure B, from reaction of 2-isothiocyanato-5-methoxybenzonitrile (33 mg, 0.16 mmol) and tryptamine (27 mg, 0.17 mmol) to afford BT2014 as a cream powder (24 mg, 42%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.10 (s br, 1H), 10.84 (s, 1H), 9.53 (s br, 1H), 7.94 (d, J=7.8 Hz, 1H), 7.71 (d, J=1.7 Hz, 1H), 7.36 (d, J=7.8 Hz, 1H), 7.29-7.17 (m, 3H), 7.08 (t, J=7.5 Hz, 1H), 7.00 (t, J=7.5 Hz, 1H), 4.92-4.80 (m, 2H), 3.83 (s, 3H), 3.20-3.09 (m, 2H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) δ 173.0, 156.1, 153.3, 136.3, 129.7, 127.4, 122.7, 121.4, 121.0, 119.1, 118.2, 117.4, 115.6, 111.5, 111.3, 108.3, 55.9, 47.7, 21.7. (+)-LRESIMS m/z (rel. int.) 351 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.19H.sub.19N.sub.4OS [M+H].sup.+ 351.1274, found 351.1281. v.sub.max 3307, 1619, 1581, 1542, 1504, 1460, 1349, 1260, 1234, 1158, 1057, 993, 814 cm.sup.−1.
Synthesis Example 5—BT2015
(183) ##STR00043##
(184) Prepared according to General Procedure B, from reaction of 2-isothiocyanato-4,5-dimethoxybenzonitrile (50 mg, 0.23 mmol) and piperonyl amine (34 mg, 0.23 mmol) to afford BT2015 as a cream powder (34 mg, 40%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.02 (s br, 1H), 9.09 (s, 1H), 7.55 (s, 1H), 6.93 (s, 1H), 6.86 (s, 1H), 6.85-6.82 (m, 1H), 6.80 (d, J=8.0 Hz, 1H), 5.95 (s, 2H), 5.78 (s br, 2H), 3.80 (s, 3H), 3.79 (s, 3H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) one signal obstructed or overlapping δ 173.8, 153.3 (br), 146.9, 146.5, 145.8, 131.3 (br), 130.6 (br), 120.5 (br), 108.1 (br), 107.8, 107.4 (br), 100.7, 98.3 (br), 56.1, 55.7, 49.1; (+)-LRESIMS m/z (rel. int.) 372 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.18H.sub.18N.sub.3O.sub.4S [M+H].sup.+ 372.1013, found 372.1021; v.sub.max 3275, 1628, 1577, 1552, 1516, 1502, 1437, 1242, 1188, 1134, 1030, 1001, 924, 821 cm.sup.−1.
Synthesis Example 6—BT2017
(185) ##STR00044##
(186) A mixture of 2-isothiocyanato-4,5-dimethoxybenzonitrile (30 mg, 0.14 mmol), 4-amino-1-Boc-piperidine (55 mg, 0.27 mmol) and triethyl amine (38 μL, 0.27 mmol) in DCM (3 mL) was stirred for 24 h. The solid was collected and washed with DCM (3 mL) to afford BT2017 as a cream powder (50 mg, 87%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.85 (s, 1H), 9.13 (s, 1H), 7.50 (s, 1H), 6.81 (s, 1H), 6.08-5.96 (m, 1H), 4.16-3.94 (m, 2H), 3.80 (s, 3H), 3.77 (s, 3H), 3.07-2.94 (m, 2H), 2.87-2.61 (m, 2H), 1.60-1.52 (m, 2H), 1.42 (s, 9H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) δ 174.6, 153.9, 153.7, 152.8, 146.4, 130.3, 108.8, 107.0, 98.0, 78.5, 59.7 (br), 56.1, 55.7, 44.4-43.3 (br, 2C), 28.1 (3C), 26.4 (2C); (+)-LRESIMS m/z (rel. int.) 421 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.20H.sub.29N.sub.4O.sub.4S [M+H].sup.+ 421.1904, found 421.1900; v.sub.max 1690, 1638, 1542, 1511, 1418, 1234, 1136, 1126, 1010 cm.sup.−1.
Synthesis Example 7—BT2018
(187) ##STR00045##
(188) Prepared according to General Procedure B, from reaction of 2-isothiocyanato-4,5-dimethoxybenzonitrile (50 mg, 0.23 mmol) and n-butylamine (68 μL, 0.68 mmol) to afford BT2018 as a white powder (39 mg, 87%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.84 (s br, 1H), 9.00 (s br, 1H), 7.56 (s, 1H), 6.82 (s, 1H), 4.62-4.49 (m, 2H), 3.82 (s, 3H), 3.78 (s, 3H), 1.73-1.62 (m, 2H), 1.39-1.28 (m, 2H), 0.92 (t, J=7.3 Hz, 3H); (+)-LRESIMS m/z (rel. int.) 294 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.14H.sub.20N.sub.3O.sub.2S [M+H].sup.+ 294.1271, found 294.1274; v.sub.max 3272, 1629, 1577, 1516, 1436, 1354, 1258, 1239, 1110, 1022, 822, 764 cm.sup.−1.
Synthesis Example 8—BT2020
(189) ##STR00046##
(190) Prepared according to General Procedure B, from reaction of 5-iodo-2-isothiocyanatobenzonitrile (50 mg, 0.17 mmol) and piperonylamine (26 mg, 0.17 mmol) to afford BT2020 as a white powder (37 mg, 48%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.27 (s br, 1H), 9.63 (s br, 1H), 8.48 (s, 1H), 7.86 (dd, J=8.5, 1.8 Hz, 1H), 7.06 (d, J=8.5 Hz, 1H), 6.92 (s, 1H), 6.83 (dd, J=8.2, 0.8 Hz, 1H), 6.79 (d, J=8.2 Hz, 1H), 5.95 (s, 2H), 5.73 (s br, 2H); (+)-LRESIMS m/z (rel. int.) 438 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.16H.sub.13N.sub.3O.sub.2SI [M+H].sup.+ 437.9768, found 437.9774; v.sub.max 3279, 1608, 1538, 1503, 1446, 1249, 1187, 1042, 952, 814, 620 cm.sup.−1.
Synthesis Example 9—BT2021
(191) ##STR00047##
(192) Prepared according to General Procedure B, from reaction of 2-isothiocyanato-4,5-dimethoxybenzonitrile (50 mg, 0.23 mmol) and 2-phenylethan-1-amine (28 mg, 0.23 mmol) to afford BT2021 as a white powder (32 mg, 36%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.90 (s br, 1H), 9.12 (s br, 1H), 7.60 (s, 1H), 7.37-7.29 (m, 4H), 7.26-7.21 (m, 1H), 6.84 (s, 1H), 4.84-4.71 (m, 2H), 3.83 (s, 3H), 3.80 (s, 3H); 3.00 (t, J=8.2 Hz, 2H); (+)-LRESIMS m/z (rel. int.) 342 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.18H.sub.20N.sub.3O.sub.2S [M+H].sup.+ 342.1271, found 342.1270; v.sub.max 3274, 1630, 1577, 1552, 1516, 1436, 1351, 1279, 1238, 1253, 1010, 820, 704 cm.sup.−1.
Synthesis Example 10—BT2022
(193) ##STR00048##
(194) A mixture of 2-isothiocyanato-4,5-dimethoxybenzonitrile (30 mg, 0.14 mmol), 2,5-dimethoxybenzyl amine (46 mg, 0.27 mmol) and triethyl amine (38 μL, 0.27 mmol) in DCM (3 mL) was stirred for 24 h. The solid was collected and washed with DCM (3 mL) to afford BT2022 as a white powder (47 mg, 87%). .sup.1H NMR poor resolution of signals (DMSO-d.sub.6, 400 MHz) δ 11.93 (s br, 1H), 9.03 (s br, 1H), 7.58 (s br, 1H), 6.91 (s br, 2H), 6.74 (s br, 1H), 6.18 (s br, 1H), 5.74 (s br, 2H), 3.81 (m, 9H), 3.57 (s, 3H); (+)-LRESIMS m/z (rel. int.) 388 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.19H.sub.22N.sub.3O.sub.4S [M+H].sup.+ 388.1326, found 388.1325; v.sub.max 1642, 1550, 1505, 1433, 1396, 1280, 1226, 1174, 1118, 1050, 997, 820 cm.sup.−1.
Synthesis Example 11—BT2023
(195) ##STR00049##
(196) Prepared according to General Procedure B, from reaction of 2-isothiocyanato-4,5-dimethoxybenzonitrile (50 mg, 0.23 mmol) and hydrazine monohydrate (34 mg, 0.68 mmol) to afford BT2023 as a white powder (55 mg, 96%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 10.74-8.64 (br m, 2H), 7.76 (s, 1H), 6.92 (s, 1H), 6.38 (s, 2H), 3.89 (s, 3H), 3.85 (s, 3H); (+)-LRESIMS m/z (rel. int.) 253 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.10H.sub.13N.sub.4O.sub.2S [M+H].sup.+ 253.0754, found 253.0754; v.sub.max 1622, 1554, 1502, 1402, 1337, 1289, 1236, 1199, 1105, 991, 847, 755, 644 cm.sup.−1.
Synthesis Example 12—BT2024
(197) ##STR00050##
(198) A magnetically stirred solution of tert-butyl (2-(1H-indol-3-yl)ethyl)carbamate (200 mg, 0.77 mmol) and potassium hydroxide (101 mg, 1.54 mmol) in DMSO (3 mL) was treated with methyl iodide (72 μL, 1.15 mmol) and stirred for 18 h. The mixture was diluted with water (10 mL) and extracted with EtOAc (3×15 mL) and the combined organic layers washed with brine (15 mL). The solvent was removed in vacuo and the residue subjected to flash column chromatography [silica, 1:3 v/v EtOAc/Pet spirit elution] to give, after concentration of the appropriate fractions, tert-butyl (2-(1-methyl-1H-indol-3-yl)ethyl)carbamate (154 mg, 73%) as a thick pale yellow oil. 1H NMR (CDCl.sub.3, 400 MHz) δ 7.60 (d, J=7.9 Hz, 1H), 7.34-7.27 (m, 1H), 7.23 (ddd, J=8.1, 6.9, 1.1 Hz, 1H), 7.11 (ddd, J=7.9, 7.0, 1.0 Hz, 1H), 6.89 (s, 1H), 3.76 (s, 2H), 3.46 (d, J=6.3 Hz, 2H), 2.94 (t, J=6.8 Hz, 1H), 1.44 (s, 9H). Spectroscopic data were identical with those reported. t-Butyl (2-(1-methyl-1H-indol-3-yl)ethyl)carbamate (154 mg, 0.56 mmol) was treated with a solution of hydrogen chloride (2 mL, 4M in dioxane) and magnetically stirred for 3 h. The solvent was removed with a gentle stream of nitrogen and the remaining gum was triturated with ether (3×10 mL) then placed under high vacuum for 1 h to afford the amine hydrochloride as a cream coloured gum and used directly without further purification. A mixture of triethylamine (88 μL, 0.64 mmol), the crude amine hydrochloride formed directly above, and DCM (1 mL) was added to a magnetically stirred solution of 2-isothiocyanato-4,5-dimethoxybenzonitrile (35 mg, 0.16 mmol) in a mixture of DCM and pet. spirit (0.75 mL:0.75 mL). The mixture was stirred for 0.5 h and then pet. spirit (5 mL) was added. The solid was allowed to settle and the solvent was decanted and then ethanol (5 mL) was added and the mixture heated at 70° C. for 2 h. The mixture was cooled to rt and the solid was collected by vacuum filtration, washed with ethanol (5 mL) and dried at the pump (1 mmHg) to afford BT2024 as a cream powder (43 mg, 69%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 10.90 (s br, 2H), 7.89 (d, J=7.8 Hz, 1H), 7.81 (s, 1H), 7.39 (d, J=8.2 Hz, 1H), 7.25 (s, 1H), 7.15 (t, J=7.6 Hz, 1H), 7.03 (t, J=7.8 Hz, 1H), 6.91 (s, 1H), 4.93-4.79 (m, 2H), 3.86 (s, 3H), 3.84 (s, 3H), 3.76 (s, 3H), 3.12 (app. t, J=8.4 Hz, 2H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) δ 172.4, 154.5, 153.5, 146.9, 136.6, 132.6, 127.7, 127.6, 121.1, 119.1, 118.3, 110.1, 109.5, 106.9, 105.6, 98.2, 56.4, 55.9, 48.1, 32.2, 21.3; (+)-LRESIMS m/z (rel. int.) 395 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.21H.sub.23N.sub.4O.sub.2S [M+H].sup.+ 395.1536, found 395.1544; v.sub.max 1628, 1579, 1541, 1515, 1441, 1400, 1263, 1246, 1129, 1015, 738 cm.sup.−1.
Synthesis Example 13—BT2025
(199) ##STR00051##
(200) Prepared according to General Procedure B, from reaction of 2-isothiocyanato-4,5-dimethoxybenzonitrile (30 mg, 0.14 mmol) and phenyl hydrazine monohydrate (15 mg, 0.14 mmol) to afford BT2025 as a white powder (21 mg, 47%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 9.15 (s br, 1H), 8.99 (s br, 1H), 8.61 (s br, 1H), 7.60 (s, 1H), 7.22 (t, J=7.7 Hz, 2H), 6.90 (t, J=7.4 Hz, 1H), 6.83 (s, 1H), 6.65 (d, J=8.0 Hz, 2H), 3.88 (s, 3H), 3.83 (s, 3H); (+)-LRESIMS m/z (rel. int.) 329 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.16H.sub.17N.sub.4O.sub.2S [M+H].sup.+ 329.1067, found 329.1067; v.sub.max 3415, 3070, 1625, 1551, 1503, 1495, 1434, 1285, 1231, 990, 844 cm.sup.−1.
Synthesis Example 14—BT2026
(201) ##STR00052##
(202) Prepared according to General Procedure B, from reaction of 5-bromo-2-isothiocyanatobenzonitrile (300 mg, 1.25 mmol) and tryptamine (201 mg, 1.25 mmol) to afford BT2026 as a cream powder. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ; (+)-LRESIMS m/z (rel. int.) 399 (100), 401 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.18H.sub.16N.sub.4S.sup.79Br [M+H].sup.+ 399.0274, found 399.0274; v.sub.max 3468, 3249, 1633, 1603, 1538, 1450, 1329, 1156, 1146, 848, 803, 750 cm.sup.−1.
Synthesis Example 15—BT2027
(203) ##STR00053##
(204) Prepared according to General Procedure B, from reaction of 2-isothiocyanato-4,5-dimethoxybenzonitrile (35 mg, 0.16 mmol), 2,4-dimethoxybenzylamine hydrochloride (32 mg, 0.16 mmol) and triethylamine (22 μL, 0.16 mmol) to afford BT2027 as a white powder (21 mg, 47%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 10.76 (s br, 2H), 7.88 (s, 1H), 6.99 (s, 1H), 6.66 (d, J=8.4 Hz, 1H), 6.62 (d, J=2.4 Hz, 1H), 6.39 (dd, J=8.4, 2.4 Hz, 1H), 5.72 (s br, 2H), 3.88 (s, 3H), 3.86 (s, 6H), 3.73 (s, 3H); (+)-LRESIMS m/z (rel. int.) 388 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.19H.sub.22N.sub.3O.sub.4S [M+H].sup.+ 388.1326, found 388.1333; v.sub.max 1676, 1642, 1617, 1547, 1508, 1434, 1244, 1209, 1156, 1117, 1031, 823 cm.sup.−1.
Synthesis Example 16—BT2028
(205) ##STR00054##
(206) Prepared according to General Procedure B, from reaction of 2-isothiocyanato-4,5-dimethoxybenzonitrile (35 mg, 0.16 mmol), tyramine hydrochloride (28 mg, 0.16 mmol) and triethylamine (22 μL, 0.16 mmol) to afford a residue that was further triturated with ethanol (2 mL) to provide BT2028 as a cream powder (13 mg, 23%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.95 (s br, 1H), 9.20 (s, 1H), 9.10 (s br, 1H), 7.60 (s, 1H), 7.12 (d, J=8.0 Hz, 2H), 6.84 (s, 1H), 6.71 (d, J=8.0 Hz, 2H), 4.76-4.65 (m, 2H), 3.83 (s, 3H), 3.79 (s, 3H), 2.87 (app. t, J=8.2 Hz, 2H); (+)-LRESIMS m/z (rel. int.) 358 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.18H.sub.20N.sub.3O.sub.3S [M+H].sup.+ 358.1220, found 358.1219; v.sub.max 1629, 1580, 1550, 1512, 1438, 1354, 1248, 1235, 1212, 1126, 1008, 823, 767 cm.sup.−1.
Synthesis Example 17—BT2032
(207) ##STR00055##
(208) Prepared according to General Procedure B, from reaction of 2-isothiocyanato-5-iodobenzonitrile (300 mg, 1.1 mmol) with tryptamine (168 mg, 1.1 mmol) to afford BT2032 as a cream powder (235 mg, 66%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.19 (s, 1H), 10.84 (s, 1H), 9.70 (s, 1H), 8.55 (s, 1H), 7.90-7.85 (m, 2H), 7.35 (d, J=8.0 Hz, 1H), 7.19 (s, 1H), 7.11-7.05 (m, 2H), 6.99 (t, J=7.4 Hz, 1H), 4.86-4.76 (m, 2H), 3.11 (t, J=8.3 Hz, 2H); (+)-LRESIMS m/z (rel. int.) 447 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.18H.sub.16N.sub.4SI [M+H].sup.+ 447.0135, found 447.0140; v.sub.max 3451, 3279, 1621, 1599, 1582, 1528, 1460, 1381, 1323, 1286, 1154, 1145, 1038, 817, 741 cm.sup.−1.
Synthesis Example 18—BT2033
(209) ##STR00056##
(210) Prepared according to General Procedure B, from reaction of 2-isothiocyanato-4,5-dimethoxybenzonitrile (200 mg, 0.91 mmol) with 2-aminoethan-1-ol (55 μL, 0.91 mmol) to afford BT2033 as a white powder (179 mg, 23%). .sup.1H (DMSO-d.sub.6, 400 MHz) δ 11.90 (s br, 1H), 9.02 (s br, 1H), 7.57 (s, 1H), 6.82 (s, 1H), 4.95 (s br, 1H), 4.75-4.63 (m, 2H), 3.81 (s, 3H), 3.79 (s, 3H), 3.73-3.65 (m, 2H); (−)-LRESIMS m/z (rel. int.) 280 (100) [M−H]−; (−)-HRESIMS calcd. for C.sub.12H.sub.14N.sub.3O.sub.3S [M−H]− 280.0761, found 280.0757; v.sub.max 3514, 3266, 1631, 1584, 1549, 1518, 1437, 1396, 1280, 1263, 1232, 1096, 1058, 1012, 821 cm.sup.−1. A portion of the product (20 mg) was suspended in DCM (2 mL) and trifluoroacetic acid (100 μL) added. The mixture was concentrated under a gentle stream of nitrogen over 24 h to afford the trifluoroacetate salt of BT2033. .sup.1H (DMSO-d.sub.6, 400 MHz) TFA salt δ 13.75 (s br, 1H), 10.36 (s, 1H), 9.69 (s br, 1H), 8.68 (s br, 1H), 7.80 (s, 1H), 6.99 (s, 1H), 4.88-4.64 (m, 2H), 3.90 (s, 3H), 3.88-3.84 (m, 2H), 3.85 (s, 3H); .sup.13C (DMSO-d.sub.6, 100 MHz) TFA salt δ 171.4, 158.5 (q, JC-F=35.3 Hz) 157.2, 155.7, 147.7, 135.3, 116.0 (q, JC-F=292.9 Hz), 105.4, 102.2, 97.7, 57.0, 56.4, 56.4, 51.2.
Synthesis Example 19—BT2034
(211) ##STR00057##
(212) Prepared according to General Procedure B, from reaction of 2-isothiocyanato-4,5-dimethoxybenzonitrile (35 mg, 0.16 mmol) with 4-phenylbutan-1-amine (34 mg, 0.23 mmol) to afford BT2034 as a white powder (11 mg, 13%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.81 (s br, 1H), 8.99 (s br, 1H), 7.55 (s, 1H), 7.32-7.11 (m, 5H), 6.82 (s, 1H), 4.68-4.53 (m, 2H), 3.82 (s, 3H), 3.78 (s, 3H), 2.63 (app. t, J=7.6 Hz, 2H), 1.78-1.69 (m, 2H), 1.67-1.57 (m, 2H); (+)-LRESIMS m/z (rel. int.) 370 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.20H.sub.24N.sub.3O.sub.2S [M+H].sup.+ 370.1584, found 370.1584; v.sub.max 3270, 2981, 1629, 1579, 1551, 1517, 1462, 1267, 1240, 1212, 1123, 1021, 820, 697 cm.sup.−1.
Synthesis Example 20—BT2035
(213) ##STR00058##
(214) Prepared according to General Procedure B, from reaction of 2-isothiocyanato-5-methoxybenzonitrile (33 mg, 0.16 mmol) with piperonyl amine (25 mg, 0.17 mmol) to afford BT2035 as a white powder (35 mg, 64%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.14 (s br, 1H), 9.45 (s, 1H), 7.64 (s, 1H), 7.26 (s, 1H), 7.19 (dd, J=8.9, 2.6 Hz, 1H), 6.94 (s, 1H), 6.85 (d, J=7.9 Hz, 1H), 6.80 (d, J=8.0 Hz, 1H), 5.95 (s, 2H), 5.78 (s, 2H), 3.80 (s, 3H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) δ 173.6, 156.2, 153.4, 146.8, 145.8, 131.3, 129.6, 121.5, 120.6, 117.4, 115.5, 108.3, 108.2, 107.8, 100.7, 55.9, 49.2; (+)-LRESIMS m/z (rel. int.) 342 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.17H.sub.16N.sub.3O.sub.3S [M+H].sup.+ 342.0907, found 342.0911; v.sub.max 3259, 1621, 1544, 1502, 1443, 1238, 1164, 1040, 969, 813, 723, 621 cm.sup.−1.
Synthesis Example 21—BT2037
(215) ##STR00059##
(216) Prepared according to General Procedure B, from reaction of 2-isothiocyanato-4,5-dimethoxybenzonitrile (35 mg, 0.16 mmol) with methylamine (159 μL, 0.32 mmol, 2M solution in MeOH) to afford BT2037 as a white powder (19 mg, 48%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.91 (s, 1H), 9.06 (s, 1H), 7.58 (s, 1H), 6.83 (s, 1H), 3.82 (s, 3H), 3.82 (s, 3H), 3.80 (s, 3H); (+)-LRESIMS m/z (rel. int.) 252 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.11H.sub.14N.sub.3O.sub.2S [M+H].sup.+ 252.0801, found 252.0804; v.sub.max 3276, 1631, 1584, 1552, 1518, 1438, 1277, 1264, 1235, 1090, 820, 764, 661 cm.sup.−1.
Synthesis Example 22—BT2038
(217) ##STR00060##
(218) Prepared according to General Procedure B, from reaction of 5-iodo-2-isothiocyanatobenzonitrile (200 mg, 0.70 mmol) with 4-methoxybenzylamine (96 mg, 0.70 mmol) to afford BT2038 as a white powder (202 mg, 68%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.26 (s, 1H), 9.63 (s, 1H), 8.50 (s, 1H), 7.86 (dd, J=8.5, 1.8 Hz, 1H), 7.28 (d, J=8.7 Hz, 2H), 7.07 (d, J=8.5 Hz, 1H), 6.82 (d, J=8.7 Hz, 2H), 5.76 (s, 2H), 3.70 (s, 3H); (+)-LRESIMS m/z (rel. int.) 424 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.16H.sub.15N.sub.3OSI [M+H].sup.+ 423.9975, found 423.9972; v.sub.max 1606, 1580, 1512, 1387, 1250, 1183, 1177, 1026, 963, 949, 818, 604 cm.sup.−1.
Synthesis Example 23—BT2039
(219) ##STR00061##
(220) Prepared according to General Procedure B, from reaction of 5-bromo-2-isothiocyanatobenzonitrile (100 mg, 0.42 mmol) with but-3-yn-1-amine (29 mg, 0.42 mmol) to afford BT2039 as a tan powder (32 mg, 25%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.24 (s, 1H), 9.65 (s, 1H), 8.41 (d, J=2.1 Hz, 1H), 7.71 (dd, J=8.7, 2.1 Hz, 1H), 7.17 (d, J=8.7 Hz, 1H), 4.71-4.62 (m, 2H), 2.88 (t, J=2.7 Hz, 1H), 2.59 (ddd, J=9.3, 7.0, 2.7 Hz, 2H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) δ 174.1, 151.8, 135.8, 134.6, 128.5, 117.9, 116.5, 116.0, 81.3, 72.6, 45.4, 15.1; (+)-LRESIMS m/z (rel. int.) 308 (100), 310 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.12H.sub.11N.sub.3S.sup.79Br [M+H].sup.+ 307.9852, found 307.9861; v.sub.max 3274, 3258, 1603, 1586, 1533, 1463, 1381, 1278, 1150, 822, 657 cm.sup.−1.
Synthesis Example 24—BT2040
(221) ##STR00062##
(222) Prepared according to General Procedure B, from reaction of 2-isothiocyanato-5-methoxybenzonitrile (33 mg, 0.16 mmol) with 4-methoxybenzylamine (23 mg, 0.17 mmol) to afford BT2040 as a white powder (16 mg, 31%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.14 (s, 1H), 9.44 (s, 1H), 7.64 (s, 1H), 7.30 (d, J=8.3 Hz, 2H), 7.27-7.16 (m, 2H), 6.82 (d, J=8.3 Hz, 2H), 5.81 (s, 2H), 3.80 (s, 3H), 3.70 (s, 3H); (+)-LRESIMS m/z (rel. int.) 328 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.17H.sub.18N.sub.3O.sub.2S [M+H].sup.+ 328.1114, found 328.1116; v.sub.max 3267, 1617, 1589, 1542, 1507, 1440, 1276, 1241, 1176, 1033, 967, 812, 724, 613 cm.sup.−1.
Synthesis Example 25—BT2041
(223) ##STR00063##
(224) Prepared according to General Procedure B, from reaction of 2-isothiocyanato-5-methoxybenzonitrile (33 mg, 0.16 mmol) with 2-phenylethan-1-amine (20 mg, 0.17 mmol) to afford BT2041 as a white powder (15 mg, 30%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.10 (s, 1H), 9.47 (s, 1H), 7.68 (s, 1H), 7.36-7.31 (m, 4H), 7.27-7.16 (m, 3H), 4.77 (app. t, J=8.1 Hz, 2H), 3.82 (s, 3H), 3.01 (app. t, J=8.2 Hz, 2H); v.sub.max 3279, 1628, 1583, 1538, 1496, 1438, 1354, 1250, 1238, 1163, 1143, 1033, 991, 820, 750 cm.sup.−1.
Synthesis Example 26—BT2042
(225) ##STR00064##
(226) Prepared according to General Procedure B, from reaction of 2-isothiocyanato-4,5-dimethoxybenzonitrile (30 mg, 0.14 mmol) with tert-butyl (2-aminoethyl)carbamate (23 mg, 0.14 mmol) to afford BT2042 as a white powder (8 mg, 16%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.89 (s, 1H), 9.05 (s, 1H), 7.59 (s, 1H), 6.82 (s, 1H), 4.75-4.66 (m, 2H), 3.82 (s, 3H), 3.78 (s, 3H), 3.35-3.29 (m, 2H), 1.34 (s, 9H); (+)-LRESIMS m/z (rel. int.) 381 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.17H.sub.25N.sub.4O.sub.4S [M+H].sup.+ 381.1591, found 381.1599; v.sub.max 3381, 3302, 1688, 1635, 1600, 1506, 1460, 1438, 1339, 1279, 1238, 1203, 1164, 1127, 1088, 824, 759, 629 cm.sup.−1.
Synthesis Example 27—BT2043
(227) ##STR00065##
(228) t-Butyl (2-(1-(morpholine-4-carbonyl)-1H-indol-3-yl)ethyl)carbamate: A magnetically stirred solution of Boc-tryptamine (179 mg, 0.69 mmol) in THF (5 mL) at 0° C. was treated with a solution of lithium bis(trimethylsilyl)amide (760 μL, 0.76 mmol, 1M in THF) dropwise. The reaction was stirred for 0.5 h and was then treated in one portion with morpholine-4-carbamoyl cyanide see bottom of document (96 mg, 0.69 mmol) and stirred for 1 h. The reaction was poured onto ice cold sodium hydrogen carbonate (20 mL, half saturated) and the mixture extracted with EtOAc (3×15 mL). The combined organic layers were washed with brine, dried (Na.sub.2SO.sub.4) and concentrated in vacuo to afford a residue that was subjected to flash column chromatography [silica, 1:3 v/v EtOAc/pet. spirit elution] to give, after concentration of the appropriate fractions the morpholine-indoleamide (196 mg, 72%) as a white powder. .sup.1H NMR (CDCl.sub.3, 400 MHz) δ 7.69 (d, J=8.2 Hz, 1H), 7.57 (d, J=7.8 Hz, 1H), 7.34-7.29 (m, 1H), 7.22 (t, J=7.5 Hz, 1H), 7.15 (s, 1H), 4.62 (s, 1H), 3.84-3.74 (m, 4H), 3.67-3.53 (m, 4H), 3.50-3.43 (m, 2H), 2.92 (app. t, J=6.9 Hz, 2H), 1.44 (s, 9H); .sup.13C NMR (CDCl.sub.3, 100 MHz) δ 155.9, 154.3, 135.7, 129.5, 124.0, 123.5, 121.9, 119.2, 117.0, 113.5, 79.3, 66.7 (2C), 47.1 (2C), 40.3, 28.4, 25.7; (+)-LRESIMS m/z (rel. int.) 396 (100) [M+Na].sup.+; (+)-HRESIMS calcd. for C.sub.20H.sub.27N.sub.3O.sub.4Na [M+Na].sup.+396.1894, found 396.1907; v.sub.max 3352, 2971, 2927, 1703, 1674, 1517, 1452, 1413, 1365, 1270, 1241, 1166, 1114, 993, 746 cm.sup.−1. A sample of the above compound (186 mg) was treated with a solution of hydrogen chloride (2 mL, 4M in dioxane) and magnetically stirred for 1 h. The solvent was removed with a gentle stream of nitrogen and the remaining gum was triturated with ether (3×10 mL) then placed under high vacuum for 1 h to afford the amine hydrochloride as a cream coloured gum and a portion of which used directly without further purification. A mixture of triethylamine (44 μL, 0.32 mmol), the amine hydrochloride formed directly above (49 mg, 0.16 mmol, crude) and DCM (1.5 mL) was added to a magnetically stirred solution of 2-isothiocyanato-4,5-dimethoxybenzonitrile (35 mg, 0.16 mmol) in a mixture of DCM and pet. spirit (1.5 mL:1.5 mL). The mixture was stirred for 0.5 h and then pet. spirit (5 mL) was added. The solid was allowed to settle and the solvent was decanted and then ethanol (5 mL) was added and the mixture heated at 70° C. for 2 h. The mixture was cooled to rt and the solid was collected by vacuum filtration, washed with ethanol (5 mL) and dried at the pump (1 mmHg) to afford BT2043 as a cream powder (32 mg, 41%). .sup.1H NMR (DMSO-d.sub.6) δ 11.78 (s br, 1H), 9.60 (s br, 1H), 7.99 (d, J=7.7 Hz, 1H), 7.71 (s, 1H), 7.66 (d, J=8.2 Hz, 1H), 7.44 (s, 1H), 7.32-7.24 (m, 1H), 7.24-7.16 (m, 1H), 6.88 (s, 1H), 4.95-4.85 (m, 2H), 3.85 (s, 3H), 3.82 (s, 3H), 3.71-3.62 (m, 4H), 3.57-3.48 (m, 4H), 3.15 (app. t, J=8.1 Hz, 2H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) three signals obstructed, overlapping or not resolved δ 172.8, 153.9 (br), 153.3, 153.2, 146.7, 135.3, 129.2, 124.3, 123.5, 121.3, 119.7, 115.6, 107.1, 106.3 (br), 98.4, 65.9 (2C), 56.3, 55.8, 47.1, 46.4 (2C), 21.2; (+)-LRESIMS m/z (rel. int.) 494 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.25H.sub.28N.sub.5O.sub.4S [M+H]+ 494.1857, found 494.1862; v.sub.max 1682, 1630, 1575, 1548, 1517, 1452, 1398, 1281, 1256, 1234, 1120, 1008, 823, 737 cm.sup.−1.
Synthesis Example 28—BT2045
(229) ##STR00066##
(230) Prepared according to General Procedure B, from reaction of 2-isothiocyanato-4,5-dimethoxybenzonitrile (50 mg, 0.23 mmol) with pyridin-4-ylmethanamine (24.5 mg, 0.23 mmol) to afford BT2045 as a white powder (60 mg, 80%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.15 (s br, 1H), 9.09 (s, 1H), 8.44 (d, J=5.8 Hz, 2H), 7.60 (s, 1H), 7.20 (d, J=5.8 Hz, 2H), 6.89 (s, 1H), 5.86 (s br, 2H), 3.81 (m, 6H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) one signal obstructed, overlapping or not resolved δ 173.8, 153.3, 153.1 (br), 149.3 (2C), 146.6, 130.8 (br), 121.7 (2C), 107.4, 107.0, 98.6, 56.2, 55.7, 49.2; (+)-LRESIMS m/z (rel. int.) 329 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.16H.sub.17N.sub.4O.sub.2S [M+H].sup.+329.1067, found 329.1081; v.sub.max 1634, 1552, 1514, 1550, 1279, 1240, 998, 936, 847, 820, 763, 643 cm.sup.−1.
Synthesis Example 29—BT2046
(231) ##STR00067##
(232) Prepared according to General Procedure B, from reaction of 2-isothiocyanato-4,5-dimethoxybenzonitrile (50 mg, 0.23 mmol) with 5-methoxytryptamine (43 mg, 0.23 mmol) to afford BT2046 as a white powder (51 mg, 80%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.93 (s, 1H), 10.65 (s, 1H), 9.18 (s, 1H), 7.64 (s, 1H), 7.55 (s, 1H), 7.23 (d, J=8.7 Hz, 1H), 7.14 (s, 1H), 6.87 (s, 1H), 6.72 (dd, J=8.7, 1.9 Hz, 1H), 4.88-4.78 (m, 2H), 3.85 (s, 3H), 3.80 (s, 3H), 3.79 (s, 3H), 3.15-3.05 (m, 2H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) two signals obstructed, overlapping or not resolved δ 173.2, 153.2 (br), 152.9, 146.5, 131.4, 130.8 (br), 127.8, 123.4, 111.9, 111.2, 111.1, 107.3 (br), 101.1, 98.3 (br), 56.2, 55.7, 55.3, 47.4, 21.9; (+)-LRESIMS m/z (rel. int.) 411 (100) [M+H].sup.+; 433 (13) [M+Na].sup.+; (+)-HRESIMS calcd. for C.sub.21H.sub.23N.sub.4O.sub.3S [M+H].sup.+ 411.1485, found 411.1489; v.sub.max 3394, 3326, 1630, 1545, 1514, 1439, 1347, 1234, 1211, 1125, 1006, 792, 772 cm.sup.−1.
Synthesis Example 30—BT2049
(233) ##STR00068##
(234) Prepared according to General Procedure B, from reaction of 2-isothiocyanato-4,5-dimethoxybenzonitrile (50 mg, 0.23 mmol) with 2-morpholinoethan-1-amine (31 mg, 0.24 mmol) to afford a residue that was subjected to flash column chromatography [silica, 1:20 v/v MeOH/DCM elution] to give, after concentration of the appropriate fractions BT2049 as a white powder (38 mg, 48%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.90 (s br, 1H), 9.04 (s, 1H), 7.57 (s, 1H), 6.82 (s, 1H), 4.77-4.68 (m, 2H), 3.81 (s, 3H), 3.78 (s, 3H), 3.57 (app. t, J=4.6 Hz, 4H), 2.63 (t, J=7.4 Hz, 2H), 2.54-2.46 (m, 4H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) two signals obstructed, overlapping or not resolved δ 173.3 (br), 153.1 (br), 146.4, 130.6 (br), 107.3 (br), 98.1 (br), 66.2 (2C), 56.1, 55.7, 53.8 (br), 53.6 (2C), 43.9 (br); (+)-LRESIMS m/z (rel. int.) 351 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.16H.sub.23N.sub.4O.sub.3S [M+H].sup.+ 351.1485, found 351.1492; v.sub.max 3318, 1619, 1592, 1532, 1510, 1435, 1344, 1275, 1230, 1113, 1068, 1005, 868, 848, 776 cm.sup.−1.
Synthesis Example 31—BT2050
(235) ##STR00069##
(236) Prepared according to General Procedure B, from reaction of 2-isothiocyanato-4,5-dimethoxybenzonitrile (50 mg, 0.23 mmol) with methyl 4-(aminomethyl)benzoate hydrochloride (48 mg, 0.24 mmol) and triethylamine (33 μL, 0.24 mmol) to afford BT2050 as a white powder (52 mg, 59%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.11 (s br, 1H), 9.09 (s, 1H), 7.88 (d, J=8.3 Hz, 2H), 7.59 (s, 1H), 7.37 (d, J=8.3 Hz, 2H), 6.88 (s, 1H), 5.92 (s br, 2H), 3.83 (s, 3H), 3.81 (m, 6H); (+)-LRESIMS m/z (rel. int.) 386 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.19H.sub.20N.sub.3O.sub.4S [M+H].sup.+ 386.1169, found 386.1175; v.sub.max 3326, 1706, 1634, 1594, 1551, 1516, 1437, 1274, 1240, 1194, 1106, 1002, 937, 820, 757 cm.sup.−1.
Synthesis Example 32—BT2055
(237) ##STR00070##
(238) A magnetically stirred solution of tert-butyl (2-(5-hydroxy-1H-indol-3-yl)ethyl)carbamate (600 mg, 2.20 mmol) in DCM (12 mL) at −78° C. was treated sequentially with triethylamine (TEA) (370 μL, 2.64 mmol) and trifluoromethanesulfonic anhydride (425 μL, 2.53 mmol). The reaction was stirred for 6 h and then quenched with half saturated aqueous solution of sodium bicarbonate (10 mL). The mixture was extracted with DCM (3×30 mL) and the combined organic layers dried (Na.sub.2SO.sub.4) and concentrated in vacuo to afford the desired triflate as a thick colourless oil (550 mg, 61%) which was used directly in the next step without further purification. .sup.1H NMR (CDCl.sub.3, 400 MHz) δ 8.40 (s, 1H), 7.47 (d, J=2.4 Hz, 1H), 7.37 (d, J=8.8 Hz, 1H), 7.13 (s, 1H), 7.09 (dd, J=8.8, 2.4 Hz, 1H), 4.63 (s, 1H), 3.52-3.40 (m, 2H), 2.92 (t, J=6.9 Hz, 2H), 1.44 (s, 9H); .sup.13C NMR (CDCl.sub.3, 100 MHz) δ 156.0, 143.5, 135.1, 127.8, 124.5, 120.5, 118.86 (q, JC-F=320.8 Hz), 117.3, 115.3, 114.2, 112.2, 111.3, 79.4, 40.9, 28.4 (3C), 25.7, 21.1; (+)-LRESIMS m/z (rel. int.) 431 (100) [M+Na].sup.+; (+)-HRESIMS calcd. for C.sub.16H.sub.19N.sub.2O.sub.5F.sub.3NaS [M+Na].sup.+ 431.0859, found 431.0866; v.sub.max 3425, 3258, 1691, 1520, 1413, 1244, 1204, 1139, 931, 887, 799 cm.sup.−1. A portion of the above compound (75 mg, 0.18 mmol), was subjected to a palladium catalysed Suzuki-Miyaura reaction and reacted with 3-pyridineboronic acid pinacol ester (56 mg, 0.28 mmol) according to General Procedure D to afford a residue that was subjected to flash column chromatography [silica, 1:1 v/v EtOAc/Pet. spirit elution] to give, after concentration of the appropriate fractions tert-butyl (2-(5-(pyridin-3-yl)-1H-indol-3-yl)ethyl)carbamate (55 mg, 89%) as a beige powder. .sup.1H NMR (CDCl.sub.3, 400 MHz) δ 8.91 (d, J=2.0 Hz, 1H), 8.56 (dd, J=4.8, 1.6 Hz, 1H), 8.27 (s, 1H), 7.94 (dt, J=7.9, 2.0 Hz, 1H), 7.79 (s, 1H), 7.47 (d, J=8.4 Hz, 1H), 7.42 (dd, J=8.4, 1.7 Hz, 1H), 7.36 (ddd, J=7.9, 4.8, 0.9 Hz, 1H), 7.11 (d, J=1.4 Hz, 1H), 4.66 (s, 1H), 3.57-3.43 (m, 2H), 3.00 (t, J=6.7 Hz, 2H), 1.41 (s, 9H). A portion of the above material (48 mg, 0.14 mmol) was treated with a solution of hydrogen chloride (3 mL, 4M in dioxane) and magnetically stirred at 0° C. for 2 then 3 h at rt. Ether (10 mL) was added and the solvent decanted to afford a gum which was triturated with ether (5 mL). The residue was placed under high vacuum for 3 h to afford 2-(5-(pyridin-3-yl)-1H-indol-3-yl)ethan-1-amine dihydrochloride (37 mg, 84%) as a cream coloured gum and used directly without further purification. A mixture of triethylamine (133 μL, 0.81 mmol), amine-dihydrochloride (37 mg, 0.14 mmol, crude from above), DCM (2 mL) and ethanol (1 mL) was added to a magnetically stirred solution of 2-isothiocyanato-4,5-dimethoxybenzonitrile (30 mg, 0.14 mmol) in a mixture of DCM and pet. spirit (1 mL:1 mL). The mixture was stirred for 1 h and then pet. spirit (5 mL) was added. The solid was collected by filtration then dissolved in ethanol (2.5 mL) was added and the mixture heated at 70° C. for 2 h. The mixture was cooled to rt and the solid was collected by vacuum filtration, washed with ethanol (5 mL) and dried at the pump (1 mmHg) to afford BT2055 as a cream powder (22 mg, 36%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.06 (s br, 1H), 11.02 (s, 1H), 9.44 (s br, 1H), 8.90 (s, 1H), 8.51 (d, J=4.0 Hz, 1H), 8.22 (s, 1H), 8.03 (d, J=7.9 Hz, 1H), 7.70 (s, 1H), 7.50-7.40 (m, 2H), 7.30 (s, 1H), 6.87 (s, 1H), 4.98-4.86 (m, 2H), 3.85 (s, 3H), 3.82 (s, 3H), 3.20 (t, J=8.1 Hz, 2H); (+)-LRESIMS m/z (rel. int.) 458 (100) [M+H].sup.+; 480 (17) [M+Na].sup.+; (+)-HRESIMS calcd. for C.sub.25H.sub.24N.sub.5O.sub.2S [M+H].sup.+ 458.1645, found 458.1650. v.sub.max 1627, 1585, 1563, 1515, 1442, 1400, 1280, 1251, 1212, 1129, 1005, 788, 771, 634 cm.sup.−1.
Synthesis Example 33—BT2058
(239) ##STR00071##
(240) A magnetically stirred solution of 1H-indole-2-carbonyl chloride, prepared according to Kumarb et. al. (Eur. J. Chem. 2012, 3(2), 214), (350 mg, 1.95 mmol) in DCM (5 mL) at −10° C. was treated dropwise with a solution of pyridine (1 mL), DIPEA (272 μL, 1.95 mmol) and tert-butyl (2-aminoethyl)carbamate (312 mg, 1.95 mmol) in DCM (20 mL). The mixture was allowed to warm to rt over 18 h and then the mixture was diluted with DCM (30 mL) and washed with aqueous HCl (2×10 mL, 0.2M) followed by a half saturated aqueous solution of sodium bicarbonate (2×15 mL) then brine (10 mL). The organic layer was dried (Na.sub.2SO.sub.4) and concentrated in vacuo to afford a solid that was subjected to flash column chromatography [silica, 1:20 v/v MeOH/DCM elution] to give, after concentration of the appropriate fractions, tert-butyl (2-(1H-indole-2-carboxamido)ethyl)carbamate (296 mg, 50%) as a tan coloured solid. .sup.1H NMR (CDCl.sub.3, 400 MHz) one NH not observed δ 9.19 (s, 1H), 7.65 (d, J=8.0 Hz, 1H), 7.43 (d, J=8.2 Hz, 1H), 7.29 (d, J=7.2 Hz, 1H), 7.14 (t, J=7.5 Hz, 1H), 6.90 (s, 1H), 5.02-4.93 (m, 1H), 3.63-3.54 (m, 2H), 3.50-3.39 (m, 2H), 1.45 (s, 9H); (+) LRESIMS m/z (rel. int.) 326 (100) [M+Na].sup.+; (+)-HRESIMS calcd. for C.sub.16H.sub.21N.sub.3NaO.sub.3 [M+Na].sup.+ 326.1475, found 326.1481; v.sub.max 3308, 3226, 1694, 1609, 1541, 1422, 1280, 1254, 1152, 1018, 825, 745 cm.sup.−1. tert-Butyl (2-(1H-indole-2-carboxamido)ethyl)carbamate (100 mg, 0.33 mmol) was treated with a solution of hydrogen chloride (2 mL, 4M in dioxane) and magnetically stirred for 1 h at 4° C. then a further 0.5 h at rt. Ether (5 mL) was added to precipitate the product and the remaining gum was triturated with ether (3×5 mL) then placed under high vacuum for 1 h to afford N-(2-aminoethyl)-1H-indole-2-carboxamide hydrochloride as a cream coloured gum and used directly without further purification. A mixture of triethylamine (66 μL, 0.47 mmol), N-(2-aminoethyl)-1H-indole-2-carboxamide hydrochloride (57 mg, 0.24 mmol, crude from above) and DCM (2 mL) was added to a magnetically stirred solution of 2-isothiocyanato-4,5-dimethoxybenzonitrile (50 mg, 0.27 mmol) in a mixture of DCM and pet. spirit (1 mL:1 mL). The mixture was stirred for 1 h and then pet. spirit (5 mL) was added. The solid was collected by vacuum filtration then suspended in ethanol (5 mL) and the mixture heated at 70° C. for 1 h. The mixture was cooled to rt and the solid was collected by vacuum filtration, washed with ethanol (5 mL) and dried at the pump (1 mmHg) to afford BT2058 as a cream powder (32 mg, 33%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.93 (s br, 1H), 11.56 (s, 1H), 9.78 (s br, 1H), 8.80 (s, 1H), 7.73 (s, 1H), 7.60 (d, J=7.9 Hz, 1H), 7.42 (d, J=8.2 Hz, 1H), 7.17 (t, J=7.6 Hz, 1H), 7.07 (s, 1H), 7.03 (t, J=7.5 Hz, 1H), 6.89 (s, 1H), 5.01-4.73 (m, 2H), 3.84 (s, 6H), 3.76-3.65 (m, 2H); (+)-LRESIMS m/z (rel. int.) 424 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.21H.sub.22N.sub.5O.sub.3S [M+H].sup.+ 424.1438, found 424.1442. v.sub.max 3243, 1623, 1557, 1545, 1512, 1282, 1189, 1020, 766, 738 cm.sup.−1.
Synthesis Example 34—BT2061
(241) ##STR00072##
(242) tert-Butyl (2-(5-(benzo[c][1,2,5]oxadiazol-5-yl)-1H-indole-2-carboxamido)ethyl)carbamate was prepared and purified according to General Procedure D from reaction of benzo[c][1,2,5]oxadiazole-5-boronic acid pinacol ester (96 mg, 0.39 mmol) and tert-butyl (2-(5-bromo-1H-indole-2-carboxamido)ethyl)carbamate (100 mg, 0.26 mmol) to afford after flash chromatography, tert-butyl (2-(5-(benzo[c][1,2,5]oxadiazol-5-yl)-1H-indole-2-carboxamido)ethyl)carbamate (94 mg, 85%) as a white powder. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.80 (s, 1H), 8.60 (t, J=5.3 Hz, 1H), 8.25 (t, J=1.3 Hz, 1H), 8.20-8.15 (m, 1H), 8.12-8.11 (m, 2H), 7.71 (dd, J=8.6, 1.9 Hz, 1H), 7.56 (d, J=8.6 Hz, 1H), 7.22 (s, 1H), 6.94 (t, J=5.8 Hz, 1H), 3.37-3.39 (m, 2H), 3.18-3.10 (m, 2H), 1.07 (s, 9H); (+)-LRESIMS m/z (rel. int.) 444 (100) [M+Na].sup.+; (+)-HRESIMS calcd. for C.sub.22H.sub.23N.sub.5NaO.sub.4 [M+Na].sup.+444.1642, found 444.1642; v.sub.max 3360, 3295, 2979, 1683, 1615, 1535, 1519, 1251, 1166, 1152, 1004, 795, 773 cm.sup.−1. A portion of the above material (98 mg, 0.23 mmol) was treated with a solution of hydrogen chloride (3 mL, 4M in dioxane) and magnetically stirred at rt for 1 h. Ether (5 mL) was added and the solvent decanted to afford a gum which was triturated with ether (5 mL). The residue was placed under high vacuum for 3 h to afford the amine hydrochloride (75 mg, 91%) as a cream solid and used directly without further purification. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.89 (s, 1H), 8.94 (t, J=5.6 Hz, 1H), 8.28-8.23 (m, 1H), 8.20 (d, J=1.8 Hz, 1H), 8.14-8.08 (m, 2H), 7.73 (dd, J=8.7, 1.8 Hz, 1H), 7.57 (d, J=8.6 Hz, 1H), 7.35-7.33 (m, 1H), 4.76 (s br, 3H), 3.62-3.53 (m, 2H), 3.09-2.96 (m, 2H); (+)-LRESIMS m/z (rel. int.) 322 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.17H.sub.16N.sub.5O.sub.2 [M+H].sup.+322.1299, found 322.1304. A portion of the crude amine hydrochloride formed above (50 mg, 0.14 mmol) was then reacted with triethylamine (58 μL, 0.42 mmol) and 2-isothiocyanato-4,5-dimethoxybenzonitrile (31 mg, 0.14 mmol) according to General Procedure C to afford BT2061 as a white powder (35 mg, 47%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) 1H not observed or obscured δ 11.76 (s br, 2H), 9.14 (s, 1H), 8.70 (s, 1H), 8.24 (s, 1H), 8.17-8.08 (m, 2H), 7.71 (d, J=7.9 Hz, 1H), 7.65-7.52 (m, 2H), 7.19 (s, 1H), 6.85 (s, 1H), 4.95-4.85 (m, 2H), 3.82 (s, 3H), 3.79 (s, 3H), 3.77-3.70 (m, 2H); (+)-LRESIMS m/z (rel. int.) 542 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.27H.sub.24N.sub.7O.sub.4S [M+H]+ 542.1605, found 542.1611. v.sub.max 3206, 1622, 1542, 1513, 1439, 1344, 1278, 1234, 1020, 770 cm.sup.−1.
Synthesis Example 35—BT2063
(243) ##STR00073##
(244) tert-Butyl (2-(5-(4-((2-(dimethylamino)ethyl)carbamoyl)phenyl)-1H-indole-2-carboxamido)ethyl)carbamate was prepared according to General Procedure D from reaction of 4-(2-(dimethylamino)ethylcarbamoyl)phenylboronic acid (124 mg, 0.39 mmol) and tert-butyl (2-(5-bromo-1H-indole-2-carboxamido)ethyl)carbamate (100 mg, 0.26 mmol) to afford, after flash column chromatography [silica, 1:10-1:5 v/v ammoniacal MeOH/DCM elution] and concentration of the appropriate fractions, the desired product (90 mg, 70%) as a white powder. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.68 (s, 1H), 8.55 (t, J=5.2 Hz, 1H), 8.40 (t, J=5.6 Hz, 1H), 7.98 (s, 1H), 7.91 (d, J=8.4 Hz, 2H), 7.78 (d, J=8.4 Hz, 2H), 7.56 (dd, J=8.6, 1.6 Hz, 1H), 7.51 (d, J=8.6 Hz, 1H), 7.17 (s, 1H), 6.93 (t, J=5.4 Hz, 1H), 3.41-3.29 (m, 4H), 3.18-3.10 (m, 2H), 2.42 (t, J=6.9 Hz, 2H), 2.19 (s, 6H), 1.38 (s, 9H); (+)-LRESIMS m/z (rel. int.) 494 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.27H.sub.36N.sub.5O.sub.4 [M+H].sup.+ 494.2762, found 494.2766; v.sub.max 3381, 3229, 1693, 1635, 1607, 1552, 1272, 1166, 764 cm.sup.−1. The above material (90 mg, 0.18 mmol) was deprotected according to General Procedure E to afford N-(2-aminoethyl)-5-(4-((2-(dimethylamino)ethyl)carbamoyl)phenyl)-1H-indole-2-carboxamide dihydrochloride (76 mg, 91%) as a cream solid and used directly without further purification. A portion of the crude hydrochloride formed above (76 mg, 0.16 mmol) was then reacted with triethylamine (120 μL, 0.86 mmol) and 2-isothiocyanato-4,5-dimethoxybenzonitrile (36 mg, 0.16 mmol) according to General Procedure C to afford BT2063 as a cream powder (31 mg, 32%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.86 (s br, 1H), 11.68 (s, 1H), 9.33 (s br, 1H), 8.80 (s br, 1H), 8.74 (t, J=5.0 Hz, 1H), 7.98 (s, 1H), 7.97 (d, J=8.1 Hz, 2H), 7.81 (d, J=8.1 Hz, 2H), 7.66 (s, 1H), 7.58 (dd, J=8.6, 1.3 Hz, 1H), 7.52 (d, J=8.6 Hz, 1H), 7.16 (s, 1H), 6.86 (s, 1H), 4.98-4.81 (m, 2H), 3.83 (s, 3H), 3.81 (s, 3H), 3.73 (app. q, J=6.1 Hz, 2H), 3.61 (app. q, J=6.0 Hz, 2H), 3.18-3.08 (m, 2H), 2.73 (s, 6H); (+)-LRESIMS m/z (rel. int.) 614 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.32H.sub.36N.sub.7O.sub.4S [M+H].sup.+ 614.2544, found 614.2545; v.sub.max 1678, 1626, 1554, 1504, 1441, 1317, 1279, 1234, 1160, 1072, 813, 769 cm.sup.−1.
Synthesis Example 36—BT2066
(245) ##STR00074##
(246) tert-Butyl (2-(5-(6-methoxypyridin-3-yl)-1H-indole-2-carboxamido)ethyl)carbamate was prepared and purified according to General Procedure D from reaction of 2-methoxy-5-pyridine boronic acid (60 mg, 0.39 mmol) and tert-butyl (2-(5-bromo-1H-indole-2-carboxamido)ethyl)carbamate (100 mg, 0.26 mmol) to afford the desired compound (75 mg, 70%) as a white powder. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.64 (s, 1H), 8.53 (t, J=5.5 Hz, 1H), 8.47 (s, 1H), 8.06-7.98 (m, 1H), 7.87 (s, 1H), 7.50 (d, J=8.5 Hz, 1H), 7.46 (d, J=8.5 Hz, 1H), 7.15 (s, 1H), 6.97-6.89 (m, 1H), 6.89 (d, J=8.6 Hz, 1H), 3.89 (s, 3H), 3.38-3.27 (m, 2H), 3.13 (app. q, J=6.4 Hz, 2H), 1.38 (s, 9H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) 2 signals obscured by DMSO-d.sub.6 δ 162.4, 161.1, 155.7, 144.4, 137.6, 135.9, 132.5, 130.5, 128.9, 127.7, 122.4, 119.1, 112.9, 110.3, 102.8, 77.7, 53.1, 28.2 (3C); (+)-LRESIMS m/z (rel. int.) 411 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.22H.sub.27N.sub.4O.sub.4 [M+H].sup.+ 411.2027, found 411.2029; v.sub.max 3314, 3227, 1696, 1613, 1554, 1497, 1367, 1291, 1279, 1151, 1040, 836, 812, 625 cm.sup.−1. tert-Butyl (2-(5-(6-methoxypyridin-3-yl)-1H-indole-2-carboxamido)ethyl)carbamate (100 mg, 0.41 mmol) was deprotected according to General Procedure E to afford N-(2-aminoethyl)-5-(6-methoxypyridin-3-yl)-1H-indole-2-carboxamide dihydrochloride as a white powder, and used directly without further purification. The crude dihydrochloride formed above (87 mg, 0.27 mmol) in a solution of ethanol (6 mL) was treated with triethylamine (96 μL, 0.68 mmol) and then a solution of 2-isothiocyanato-4,5-dimethoxybenzonitrile (50 mg, 0.27 mmol) in DCM (3 mL) was added. The mixture was stirred at rt for 1 h then concentrated by a gentle stream of nitrogen and suspended in ethanol (5 mL) and refluxed for 4 h. The precipitate formed was collected and washed with ethanol (15 mL) then diethyl ether (10 mL) and dried at the pump to afford BT2066 (33 mg, 27%) as a cream coloured powder. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.94 (s, 1H), 11.61 (s, 1H), 9.14 (s, 1H), 8.65 (s, 1H), 8.50-8.44 (m, 1H), 8.03-7.99 (m, 1H), 7.85 (s, 1H), 7.62 (s, 1H), 7.49 (d, J=8.5 Hz, 1H), 7.46 (d, J=8.0 Hz, 1H), 7.11 (s, 1H), 6.89 (d, J=8.6 Hz, 1H), 6.84 (s, 1H), 4.94-4.84 (s, 2H), 3.89 (s, 3H), 3.82 (s, 3H), 3.79 (s, 3H), 3.79-3.66 (m, 2H); (+)-LRESIMS m/z (rel. int.) 531 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.27H.sub.27N.sub.6O.sub.4S [M+H]+ 531.1809, found 542.1816. v.sub.max 3209, 1625, 1551, 1513, 1438, 1280, 1238, 1020, 804, 765, 738 cm.sup.−1.
Synthesis Example 37—BT2068
(247) ##STR00075##
(248) tert-Butyl (2-(5-(4-(4-methylpiperazine-1-carbonyl)phenyl)-1H-indole-2-carboxamido)ethyl)carbamate was prepared and purified according to General Procedure D from reaction of [4-(4-methylpiperazine-1-carbonyl)phenyl]boronic acid pinacol ester (130 mg, 0.39 mmol) and tert-butyl (2-(5-bromo-1H-indole-2-carboxamido)ethyl)carbamate (100 mg, 0.26 mmol) to afford the desired compound (72 mg, 54%) as a white powder. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.68 (s, 1H), 8.55 (t, J=5.5 Hz, 1H), 7.94 (s, 1H), 8.05 (d, J=8.0 Hz, 2H), 7.55-7.50 (m, 2H), 7.45 (d, J=8.0 Hz, 2H), 7.17 (s, 1H), 6.93 (t, J=5.5 Hz, 1H), 3.65-3.33 (m, 6H), 3.14 (app. q, J=5.7 Hz, 2H), 2.38-2.28 (m, 4H), 2.20 (s, 3H), 1.38 (s, 9H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) 2 signals obscured by DMSO-d.sub.6 δ 168.9, 161.1, 155.7, 142.5, 136.2, 133.7, 132.6, 131.2, 127.7, 127.6 (2C), 127.6, 126.5 (2C), 122.7, 119.6, 112.8, 102.9, 77.7, 54.6 (br, 2C), 45.6, 28.2 (3C); (+)-LRESIMS m/z (rel. int.) 506 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.28H.sub.36N.sub.5O.sub.4 [M+H].sup.+ 506.2762, found 506.2759; v.sub.max 3257, 1691, 1625, 1605, 1557, 1522, 1423, 1293, 1246, 1170, 1128, 1004, 808, 764 cm.sup.−1. The Boc protected compound (100 mg, 0.20 mmol) was deprotected according to General Procedure E to afford the corresponding amine dihydrochloride (87 mg, 91%) as a white powder, and used directly without further purification. The crude dihydrochloride formed above (87 mg, 0.18 mmol) was reacted with triethylamine (76 μL, 0.55 mmol) and 2-isothiocyanato-4,5-dimethoxybenzonitrile (40 mg, 0.18 mmol) according to General Procedure C to afford BT2068 (25 mg, 22%) as a white powder. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) 1H obscured or overlapping δ 11.95 (s br, 1H), 11.66 (s br, 1H), 9.14 (s br, 1H), 8.68 (s br, 1H), 7.93 (s, 1H), 7.74 (d, J=7.6 Hz, 2H), 7.61 (s br, 1H), 7.56-7.49 (m, 2H), 7.45 (d, J=7.6 Hz, 2H), 7.15 (s br, 1H), 4.97-4.84 (m, 2H), 3.82 (s, 3H), 3.79 (s, 3H), 3.78-3.71 (m, 2H), 3.66-3.48 (m, 4H), 2.38-2.27 (s br, 4H), 2.21 (s, 3H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) complex spectrum due to slow rotation δ 173.8 (br), 168.9, 161.0 (br), 154.1 (br), 153.1 (br), 146.4, 142.5, 136.2, 133.7 (br), 132.9, 131.3, 130.7 (br), 127.7, 127.6 (br), 126.5 (br), 122.7 (br), 119.6, 112.9, 107.4 (br), 102.9 (br), 98.2 (br), 56.2, 55.7, 54.6 (br), 46.1 (br), 45.6, 36.6 (br). (+)-LRESIMS m/z (rel. int.) 626 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.33H.sub.36N.sub.7O.sub.4S [M+H].sup.+ 626.2544, found 626.2552; v.sub.max 3216, 1624, 1545, 1513, 1436, 1278, 1235, 1188, 1021, 851, 805, 762 cm.sup.−1.
Synthesis Example 38—BT2070
(249) ##STR00076##
(250) tert-Butyl (2-(5-(pyridin-3-yl)-1H-indole-2-carboxamido)ethyl)carbamate was prepared and purified according to General Procedure D from reaction of 3-pyridylboronic acid pinacol ester (80 mg, 0.39 mmol) and tert-butyl (2-(5-bromo-1H-indole-2-carboxamido)ethyl)carbamate (100 mg, 0.26 mmol) to afford after flash chromatography, the desired compound (62 mg, 62%) as a white powder. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.71 (s, 1H), 8.92 (d, J=2.2 Hz, 1H), 8.56 (t, J=5.5 Hz, 1H), 8.51 (dd, J=4.7, 1.4 Hz, 1H), 8.08 (dt, J=8.0, 2.2 Hz, 1H), 7.98 (s, 1H), 7.54 (s, 2H), 7.45 (dd, J=8.0, 4.7 Hz, 1H), 7.18 (d, J=1.4 Hz, 1H), 6.94 (t, J=5.5 Hz, 1H), 3.33 (app. q, J=6.7 Hz, 2H), 3.14 (app. q, J=6.7 Hz, 2H), 1.38 (s, 9H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) 2 signals obscured by DMSO-d.sub.6δ 161.1, 155.7, 147.7, 147.5, 136.7, 136.2, 133.9, 132.7, 128.9, 127.8, 123.7, 122.6, 119.9, 113.0, 102.9, 77.7, 28.2 (3C); (+)-LRESIMS m/z (rel. int.) 381 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.21H.sub.25N.sub.4O.sub.3 [M+H].sup.+ 381.1921, found 381.1926; v.sub.max 3371, 3222, 2974, 1686, 1634, 1564, 1528, 1330, 1267, 1168, 795 cm.sup.−1. The Boc protected compound (97 mg, 0.25 mmol) was deprotected according to General Procedure E to afford the corresponding amine dihydrochloride (75 mg, 85%) as a white powder, and used directly without further purification. The crude dihydrochloride formed above (75 mg, 0.21 mmol) was reacted with triethylamine (118 μL, 0.85 mmol) and 2-isothiocyanato-4,5-dimethoxybenzonitrile (47 mg, 0.21 mmol) according to General Procedure C to afford BT2070 (18 mg, 17%) as a white powder. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.96 (s br, 1H), 11.68 (s, 1H), 9.17 (s br, 1H), 8.91 (s, 1H), 8.72 (s br, 1H), 8.52 (d, J=4.0 Hz, 1H), 8.09 (dt, J=7.9, 4.0 Hz, 1H), 7.96 (s, 1H), 7.62 (s, 1H), 7.55-7.53 (m, 2H), 7.46 (dd, J=7.9, 4.7 Hz, 1H), 7.15 (s, 1H), 6.85 (s, 1H), 4.89 (s br, 2H), 3.82 (s, 3H), 3.80 (s, 3H), 3.76-3.69 (m, 2H); (+)-LRESIMS m/z (rel. int.) 501 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.26H.sub.25N.sub.6O.sub.3S [M+H].sup.+ 501.1703, found 501.1709. v.sub.max 3202, 1623, 1546, 1512, 1437, 1396, 1279, 1233, 1186, 1018, 796, 762 cm.sup.−1.
Synthesis Example 39—BT2071
(251) ##STR00077##
(252) Following a procedure analogous to that used in General Procedure H, 1H-indole-3-carboxylic acid (400 mg, 2.48 mmol) was converted to 1H-indole-3-carbonyl chloride and reacted with tert-butyl (2-aminoethyl)carbamate (397 mg, 2.48 mmol) to afford tert-butyl (2-(1H-indole-3-carboxamido)ethyl)carbamate (352 mg, 47%) as a white solid. .sup.1H NMR (CD.sub.3OD, 400 MHz) NH protons not observed δ 8.13-8.05 (m, 1H), 7.85 (s, 1H), 7.45-7.38 (m, 1H), 7.21-7.11 (m, 2H), 3.47 (app. t, J=6.1 Hz, 2H), 3.29 (app. t, J=6.1 Hz, 2H), 1.41 (s, 9H); .sup.13C NMR (CD.sub.3OD, 100 MHz) δ 168.8, 158.9, 138.1, 129.2, 126.9, 123.4, 121.9, 121.7, 112.8, 111.8, 80.2, 41.3, 40.8, 28.7; (+)-LRESIMS m/z (rel. int.) 326 (100) [M+Na].sup.+; (+)-HRESIMS calcd. for C.sub.16H.sub.21N.sub.3O.sub.3Na [M+Na].sup.+ 326.1475, found 326.1481; v.sub.max 3357, 3222, 1684, 1611, 1547, 1495, 1441, 1321, 1234, 1210, 1151, 1105, 777, 739 cm.sup.−1. A portion of the Boc protected compound (136 mg, 0.45 mmol) was deprotected according to General Procedure E to afford (100 mg, 93%) as a grey gum, and used directly without further purification. The crude hydrochloride formed above (65 mg, 0.27 mmol) was reacted with triethylamine (118 μL, 0.85 mmol) and 2-isothiocyanato-4,5-dimethoxybenzonitrile (47 mg, 0.21 mmol) according to General Procedure C to afford BT2071 (41 mg, 36%) as a pale yellow solid. 1H NMR (DMSO-d.sub.6, 400 MHz) 1H obscured or overlapping δ 11.97 (s br, 1H), 11.56 (s br, 1H), 9.23 (s br, 1H), 8.12 (d, J=7.7 Hz, 1H), 7.99 (s, 1H), 7.60 (s, 1H), 7.43 (d, J=7.7 Hz, 1H), 7.19-7.07 (m, 2H), 6.83 (s, 1H), 4.86 (s br, 2H), 3.83 (s, 3H), 3.81 (s, 3H), 3.70-3.62 (m, 2H); (+)-LRESIMS m/z (rel. int.) 424 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.21H.sub.22N.sub.5O.sub.3S [M+H].sup.+ 424.1438, found 424.1445; v.sub.max 1704, 1620, 1503, 1440, 1207, 1064, 743 cm.sup.−1.
Synthesis Example 40—BT2072
(253) ##STR00078##
(254) tert-Butyl (2-(6-(pyridin-3-yl)-1H-indole-2-carboxamido)ethyl)carbamate was prepared and purified according to General Procedure D from reaction of 3-pyridylboronic acid pinacol ester (80 mg, 0.39 mmol) and tert-butyl (2-(5-bromo-1H-indole-2-carboxamido)ethyl)carbamate (100 mg, 0.26 mmol) to afford after flash chromatography the desired compound (53 mg, 53%) as a white powder. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.73 (s, 1H), 8.87 (d, J=1.7 Hz, 1H), 8.62-8.49 (m, 2H), 8.09-8.00 (m, 1H), 7.74 (d, J=8.3 Hz, 1H), 7.68 (s, 1H), 7.48 (dd, J=8.0, 4.7 Hz, 1H), 7.39 (d, J=8.4 Hz, 1H), 7.14 (s, 1H), 6.94 (t, J=5.2 Hz, 1H), 3.36-3.29 (m, 2H), 3.13 (q, J=6.5 Hz, 2H), 1.38 (s, 9H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) two signals obscured by DMSO-d.sub.6 δ 161.0, 155.7, 147.9, 147.7, 136.9, 136.7, 134.1, 132.8, 132.3, 127.0, 123.8, 122.2, 119.2, 110.4, 102.4, 77.7, 28.2 (3C); (+)-LRESIMS m/z (rel. int.) 381 (100) [M+H].sup.+, 403 (80) [M+Na].sup.+; (+)-HRESIMS calcd. for C.sub.21H.sub.25N.sub.4O.sub.3 [M+H].sup.+ 381.1921, found 381.1927; v.sub.max 3355, 3310, 1684, 1640, 1533, 1270, 1239, 1165, 991, 791 cm.sup.−1. Boc-protected compound (96 mg, 0.25 mmol) was deprotected according to General Procedure E to afford (78 mg, 88%) as a white powder, and used directly without further purification. (+)-LRESIMS m/z (rel. int.) 281 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.16H.sub.17N.sub.4O [M+H].sup.+ 281.1397, found 281.1404. A suspension of the crude hydrochloride formed above (72 mg, 0.20 mmol) was suspended in ethanol (4 mL) and treated with triethylamine (113 μL, 0.82 mmol) and sonicated until the mixture became clear. 2-Isothiocyanato-4,5-dimethoxybenzonitrile (45 mg, 0.20 mmol) was added in portion and stirred for 0.5 h at rt and then heated at 70° C. for 4 h. The precipitate was collected by vacuum filtration and was subjected to flash column chromatography [silica, 1:10 v/v MeOH/DCM elution] to give, after concentration of the appropriate fractions BT2072 (15 mg, 15%) as a cream powder. H NMR (DMSO-de, 400 MHz) δ 11.96 (s br, 1H), 11.71 (s br, 1H), 9.15 (s, 1H), 8.87 (s br, 1H), 8.68 (s, 1H), 8.55 (dd, J=4.7, 1.2 Hz, 1H), 8.09-8.01 (m, 1H), 7.73 (d, J=8.3 Hz, 1H), 7.68 (s, 1H), 7.62 (s, 1H), 7.49 (dd, J=8.3, 4.7 Hz, 1H), 7.39 (d, J=8.5 Hz, 1H), 7.10 (s, 1H), 6.84 (s, 1H), 4.89 (s br, 2H), 3.82 (s, 3H), 3.79 (s, 3H), 3.78-3.67 (m, 2H); (+)-LRESIMS m/z (rel. int.) 501 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.26H.sub.25N.sub.6O.sub.3S [M+H].sup.+501.1703, found 501.1707; v.sub.max 3215, 1625, 1550, 1514, 1459, 1436, 1282, 1237, 1022, 809, 633 cm.sup.−1.
Synthesis Example 41—BT2073
(255) ##STR00079##
(256) tert-Butyl (2-(6-(4-(4-methylpiperazine-1-carbonyl)phenyl)-1H-indole-2-carboxamido)ethyl)carbamate was prepared and purified according to General Procedure D from reaction of [4-(4-Methylpiperazine-1-carbonyl)phenyl]boronic acid pinacol ester (130 mg, 0.39 mmol) and tert-butyl (2-(5-bromo-1H-indole-2-carboxamido)ethyl)carbamate (100 mg, 0.26 mmol) to afford the desired compound (88 mg, 66%) as a white powder. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.72 (s, 1H), 8.55 (t, J=5.9 Hz, 1H), 7.74-7.66 (m, 1H), 7.71 (d, J=8.0 Hz, 2H), 7.68 (s, 1H), 7.48 (d, J=8.1 Hz, 2H), 7.38 (d, J=8.0 Hz, 1H), 7.12 (s, 1H), 6.94 (t, J=5.5 Hz, 1H), 3.69-3.26 (m, 6H), 3.13 (app. q, J=6.3 Hz, 2H), 2.41-2.28 (m, 4H), 2.20 (s, 3H), 1.38 (s, 9H); (+)-LRESIMS m/z (rel. int.) 506 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.28H.sub.36N.sub.5O.sub.4 [M+H].sup.+ 506.2762, found 506.2769; v.sub.max 3229, 2788, 1716, 1618, 1493, 1434, 1291, 1266, 1166, 1000, 829, 779 cm.sup.−1. The Boc-protected compound (95 mg, 0.19 mmol) was deprotected according to General Procedure E to afford a white powder, and used directly without further purification. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.83 (s, 1H), 11.30 (s, 1H), 8.95 (t, J=5.6 Hz, 1H), 8.16 (s, 3H), 7.76 (d, J=8.0 Hz, 2H), 7.74 (d, J=8.2 Hz, 1H), 7.70 (s, 1H), 7.56 (d, J=8.0 Hz, 2H), 7.40 (dd, J=8.2, 1.5 Hz, 1H), 7.28 (d, J=1.5 Hz, 1H), 3.58 (app. q, J=5.9 Hz, 2H), 3.51-3.35 (m, 4H), 3.17-3.06 (m, 4H), 3.06-2.99 (m, 2H), 2.77 (s, 3H); (+)-LRESIMS m/z (rel. int.) 406 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.23H.sub.28N.sub.5O.sub.2 [M+H].sup.+ 406.2238, found 406.2244. The crude dihydrochloride formed above (78 mg, 0.16 mmol) was reacted with triethylamine (91 μL, 0.65 mmol) and 2-isothiocyanato-4,5-dimethoxybenzonitrile (36 mg, 0.16 mmol) according to General Procedure C to afford BT2073 (81 mg, 81%) as a pale yellow solid. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.90 (s br, 1H), 11.68 (s, 1H), 9.15 (s, 1H), 8.66 (s, 1H), 7.72 (d, J=8.0 Hz, 2H), 7.71-7.66 (m, 2H), 7.62 (s, 1H), 7.48 (d, J=8.0 Hz, 2H), 7.38 (d, J=8.3 Hz, 1H), 7.08 (s, 1H), 6.84 (s, 1H), 4.97-4.83 (m, 2H), 3.82 (s, 3H), 3.79 (s, 3H), 3.77-3.70 (m, 2H), 3.69-3.48 (m, 4H), 2.43-2.26 (m, 4H), 2.20 (s, 3H); (+)-LRESIMS m/z (rel. int.) 626 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.33H.sub.36N.sub.7O.sub.4S [M+H].sup.+ 626.2544, found 626.2547; v.sub.max 3195, 1622, 1562, 1542, 1515, 1442, 1365, 1235, 1192, 1018, 834 cm.sup.−1.
Synthesis Example 42—BT2075
(257) ##STR00080##
(258) tert-Butyl (2-(5-(6-(4-methylpiperazin-1-yl)pyridin-3-yl)-1H-indole-2-carboxamido)ethyl)carbamate was prepared according to General Procedure D from reaction of 2-(piperazin-1-yl)pyridine-5-boronic acid pinacol ester (119 mg, 0.39 mmol) and tert-butyl (2-(5-bromo-1H-indole-2-carboxamido)ethyl)carbamate (100 mg, 0.26 mmol) to afford after flash column chromatography [silica, 1:10 v/v ammoniacal MeOH/DCM elution] and concentration of the appropriate fractions, the desired compound (62 mg, 49%) as a white powder. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) b 11.58 (s, 1H), 8.50 (t, J=5.6 Hz, 1H), 8.44 (d, J=2.5 Hz, 1H), 7.85 (dd, J=8.8, 2.5 Hz, 1H), 7.81 (s, 1H), 7.47 (d, J=8.6 Hz, 1H), 7.43 (dd, J=8.6, 1.5 Hz, 1H), 7.12 (s, 1H), 6.94 (d, J=5.3 Hz, 1H), 6.90 (d, J=8.8 Hz, 1H), 3.51 (t, J=5.1 Hz, 4H), 3.38-3.27 (m, 2H), 3.12 (app. q, J=6.0 Hz, 2H), 2.45-2.37 (m, 4H), 2.22 (s, 3H), 1.38 (s, 9H); v.sub.max 3376, 3319, 2933, 1683, 1633, 1529, 1241, 1168, 810 cm.sup.−1. A magnetically stirred suspension of tert-butyl (2-(5-(6-(4-methylpiperazin-1-yl)pyridin-3-yl)-1H-indole-2-carboxamido)ethyl)carbamate (86 mg, 1.80 mmol) in DCM (4 mL) maintained at 0° C. was treated with trifluoroacetic acid (1 mL) and magnetically stirred for 2 h. The cold bath was removed and the mixture stirred at rt for 1 h and then the solvent was removed with a gentle stream of nitrogen and the remaining gum was triturated with ether (3×10 mL) then placed under high vacuum for 1 h to afford the deprotected TFA salt as a beige powder and used directly without further purification. (+)-LRESIMS m/z (rel. int.) 379 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.21H.sub.27N.sub.6O [M+H].sup.+ 379.2241, found 379.2245; v.sub.max 1673, 1647, 1548, 1422, 1177, 1131, 1025, 799 cm.sup.−1. The powder so formed was reacted with triethylamine (150 μL, 1.07 mmol) and 2-isothiocyanato-4,5-dimethoxybenzonitrile (39 mg, 0.18 mmol) according to General Procedure C to afford BT2075 (83 mg, 78%) as a beige powder. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.93 (s, 1H), 11.55 (s, 1H), 9.15 (s, 1H), 8.62 (s, 1H), 8.44 (s, 1H), 7.85 (d, J=8.7 Hz, 1H), 7.79 (s, 1H), 7.62 (s, 1H), 7.50-7.40 (m, 2H), 7.07 (s, 1H), 6.90 (d, J=8.7 Hz, 1H), 6.84 (s, 1H), 4.96-4.83 (m, 2H), 3.82 (s, 3H), 3.79 (s, 3H), 3.77-3.69 (m, 2H), 3.56-3.47 (m, 4H), 2.44-2.39 (m, 4H), 2.22 (s, 3H); (+)-LRESIMS m/z (rel. int.) 599 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.31H.sub.35N.sub.8O.sub.3S [M+H].sup.+ 599.2547, found 599.2553; v.sub.max 3205, 1610, 1582, 1548, 1514, 1397, 1186, 1019, 859, 805 cm.sup.−1.
Synthesis Example 43—BT2076
(259) ##STR00081##
(260) tert-Butyl (2-(6-(6-(4-methylpiperazin-1-yl)pyridin-3-yl)-1H-indole-2-carboxamido)ethyl)carbamate was prepared and purified according to General Procedure D from reaction of 2-(piperazin-1-yl)pyridine-5-boronic acid pinacol ester (81 mg, 0.27 mmol) and tert-butyl (2-(6-bromo-1H-indole-2-carboxamido)ethyl)carbamate (100 mg, 0.26 mmol) to afford the desired product (62 mg, 49%) as a cream coloured powder. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.63 (s, 1H), 8.52 (t, J=5.3 Hz, 1H), 8.42 (d, J=2.5 Hz, 1H), 7.82 (dd, J=9.0, 2.5 Hz, 1H), 7.65 (d, J=8.4 Hz, 1H), 7.56 (s, 1H), 7.29 (dd, J=8.4, 1.5 Hz, 1H), 7.09 (s, 1H), 6.95 (d, J=5.5 Hz, 1H), 6.92 (d, J=9.0 Hz, 1H), 3.55-3.49 (m, 4H), 3.35-3.29 (m, 2H), 3.12 (q, J=6.1 Hz, 2H), 2.43-2.39 (m, 4H), 2.22 (s, 3H), 1.38 (s, 9H); (+)-LRESIMS m/z (rel. int.) 479 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.26H.sub.35N.sub.6O.sub.3 [M+H].sup.+ 479.2765, found 479.2771; v.sub.max 3322, 1689, 1633, 1535, 1246, 1167, 1005, 812 cm.sup.−1. The Boc-protected compound formed directly above (80 mg, 0.17 mmol) was deprotected according to General Procedure F to afford a tan powder, and used directly without further purification. The powder so formed was reacted with triethylamine (232 μL, 1.65 mmol) and 2-isothiocyanato-4,5-dimethoxybenzonitrile (46 mg, 0.21 mmol) according to the procedure used for General Procedure C to afford BT2076 (67 mg, 65%) as a cream powder. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.94 (s br, 1H), 11.55 (s, 1H), 9.15 (s, 1H), 8.62 (s, 1H), 8.42 (s, 1H), 7.82 (d, J=8.7 Hz, 1H), 7.69-7.59 (m, 2H), 7.56 (s, 1H), 7.29 (d, J=8.2 Hz, 1H), 7.05 (s, 1H), 6.92 (d, J=8.7 Hz, 1H), 6.84 (s, 1H), 4.94-4.84 (m, 2H), 3.82 (s, 3H), 3.79 (s, 3H), 3.76-3.67 (m, 2H), 3.58-3.47 (m, 4H), 2.45-2.37 (m, 4H), 2.22 (s, 3H); (+)-LRESIMS m/z (rel. int.) 599 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.31H.sub.35N.sub.8O.sub.3S [M+H].sup.+ 599.2547, found 599.2551. A portion of the product (20 mg) was suspended in DCM (2 mL) and trifluoroacetic acid (100 μL) added. The mixture was concentrated under a gentle stream of nitrogen over 24 h to afford the trifluoroacetic acid salt of BT2076. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) TFA salt δ 13.77 (s, 1H), 11.69-11.60 (m, 1H), 10.57 (s, 1H), 10.31 (s, 1H), 10.05 (s, 1H), 9.01 (t, J=5.9 Hz, 1H), 8.51 (d, J=2.5 Hz, 1H), 8.01 (dd, J=9.0, 2.5 Hz, 1H), 7.88 (s, 1H), 7.85 (s, 1H), 7.52-7.47 (m, 2H), 7.12 (d, J=2.0 Hz, 2H), 7.10 (d, J=9.0 Hz, 1H), 7.01 (s, 1H), 5.60-4.56 (m br, 2H), 4.54-4.35 (m, 2H), 3.91 (s, 3H), 3.86 (s, 3H), 3.81-3.69 (m, 2H), 3.62-3.49 (m, 2H), 3.30-3.02 (m, 4H), 2.87 (s, 3H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) TFA salt δ 171.6, 162.4, 158.5 (q, JC-F=35.4 Hz), 157.3, 156.2, 155.3, 147.8, 144.3, 136.9, 136.0, 135.4, 131.6, 129.1, 127.7, 127.6, 122.5, 118.7, 116.0 (q, JC-F=292.8 Hz), 113.0, 108.3, 105.4, 103.6, 102.4, 97.8, 56.4 (2C), 52.0, 47.7, 42.4, 42.2, 35.0.
Synthesis Example 44—BT2077
(261) ##STR00082##
(262) tert-Butyl (3-(6-(pyridin-3-yl)-1H-indole-2-carboxamido)propyl)carbamate was prepared according to General Procedure D from reaction of 3-pyridylboronic acid pinacol ester (80 mg, 0.39 mmol) and tert-butyl (3-(6-bromo-1H-indole-2-carboxamido)propyl)carbamate (104 mg, 0.26 mmol) and purified as follows: After microwave irradiation the solvent was removed in vacuo and water (20 mL) was added. The precipitate which formed was collected and subjected to flash column chromatography [silica, 1:10 v/v MeOH/DCM elution] to give, after concentration of the appropriate fractions the desired compound (66 mg, 64%) as a white solid. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.73 (s, 1H), 8.88 (d, J=1.5 Hz, 1H), 8.55 (dd, J=4.6, 1.0 Hz, 1H), 8.51 (t, J=5.7 Hz, 1H), 8.05 (dt, J=7.8, 1.5 Hz, 1H), 7.74 (d, J=8.3 Hz, 1H), 7.69 (s, 1H), 7.48 (dd, J=7.8, 4.8 Hz, 1H), 7.39 (dd, J=8.3, 1.0 Hz, 1H), 7.14 (s, 1H), 6.85 (t, J=5.8 Hz, 1H), 3.29 (app. q, J=7.0 Hz, 2H), 3.00 (app. q, J=6.6 Hz, 2H), 1.70-1.62 (m, 2H), 1.38 (s, 9H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) δ160.9, 155.6, 147.9, 147.7, 136.9, 136.7, 134.1, 132.9, 132.3, 127.0, 123.9, 122.2, 119.2, 110.4, 102.2, 77.5, 37.7, 36.6, 29.7, 28.2 (3C); (+)-LRESIMS m/z (rel. int.) 395 (100) [M+H].sup.+, 417 (14) [M+Na].sup.+, 789 (15) [2M+H].sup.+; (+)-HRESIMS calcd. for C.sub.22H.sub.27N.sub.4O.sub.3 [M+H].sup.+ 395.2078, found 395.2084; v.sub.max 3355, 1684, 1640, 1534, 1367, 1270, 1239, 1165, 991, 791 cm.sup.−1. The Boc-protected amine (82 mg, 0.21 mmol) was deprotected following General Procedure F to afford a white powder, and used directly without further purification. The powder so formed was reacted with triethylamine (232 μL, 1.66 mmol) and 2-isothiocyanato-4,5-dimethoxybenzonitrile (46 mg, 0.21 mmol) according to General Procedure C to afford BT2077 (47 mg, 44%) as a white powder. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.89 (s, 1H), 11.74 (s, 1H), 9.11 (s, 1H), 8.88 (d, J=2.0 Hz, 1H), 8.68 (t, J=5.7 Hz, 1H), 8.54 (dd, J=4.7, 1.5 Hz, 1H), 8.05 (ddd, J=7.9, 2.0, 1.7 Hz, 1H), 7.75 (d, J=8.3 Hz, 1H), 7.70 (s, 1H), 7.61 (s, 1H), 7.48 (dd, J=7.9, 4.7 Hz, 1H), 7.39 (dd, J=8.3, 1.7 Hz, 1H), 7.15 (s, 1H), 6.84 (s, 1H), 4.70 (s, 2H), 3.83 (s, 3H), 3.79 (s, 3H), 3.44-3.35 (m, 2H), 2.09-1.98 (m, 2H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) δ 173.2, 160.7, 153.6 (br), 153.1 (br), 147.9, 147.7, 146.5, 136.9, 136.7, 134.1, 133.0, 132.3, 130.7, 127.1, 123.9, 122.2, 119.2, 110.5, 107.4 (br), 107.2 (br), 102.1, 98.2, 56.2, 55.7, 44.8 (br), 36.6, 26.6; (+)-LRESIMS m/z (rel. int.) 515 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.27H.sub.27N.sub.6O.sub.3S [M+H].sup.+ 515.1860, found 515.1866; v.sub.max 3144, 1626, 1555, 1517, 1442, 1404, 1284, 1260, 1242, 1107, 1024, 1016, 782, 710 cm−1; A portion of the product (20 mg) was suspended in DCM (2 mL) and trifluoroacetic acid (100 μL) added. The mixture was concentrated under a gentle stream of nitrogen over 24 h to afford the trifluoroacetic acid salt of BT2077. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) TFA salt δ 13.73 (s, 1H), 11.89 (s, 1H), 10.87 (s, 1H), 10.36 (s, 1H), 9.81 (s, 1H), 9.17 (s, 1H), 8.80 (d, J=5.1 Hz, 1H), 8.71 (t, J=5.7 Hz, 1H), 8.66 (d, J=8.2 Hz, 1H), 7.96 (dd, J=8.2, 5.1 Hz, 1H), 7.85-7.78 (m, 3H), 7.49 (dd, J=8.3, 1.6 Hz, 1H), 7.17 (d, J=1.6 Hz, 1H), 6.98 (s, 1H), 5.71-4.13 (m, 2H), 3.88 (s, 3H), 3.86 (s, 3H), 3.51 (q, J=6.5 Hz, 2H), 2.11-1.99 (m, 2H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) TFA salt δ 171.0, 160.9, 158.5 (q, JC-F=35.5 Hz), 157.2, 154.7, 147.7, 142.1, 140.8, 139.1, 136.8, 135.3, 133.7, 129.7, 127.9, 126.4, 122.6, 119.2, 116.0 (q, JC-F=292.2 Hz), 111.1, 105.5, 102.5, 102.2, 97.7, 56.4, 56.4, 47.1, 36.2, 25.3.
Synthesis Example 45—BT2078
(263) ##STR00083##
(264) tert-Butyl (2-(5-(1-methyl-1H-pyrazol-4-yl)-1H-indole-2-carboxamido)ethyl)carbamate was prepared and purified according to General Procedure D from reaction of 1-methylpyrazole-4-boronic acid pinacol ester (81 mg, 0.39 mmol) and tert-butyl (2-(5-bromo-1H-indole-2-carboxamido)ethyl)carbamate (100 mg, 0.26 mmol) to afford the desired product (95 mg, 95%) as a white powder. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.52 (s, 1H), 8.48 (t, J=5.5 Hz, 1H), 8.05 (s, 1H), 7.81 (s, 1H), 7.76 (s, 1H), 7.39 (s, 2H), 7.05 (s, 1H), 6.93 (t, J=5.4 Hz, 1H), 3.85 (s, 3H), 3.33-3.28 (m, 2H), 3.12 (q, J=6.1 Hz, 2H), 1.38 (s, 9H); (+)-LRESIMS m/z (rel. int.) 384 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.20H.sub.26N.sub.5O.sub.3 [M+H]+ 384.2030, found 384.2032; v.sub.max 1687, 1639, 1527, 1365, 1269, 1232, 1166, 998, 795 cm.sup.−1. The Boc-protected compound formed directly above (70 mg, 0.18 mmol) was deprotected according to General Procedure F to afford a white powder, and used directly without further purification. The powder so formed was reacted with triethylamine (202 μL, 1.46 mmol) and 2-isothiocyanato-4,5-dimethoxybenzonitrile (40 mg, 0.18 mmol) according to General Procedure C to afford BT2078 (74 mg, 80%) as a beige solid. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.91 (s br, 1H), 11.52 (s, 1H), 9.15 (s br, 1H), 8.68 (s br, 1H), 8.04 (s, 1H), 7.82 (s, 1H), 7.76 (s, 1H), 7.60 (s, 1H), 7.42-7.37 (m, 2H), 7.05 (s, 1H), 6.83 (s, 1H), 5.02-4.74 (m, 2H), 3.86 (s, 3H), 3.82 (s, 3H), 3.79 (s, 3H), 3.78-3.68 (m, 2H); (+)-LRESIMS m/z (rel. int.) 504 (100) [M+H].sup.+; v.sub.max 3224, 1623, 1545, 1510, 1437, 1395, 1280, 1233, 1191, 1020, 851, 797, 762 cm.sup.−1.
Synthesis Example 46—BT2079
(265) ##STR00084##
(266) A 10 mL snap-cap microwave vessel containing a magnetic stirring-bar was charged with a mixture of phenyl (2-cyano-4,5-dimethoxyphenyl)carbamate (100 mg, 0.34 mmol), tryptamine (54 mg, 0.34 mmol) and acetonitrile (3 mL). The sealed vessel was then subjected to microwave irradiation (100° C./1 h, ramp time 1 minute, maximum power 300 W with N.sub.2 cooling during reaction (Powermax)). The vessel was cooled in ice and the resulting crystals collected by vacuum filtration to afford 1-(2-(1H-indol-3-yl)ethyl)-3-(2-cyano-4,5-dimethoxyphenyl)urea as a white solid (92 mg, 76%) and transferred to a 10 mL snap-cap microwave vessel containing a magnetic sirring-bar, ethanol (4 mL) and triethylamine (200 μL). The sealed vessel was then subjected to microwave irradiation (100° C./0.2 h, ramp time 1 minute, maximum power 300 W with N.sub.2 cooling during reaction (Powermax)) and after this time TLC analysis revealed incomplete conversion. The mixture was re-subjected to the above microwave conditions for a further 0.17 h and the residue obtained after concentration in vacuo, was subjected to flash column chromatography [silica, 1:10 v/v MeOH/DCM elution] to give, after concentration of the appropriate fractions BT2079 (35 mg, 38%) as a white powder. H NMR (DMSO-d.sub.6, 400 MHz) δ 10.80 (s, 1H), 10.51 (s br, 1H), 8.79 (s br, 1H), 7.82 (d, J=8.1 Hz, 1H), 7.61 (s, 1H), 7.34 (d, J=8.1 Hz, 1H), 7.18 (d, J=1.8 Hz, 1H), 7.07 (t, J=7.3 Hz, 1H), 6.98 (t, J=7.3 Hz, 1H), 6.61 (s, 1H), 4.35-4.23 (m, 2H), 3.81 (s, 3H), 3.79 (s, 3H), 3.03-2.95 (m, 2H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) δ 155.1, 153.2, 150.0, 145.0, 136.3, 132.1, 127.5, 122.7, 121.0, 118.8, 118.2, 111.7, 111.4, 108.0, 105.0, 97.9, 56.2, 55.7, 41.4, 22.8; (+)-LRESIMS m/z (rel. int.) 365 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.20H.sub.21N.sub.4O.sub.3 [M+H].sup.+ 365.1608, found 365.1615; v.sub.max1683, 1619, 1512, 1466, 1440, 1237, 1112, 1013, 841, 785 cm.sup.−1.
Synthesis Example 47—BT2080
(267) ##STR00085##
(268) tert-Butyl (2-(5-(azetidine-1-carbonyl)-1H-indole-2-carboxamido)ethyl)carbamate was prepared according to General Procedure G from 2-((2-((tert-butoxycarbonyl)amino)ethyl)carbamoyl)-1H-indole-5-carboxylic acid (100 mg, 0.29 mmol) and azetidine hydrochloride (27 mg, 0.29 mmol). The residue obtained after concentration in vacuo, was subjected to flash column chromatography [silica, 1:10 v/v MeOH/DCM elution] to give, after concentration of the appropriate fractions the desired azetidine amide (35 mg, 31%) as a white powder. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.81 (s, 1H), 8.55 (t, J=5.7 Hz, 1H), 7.92 (s, 1H), 7.47 (dd, J=8.6, 1.4 Hz, 1H), 7.43 (d, J=8.6 Hz, 1H), 7.17 (s, 1H), 6.92 (t, J=5.7 Hz, 1H), 4.42-4.25 (m, 2H), 4.12-3.97 (m, 2H), 3.36-3.27 (m, 2H), 3.12 (app. q, J=6.3 Hz, 2H), 2.32-2.19 (m, 2H), 1.37 (s, 9H); (+)-LRESIMS m/z (rel. int.) 387 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.20H.sub.27N.sub.4O.sub.4 [M+H].sup.+ 387.2027, found 387.2029; v.sub.max 1673, 1643, 1594, 1560, 1528, 1422, 1287, 1175, 985, 813, 751 cm.sup.−1. The Boc-protected compound formed directly above (32 mg, 83 μmol) was deprotected following General Procedure F to afford a white powder, and used directly without further purification. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) TFA salt δ 11.87 (s, 1H), 8.72 (t, J=5.2 Hz, 1H), 7.95 (s, 1H), 7.88 (s, 3H), 7.48 (d, J=8.6 Hz, 1H), 7.45 (d, J=8.6 Hz, 1H), 7.20 (s, 1H), 4.41-4.27 (m, 2H), 4.14-3.97 (m, 2H), 3.62-3.47 (m, 2H), 3.07-2.95 (m, 2H), 2.32-2.20 (m, 2H). A 10 mL snap-cap microwave vessel containing a magnetic stirring-bar was charged with a mixture of phenyl (2-cyano-4,5-dimethoxyphenyl)carbamate (18.2 mg, 60 μmol), the crude trifluoroacetic acid salt, formed directly above (83 μmol), ethanol (1 mL) and triethylamine (150 μL) then subjected to microwave irradiation (100° C./1 h, ramp time 1 minute, maximum power 300 W with N.sub.2 cooling during reaction (Powermax)). The vessel was cooled in ice and the resulting crystals collected by vacuum filtration to afford BT2080 (12 mg, 41%) as a white solid. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.78 (s, 1H), 10.50 (s, 1H), 8.75-8.61 (m, 2H), 7.90 (s, 1H), 7.58 (s, 1H), 7.46 (d, J=8.7 Hz, 1H), 7.42 (d, J=8.7 Hz, 1H), 7.11 (s, 1H), 6.55 (s, 1H), 4.40-4.30 (m, 2H), 4.30-4.19 (m, 2H), 4.11-3.99 (m, 2H), 3.79 (s, 3H), 3.77 (s, 3H), 3.63-3.54 (m, 2H), 2.31-2.21 (m, 2H); (+)-LRESIMS m/z (rel. int.) 491 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.25H.sub.27N.sub.6O.sub.5 [M+H].sup.+ 491.2037, found 491.2039; v.sub.max 3612, 3179, 1690, 1614, 1561, 1513, 1443, 1276, 1213, 1046, 756 cm.sup.−1.
Synthesis Example 48—BT2081
(269) ##STR00086##
(270) tert-Butyl (2-(5-(morpholine-4-carbonyl)-1H-indole-2-carboxamido)ethyl)carbamate was prepared according to General Procedure G from 2-((2-((tert-butoxycarbonyl)amino)ethyl)carbamoyl)-1H-indole-5-carboxylic acid (100 mg, 0.29 mmol) and morpholine (25 mg, 0.29 mmol). The residue obtained after concentration in vacuo, was subjected to flash column chromatography [silica, 1:20 v/v ammoniacal MeOH/DCM elution] to give, after concentration of the appropriate fractions the desired morpholine amide (85 mg, 71%) as a white powder. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.78 (s, 1H), 8.55 (t, J=5.2 Hz, 1H), 7.71 (s, 1H), 7.46 (d, J=8.4 Hz, 1H), 7.23 (d, J=8.4 Hz, 1H), 7.16 (s, 1H), 6.92 (t, J=5.6 Hz, 1H), 3.66-3.57 (m, 4H), 3.56-3.48 (m, 4H), 3.37-3.29 (m, 2H), 3.18-3.09 (m, 2H), 1.37 (s, 9H); (+)-LRESIMS m/z (rel. int.) 417 (50) [M+H].sup.+, 439 (100) [M+Na].sup.+; (+)-HRESIMS calcd. for C.sub.21H.sub.29N.sub.4O.sub.5 [M+H].sup.+ 417.2132, found 417.2137; v.sub.max 1704, 1624, 1556, 1421, 1252, 1168, 1114 cm.sup.−1. The Boc-protected compound formed directly above (50 mg, 0.12 mmol) was deprotected following General Procedure F to afford a white powder, and used directly without further purification. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.84 (s, 1H), 8.71 (t, J=4.9 Hz, 1H), 7.86 (s, 3NH), 7.74 (s, 1H), 7.47 (d, J=8.4 Hz, 1H), 7.25 (d, J=8.4 Hz, 1H), 7.17 (s, 1H), 3.68-3.46 (m, 10H), 3.06-2.97 (m, 2H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) TFA salt, 1 signal obscured or overlapping δ 170.2, 161.5, 158.25 (q, J=35.8 Hz), 136.8, 132.5, 127.1, 126.3, 122.9, 121.2, 115.89 (q, J=293.2 Hz), 112.2, 103.5, 66.2 (2C), 38.7, 36.7. A 10 mL snap-cap microwave vessel containing a magnetic stirring-bar was charged with a mixture of phenyl (2-cyano-4,5-dimethoxyphenyl)carbamate (26.4 mg, 89 μmol), the crude TFA salt formed directly above (120 μmol), ethanol (2 mL) and triethylamine (167 μL) then subjected to microwave irradiation (100° C./1 h, ramp time 1 minute, maximum power 300 W with N.sub.2 cooling during reaction (Powermax)). The vessel was cooled in ice and the resulting crystals collected by vacuum filtration to BT2081 (22 mg, 35%) as a white solid. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.76 (s, 1H), 10.50 (s, 1H), 8.77-8.59 (m, 2H), 7.69 (s, 1H), 7.58 (s, 1H), 7.44 (d, J=8.4 Hz, 1H), 7.23 (d, J=8.4 Hz, 1H), 7.09 (s, 1H), 6.56 (s, 1H), 4.32-4.22 (m, 2H), 3.79 (s, 3H), 3.77 (s, 3H), 3.67-3.47 (m, 10H); (+)-LRESIMS m/z (rel. int.) 521 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.26H.sub.29N.sub.6O.sub.6 [M+H].sup.+ 521.2143, found 521.2150; v.sub.max 1691, 1623, 1558, 1512, 1462, 1419, 1288, 1212, 1114, 1041, 1019, 820, 758 cm.sup.−1.
Synthesis Example 49—BT2082
(271) ##STR00087##
(272) tert-Butyl (2-(5-(morpholine-4-carbonyl)-1H-indole-2-carboxamido)ethyl)carbamate (30 mg, 72 μmol) (prepared in Example 48) was deprotected following General Procedure F to afford a white powder, and used directly without further purification. The powder so formed was reacted with triethylamine (60 μL, 0.43 mmol) and 2-isothiocyanato-4,5-dimethoxybenzonitrile (16 mg, 72 μmol) according to General Procedure C to afford BT2082 (31 mg, 80%) as a white solid. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ11.94 (s, 1H), 11.76 (s, 1H), 9.13 (s, 1H), 8.68 (s, 1H), 7.70 (s, 1H), 7.60 (s, 1H), 7.45 (d, J=8.3 Hz, 1H), 7.23 (d, J=8.3 Hz, 1H), 7.12 (s, 1H), 6.83 (s, 1H), 4.96-4.82 (m, 2H), 3.82 (s, 3H), 3.79 (s, 3H), 3.76-3.70 (m, 2H), 3.66-3.48 (m, 8H); (+)-LRESIMS m/z (rel. int.) 537 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.26H.sub.29N.sub.6O.sub.5S [M+H].sup.+ 537.1915, found 537.1920; v.sub.max 3213, 1622, 1557, 1514, 1436, 1280, 1016, 756, 580 cm.sup.−1.
Synthesis Example 50—BT2083
(273) ##STR00088##
(274) tert-Butyl (2-(5-(4-ethylpiperazine-1-carbonyl)-1H-indole-2-carboxamido)ethyl)carbamate was prepared according to the General Procedure G from 2-((2-((tert-butoxycarbonyl)amino)ethyl)carbamoyl)-1H-indole-5-carboxylic acid (100 mg, 0.29 mmol) and 1-ethylpiperazine (36 μL, 0.29 mmol). The residue obtained after concentration in vacuo, was subjected to flash column chromatography [silica, 1:10 v/v ammoniacal MeOH/DCM elution] to give, after concentration of the appropriate fractions the desired amide (36 mg, 29%) as a white powder. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.77 (s, 1H), 8.54 (t, J=5.6 Hz, 1H), 7.67 (s, 1H), 7.45 (d, J=8.4 Hz, 1H), 7.20 (dd, J=8.4, 1.6 Hz, 1H), 7.15 (s, 1H), 6.92 (t, J=5.8 Hz, 1H), 3.61-3.43 (m, 4H), 3.35-3.28 (m, 2H), 3.12 (app. q, J=6.4 Hz, 2H), 2.43-2.30 (m, 4H), 2.36 (q, J=7.1 Hz, 2H) 1.38 (s, 9H), 1.00 (t, J=7.1 Hz, 3H); (+)-LRESIMS m/z (rel. int.) 444 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.23H.sub.34N.sub.5O.sub.4 [M+H].sup.+ 444.2605, found 444.2607; v.sub.max 1704, 1621, 1556, 1424, 1324, 1299, 1252, 1166 cm.sup.−1. The Boc-protected compound formed directly above (35 mg, 79 μmol) was deprotected following General Procedure F to afford a white powder, and used directly without further purification. The powder so formed was reacted with triethylamine (88 μL, 0.63 mmol) and 2-isothiocyanato-4,5-dimethoxybenzonitrile (17 mg, 79 μmol) according to General Procedure C to afford BT2083 (20 mg, 80%) as a beige solid. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.94 (s br, 1H), 11.75 (s, 1H), 9.13 (s br, 1H), 8.68 (s br, 1H), 7.66 (s, 1H), 7.60 (s, 1H), 7.44 (d, J=8.3 Hz, 1H), 7.20 (d, J=8.3 Hz, 1H), 7.11 (s, 1H), 6.83 (s, 1H), 4.95-4.79 (m, 2H), 3.81 (s, 3H), 3.79 (s, 3H), 3.74-3.69 (m, 2H), 3.55-3.46 (m, 4H), 2.40-2.34 (m, 4H), 2.35 (q, J=7.1 Hz, 2H), 1.00 (t, J=7.1 Hz, 3H); (+)-LRESIMS m/z (rel. int.) 564 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.28H.sub.33N.sub.7O.sub.4S [M+H].sup.+ 564.2387, found 564.2393; v.sub.max 3197, 1629, 1563, 1549, 1514, 1279, 1016, 754 cm.sup.−1.
Synthesis Example 51—BT2084
(275) ##STR00089##
(276) tert-Butyl (2-(5-(pyridin-4-yl)-1H-indole-2-carboxamido)ethyl)carbamate was prepared and purified according to General Procedure D from reaction of pyridine-4-boronic acid (48 mg, 0.39 mmol) and tert-butyl (2-(5-bromo-1H-indole-2-carboxamido)ethyl)carbamate (100 mg, 0.26 mmol) to afford the desired product (30 mg, 30%) as a white powder. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.77 (s, 1H), 8.63-8.55 (m, 3H), 8.11 (s, 1H), 7.73 (d, J=5.8 Hz, 2H), 7.63 (dd, J=8.6, 1.5 Hz, 1H), 7.55 (d, J=8.6 Hz, 1H), 7.21 (s, 1H), 6.94 (t, J=5.3 Hz, 1H), 3.37-3.30 (m, 2H), 3.18-3.10 (m, 2H), 1.38 (s, 9H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) 2C obscured by DMSO-d.sub.6 δ 161.0, 155.7, 150.0 (2C), 148.1, 136.9, 132.9, 128.8, 127.7, 122.2, 121.1 (2C), 120.1, 113.0, 103.1, 77.7, 28.2 (3C); (+)-LRESIMS m/z (rel. int.) 381 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.21H.sub.25N.sub.4O.sub.3 [M+H].sup.+ 381.1921, found 381.1925. The Boc-protected compound (50 mg, 0.13 mmol) was deprotected following General Procedure F to afford a white powder, and used directly without further purification. The powder so formed was reacted with triethylamine (109 μL, 0.79 mmol) and 2-isothiocyanato-4,5-dimethoxybenzonitrile (29 mg, 0.13 mmol) according to General Procedure C to afford BT2084 (52 mg, 79%) as a beige solid. 1H NMR (DMSO-d.sub.6, 400 MHz) 1H obscured or overlapping δ 11.75 (s, 1H), 9.09 (s, 1H), 8.76 (s, 1H), 8.59 (d, J=4.8 Hz, 2H), 8.09 (s, 1H), 7.74 (d, J=4.8 Hz, 2H), 7.63 (d, J=8.3 Hz, 1H), 7.60 (s, 1H), 7.53 (d, J=8.3 Hz, 1H), 7.17 (s, 1H), 6.83 (s, 1H), 4.97-4.83 (m, 2H), 3.82 (s, 3H), 3.79 (s, 3H), 3.76-3.68 (m, 2H); (+)-LRESIMS m/z (rel. int.) 501 (100) [M+H].sup.+; v.sub.max 3194, 1622, 1626, 1597, 1562, 1558, 1538 cm.sup.−1.
Synthesis Example 52—BT2085
(277) ##STR00090##
(278) tert-Butyl (1-(5-(pyridin-3-yl)-1H-indole-2-carbonyl)azetidin-3-yl)carbamate was prepared according to General Procedure D from reaction of 3-pyridylboronic acid pinacol ester (80 mg, 0.39 mmol) and tert-butyl (1-(5-bromo-1H-indole-2-carbonyl)azetidin-3-yl)carbamate (100 mg, 0.25 mmol) to afford after flash column chromatography [silica, 1:10 v/v ammoniacal MeOH/DCM elution] and concentration of the appropriate fractions the desired compound (35 mg, 30%) as a white powder. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.76 (s, 1H), 8.90 (d, J=2.4 Hz, 1H), 8.53 (dd, J=4.8, 1.7 Hz, 1H), 8.07 (dt, J=7.9, 1.7 Hz, 1H), 7.97 (s, 1H), 7.67 (d, J=7.4 Hz, 1H), 7.62-7.52 (m, 2H), 7.47 (dd, J=7.9, 4.8 Hz, 1H), 6.88 (s, 1H), 4.81-4.71 (m, 1H), 4.54-4.40 (m, 1H), 4.39-4.27 (m, 2H), 4.00-3.91 (m, 1H), 1.41 (s, 9H); (+)-LRESIMS m/z (rel. int.) 393 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.22H.sub.25N.sub.4O.sub.3 [M+H]+ 393.1921, found 393.1930; v.sub.max 3363, 3223, 1682, 1604, 1527, 1455, 1353, 1270, 1169, 877, 793, 767, 641 cm.sup.−1. The Boc-protected compound (152 mg, 0.39 mmol) was deprotected following General Procedure F to afford a hygroscopic cream coloured viscous paste, and used directly without further purification. The product so formed was reacted with triethylamine (431 μL, 3.09 mmol) and 2-isothiocyanato-4,5-dimethoxybenzonitrile (85 mg, 0.39 mmol) according to General Procedure C to afford BT2085 (138 mg, 70%) as a cream solid. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) 1H obscured or overlapping δ 12.08 (s, 1H), 11.75 (s, 1H), 9.34 (s, 1H), 8.90 (d, J=1.9 Hz, 1H), 8.51 (dd, J=4.6, 1.1 Hz, 1H) 8.06 (dt, J=8.1, 1.9 Hz, 1H), 7.99 (s, 1H), 7.62-7.53 (m, 3H), 7.46 (dd, J=8.0, 4.6 Hz, 1H), 6.87 (d, J=1.9 Hz, 1H), 6.81 (s, 1H), 5.98-5.90 (m, 1H), 5.02-4.98 (m, 1H), 4.89-4.81 (m, 1H), 4.67-4.59 (m, 1H), 4.44-4.34 (m, 1H), 3.80 (s, 3H), 3.78 (s, 3H); (+)-LRESIMS m/z (rel. int.) 513 (100) [M+H].sup.+; (+)-HRESIMS calcd. for C.sub.27H.sub.25N.sub.6O.sub.3S [M+H].sup.+ 513.1703, found 513.1702; v.sub.max 1623, 1596, 1537, 1514, 1447, 1433, 1389, 1283, 1253, 1235, 1206, 1132, 1011, 791, 730 626 cm.sup.−1.
Synthesis Example 53—BT2100
(279) ##STR00091##
(280) A magnetically stirred mixture of 1H-indazole-3-carboxylic acid (100 mg, 0.62 mmol) in DMF (5 mL) was treated sequentially with N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC) (142 mg, 0.74 mmol), 1-hydroxybenzotriazole hydrate (100 mg, 0.74 mmol) and triethylamine (103 μL, 0.74 mmol) and stirred at rt 0.5 h. tert-Butyl (2-aminoethyl)carbamate (118 mg, 0.74 mmol) was added and the mixture stirred for 60 h then concentrated in vacuo and purified by flash column chromatography [silica, 1:20 v/v MeOH/DCM elution] to give, after concentration of the appropriate fractions, tert-butyl (2-(1H-indazole-3-carboxamido)ethyl)carbamate (32 mg, 14%) as a white powder. .sup.1H NMR (400 MHz, Chloroform-d) δ 10.65 (s, 1H), 8.40 (dt, J=8.2, 1.1 Hz, 1H), 7.56 (br s, 1H), 7.53 (d, J=8.4 Hz, 1H), 7.44 (ddd, J=8.4, 6.9, 1.0 Hz, 1H), 7.30 (ddd, J=7.9, 6.9, 1.0 Hz, 1H), 5.06-4.94 (m, 1H), 3.64 (app. q, J=6.1 Hz, 2H), 3.44 (app. q, J=6.0 Hz, 2H), 1.42 (s, 9H); (+)-LRESIMS m/z (rel. int.) 305 (30) [M+H].sup.+, 327 (100) [M+Na].sup.+, 631 (40) [2M+Na].sup.+. The Boc-protected compound formed directly above (32 mg, 0.11 mmol) was deprotected following General Procedure F with TFA (1.0 mL) and DCM (4.0 mL) to afford after trituration with ether (10 mL) TFA salt of N-(2-aminoethyl)-1H-indazole-3-carboxamide as a gum that was used directly in the next step without further purification. The amine TFA-salt (0.11 mmol) was then suspended in ethanol (5 mL) and treated with triethylamine (175 μL, 1.26 mmol) and magnetically stirred for 5 min. The reaction mixture was then treated with a solution of 2-isothiocyanato-4,5-dimethoxybenzonitrile (23 mg, 0.11 mmol) in DCM (3 mL). The mixture was then stirred for 1 h followed by stirring at 75° C. for 1 h. The reaction mixture was cooled and then the precipitate was collected by vacuum filtration and washed with ether (5 mL) to afford BT2100 (24 mg, 51%) as a pale-yellow solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 13.48 (s, 1H), 11.95 (s, 1H), 9.12 (s, 1H), 8.51 (s, 1H), 8.13 (d, J=8.6 Hz, 1H), 7.67-7.54 (m, 2H), 7.45-7.33 (m, 1H), 7.28-7.18 (m, 1H), 6.83 (s, 1H), 4.98-4.84 (m, 2H), 3.81 (s, 3H), 3.78 (s, 3H), 3.73-3.78 (m, 2H); (+)-LRESIMS m/z (rel. int.) 425 (100) [M+H].sup.+, v.sub.max 2134, 1684, 1647, 1560, 1532, 1500, 1438, 1401, 1233, 1211, 1159, 1211, 1159, 1063 cm.sup.−1.
Synthesis Example 54—BT2103
(281) ##STR00092##
7-(benzyloxy)-1H-indole-3-carboxylic acid
(282) A magnetically stirred solution of 7-(benzyloxy)-1H-indole (2.0 g, 8.9 mmol) in anhydrous DMF (8 mL) maintained at 0° C. under an atmosphere of nitrogen was treated with trifluoroacetic acid anhydride (1.8 mL, 12.7 mmol) dropwise over 0.17 h. The mixture was then stirred for 2 h and poured into water (150 mL) and stirred vigorously for 0.17 h. The solid was collected by vacuum filtration and added to flask containing an aqueous solution of sodium hydroxide (50 mL, 4M) and refluxed for 2 h. The mixture was cooled over 18 h then diluted with water (150 mL) and extracted with ether (2×50 mL). The aqueous layer was then acidified to pH 0-1 with aqueous HCl (6M) and the precipitate collected by vacuum filtration and held under high vacuum for several hours to afford 7-(benzyloxy)-1H-indole-3-carboxylic acid as a brown powder (1.43 g, 60%) and used without further purification. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.00 (s, 1H), 11.92 (s, 1H), 7.85 (d, J=3.1 Hz, 1H), 7.61-7.55 (m, 3H), 7.44-7.38 (m, 2H), 7.36-7.31 (m, 1H), 7.05 (t, J=7.8 Hz, 1H), 6.84 (d, J=7.8 Hz, 1H), 5.28 (s, 2H); .sup.13C NMR (100 MHz, DMSO-d.sub.6) δ 165.9, 145.3, 137.1, 131.5, 128.4 (2C), 127.8, 127.7, 127.6 (2C), 126.7, 121.6, 113.4, 108.0, 104.1, 69.3; (+)-LRESIMS m/z (rel. int.) 290 (100) [M+Na].sup.+, 557 (90) [2M+Na].sup.+; v.sub.max 3427, 1652, 1629, 1529, 1506, 1276, 1251, 1194, 1090, 1004, 759 cm.sup.−1.
tert-butyl (2-(7-(benzyloxy)-1H-indole-3-carboxamido)ethyl)carbamate
(283) Following a procedure analogous to General Procedure H, 7-(benzyloxy)-1H-indole-3-carboxylic acid (300 mg, 1.12 mmol) was converted to 7-(benzyloxy)-1H-indole-3-carbonyl chloride with oxalyl chloride (118 μL, 1.40 mmol) and reacted with tert-butyl (2-aminoethyl)carbamate (180 mg, 1.12 mmol) to afford tert-butyl (2-(7-(benzyloxy)-1H-indole-3-carboxamido)ethyl)carbamate (252 mg, 55%) as a tan solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.65 (s, 1H), 7.91 (d, J=2.7 Hz, 1H), 7.87 (app. t, J=5.3 Hz, 1H), 7.71 (d, J=8.0 Hz, 1H), 7.57 (d, J=7.3 Hz, 2H), 7.41 (t, J=7.3 Hz, 2H), 7.37-7.32 (m, 1H), 6.99 (t, J=8.0 Hz, 1H), 6.89 (t, J=5.4 Hz, 1H), 6.80 (d, J=8.0 Hz, 1H), 5.26 (s, 2H), 3.27 (app. q, J=6.2 Hz, 2H), 3.09 (app. q, J=6.2 Hz, 2H), 1.38 (s, 9H); .sup.13C NMR (100 MHz, DMSO-d.sub.6) one CH.sub.2N obscured by DMSO-d.sub.6 δ 164.7, 155.7, 145.1, 137.2, 128.4 (2C), 127.8, 127.8, 127.6 (2C), 127.3, 126.4, 120.8, 113.9, 111.1, 103.6, 77.6, 69.2, 38.8, 28.2 (3C). (+)-LRESIMS m/z (rel. int.) 432 (100) [M+Na].sup.+; v.sub.max 3296, 1695, 1614, 1537, 1437, 1284, 1215, 1170, 785, 723, 689 cm.sup.−1.
tert-butyl (2-(7-hydroxy-1H-indole-3-carboxamido)ethyl)carbamate
(284) A mixture of tert-butyl (2-(7-(benzyloxy)-1H-indole-3-carboxamido)ethyl)carbamate (200 mg, 0.49 mmol) and 10% palladium on carbon (10 mg), in a mixture of methanol (15 mL) and ethyl acetate (5 mL) was stirred under a balloon of hydrogen for 2 h. The mixture was treated with celite (5 mL) and then the reaction mixture filtered and the celite washed with EtOAc (10 mL). The filtrate was concentrated in vacuo to afford a residue that was purified by flash column chromatography [silica, 1:10 v/v MeOH/DCM elution] to give, after concentration of the appropriate fractions the debenzylated product as a tan powder (140 mg, 90%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.41 (s, 1H), 9.70 (s, 1H), 7.89-7.82 (m, 1H), 7.80 (t, J=5.6 Hz, 1H), 7.55 (d, J=8.0 Hz, 1H), 6.91-6.87 (m, 1H), 6.86 (dd, J=9.3, 6.6 Hz, 1H), 6.54 (d, J=7.5 Hz, 1H), 3.26 (app. q, J=6.5 Hz, 2H), 3.09 (app. q, J=6.5 Hz, 2H), 1.38 (s, 9H); (+)-LRESIMS m/z (rel. int.) 342 (100) [M+Na].sup.+; v.sub.max 3345, 3270, 1688, 1601, 1601, 1550, 1524, 1449, 1242, 1157, 630 cm.sup.−1. The product formed directly above (140 mg, 0.44 mmol) was deprotected at 0° C. using a method analogous to General Procedure F to afford the deprotected, TFA-salt as a brown paste that was used without further purification. The TFA salt was then dissolved in ethanol (8 mL) and treated with triethylamine (244 μL, 1.76 mmol) and magnetically stirred for 5 min then treated with a solution of 2-isothiocyanato-4,5-dimethoxybenzonitrile (97 mg, 0.44 mmol) in DCM (3 mL). The mixture was then stirred for 1 h at rt and then heated to 72° C. and stirred for 1 h. The precipitate was collected by vacuum filtration which was washed with ether (5 mL) to afford BT2103 (102 mg, 53%) as a tan powder. (+)-LRESIMS m/z (rel. int.) 440 (100) [M+H].sup.+, 462 [M+Na].sup.+; v.sub.max 1614, 1583, 1505, 1440, 1293, 1233, 1175, 1073, 1046, 989, 736 cm.sup.−1. .sup.1H NMR analyses of BT2103 resulted in severe broadening. To overcome this, the trifluoroacetic acid salt of BT2103 was prepared. Accordingly, a portion of BT2103 (20 mg) suspended in CDCl.sub.3 (1 mL) was treated with TFA (100 μL) and then after 1 min the solution was concentrated under a gentle stream of N.sub.2 to furnish the TFA salt as a tan powder. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 13.75 (s, 1H), 11.65 (d, J=3.1 Hz, 1H), 10.90 (s, 1H), 10.59 (s, 1H), 10.34-9.06 (br s, 1H), 8.74 (t, J=5.6 Hz, 1H), 7.94 (d, J=3.1 Hz, 1H), 7.77 (s, 1H), 7.51 (d, J=8.0 Hz, 1H), 6.96 (s, 1H), 6.90 (t, J=8.0 Hz, 1H), 6.58 (d, J=7.5 Hz, 1H), 5.74-4.95 (br m, 2H), 3.91 (s, 3H), 3.86 (s, 3H), 3.64 (app. q, J=7.1 Hz, 2H); .sup.13C NMR (DMSO-d.sub.6, 100 MHz) δ 171.3, 167.6, 158.4 (q, .sup.2J.sub.C-F=35.6 Hz), 157.2, 155.4, 147.7, 143.9, 135.2, 128.3, 127.7, 126.3, 121.6, 116.0 (q, .sup.1J.sub.C-F=292.8 Hz), 111.5, 109.3, 106.5, 105.1, 102.4, 97.8, 56.4, 56.4, 47.8, 34.7.
Synthesis Example 55—BT2104
(285) ##STR00093##
N-(2-morpholinoethyl)-1H-indole-3-carboxamide
(286) Following General Procedure H, 1H-indole-3-carbonyl chloride (499 mg, 2.78 mmol) was reacted with 2-morpholinoethan-1-amine (361 mg, 2.78 mmol) to afford N-(2-morpholinoethyl)-1H-indole-3-carboxamide (442 mg, 58%) as a cream solid and used without further purification. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.51 (s, 1H), 8.14-8.09 (m, 1H), 7.97 (s, 1H), 7.78 (t, J=5.7 Hz, 1H), 7.43-7.39 (m, 1H), 7.16-7.06 (m, 2H), 3.60-3.56 (m, 4H), 3.38 (q, J=6.6 Hz, 2H), 2.49-2.40 (m, 6H); (+)-LRESIMS m/z (rel. int.) 274 (100) [M+H].sup.+, 296 (80) [M+Na].sup.+; v.sub.max 3379, 2818, 1635, 1546, 1525, 1212, 1114, 1005, 866, 800, 753 cm.sup.−1.
tert-butyl (2-(3-((2-morpholinoethyl)carbamoyl)-1H-indol-1-yl)ethyl)carbamate
(287) A magnetically stirred solution of N-(2-morpholinoethyl)-1H-indole-3-carboxamide (150 mg, 0.55 mmol) in DMF at 0° C. was treated in one portion with sodium hydride (26 mg, 0.66 mmol, 60% dispersion in mineral oil) and stirred for 1 h at rt. tert-Butyl (2-bromoethyl)carbamate (122 mg, 0.55 mmol) was added and the mixture stirred for 24 h at rt. The mixture was then quenched by the addition of methanol (1 mL) and concentrated in vacuo to afford a residue which was subjected to flash column chromatography [silica, 1:10 v/v MeOH/DCM elution] to give, after concentration of the appropriate fractions tert-butyl (2-(3-((2-morpholinoethyl)carbamoyl)-1H-indol-1-yl)ethyl)carbamate (65 mg, 28%) as a white foam. .sup.1H NMR (400 MHz, Chloroform-d) δ 8.01-7.93 (m, 1H), 7.72 (s, 1H), 7.47-7.39 (m, 1H), 7.33-7.22 (m, 2H), 6.77 (t, J=4.8 Hz, 1H), 4.90 (t, J=6.2 Hz, 1H), 4.29 (t, J=6.1 Hz, 2H), 3.75 (t, J=4.6 Hz, 4H), 3.57 (q, J=5.6 Hz, 2H), 3.49 (q, J=6.1 Hz, 2H), 2.62 (t, J=6.0 Hz, 2H), 2.53 (t, J=4.6 Hz, 4H), 1.43 (s, 9H); (+)-LRESIMS m/z (rel. int.) 417 (100) [M+H].sup.+, 439 (80) [M+Na].sup.+; v.sub.max 3371, 1680, 1611, 1539, 1517, 1392, 1286, 1115, 984, 859, 751 cm.sup.−1.
(288) BT2104
(289) tert-Butyl (2-(3-((2-morpholinoethyl)carbamoyl)-1H-indol-1-yl)ethyl)carbamate (60 mg, 0.14 mmol) was deprotected following General Procedure F to afford a pale-orange oil, and used directly without further purification. The powder so formed was reacted with triethylamine (174 μL, 1.73 mmol) and 2-isothiocyanato-4,5-dimethoxybenzonitrile (32 mg, 0.14 mmol) according to General Procedure C to afford BT2104 (32 mg, 43%) as a cream-coloured powder. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.09 (s, 1H), 9.25 (s, 1H), 8.16 (d, J=7.8 Hz, 1H), 8.03 (s, 1H), 7.91 (t, J=5.6 Hz, 1H), 7.87 (d, J=8.2 Hz, 1H), 7.66 (s, 1H), 7.24-7.18 (m, 1H), 7.17-7.12 (m, 1H), 6.86 (s, 1H), 5.03-4.92 (m, 2H), 4.58-4.49 (m, 2H), 3.84 (s, 3H), 3.80 (s, 3H), 3.58 (app. t, J=4.7 Hz, 4H), 3.37 (app. q, J=6.5 Hz, 2H), 2.48-2.40 (m, 6H); (+)-LRESIMS m/z (rel. int.) 537 (100) [M+H]+, 559 (50) [M+Na].sup.+; v.sub.max 1615, 1537, 1514, 1438, 1437, 1230, 1113, 1010, 863, 728 cm.sup.−1.
Synthesis Example 56—BT2105
(290) ##STR00094##
(291) Following a procedure analogous to General Procedure H, 1H-indole-2-carboxylic acid (500 mg, 3.10 mmol) was converted to 1H-indole-2-carbonyl chloride with oxalyl chloride (302 μL, 3.57 mmol) and reacted with tert-butyl azetidin-3-ylcarbamate hydrochloride (647 mg, 3.10 mmol) to afford tert-butyl (1-(1H-indole-2-carbonyl)azetidin-3-yl)carbamate (792 mg, 81%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.60 (s, 1H), 7.66 (d, J=7.3 Hz, 1H), 7.62 (d, J=8.0 Hz, 1H), 7.44 (d, J=8.2 Hz, 1H), 7.22-7.16 (m, 1H), 7.04 (t, J=7.3 Hz, 1H), 6.82-6.78 (m, 1H), 4.78-4.68 (m, 1H), 4.50-4.36 (m, 1H), 4.34-4.24 (m, 2H), 3.99-3.87 (m, 1H), 1.40 (s, 9H); .sup.13C NMR (101 MHz, DMSO-d.sub.6) δ 161.5, 154.8, 136.0, 129.2, 127.4, 123.6, 121.7, 119.8, 112.3, 104.4, 78.3, 59.6, 55.6, 40.7, 28.2 (3C). (+)-LRESIMS m/z (rel. int.) 338 (100) [M+Na].sup.+, 653 (80) [2M+Na].sup.+; v.sub.max 3229, 1686, 1607, 1530, 1458, 1415, 1345, 1172, 803, 741, 645 cm.sup.−1. A portion of the Boc-protected compound formed directly above (200 mg, 0.63 mmol) was deprotected following General Procedure F with TFA (1.5 mL) and DCM (4.0 mL) to afford after trituration with ether (10 mL) TFA salt of (3-aminoazetidin-1-yl)(1H-indol-2-yl)methanone as a gum that was used directly in the next step without further purification. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.68 (s, 1H), 8.61 (s, 3H), 7.64 (d, J=8.2 Hz, 1H), 7.45 (d, J=8.2 Hz, 1H), 7.21 (t, J=7.3 Hz, 1H), 7.06 (t, J=7.3 Hz, 1H), 6.84 (d, J=1.5 Hz, 1H), 4.87-4.70 (m, 1H), 4.54-4.41 (m, 1H), 4.39-4.25 (m, 1H), 4.22-4.04 (m 2H). A portion of the amine TFA-salt (150 mg, 0.46 mmol) was then suspended in ethanol (18 mL) and treated with triethylamine (380 μL, 2.73 mmol) and magnetically stirred for 5 min. The reaction mixture was then treated with a solution of 2-isothiocyanato-4,5-dimethoxybenzonitrile (100 mg, 0.46 mmol) in DCM (5 mL). The mixture was then stirred for 2 h followed by stirring at 70° C. for 1 h. The reaction mixture was cooled and then the precipitate was collected by vacuum filtration and washed with ether (5 mL) to afford BT2105 (168 mg, 85%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.85 (br s, 1H), 11.59 (s, 1H), 9.34 (s, 1H), 7.63 (d, J=8.0 Hz, 1H), 7.57 (s, 1H), 7.43 (d, J=8.3 Hz, 1H), 7.19 (t, J=7.6 Hz, 1H), 7.04 (t, J=7.6 Hz, 1H), 6.81 (s, 1H), 6.79 (d, J=1.6 Hz, 1H), 6.00-5.87 (m, 1H), 5.04-4.95 (m, 1H), 4.89-4.77 (m, 1H), 4.68-4.56 (m, 1H), 4.43-4.31 (m, 1H), 3.80 (m, 3H), 3.78 (s, 3H). (+)-LRESIMS m/z (rel. int.) 436 (100) [M+H].sup.+; v.sub.max 1625, 1596, 1537, 1510, 1434, 1229, 1006, 735 cm.sup.−1.
Synthesis Example 57—BT2113
(292) ##STR00095##
(293) Following a procedure analogous to that used by Trost et. al. (Chem. Eur. J. 2009, 15, 6910), magnetically stirred solution of methyl 1H-pyrrole-2-carboxylate (1.00 g, 7.99 mmol) in THF (80 mL) and methanol (40 mL) maintained at 0° C. was treated with N-bromosuccinimide (245 mg, 1.38 mmol) and followed by addition of portions of (315 mg, 1.77 mmol), (255 mg, 1.43 mmol), (635 mg, 3.57 mmoL) 0.5 h apart. The mixture was then and held at 2° C. without stirring for 18 h and then concentrated in vacuo and purified by gradient flash column chromatography [silica, 1:10.fwdarw.1:5 v/v EtOAc/pet spirit elution] to give, after concentration of the appropriate fractions, methyl 5-bromo-1H-pyrrole-2-carboxylate (562 mg, 34%) as a white powder. Spectral data were consistent with those reported by Trost et. al. .sup.1H NMR (400 MHz, Chloroform-d) δ 9.20 (s, 1H), 6.82 (dd, J=3.9, 2.7 Hz, 1H), 6.22 (dd, J=3.9, 2.7 Hz, 1H), 3.86 (s, 3H).
(294) Methyl 5-bromo-1H-pyrrole-2-carboxylate (147 mg, 0.72 mmol), was subjected to a palladium catalysed Suzuki-Miyaura reaction and reacted with phenylboronic acid (132 mg, 1.08 mmol) according to General Procedure D to afford a residue that was subjected to flash column chromatography [silica, 1:4 ether:pet. spirit elution] to give, after concentration of the appropriate fractions methyl 5-phenyl-1H-pyrrole-2-carboxylate (88 mg, 75%) as a white solid. Obtained spectral data were consistent with those reported by Laha et. al. (Chem. Commun. 2016, 52, 4329). .sup.1H NMR (400 MHz, Chloroform-d) δ 9.28 (s, 1H), 7.60-7.54 (m, 2H), 7.46-7.38 (m, 2H), 7.34-7.28 (m, 1H), 6.96 (dd, J=3.9, 2.4 Hz, 1H), 6.55 (dd, J=3.9, 2.7 Hz, 1H), 3.88 (s, 3H); (+)-LRESIMS m/z (rel. int.) 224 (100) [M+Na].sup.+; v.sub.max 3291, 1675, 1466, 1441, 1339, 1268, 1152, 1005, 799, 757 cm.sup.−1. A portion of the above compound, methyl 5-phenyl-1H-pyrrole-2-carboxylate (80 mg, 0.4 mmol) was dissolved in a mixture of methanol (8 mL) and aqueous sodium hydroxide (4 mL, 3M) and held at reflux for 4 h. The clear solution was then neutralised to pH 1 with aqueous HCl (1M) and the mixture extracted with EtOAc (2×15 mL) and DCM (2×15 mL). The EtOAc and DCM extracts were separately washed with brine (10 mL) then combined and dried (Na.sub.2SO.sub.4). Concentration in vacuo afforded a purple residue (68 mg, 91%) that was used without further purification. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.29 (s, 1H), 11.96 (s, 1H), 7.84 (d, J=7.3 Hz, 2H), 7.37 (t, J=7.7 Hz, 2H), 7.27-7.22 (m, 1H), 6.81 (dd, J=3.8, 2.3 Hz, 1H), 6.62 (dd, J=3.8, 2.5 Hz, 1H); (+)-LRESIMS m/z (rel. int.) 186 (100) [M−H].sup.−; v.sub.max 3420, 1657, 1561, 1514, 1469, 1435, 1333, 1268, 1040, 910, 765, 749, 691 cm.sup.−1. A portion of the acid, 5-phenyl-1H-pyrrole-2-carboxylic acid (60 mg, 0.18 mmol) was then converted to its acid chloride with oxalyl chloride (34 μL, 0.40 mmol) and reacted with tert-butyl(2-aminoethyl)carbamate (56 mg, 0.35 mmol according to General Procedure H. The residue obtained after workup, was subjected to flash column chromatography [silica, 1:5 ether:pet. spirit elution] to give, after concentration of the appropriate fractions tert-butyl (2-(5-phenyl-1H-pyrrole-2-carboxamido)ethyl)carbamate (70 mg, 61%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.69 (s, 1H), 8.13 (t, J=4.9 Hz, 1H), 7.79 (app. d, J=7.3 Hz, 2H), 7.41-7.30 (m, 2H), 7.26-7.16 (m, 1H), 6.96-6.88 (m, 1H), 6.80 (d, J=3.8 Hz, 1H), 6.56 (d, J=3.8 Hz, 1H), 3.27 (app. q, J=6.3 Hz, 2H), 3.09 (app. q, J=6.3 Hz, 2H), 1.38 (s, 9H); (+)-LRESIMS m/z (rel. int.) 352 (100) [M+Na].sup.+, 681 (10) [2M+Na].sup.+; v.sub.max 3302, 3224, 1695, 1606, 1562, 1537, 1269, 1155, 808, 773 cm.sup.−1.
(295) A portion of the Boc-protected compound formed directly above (60 mg, 0.18 mmol) was deprotected at rt following General Procedure F with TFA (1.5 mL) and DCM (4.0 mL) to afford after trituration with ether (3×5 mL) to afford the TFA salt of N-(2-aminoethyl)-5-phenyl-1H-pyrrole-2-carboxamide as a sticky gum that was used directly in the next step without further purification. The amine TFA-salt (0.18 mmol) was then suspended in ethanol (4 mL) and treated with triethylamine (304 μL, 2.22 mmol) and magnetically stirred for 5 min. The reaction mixture was then treated with a solution of 2-isothiocyanato-4,5-dimethoxybenzonitrile (40 mg, 0.18 mmol) in DCM (3 mL). The mixture was then maintained with stirring at 70-72° C. for 1 h, cooled to rt and then the precipitate was collected by vacuum filtration and washed with ether (5 mL) to afford BT2113 (62 mg, 77%) as a pale pink powder. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.77 (s, 1H), 11.62 (s, 1H), 9.15 (s, 1H), 8.20 (s, 1H), 7.78 (d, J=7.5 Hz, 2H), 7.62 (s, 1H), 7.36 (t, J=7.5 Hz, 2H), 7.21 (t, J=7.2 Hz, 1H), 6.91-6.75 (m, 2H), 6.56 (s, 1H), 4.91-4.76 (m, 2H), 3.82 (s, 3H), 3.79 (s, 3H), 3.71-3.62 (m, 2H); (+)-LRESIMS m/z (rel. int.) 450 (100) [M+Na].sup.+; v.sub.max 1611, 1538, 1500, 1436, 1230, 1015, 857, 751 cm.sup.−1.
Synthesis Example 58—BT2115
(296) ##STR00096##
(297) tert-Butyl (2-(6-bromopicolinamido)ethyl)carbamate (102 mg, 0.30 mmol), was subjected to a palladium catalysed Suzuki-Miyaura reaction and reacted with phenylboronic acid (55 mg, 0.45 mmol) according to General Procedure D to afford a residue that was subjected to flash column chromatography [silica, 1:10 MeOH:DCM] to give, after concentration of the appropriate fractions tert-butyl (2-(6-phenylpicolinamido)ethyl)carbamate (76 mg, 75%) as a white solid. .sup.1H NMR (400 MHz, Chloroform-d) δ 8.66-8.55 (m 1H), 8.14 (dd, J=7.6, 0.6 Hz, 1H), 8.11-8.04 (m, 2H), 7.92 (t, J=7.6 Hz, 1H), 7.87 (dd, J=7.9, 1.3 Hz, 1H), 7.56-7.41 (m, 3H), 5.04-4.95 (m, 1H), 3.64 (app. q, J=5.9 Hz, 2H), 3.43 (app. q, J=5.5 Hz, 2H), 1.41 (s, 9H); (+)-LRESIMS m/z (rel. int.) 364 (100) [M+Na].sup.+; v.sub.max 3384, 1676, 1513, 1249, 1162, 747 cm.sup.−1. A portion of the Boc-protected compound formed directly above (41 mg, 0.12 mmol) was deprotected following General Procedure F with TFA (1.5 mL) and DCM (4.0 mL) to afford a clear glass that on trituration with ether afforded a white gum that was used directly without further purification. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 9.08 (app. t, J=6.1 Hz, 1H), 8.34-8.29 (m, 2H), 8.21 (dd, J=7.8, 0.9 Hz, 1H), 8.09 (t, J=7.8 Hz, 1H), 8.01 (dd, J=7.8, 0.8 Hz, 1H), 7.84 (br s, 3H) 7.59-7.46 (m, 3H), 3.62 (app. q, J=6.2 Hz, 2H), 3.10-3.02 (m, 2H); LRESIMS m/z (rel. int.) 242 (100) [M+H]; The TFA-salt (0.12 mmol) was then suspended in ethanol (5 mL) and treated with triethylamine (244 μL, 1.20 mmol) and magnetically stirred for 5 min. The reaction was then treated with a solution of 2-isothiocyanato-4,5-dimethoxybenzonitrile (26 mg, 0.12 mmol) in DCM (2 mL). The mixture was then stirred for 1 h at rt then heated to 70° C. and stirred for 2 h. The precipitate was collected by vacuum filtration and then washed with ether (5 mL) to afford BT2115 (35 mg, 63%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.98 (br s, 1H), 9.10-9.02 (m, 2H), 8.34-8.22 (m, 2H), 8.16 (d, J=7.8 Hz, 1H), 8.04 (app. t, J=7.7 Hz, 1H), 7.94 (d, J=6.8 Hz, 1H), 7.58 (s, 1H), 7.56-7.46 (m, 3H), 6.82 (s, 1H), 5.05-4.93 (m, 2H), 3.86-3.80 (m, 2H), 3.80 (s, 3H), 3.78 (s, 3H). (+)-LRESIMS m/z (rel. int.) 462 (100) [M+H].sup.+; v.sub.max 1653, 1628, 1526, 1512, 1434, 1275, 1230 1014, 862, 756, 697 cm.sup.−1.
Synthesis Example 59—BT2116 and BT2130
(298) ##STR00097##
(299) tert-Butyl (2-(6-bromopicolinamido)ethyl)carbamate (103 mg, 0.30 mmol), was subjected to a palladium catalysed Suzuki-Miyaura reaction and reacted with [4-(4-Methylpiperazine-1-carbonyl)phenyl]boronic acid pinacol ester (148 mg, 0.45 mmol) according to General Procedure D to afford a residue that was subjected to flash column chromatography [silica, 1:10 ammoniacal MeOH:DCM] to give, after concentration of the appropriate fractions the desired product (113 mg, 54%) as an amber glass. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 9.01 (t, J=5.8 Hz, 1H), 8.40 (d, J=8.0 Hz, 2H), 8.22 (d, J=7.8 Hz, 1H), 8.09 (t, J=7.8 Hz, 1H), 8.01 (d, J=7.6 Hz, 1H), 7.51 (d, J=8.0 Hz, 2H), 7.02 (t, J=5.7 Hz, 1H), 3.70-3.57 (m, 2H), 3.40 (app. q, J=5.7 Hz, 2H), 3.38-3.33 (m, 2H), 3.18 (app. q, J=5.4 Hz, 2H), 2.45-2.23 (m, 4H), 2.20 (s, 3H), 1.35 (s, 9H). (+)-LRESIMS m/z (rel. int.) 468 (20) [M+H].sup.+, 490 (100) [M+Na].sup.+; v.sub.max 3373, 2806, 1707, 1682, 1627, 1517, 1448, 1422, 1268, 1169, 832 cm.sup.−1. A portion of the Boc-protected compound formed directly above (67 mg, 0.14 mmol) was deprotected following General Procedure F with TFA (1.5 mL) and DCM (4.0 mL) to afford a white paste after trituration with ether (10 mL) that was used directly without further purification. LRESIMS m/z (rel. int.) 368 (100) [M+H].sup.+. The TFA-salt formed directly above (0.14 mmol) was then suspended in ethanol (5 mL) and treated with triethylamine (156 μL, 1.12 mmol) and magnetically stirred for 5 min. The reaction was then treated with a solution of 2-isothiocyanato-4,5-dimethoxybenzonitrile (32 mg, 0.14 mmol) in DCM (2 mL). The mixture was then stirred for 1 h at rt then heated to 70° C. and stirred for 2 h. The reaction was cooled and the precipitate was collected by vacuum filtration and washed with ether (5 mL) to afford BT2116 (47 mg, 56%) as a yellow solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.99 (br s, 1H), 9.15 (m, 2H), 8.35 (d, J=7.9 Hz, 2H), 8.21 (d, J=7.8 Hz, 1H), 8.07 (t, J=7.8 Hz, 1H), 7.97 (d, J=7.6 Hz, 1H), 7.59 (s, 1H), 7.53 (d, J=8.3 Hz, 2H), 6.83 (s, 1H), 5.23-4.73 (m, 2H), 3.85-3.81 (m, 2H), 3.79 (s, 3H), 3.79 (s, 3H), 3.70-3.59 (m, 2H), 3.42-3.34 (m, 2H), 2.45-2.27 (m, 4H), 2.22 (s, 3H). (+)-LRESIMS m/z (rel. int.) 588 (100) [M+H].sup.+, 610 (50) [M+Na].sup.+; v.sub.max 1699, 1659, 1635, 1621, cm.sup.−1. A portion of BT2116 (14 mg) was treated with aqueous HCl (3.5 mL, 2M), sonicated at rt for 5 min then stirred for 10 min before removal of water with a gentle stream of nitrogen to afford BT2130, the hydrochloride salt of BT2116. .sup.1H NMR (400 MHz, DMSO-d.sub.6) very broad —NCH.sub.2— signals δ 13.75 (s, 1H), 11.16 (s, 1H), 10.82 (s, 1H), 10.32 (s, 1H), 9.42 (t, J=6.1 Hz, 1H), 8.44 (d, J=8.3 Hz, 2H), 8.28 (d, J=7.9 Hz, 1H), 8.11 (t, J=7.8 Hz, 1H), 8.05 (s, 1H), 7.99 (d, J=7.6 Hz, 1H), 7.58 (d, J=8.3 Hz, 2H), 7.03 (s, 1H), 5.45-4.30 (br m, 2H), 3.92 (s, 3H), 3.87 (s, 3H), 3.86-3.80 (m, 2H), 3.53-3.04 (m, 8H), 2.78 (s, 3H); (+)-LRESIMS m/z (rel. int.) 588 (100) [M+H].sup.+, 610 (10) [M+Na].sup.+; v.sub.max 2972, 1623, 1512, 1439, 1401, 1271, 1242, 1180, 1089, 1019, 975, 749 cm.sup.−1
Synthesis Example 60—BT2117
(300) ##STR00098##
(301) tert-Butyl (2-(6-bromopicolinamido)ethyl)carbamate (102 mg, 0.30 mmol), was subjected to a palladium catalysed Suzuki-Miyaura reaction and reacted with 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[c][1,2,5]oxadiazole (55 mg, 0.45 mmol) according to General Procedure D to afford a residue that was subjected to flash column chromatography [silica, 1:10 MeOH:DCM] to give, after concentration of the appropriate fractions tert-butyl (2-(6-(benzo[c][1,2,5]oxadiazol-5-yl)picolinamido)ethyl)carbamate (84 mg, 74%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 9.14 (app. t, J=5.8 Hz, 1H), 9.08 (app. t, J=1.1 Hz, 1H), 8.69 (dd, J=9.5, 0.9 Hz, 1H), 8.46 (dd, J=7.8, 0.9 Hz, 1H), 8.20 (dd, J=9.5, 0.5 Hz, 1H), 8.17 (t, J=7.8 Hz, 1H), 8.11 (dd, J=7.6, 0.5 Hz, 1H), 7.02 (t, J=5.7 Hz, 1H), 3.42 (app. q, J=6.3 Hz, 2H), 3.19 (app. q, J=5.7, 2H), 1.35 (s, 9H); (+)-LRESIMS m/z (rel. int.) 406 (100) [M+Na].sup.+; v.sub.max 1668, 1511, 1449, 1253, 1167, 880 754 cm.sup.−1. A portion of the Boc-protected compound formed directly above (43 mg, 0.11 mmol) was deprotected following General Procedure F with TFA (1.5 mL) and DCM (4.0 mL) to afford a clear glass that was used directly without further purification. LRESIMS m/z (rel. int.) 284 (90) [M+H].sup.+, 306 (100) [M+Na].sup.+, 589 (30) [2M+Na].sup.+; The TFA-salt formed directly above (0.11 mmol) was then suspended in ethanol (5 mL) and treated with triethylamine (156 μL, 1.12 mmol) and magnetically stirred for 5 min. The reaction was then treated with a solution of 2-isothiocyanato-4,5-dimethoxybenzonitrile (25 mg, 0.11 mmol) in DCM (2 mL). The mixture was then stirred for 1 h at rt then heated to 70° C. and stirred for 2 h. The reaction was cooled and the precipitate was collected by vacuum filtration and washed with ether (5 mL) to afford BT2117 (42 mg, 75%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.88 (br s, 1H), 9.19 (br s, 1H), 9.09-8.97 (m, 2H), 8.64 (d, J=8.1 Hz, 1H), 8.44 (d, J=7.3 Hz, 1H), 8.19 (dd, J=9.5, 0.5 Hz, 1H), 8.15 (d, J=7.8 Hz, 1H), 8.06 (d, J=7.8 Hz, 1H), 7.53 (s, 1H), 6.81 (s, 1H), 5.07-4.92 (m, 2H), 3.87-3.79 (m, 2H), 3.77 (s, 3H), 3.76 (s, 3H); LRESIMS m/z (rel. int.) 504 (100) [M+H].sup.+; v.sub.max 1652, 1626, 1588, 1528, 1514, 1434, 1231, 1176, 1022, 857, 761 cm.sup.−1.
Synthesis Example 61—BT2125
(302) ##STR00099##
(303) Following a procedure analogous to General Procedure H, 6-bromonicotinic acid (2.00 g, 9.90 mmol) was converted to 6-bromonicotinoyl chloride with oxalyl chloride (0.88 mL, 10.40 mmol) and reacted with tert-butyl(2-aminoethyl)carbamate (1.59 g, 9.90 mmol) to afford 1.03 g of tert-butyl (2-(6-chloropicolinamido)ethyl)carbamate (major) and tert-butyl (2-(6-bromopicolinamido)ethyl)carbamate (minor) (10:3.8 mixture) as a white powder that was used without further purification. (+)-LRESIMS m/z (rel. int.) 322 (100) C.sub.13H.sub.18.sup.35CIN.sub.3NaO.sub.3 [M+Na].sup.+, 324 (40) C.sub.13H.sub.18.sup.37CIN.sub.3NaO.sub.3 [M+Na].sup.+, 366 (30) C.sub.13H.sub.18.sup.79BrN.sub.3NaO.sub.3 [M+Na].sup.+, 368 (30) C.sub.13H.sub.18.sup.81BrN.sub.3NaO.sub.3 [M+Na].sup.+; v.sub.max 3359, 2978, 1704, 1640, 1543, 1517, 1448, 1340, 1272, 1160, 1103, 970, 853, 764, 606 cm.sup.−1.
tert-butyl (2-(6-chloropicolinamido)ethyl)carbamate
(304) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 8.81 (d, J=2.1 Hz, 1H), 8.72 (t, J=5.3 Hz, 1H), 8.21 (dd, J=8.3, 2.4 Hz, 1H), 7.64 (d, J=8.3 Hz, 1H), 6.92 (t, J=5.9 Hz, 1H), 3.28 (app. q, J=6.1 Hz, 2H), 3.11 (app. q, J=6.2 Hz, 2H), 1.36 (s, 9H).
tert-butyl (2-(6-bromopicolinamido)ethyl)carbamate
(305) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 8.78 (d, J=2.2 Hz, 1H), 8.72 (t, J=5.3 Hz, 1H), 8.10 (dd, J=8.3, 2.6 Hz, 1H), 7.78 (d, J=8.3 Hz, 1H), 6.92 (t, J=5.9 Hz, 1H), 3.28 (app. q, J=6.1 Hz, 2H), 3.11 (q, J=6.2 Hz, 2H), 1.36 (s, 9H).
(306) A portion of the mixture formed above (103 mg, 0.33 mmol), was subjected to a palladium catalysed Suzuki-Miyaura reaction with the addition of sodium bromide (61 mg, 0.60 mmol) and reacted with phenylboronic acid (73 mg, 0.60 mmol) according to General Procedure D, to afford tert-butyl (2-(6-phenylnicotinamido)ethyl)carbamate as a white solid (87 mg, 77%) that was used without further purification. 1H NMR (400 MHz, DMSO-d.sub.6) δ 9.07 (s, 1H), 8.68 (app. t, J=5.2 Hz, 1H), 8.26 (dd, J=8.5, 1.5 Hz, 1H), 8.17-8.13 (m, 2H), 8.08 (d, J=8.3 Hz, 1H), 7.58-7.43 (m, 3H), 6.94 (app. t, J=5.5 Hz, 1H), 3.37-3.28 (m, 2H), 3.13 (app. q, J=6.0 Hz, 2H), 1.37 (s, 9H). A portion of the Boc-protected compound formed directly above (80 mg, 0.23 mmol) was deprotected following General Procedure F with TFA (1.5 mL) and DCM (4.0 mL) to afford a gum after trituration with ether (10 mL) to afford the TFA salt of tert-butyl (2-(6-phenylnicotinamido)ethyl)carbamate that was used directly in the next step without further purification. The amine TFA-salt (0.23 mmol) was then suspended in ethanol (4 mL) and treated with triethylamine (156 μL, 1.12 mmol) and magnetically stirred for 5 min. The reaction mixture was then treated with a solution of 2-isothiocyanato-4,5-dimethoxybenzonitrile (52 mg, 0.23 mmol) in DCM (2 mL). The mixture was then stirred for 2 h at rt then the precipitate was collected by vacuum filtration and washed with ether (5 mL) to afford BT2125 (83 mg, 78%) as a yellow solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.93 (br s, 1H), 9.13 (br s, 1H), 9.02 (s, 1H), 8.82 (br s, 1H), 8.21 (d, J=7.6 Hz, 1H), 8.14 (d, J=7.1 Hz, 2H), 8.05 (d, J=8.1 Hz, 1H), 7.61 (s, 1H), 7.57-7.44 (m, 3H), 6.84 (s, 1H), 4.93-4.84 (m, 2H), 3.82 (s, 3H), 3.79 (s, 3H), 3.77-3.70 (m, 2H); (+)-LRESIMS m/z (rel. int.) 462 (100) [M+H].sup.+; v.sub.max 1666, 1625, 1561, 1501, 1438, 1327, 1290, 1233, 1157, 1061, 987, 856, 747, cm.sup.−1.
Synthesis Example 62—BT2132 and BT2144
(307) ##STR00100##
(308) A mixture of ethyl 5-phenyl-1,3,4-oxadiazole-2-carboxylate (500 mg, 2.29 mmol) prepared according to the procedure of Dost et. al. (J. Prakt. Chem. 1985, 327, 109) and tert-butyl (2-aminoethyl)carbamate (1.50 mL) in ethanol (15 mL) was stirred for 18 h The mixture was then concentrated in vacuo and purified by flash column chromatography [silica, 1:10 v/v ether/DCM elution] to give, after concentration of the appropriate fractions, tert-butyl (2-(5-phenyl-1,3,4-oxadiazole-2-carboxamido)ethyl)carbamate as a white powder (401 mg, 52%). .sup.1H NMR (400 MHz, Chloroform-d) δ 8.19-8.11 (m, 2H), 7.94 (s, 1H), 7.63-7.48 (m, 3H), 5.06 (s, 1H), 3.64 (app. q, J=5.7 Hz, 2H), 3.43 (app. q, J=5.8 Hz, 2H), 1.43 (s, 9H); 13C NMR (101 MHz, CDCl.sub.3) δ 166.4, 158.4, 156.6, 153.9, 132.6, 129.2 (2C), 127.5 (2C), 122.8, 80.0, 40.9, 40.0, 28.3 (3C). (+)-LRESIMS m/z (rel. int.) 355 (100) [M+Na].sup.+; v.sub.max 3350, 1683, 1535, 1450, 1276, 1169, 985, 849, 710 cm.sup.−1. A portion of the Boc-protected compound formed directly above (72 mg, 0.22 mmol) was deprotected at 0° C. following General Procedure F with TFA (1.5 mL) and DCM (5.0 mL) to afford after trituration with ether (3×5 mL) to afford the TFA salt of N-(2-aminoethyl)-5-phenyl-1,3,4-oxadiazole-2-carboxamide as a sticky gum that was used directly in the next step without further purification. The amine TFA-salt (0.22 mmol) was then suspended in ethanol (5 mL) and treated with triethylamine (181 μL, 1.30 mmol) and magnetically stirred for 5 min. The reaction mixture was then treated with a solution of 2-isothiocyanato-4,5-dimethoxybenzonitrile (48 mg, 0.22 mmol) in DCM (2 mL). The mixture was then maintained at rt with stirring for 2 h and further at 70° C. for 1 h. The mixture was cooled to rt and the precipitate collected by vacuum filtration and washed with ether (5 mL) to afford BT2132 (79 mg, 80%) as a white powder. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.95 (s, 1H), 9.49 (s, 1H), 9.12 (s, 1H), 8.10-8.03 (m, 2H), 7.73-7.59 (m, 4H), 6.84 (s, 1H), 4.95-4.87 (m, 2H), 3.80 (s, 3H), 3.79 (s, 3H), 3.78-3.70 (m, 2H). (+)-LRESIMS m/z (rel. int.) 453 (100) [M+H].sup.+; v.sub.max 3310, 1678, 1634, 1549, 1520, 1508, 1456, 1442, 1284, 1240, 1205, 1082, 1016, 867, 710 cm.sup.−1.
(309) BT2144
(310) A portion of BT2132 (15 mg) at rt was treated with aqueous HCl (1.5 mL, 1.5 M), sonicated for 5 min then stirred for 10 min before removal of water with a gentle stream of nitrogen to afford BT2144 (16 mg, 99%) as a white solid, the hydrochloride salt of BT2132. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 13.75 (s, 1H), 10.73 (s, 1H), 9.95 (s, 1H), 9.50 (t, J=6.2 Hz, 1H), 8.08 (dt, J=9.0, 1.6 Hz, 3H), 7.76-7.60 (m, 3H), 7.03 (s, 1H), 5.00 (s, 2H), 3.93 (s, 3H), 3.90 (s, 3H), 3.77 (q, J=6.3 Hz, 2H). (+)-LRESIMS m/z (rel. int.) 453 (100) [M+H]f; v.sub.max 1686, 1665, 1626, 1545, 1513, 1402, 1281, 1239, 1210, 1175, 1084, 847, 685 cm.sup.−1.
Synthesis Example 63—BT2133
(311) ##STR00101##
(312) A mixture of ethyl 5-phenylisoxazole-3-carboxylate (100 mg, 0.46 mmol) prepared according to the procedure of Watterson et. al. (J. Med. Chem. 2016, 59, 2820) and tert-butyl (2-aminoethyl)carbamate (100 mg, 0.62 mmol) in ethanol (2 mL) was stirred for 72 h at rt. The mixture was then concentrated in vacuo and purified by flash column chromatography [silica, 1:10 v/v ether/DCM elution] to give, after concentration of the appropriate fractions, tert-butyl (2-(5-phenylisoxazole-3-carboxamido)ethyl)carbamate as a white powder (32 mg, 21%). .sup.1H NMR (400 MHz, Chloroform-d) δ 7.84-7.76 (m, 2H), 7.54-7.44 (m, 3H), 7.33-7.25 (m, 1H), 6.96 (s, 1H), 4.89 (s, 1H), 3.59 (app. q, J=5.8 Hz, 2H), 3.39 (app. q, J=5.9 Hz, 2H), 1.44 (s, 9H). .sup.13C NMR (100 MHz, CDCl.sub.3) δ 171.6, 159.5, 159.0, 156.4, 130.7, 129.1, 126.8, 125.9, 99.0, 79.8, 40.2, 40.2, 28.3; (+)-LRESIMS m/z (rel. int.) 354 (100) [M+Na].sup.+; v.sub.max 3382, 3343, 1685, 1665, 1548, 1524, 1446, 1274, 1262, 1154, 1174, 969, 766 cm.sup.−1. The Boc-protected compound tert-butyl (2-(5-phenylisoxazole-3-carboxamido)ethyl)carbamate (62 mg, 0.19 mmol) was deprotected at 0° C. following General Procedure F with TFA (1.5 mL) and DCM (6.0 mL) to afford after trituration with ether (3×5 mL) to afford the TFA salt of N-(2-aminoethyl)-5-phenylisoxazole-3-carboxamide as a sticky gum that was used directly in the next step without further purification. The amine TFA-salt (0.19 mmol) was then suspended in ethanol (4 mL) and treated with triethylamine (156 μL, 1.12 mmol) and magnetically stirred for 5 min. The reaction mixture was then treated with a solution of 2-isothiocyanato-4,5-dimethoxybenzonitrile (41 mg, 0.19 mmol) in DCM (2 mL). The mixture was then maintained at rt with stirring for 2 h and further at 70° C. for 1 h. The mixture was cooled to rt and the precipitate collected by vacuum filtration and washed with ether (5 mL) to afford BT2133 (38 mg, 47%) as a white powder. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.93 (s, 1H), 9.13 (s, 1H), 8.99 (s, 1H), 7.96-7.88 (m, 2H), 7.63 (s, 1H), 7.60-7.50 (m, 3H), 7.32 (s, 1H), 6.83 (s, 1H), 4.96-4.82 (m, 2H), 3.82 (s, 3H), 3.79 (s, 3H), 3.75-3.68 (m, 2H); (+)-LRESIMS m/z (rel. int.) 452 (100) [M+H].sup.+; v.sub.max 1653, 1553, 1535, 1512, 1434, 1228, 1193, 1020, 859, 761, 592 cm.sup.−1.
Synthesis Example 64—BT2134
(313) ##STR00102##
(314) A mixture of BT2058 (50 mg, 0.12 mmol), potassium carbonate (32 mg, 0.23 mmol) in acetone at rt was treated with methyl iodide (29 μL, 0.47 mmol) in one portion. The mixture was stirred for 48 h and then water (4 mL) was added and the mixture stirred for 1 min. The mixture was filtered by vacuum filtration to afford a solid that was washed with acetone (3 mL) to afford a BT2134 as a white powder (23 mg, 45%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.56 (s, 1H), 8.81 (app. t, J=5.6 Hz, 1H), 8.46 (s, 1H), 7.64 (s, 1H), 7.60 (d, J=8.0 Hz, 1H), 7.41 (d, J=8.0 Hz, 1H), 7.17 (t, J=7.6 Hz, 1H), 7.04 (s, 1H), 7.01 (d, J=7.6 Hz, 1H), 6.83 (s, 1H), 4.39-4.32 (m, 2H), 3.85 (s, 3H), 3.84 (s, 3H), 3.72-3.64 (m, 2H), 2.48 (s, 3H). .sup.13C NMR (DMSO-d.sub.6, 100 MHz) δ 161.4, 155.2, 152.8, 147.5, 139.3, 136.4, 131.8, 127.1, 123.2, 121.4, 119.6, 112.3, 111.0 (br), 107.5, 106.1, 102.3, 56.1, 55.7, 44.7, 39.5, 36.6, 14.6. (+)-LRESIMS m/z (rel. int.) 438 (100) [M+H].sup.+; v.sub.max 3409, 3250, 1708, 1636, 1607, 1549, 1503, 1230, 1015, 744 cm.sup.−1.
Synthesis Example 65—BT2135
(315) ##STR00103##
(316) A mixture of BT2058 (50 mg, 0.12 mmol), potassium carbonate (32 mg, 0.23 mmol) in acetone at rt was treated with benzyl bromide (56 μL, 0.47 mmol) in one portion. The mixture was stirred for 18 h and then water (5 mL) was added and the mixture stirred for 1 min. The mixture was filtered by vacuum filtration to afford a solid that was washed with acetone (5 mL) to provide BT2135 as a white powder (36 mg, 59%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.57 (s, 1H), 8.78 (s, 1H), 8.46 (s, 1H), 7.65 (s, 1H), 7.61 (d, J=7.8 Hz, 1H), 7.43 (d, J=8.2 Hz, 1H), 7.27-7.00 (m, 8H), 6.88 (s, 1H), 4.32 (d, J=9.0 Hz, 4H), 3.86 (s, 3H), 3.85 (s, 3H), 3.72-3.62 (m, 2H); (+)-LRESIMS m/z (rel. int.) 514 (100) [M+H].sup.+; v.sub.max 3419, 3264, 1711, 1637, 1609, 1547, 1501, 1439, 1339, 1228, 1015, 771, 743 cm.sup.−1.
Synthesis Example 66—BT2136
(317) ##STR00104##
(318) BT2136 was prepared and purified using General Procedure J from BT2058 (50 mg, 0.12 mmol), potassium carbonate (32 mg, 0.23 mmol) and ethyl 2-bromoacetate (53 μL, 0.47 mmol) in acetone (2.0 mL) to afford BT2136 as a white powder (23 mg, 39%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.55 (s, 1H), 8.81 (t, J=5.4 Hz, 1H), 8.53 (s, 1H), 7.65 (s, 1H), 7.60 (d, J=8.0 Hz, 1H), 7.41 (d, J=8.0 Hz, 1H), 7.17 (t, J=7.5 Hz, 1H), 7.06 (d, J=1.2 Hz, 1H), 7.02 (t, J=7.5 Hz, 1H), 6.67 (s, 1H), 4.35 (app. t, J=6.0 Hz, 2H), 4.09 (q, J=7.1 Hz, 2H), 3.96 (s, 2H), 3.85 (s, 3H), 3.82 (s, 3H), 3.74-3.65 (app. q, J=6.0 Hz, 2H), 1.17 (t, J=7.1 Hz, 3H). (+)-LRESIMS m/z (rel. int.) 510 (100) [M+H].sup.+; v.sub.max 3295, 1739, 1631, 1600, 1543, 1501, 1232, 1152, 1017, 744 cm.sup.−1.
Synthesis Example 67—BT2137
(319) ##STR00105##
(320) Prepared and purified using a method analogous to General Procedure J from BT2072 (50 mg, 0.10 mmol), potassium carbonate (28 mg, 0.20 mmol) and 1-(bromomethyl)-3-(trifluoromethyl)benzene (48 mg, 0.20 mmol) in acetone (1.5 mL) to afford BT2137 as a white powder (13 mg, 20%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.74 (s, 1H), 8.89 (d, J=1.6 Hz, 1H), 8.83 (t, J=5.8 Hz, 1H), 8.55 (dd, J=4.6, 0.9 Hz, 1H), 8.49 (br s, 1H), 8.06 (dt, J=8.0, 2.0 Hz, 1H), 7.88 (s, 1H), 7.74 (d, J=8.3 Hz, 1H), 7.69 (s, 1H), 7.64 (s, 1H), 7.53-7.45 (m, 3H), 7.41 (dd, J=8.3, 1.6 Hz, 1H), 7.25 (t, J=7.8 Hz, 1H), 7.07 (d, J=0.8 Hz, 1H), 6.89 (s, 1H), 4.43 (s, 2H), 4.34-4.26 (m, 2H), 3.86 (s, 3H), 3.85 (s, 3H), 3.69-3.63 (m, 2H); (+)-LRESIMS m/z (rel. int.) 659 (100) [M+H].sup.+; v.sub.max 1603, 1547, 1501, 1439, 1329 1231, 1161, 1114, 1070, 1017, 803, 698 cm.sup.−1.
Synthesis Example 68—BT2138
(321) ##STR00106##
(322) Prepared and purified using a method analogous to General Procedure J from BT2072 (50 mg, 0.10 mmol), potassium carbonate (28 mg, 0.20 mmol) and 1-(bromomethyl)-4-(trifluoromethyl)benzene (48 mg, 0.20 mmol) in acetone (1.5 mL) to afford, after flash column chromatography [silica, 1:10 v/v ammoniacal MeOH/DCM elution] BT2138 as a white powder (11 mg, 17%). .sup.1H NMR (DMSO-de, 400 MHz) δ 11.75 (s, 1H), 8.88 (d, J=1.5 Hz, 1H), 8.83 (app. t, J=5.8 Hz, 1H), 8.55 (d, J=4.7 Hz, 1H), 8.48 (br s, 1H), 8.05 (dt, J=8.0, 1.5 Hz, 1H), 7.75 (d, J=8.4 Hz, 1H), 7.69 (s, 1H), 7.65 (s, 1H), 7.50 (dd, J=7.8, 4.8 Hz, 1H), 7.42 (dd, J=8.4, 1.5 Hz, 1H), 7.38 (d, J=7.8 Hz, 2H), 7.28 (d, J=7.9 Hz, 2H), 7.10 (br s, 1H), 6.86 (s, 1H), 4.42 (s, 2H), 4.37-4.25 (m, 2H), 3.86 (s, 3H), 3.85 (s, 3H), 3.74-3.63 (m, 2H); v.sub.max 1644, 1601, 1545, 1500, 1438, 1325, 1229, 1156, 1110, 1066, 1017, 810 cm.sup.−1.
Synthesis Example 69—BT2139 and BT2140
(323) ##STR00107##
(324) Prepared and purified using a method analogous to General Procedure J from BT2154 (40 mg, 78 μmol), potassium carbonate (22 mg, 0.16 mmol) and 1-(bromomethyl)-4-(trifluoromethyl)benzene (19 mg, 86 μmol) in acetone (1.5 mL) to afford BT2139 as a white powder (38 mg, 73%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.79 (s, 1H), 8.88 (d, J=2.0 Hz, 1H), 8.79 (s, 1H), 8.55 (dd, J=4.7, 1.5 Hz, 2H), 8.05 (dt, J=8.0, 1.8 Hz, 2H), 7.78-7.73 (m, 2H), 7.72-7.69 (m, 2H), 7.58 (s, 1H), 7.49 (dd, J=7.7, 4.7 Hz, 1H), 7.40 (dd, J=8.4, 1.7 Hz, 1H), 6.92 (s, 1H), 6.87-6.80 (d, J=1.4 Hz, 1H), 5.41-5.32 (m, 1H), 5.32-5.24 (m, 1H), 4.95-4.85 (m, 1H), 4.84-4.75 (m, 1H), 4.59 (s, 2H), 4.39-4.28 (m, 1H), 3.87 (s, 3H), 3.81 (s, 3H). (+)-LRESIMS m/z (rel. int.) 336 (100) [M+2H].sup.2+; 671 (40) [M+H].sup.+; v.sub.max 3239, 1604, 1553, 1507, 1454, 1322, 1220, 1114, 1065, 1011, 796 cm.sup.−1.
(325) BT2140
(326) A portion of BT2139 (7.3 mg) in methanol (300 μL) at rt was treated with aqueous HCl (1.5 mL, 1.5 M), sonicated for 5 min then stirred for 10 min before removal of water with a gentle stream of nitrogen to afford BT2140 (7.5 mg), the hydrochloride salt of BT2139. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.05 (s, 1H), 10.69 (br s, 1H), 9.64 (br s, 1H), 9.15 (s, 1H), 8.79 (d, J=5.2 Hz, 1H), 8.65 (d, J=8.2 Hz, 1H), 8.14 (s, 1H), 7.96 (dd, J=7.9, 5.5 Hz, 1H), 7.89 (d, J=8.4 Hz, 1H), 7.82 (d, J=8.2 Hz, 2H), 7.80 (s, 1H), 7.69 (d, J=8.2 Hz, 2H), 7.54 (dd, J=8.4, 1.5 Hz, 1H), 7.29 (s, 1H), 6.89-6.87 (m, 1H), 5.49-5.37 (m, 1H), 5.27-5.15 (m, 1H), 4.87-4.77 (m, 2H), 4.76 (s, 2H), 4.49-4.38 (m, 1H), 4.01 (s, 3H), 3.94 (s, 3H); (+)-LRESIMS m/z (rel. int.) 671 (100) [M+H].sup.+; v.sub.max 1601, 1503, 1440, 1392, 1326, 1299, 1169, 1100, 1066, cm.sup.−1.
Synthesis Example 70—BT2141 and BT2143
(327) ##STR00108##
(328) Prepared and purified using a method analogous to General Procedure J from BT2154 (80 mg, 0.16 mmol), potassium carbonate (43 mg, 0.32 mmol) and methyl iodide (11 μL, 17 mmol) in acetone (2.0 mL) to afford BT2141 as a white powder (20 mg, 24%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.80 (s, 1H), 8.88 (d, J=2.0 Hz, 1H), 8.74 (s, 1H), 8.55 (dd, J=4.7, 1.3 Hz, 1H), 8.06 (ddd, J=7.9, 1.8 Hz, 1H), 7.76 (d, J=8.4 Hz, 1H), 7.71 (s, 1H), 7.57 (s, 1H), 7.49 (dd, J=7.8, 4.8 Hz, 1H), 7.41 (dd, J=8.3, 1.8 Hz, 1H), 6.86 (d, J=1.3 Hz, 1H), 6.84 (s, 1H), 5.48-5.36 (m, 1H), 5.38-5.27 (m, 1H), 4.98-4.86 (m, 1H), 4.88-4.78 (m, 1H), 4.37 (t, J=8.9 Hz, 1H), 3.84 (s, 3H), 3.81 (s, 3H), 2.58 (s, 3H); (+)-LRESIMS m/z (rel. int.) 527 (100) [M+H].sup.+; v.sub.max 3266, 1597, 1506, 1439, 1374, 1231, 1014, 798, 622 cm.sup.−1.
(329) BT2143
(330) A portion of BT2141 (5.0 mg) at rt was treated with aqueous HCl (1.5 mL, 1.5 M), sonicated for 5 min then stirred for 10 min before removal of water with a gentle stream of nitrogen to afford BT2143 (4 mg, 70%), the hydrochloride salt of BT2141. H NMR (400 MHz, DMSO-d.sub.6) δ 12.03 (s, 1H), 10.55 (br s, 1H), 9.54 (br s, 1H), 9.11 (d, J=1.7 Hz, 1H), 8.75 (d, J=4.7 Hz, 1H), 8.57 (d, J=7.8 Hz, 1H), 8.09 (s, 1H), 7.91-7.88 (m, 1H), 7.86 (d, J=8.3 Hz, 1H), 7.78 (s, 1H), 7.51 (dd, J=8.4, 1.4 Hz, 1H), 7.19 (s, 1H), 6.94-6.93 (m, 1H), 5.48-5.40 (m, 1H), 5.28-5.16 (m, 1H), 5.00-4.87 (m, 1H), 4.85-4.75 (m, 1H), 4.56-4.42 (m, 1H), 3.98 (s, 3H), 3.94 (s, 3H), 2.75 (s, 3H); (+)-LRESIMS m/z (rel. int.) 527 (100) [M+H].sup.+; v.sub.max 3258, 1678, 1592, 1531, 1507, 1468, 1441, 1393, 1300, 1251, 1119, 998, 861 cm.sup.−1.
Synthesis Example 71—BT2142 and BT2145
(331) ##STR00109##
(332) Prepared using a method analogous to General Procedure J from BT2154 (40 mg, 78 μmol), potassium carbonate (22 mg, 0.16 mmol) and 1-(bromomethyl)-4-(trifluoromethoxy)benzene (14 μL, 86 μmol) in acetone (1.0 mL) and purified by flash column chromatography [silica, 1:20 v/v ammoniacal MeOH/DCM elution] to give, after concentration of the appropriate fractions, BT2142 as a white powder (35 mg, 65%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.79 (s, 1H), 8.88 (d, J=1.9 Hz, 1H), 8.78 (s, 1H), 8.55 (dd, J=4.7, 1.5 Hz, 1H), 8.05 (ddd, J=8.0, 2.2, 1.7 Hz, 18H), 7.75 (d, J=8.4 Hz, 1H), 7.70 (s, 1H), 7.65 (d, J=8.7 Hz, 2H), 7.58 (s, 1H), 7.49 (ddd, J=8.0, 4.8, 0.5 Hz, 1H), 7.40 (dd, J=8.4, 1.6 Hz, 1H), 7.34 (d, J=8.0 Hz, 2H), 6.83 (d, J=1.4 Hz, 1H), 5.41-5.33 (m, 1H), 5.33-5.27 (m, 1H), 4.95-4.87 (m, 1H), 4.84-4.74 (m, 1H), 4.54 (s, 2H), 4.37-4.28 (m, 1H), 3.87 (s, 6H), 3.81 (s, 6H); (+)-LRESIMS m/z (rel. int.) 687 (100) [M+H].sup.+; v.sub.max 3242, 1607, 1550, 1506, 1454, 1269, 1220, 1165, 1010, 797 cm.sup.−1.
(333) BT2145
(334) A portion of BT2142 (5.0 mg) at rt was treated with aqueous HCl (1.5 mL, 1.5 M), sonicated for 5 min then stirred for 10 min before removal of water with a gentle stream of nitrogen to afford BT2145 (5 mg), the hydrochloride salt of BT2142. H NMR (400 MHz, DMSO-d.sub.6) δ 12.01 (s, 1H), 10.60 (br s, 1H), 9.55 (br s, 1H), 9.10 (s, 1H), 8.75 (d, J=5.1 Hz, 1H), 8.56 (d, J=8.1 Hz, 1H), 8.11 (s, 1H), 7.91-7.84 (m, 2H), 7.78 (s, 1H), 7.71 (d, J=8.6 Hz, 2H), 7.51 (dd, J=8.4, 1.5 Hz, 1H), 7.32 (d, J=8.1 Hz, 2H), 7.29 (s, 1H), 6.88-6.85 (m, 1H), 5.47-5.35 (m, 1H), 5.20 (s, 1H), 4.76 (s, 2H), 4.70 (s, 2H), 4.43 (s, 1H), 4.00 (s, 3H), 3.94 (s, 3H). (+)-LRESIMS m/z (rel. int.) 687 (100) [M+H].sup.+; v.sub.max 1600, 1502, 1441, 1300, 1237, 1219, 1167, 999, 739 cm.sup.−1.
Synthesis Example 72—BT2153
(335) ##STR00110##
(336) Prepared and purified using a method analogous to General Procedure J from BT2070 (50 mg, 0.10 mmol), potassium carbonate (28 mg, 0.20 mmol) and 1-(bromomethyl)-4-(trifluoromethoxy)benzene (26 mg, 0.10 mmol) in acetone (1.5 mL) to afford BT2153 as a white powder (36 mg, 53%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.73 (s, 1H), 8.92 (d, J=1.9 Hz, 1H), 8.84 (t, J=5.8 Hz, 1H), 8.52 (dd, J=4.9, 1.9 Hz, 1H), 8.47 (s, 1H), 8.07 (ddd, J=7.9, 1.7 Hz, 1H), 7.98 (s, 1H), 7.65 (s, 1H), 7.57 (dd, J=8.7, 1.7 Hz, 1H), 7.54 (d, J=8.7 Hz, 1H), 7.46 (dd, J=7.9, 4.9, 1H), 7.30 (d, J=8.3 Hz, 2H), 7.13 (d, J=1.5 Hz, 1H), 6.96 (d, J=8.3 Hz, 2H), 6.87 (s, 1H), 4.36 (s, 2H), 4.34-4.29 (m, 2H), 3.86 (s, 3H), 3.85 (s, 3H), 3.72-3.65 (m, 2H). .sup.13C NMR (100 MHz, DMSO-d.sub.6) δ 161.4, 155.3, 154.1, 152.8, 147.6, 147.6, 147.5, 147.2, 139.1, 136.8, 136.5, 136.2, 133.9, 132.8, 130.9, 128.9, 127.8, 123.7 (2C), 122.6, 120.6 (2C), 119.9 (q, J.sub.C-F=256.2 Hz), 119.9, 113.0, 111.2, 107.4, 106.1, 102.9, 56.2, 55.7, 45.2, 36.3, 34.5. (+)-LRESIMS m/z (rel. int.) 675 (100) [M+H].sup.+; v.sub.max 1623, 1601, 1547, 1502, 1436, 1266, 1214, 1159, 1019, 794 cm.sup.−1.
Synthesis Example 73—BT2154
(337) ##STR00111##
(338) Four 10 mL snap-cap microwave vessels, each fitted with a magnetic stirring bar, were charged with a mixture of 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (160 mg, 0.78 mmol), tert-butyl (1-(6-bromo-1H-indole-2-carbonyl)azetidin-3-yl)carbamate (205 mg, 0.52 mmol) and potassium carbonate (380 mg, 2.74 mmol) then treated with a degassed mixture of dimethoxyethane, water and ethanol (7:3:2, 7 mL). Bis(triphenylphosphine)palladium(II) dichloride (10 mg) was added and the mixture was sparged with nitrogen for 0.05 hr, sealed then separately subjected to microwave irradiation (120° C./0.33 h, ramp time 1 minute, maximum power 200 W). All vessels were poured into one conical flask containing water (35 mL) and the resulting precipitate collected via vacuum filtration. The residue obtained was then washed with ether (30 mL) to afford the desired indole derivative substituted at C-6 as a tan solid (740 mg, 91% combined yield) and used without further purification. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.78 (s, 1H), 8.87 (d, J=1.5 Hz, 1H), 8.55 (dd, J=4.4, 0.8 Hz, 1H), 8.09-8.01 (m, 1H), 7.76 (d, J=8.4 Hz, 1H), 7.69 (s, 1H), 7.67 (d, J=7.3 Hz, 1H), 7.49 (dd, J=8.0, 4.8 Hz, 1H), 7.41 (dd, J=8.4, 1.7 Hz, 1H), 7.22 (br s, 1H), 4.79-4.72 (m, 1H), 4.48-4.39 (m, 1H), 4.36-4.25 (m, 2H), 3.99-3.90 (m, 1H), 1.40 (s, 9H); (+)-LRESIMS m/z (rel. int.) 393 (100) [M+H].sup.+; v.sub.max 3260, 1700, 1594, 1532, 1506, 1456, 1365, 1161, 799 cm.sup.−1. The product formed directly above (324 mg, 0.83 mmol) was deprotected at 0° C. using a method analogous to that described in General Procedure F to afford the di-TFA-salt of the compound formed directly above as a tan solid (310 mg) that was dissolved in ethanol (10 mL), magnetically stirred and treated with triethylamine (663 μL, 4.77 mmol). The resulting solution was then treated with a solution of 2-isothiocyanato-4,5-dimethoxybenzonitrile (131 mg, 0.60 mmol) in DCM (3 mL) dropwise and stirred for 2 hours at rt. The mixture was diluted with ethanol (20 mL) then heated at 70° C. for 30 mins, cooled to rt, and the solid collected and washed with ether (10 mL) to afford BT2154 (239 mg, 78%) as a brown solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.09 (s, 1H), 11.76 (s, 1H), 9.34 (s, 1H), 8.88 (s, 1H), 8.55 (d, J=4.6 Hz, 1H), 8.06 (d, J=7.9 Hz, 1H), 7.77 (d, J=8.4 Hz, 1H), 7.70 (s, 1H), 7.57 (s, 1H), 7.49 (dd, J=7.5, 5.2 Hz, 1H), 7.40 (d, J=8.3 Hz, 1H), 6.85 (s, 1H), 6.81 (s, 1H), 6.01-5.89 (m, 1H), 5.08-4.96 (m, 1H), 4.90-4.79 (m, 1H), 4.71-4.57 (m, 1H), 4.44-4.33 (m, 1H), 3.80 (s, 3H), 3.78 (s, 3H). (+)-LRESIMS m/z (rel. int.) 513 (100) [M+H].sup.+; v.sub.max 3213, 1625, 1599, 1534, 1508, 1432, 1387, 1281, 1245, 1232, 1008, 797 cm.sup.−1.
II. BIOLOGICAL EXAMPLES
Example I—Establishment of the Adoptive Transfer of Type 1 Diabetes in Transgenic RIP-OVAhi Mice: An Acute Experimental Type 1 Diabetes Model for Rapid Drug Efficacy Profiling
(339) In NOD mice, the autoimmune disease process develops slowly because of the asynchronous destruction of the beta cells in different islets. Indeed Type 1 diabetes develops only after >90% of the total islet beta cell mass has been destroyed. The slow course of the disease in NOD mice offers realistic potential for immune intervention but, unfortunately, high throughput screening of test drugs is not feasible. A more acute model of diabetes can be induced experimentally using RIP-OVAhi transgenic mice which express a foreign ovalbumin (OVA) peptide, under the control of the rat insulin promoter (RIP), only in islet beta cells. Following adoptive transfer of naïve OVA specific CD8+ (OT-I) and activated OVA-specific CD4+ (OT-II) transgenic T cells, the transgenic T cells traffic to the host lymph nodes where the OT-I T cells become activated and then to the OVA-expressing pancreatic islets where they rapidly induce islet beta cell destruction and diabetes, all mice succumbing to rapid onset of high blood glucose levels (hyperglycaemia) in the second week after cell transfer. Using heparanase knockout donor and recipient mouse strains, it has been demonstrated that this acute model of Type 1 diabetes is heparanase-dependent (
Comparative Example II—Control of Blood Glucose Levels
(340) Using the RIP-OVAhi transgenic mice the effect of a heparanase inhibitor compound (denoted “BT-1055”) was tested for its effect on blood glucose levels. Following adoptive transfer of naïve OVA specific CD8+ (OT-I) and activated OVA-specific CD4+ (OT-II) transgenic T cells into a number of RIP-OVAhi mice blood glucose was monitored regularly over two weeks in populations of mice treated with BT-1055 and a control group administered saline (
Comparative Example III—Effect of Treatment with a Sulfated Oligosaccharide (PI-88) on Insulin Production
(341) The db/db mouse has defective leptin signalling resulting from a point mutation in the leptin receptor gene. Plasma insulin levels become elevated very early in life (10-14 days of age), and the affected animals are obese by the time they are 3-4 weeks old. The animals are insulin resistant, hypertriglyceridemic, and have impaired glucose tolerance.
(342) db/db mice were treated with saline or 50 mg/kg of PI-88 by daily intraperitoneal injection. At the cessation of treatment pancreatic islets were isolated and stained for insulin and the level of insulin expression assessed by quantitative morphometric analyses.
Comparative Example IV—Effect of Treatment with a Sulfated Oligosaccharide (PI-88) on Heparan Sulfate Levels
(343) db/db mice were treated with saline or 50 mg/kg of PI-88 daily by intraperitoneal injection. At the cessation of treatment pancreatic islets were isolated and stained with alcian blue to visualise heparan sulfate and the heparan sulfate content of the islets was assessed by quantitative morphometric analyses.
Example V—General Screening Protocol for Compounds
(344) 1. Heparanase Inhibition:
(345) Heparanase inhibitor compounds of the invention may be tested for inhibition of heparanase at 10 μM using a standardised enzymatic assay (Hammond E, et al. Development of a colorimetric assay for heparanase activity suitable for kinetic analysis and inhibitor screening. Anal Biochem. 2010; 396:112-116). Molecules exhibiting 50% or greater heparanase inhibition may be further assessed to determine the specific half-maximal inhibitor concentrations of heparanase (IC.sub.50). Molecules with heparanase inhibition IC.sub.50 within a desired range (e.g., 100 μM and below, 50 μM and below, 10 μM and below, 5 μM and below, 2 μM and below, etc) may be prioritised for further in vitro and in vivo screening as appropriate.
(346) 2. Establish Off-Target Activity Counter-Screens:
(347) A first category of counter-screens may be used to determine whether heparanase inhibition by compounds of the invention occurs via competitive inhibition and not protein aggregation. A second category of counter-screens forms part of a larger suite of biochemical characterization to identify areas that might be improved as part of lead optimisation. For example, the second category of counter-screens may be used to assess off-target activity against the following: heparanase-related glycosidases including (α,β)-glucosidase and β-glucuronidase; heparan sulfate binding proteins including VEGF, FGF-1 and FGF-2; and kinase inhibition panels to establish the ratio of on-target to off-target activity.
3. In Vitro Toxicity:
(348) Heparanase inhibitor compounds of the invention may be tested for dose dependent in vitro toxicity using the Jurkat cell line and the MTT assay which provides a measure of mitochondrial function as a surrogate for cell viability. Data may be represented as Tox50 as drug concentration at which only 50% of cells remain viable. Typically, compounds that have a Tox50 of 200 μM or greater may be prioritised for further evaluation.
(349) 4. In Vitro ADME (Absorption, Distribution, Metabolism, and Excretion):
(350) Compounds of the invention can undergo an initial assessment of ADMET (absorption, distribution, metabolism, excretion and toxicology) properties using standard methods.
(351) Initial in vitro assays may include the following:
(352) A. Solubility: Compounds may be tested for solubility in a standard kinetic solubility assay at different pH levels (pH 2.1, 4.0 and 7.4) with a typical target solubility of 100 μg/ml. B. PAMPA assays (membrane permeability/adsorption): The PAMPA assay is a well validated surrogate measure of membrane permeability which uses an artificial lipid bilayer, and is used increasingly in place of the Caco2 cell. Compounds may be compared with known standards drug compounds including Verapamil (high permeability) and Theophyline (low permeability). High permeability may be a criterion for prioritising compounds for further testing. C. Liver microsome assay: The liver microsome assay is a well-established method for determining the oxidative metabolic stability of drug candidates. Mouse and human microsomes may be used to support decision making when selecting candidate compounds for in vivo studies in mouse models. The percentage of compound remaining at multiple time points (0, 5, 15, 30, 45 min) may be determined in comparison with standard drug compounds including Propranolol (highly stable), Ketoprofen (intermediate stability) and Verapamil (rapid clearance). D. Protein binding: The plasma clearance of drug candidates is determined by a combination of metabolic and renal clearance. The renal clearance of drug candidates which have high protein binding is lowered, since renal clearance is size dependent and proteins exceed the upper cut-off. Excessive protein binding can however limit the availability of the drug towards the biological target. Acceptable values may typically be in the range 90-98%. Measurement of free drug concentration in plasma using equilibrium dialysis in comparison to drug standards including Propranolol (low binding), Paroxetine (intermediate binding) and Losartan (high binding). Compounds with intermediate protein binding may be prioritised.
Example VI—Heparanase Inhibition
(353) Compounds of the invention were tested over a range of concentrations in a beta cell protection assay to evaluate their capacity to protect beta cells from dying in culture, compared to untreated controls. In addition to measuring uptake of Sytox green by flow cytometry, as a measure of cell death, a beta cell survival assay using Calcein, a dye taken up only by viable cells, in combination with the DNA-binding dye propidium iodide (PI) that identifies dead and dying cells. Mouse islets were isolated from BALB/c pancreas dispersed into beta cell suspensions using a modified method of Josefsen et al (J. Endocrinol 1996 149:145-154) and the beta cells cultured for 2 days in the presence or absence of compounds of the invention (0.5-50 μg/ml). Heparin and PI-88 (Muparfostat, Progen Pharmaceuticals) were included in the assays as positive controls for protection.
(354) Heparanase inhibition was determined using standard methods and the results expressed as the half maximal inhibitory concentration (IC.sub.50) being the concentration of the compound of the invention required to achieve 50% inhibition of heparanase. The results are shown in Table 1:
(355) TABLE-US-00001 TABLE 1 Heparanase inhibition (IC.sub.50) Compound Code Structure IC.sub.50 BT2012
(356) It will be appreciated by persons skilled in the art that numerous variations and/or modifications may be made to the invention as shown in the specific embodiments without departing from the spirit or scope of the invention as broadly described. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive.