TRICYCLIC TETRAHYDROISOQUINOLINE DERIVATIVE, PREPARATION METHOD THEREFOR AND APPLICATION THEREOF IN MEDICINE

Abstract

The present disclosure relates to a tricyclic tetrahydroisoquinoline derivative, a preparation method therefor and an application thereof in medicine. In particular, the present disclosure relates to a tricyclic tetrahydroisoquinoline derivative represented by general formula (I), a preparation method therefor and a pharmaceutical composition comprising said derivative, a use thereof as an estrogen receptor modulator, and a use thereof in preparing a drug for treating estrogen receptor-mediated or dependent diseases or disorders. The substituents in general formula (I) are the same as those defined in the description.

##STR00001##

Claims

1. (canceled)

2. A compound of general formula (I) or a tautomer, mesomer, racemate, enantiomer or diastereomer thereof or a mixture thereof, or a pharmaceutically acceptable salt thereof, ##STR00130## wherein: R.sup.1a and R.sup.1b are identical or different and are each independently selected from the group consisting of H atom, deuterium atom, halogen, alkyl, deuterated alkyl, haloalkyl, alkoxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, carboxyl, aldehyde, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, alkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; ring A is heterocyclyl; Z is selected from the group consisting of O atom, S atom, NR.sup.7 and CR.sup.9R.sup.10; G.sup.1, G.sup.2, G.sup.3 and G.sup.4 are identical or different and are each independently CR.sup.8 or a N atom; R.sup.2 are identical or different and are each independently selected from the group consisting of hydrogen atom, halogen, alkyl, haloalkyl, hydroxyalkyl, alkoxy, cyano, amino, nitro, carboxyl, aldehyde, hydroxy, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, alkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.3 is selected from the group consisting of hydrogen atom, alkyl and cycloalkyl, wherein the alkyl and cycloalkyl are optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, alkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, carboxyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.4 is selected from the group consisting of hydrogen atom, alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, alkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.5 is selected from the group consisting of hydrogen atom, alkyl, haloalkyl and cycloalkyl, wherein the alkyl and cycloalkyl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, alkoxy, cyano, amino, nitro, carboxyl, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.6 are identical or different and are each independently selected from the group consisting of hydrogen atom, alkyl, deuterated alkyl, haloalkyl, alkoxy, cyano, amino, nitro, halogen, carboxyl, aldehyde, hydroxy, hydroxyalkyl, cycloalkyl and heterocyclyl, wherein the alkyl, cycloalkyl and heterocyclyl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, alkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, carboxyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.7 is selected from the group consisting of hydrogen atom, alkyl, haloalkyl, alkenyl, propargyl, cycloalkyl and heterocyclyl, wherein the alkyl, cycloalkyl and heterocyclyl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, alkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, carboxyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.8 are identical or different and are each independently selected from the group consisting of hydrogen atom, halogen, alkyl, alkoxy, haloalkyl, alkenyl, alkynyl, cyano, cycloalkyl and heterocyclyl, wherein the alkyl, cycloalkyl and heterocyclyl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, alkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, carboxyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.9 and R.sup.10 are identical or different and are each independently selected from the group consisting of hydrogen atom, halogen, alkyl, alkoxy, hydroxy, hydroxyalkyl, haloalkyl, alkenyl, alkynyl, cyano, cycloalkyl and heterocyclyl; n is 1, 2 or 3; s is 0, 1 or 2; and p is 0, 1, 2 or 3.

3. A compound of general formula (I) or a tautomer, mesomer, racemate, enantiomer or diastereomer thereof or a mixture thereof, or a pharmaceutically acceptable salt thereof according to claim 2, wherein ring A is 3- to 6-membered heterocyclyl containing 1 to 3 heteroatoms selected from the group consisting of N atom, O atom and S atom.

4. A compound of general formula (I) or a tautomer, mesomer, racemate, enantiomer, diastereomer or a mixture thereof, or a pharmaceutically acceptable salt thereof according to claim 2, wherein G.sup.1, G.sup.2, G.sup.3 and G.sup.4 are all CR.sup.8, or one of G.sup.1, G.sup.2, G.sup.3 and G.sup.4 is a N atom, and the others are CR.sup.8; R.sup.8 are as defined in claim 2.

5. The compound of general formula (I) or the tautomer, mesomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 2, wherein the compound of general formula (I) or the tautomer, mesomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof, is a compound of general formula (II), ##STR00131## wherein: r is 0, 1, 2 or 3; q is 1, 2 or 3; t is 1 or 2; Z, G.sup.1, R.sup.1a, R.sup.1b, R.sup.2-R.sup.6, R.sup.8, n and s are as defined in claim 2.

6. The compound of general formula (I) or the tautomer, mesomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 2, wherein the compound of general formula (I) or the tautomer, mesomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof, is a compound of general formula (IIG) or (IIGa), ##STR00132## wherein: R.sup.11a and R.sup.11b are identical or different and are each independently selected from the group consisting of hydrogen atom, halogen, alkyl, haloalkyl, hydroxyalkyl, alkoxy, cyano, amino, nitro, carboxyl, hydroxy, cycloalkyl, heterocyclyl, aryl and heteroaryl; k is an integer from 1 to 6; q is 1, 2 or 3; t is 1 or 2; r is 0, 1, 2 or 3; G.sup.1, Z, R.sup.1a, R.sup.1b, R.sup.2-R.sup.4, R.sup.6, R.sup.8, n and s are as defined in claim 2.

7. The compound of general formula (I) or the tautomer, mesomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 2, wherein Z is NR.sup.7; R.sup.7 is a hydrogen atom; n is 1.

8. (canceled)

9. (canceled)

10. (canceled)

11. (canceled)

12. The compound of general formula (I) or the tautomer, mesomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 2, wherein G.sup.1 is a N atom; or G.sup.1 is CR.sup.8; R.sup.8 is as defined in claim 2.

13. (canceled)

14. (canceled)

15. (canceled)

16. The compound of general formula (I) or the tautomer, mesomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 2, wherein R.sup.1a and R.sup.1b are identical or different and are each independently selected from the group consisting of H atom, deuterium atom, fluorine atom and C.sub.1-6 alkyl; R.sup.2 is a hydrogen atom; R.sup.3 is a hydrogen atom; R.sup.4 is a hydrogen atom or alkyl; R.sup.6 is C.sub.1-6 haloalkyl.

17. (canceled)

18. (canceled)

19. (canceled)

20. The compound of general formula (I) or the tautomer, mesomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 2, wherein R.sup.5 is alkyl optionally substituted with one or more substituents selected from the group consisting of halogen, amino, cyano, hydroxy, alkoxy, carboxyl and cycloalkyl.

21. (canceled)

22. The compound of general formula (I) or the tautomer, mesomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 2, wherein the compound of general formula (I) or the tautomer, mesomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt, is selected from the group consisting of the following compounds: ##STR00133## ##STR00134## ##STR00135## ##STR00136## ##STR00137##

23. (canceled)

24. A compound of general formula (IA) or a tautomer, mesomer, racemate, enantiomer or diastereomer thereof or a mixture thereof, or a pharmaceutically acceptable salt thereof, ##STR00138## wherein: R.sup.1a and R.sup.1b are identical or different and are each independently selected from the group consisting of H atom, deuterium atom, halogen, alkyl, deuterated alkyl, haloalkyl, alkoxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, carboxyl, aldehyde, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, alkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; X is Br; G.sup.1, G.sup.2, G.sup.3 and G.sup.4 are identical or different and are each independently CR.sup.8 or a N atom; R.sup.2 are identical or different and are each independently selected from the group consisting of hydrogen atom, halogen, alkyl, haloalkyl, hydroxyalkyl, alkoxy, cyano, amino, nitro, carboxyl, aldehyde, hydroxy, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, alkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.3 is selected from the group consisting of hydrogen atom, alkyl and cycloalkyl, wherein the alkyl and cycloalkyl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, alkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, carboxyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.4 is selected from the group consisting of hydrogen atom, alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, alkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.5 is selected from the group consisting of alkyl, haloalkyl and cycloalkyl, wherein the alkyl and cycloalkyl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, alkoxy, cyano, amino, nitro, carboxyl, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.8 are identical or different and are each independently selected from the group consisting of hydrogen atom, halogen, alkyl, alkoxy, haloalkyl, alkenyl, alkynyl, cyano, cycloalkyl and heterocyclyl, wherein the alkyl, cycloalkyl and heterocyclyl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, alkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, carboxyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; n is 1, 2 or 3; and s is 0, 1 or 2.

25. The compound of general formula (IA) or the tautomer, mesomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 24, wherein the compound of general formula (IA) or the tautomer, mesomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt, is selected from the group consisting of the following compounds: ##STR00139##

26. (canceled)

27. A compound of general formula (IIGA) or (IIGaA) or a tautomer, mesomer, racemate, enantiomer or diastereomer thereof or a mixture thereof, or a pharmaceutically acceptable salt thereof, ##STR00140## wherein: R.sup.1a and R.sup.1b are identical or different and are each independently selected from the group consisting of H atom, deuterium atom, halogen, alkyl, deuterated alkyl, haloalkyl, alkoxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, carboxyl, aldehyde, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, alkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.w is a hydroxy protective group; Z is selected from the group consisting of O atom, S atom, NR.sup.7 and CR.sup.9R.sup.10; G.sup.1 is CR.sup.8 or a N atom; R.sup.2 are identical or different and are each independently selected from the group consisting of hydrogen atom, halogen, alkyl, haloalkyl, hydroxyalkyl, alkoxy, cyano, amino, nitro, carboxyl, aldehyde, hydroxy, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, alkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.3 is selected from the group consisting of hydrogen atom, alkyl and cycloalkyl, wherein the alkyl and cycloalkyl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, alkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, carboxyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.4 is selected from the group consisting of hydrogen atom, alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, alkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.6 is independently selected from the group consisting of hydrogen atom, alkyl, deuterated alkyl, haloalkyl, alkoxy, cyano, amino, nitro, halogen, carboxyl, carboxylate group, aldehyde, hydroxy, hydroxyalkyl, cycloalkyl and heterocyclyl, wherein the alkyl, cycloalkyl and heterocyclyl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, alkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, carboxyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.7 is selected from the group consisting of hydrogen atom, alkyl, haloalkyl, alkenyl, propargyl, cycloalkyl and heterocyclyl, wherein the alkyl, cycloalkyl and heterocyclyl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, alkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, carboxyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.8 are identical or different and are each independently selected from the group consisting of hydrogen atom, halogen, alkyl, alkoxy, haloalkyl, alkenyl, alkynyl, cyano, cycloalkyl and heterocyclyl, wherein the alkyl, cycloalkyl and heterocyclyl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, alkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, carboxyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.9 and R.sup.10 are identical or different and are each independently selected from the group consisting of hydrogen atom, halogen, alkyl, alkoxy, hydroxy, hydroxyalkyl, haloalkyl, alkenyl, alkynyl, cyano, cycloalkyl and heterocyclyl; R.sup.11a and R.sup.11b are identical or different and are each independently selected from the group consisting of hydrogen atom, halogen, alkyl, haloalkyl, hydroxyalkyl, alkoxy, cyano, amino, nitro, carboxyl, aldehyde, hydroxy, cycloalkyl, heterocyclyl, aryl and heteroaryl; k is an integer from 1 to 6; q is 1, 2 or 3; t is 1 or 2; n is 1, 2 or 3; r is 0, 1, 2 or 3; and s is 0, 1 or 2.

28. The compound of general formula (IIGA) or (IIGaA) or the tautomer, mesomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 27, wherein the compound of general formula (IIGA) or (IIGaA) or the tautomer, mesomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt, is selected from the group consisting of the following compounds: ##STR00141## ##STR00142## ##STR00143## ##STR00144## ##STR00145## ##STR00146##

29. A method for preparing the compound of general formula (I) or the tautomer, mesomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 2, comprising: ##STR00147## subjecting a compound of general formula (IA) and a compound of general formula (IB) to a coupling reaction to give the compound of general formula (I), wherein: X is Br; ring A, Z, G.sup.1, G.sup.2, G.sup.3, G.sup.4, R.sup.1a, R.sup.1b, R.sup.2-R.sup.6, n, p and s are as defined in claim 2.

30. A method for preparing the compound of general formula (IIG) or (IIGa) or the tautomer, mesomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 6, comprising: ##STR00148## ##STR00149## removing a hydroxy protective group from a compound of general formula (IIGA) to give the compound of general formula (IIG), or removing a hydroxy protective group from a compound of general formula (IIGaA) to give the compound of general formula (IIGa), wherein: R.sup.w is a hydroxy protective group; Z, G.sup.1, R.sup.1a, R.sup.1b, R.sup.2-R.sup.4, R.sup.6, R.sup.8, R.sup.11a, R.sup.11b, q, t, k, r, n and s are as defined in claim 6.

31. A pharmaceutical composition comprising a therapeutically effective amount of the compound of general formula (I) or the tautomer, mesomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 2, and one or more pharmaceutically acceptable carriers, diluents or excipients.

32. (canceled)

33. A method of preventing and/or treating cancer in a subject in need thereof, the method comprising administrating to the subject, an effective amount of the pharmaceutical composition according to claim 31.

34. A method for preventing and/or treating an estrogen receptor-mediated or -dependent disease or condition in a subject in need thereof, the method comprising administering to the subject an effective amount of the pharmaceutical composition according to claim 31, wherein the estrogen receptor-mediated or -dependent disease or condition is selected from the group consisting of cancer, central nervous system deficit, cardiovascular system deficit, blood system deficit, immune and inflammatory disease, susceptible infection, metabolic deficit, neurologic deficit, psychiatric deficit and reproductive deficit.

35. The method of claim 34, wherein the cancer is selected from the group consisting of breast cancer, endometrial cancer, uterine cancer, cervical cancer, skin cancer, prostate cancer, ovarian cancer, fallopian tube tumor, hemophilia and leukemia.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0205] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0206] The term “alkyl” refers to a saturated aliphatic hydrocarbon group which is a linear or branched group containing 1 to 20 carbon atoms, preferably to an alkyl group containing 1 to 12 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12) carbon atoms, and more preferably to an alkyl group containing 1 to 6 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various side-chain isomers thereof, etc. More preferred is a lower alkyl having 1 to 6 carbon atoms, and non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl and the like. The alkyl may be substituted or unsubstituted, and when it is substituted, the substituent may be substituted at any available connection site, and the substituent is preferably one or more of the following groups; it is substituted with one or more substituents independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio and oxo.

[0207] The term “alkenyl” refers to an alkyl compound containing at least one carbon-carbon double bond in the molecule, wherein the alkyl is as defined above. The alkenyl is a linear or branched group containing 2 to 20 carbon atoms, preferably 2 to 12 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12) carbon atoms, and more preferably 2 to 6 carbon atoms. The alkenyl may be substituted or unsubstituted, and when it is substituted, the substituent is preferably one or more of the following groups; it is substituted with one or more substituents independently selected from the group consisting of hydrogen atom, alkyl, alkoxy, halogen, haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.

[0208] The term “alkynyl” refers to an alkyl compound containing at least one carbon-carbon triple bond in the molecule, wherein the alkyl is as defined above. The alkynyl is a linear or branched group containing 2 to 20 carbon atoms, preferably 2 to 12 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12) carbon atoms, and more preferably 2 to 6 carbon atoms. Non-limiting examples of the alkynyl include, but are not limited to —C≡CH, —CH.sub.2C≡CH, —CH.sub.2C≡CCH.sub.3, —C≡CCH.sub.2CH.sub.3, —CH.sub.2C≡CCH.sub.2CH.sub.3, —C≡CCH(CH.sub.3).sub.2, —C(CH.sub.3).sub.2C≡CH, —C(CH.sub.3).sub.2CCCH.sub.3 and the like. The alkynyl may be substituted or unsubstituted, and when it is substituted, the substituent is preferably one or more of the following groups; it is substituted with one or more substituents preferably independently selected from the group consisting of hydrogen atom, alkyl, alkoxy, halogen, haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.

[0209] The term “alkylene” refers to a saturated linear or branched aliphatic hydrocarbon group having 2 residues derived from the parent alkane by removal of two hydrogen atoms from the same carbon atom or two different carbon atoms, which is a linear or branched group containing 1 to 20 carbon atoms, preferably an alkylene group containing 1 to 12 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12) carbon atoms, and more preferably an alkylene group containing 1 to 6 carbon atoms. Non-limiting examples of the alkylene include, but are not limited to, methylene (—CH.sub.2—), 1,1-ethylene (—CH(CH.sub.3)—), 1,2-ethylene (—CH.sub.2CH.sub.2—), 1,1-propylene (—CH(CH.sub.2CH.sub.3)—), 1,2-propylene (—CH.sub.2CH(CH.sub.3)—), 1,3-propylene (—CH.sub.2CH.sub.2CH.sub.2—), 1,4-butylene (—CH.sub.2CH.sub.2CH.sub.2CH.sub.2—) and the like. The alkylene may be substituted or unsubstituted, and when it is substituted, the substituent may be substituted at any available connection site with one or more substituents preferably independently optionally selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, thiol, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio and oxo.

[0210] The term “alkoxy” refers to —O-(alkyl) and —O-(unsubstituted cycloalkyl), wherein the alkyl is as defined above. Non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy. The alkoxy may be optionally substituted or unsubstituted, and when it is substituted, the substituent is preferably one or more of the following groups; it is substituted with one or more substituents independently selected from the group consisting of hydrogen atom, halogen, alkyl, alkoxy, haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.

[0211] The term “cycloalkyl” refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent. The cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 12 (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12) carbon atoms, more preferably 3 to 8 carbon atoms, and most preferably 3 to 6 (e.g., 3, 4, 5 or 6) carbon atoms. Non-limiting examples of the monocyclic cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like, preferably cycloalkyl. Polycyclic cycloalkyl includes spiro cycloalkyl, fused cycloalkyl, and bridged cycloalkyl.

[0212] The term “spiro cycloalkyl” refers to a 5- to 20-membered polycyclic group in which monocyclic rings share one carbon atom (referred to as the spiro atom), wherein the spiro cycloalkyl may contain one or more double bonds. The spiro cycloalkyl is preferably 6-to 14-membered, more preferably 7- to 10-membered (e.g., 7, 8, 9 and 10-membered). According to the number of the spiro atoms shared among the rings, the spiro cycloalkyl may be monospiro cycloalkyl, bispiro cycloalkyl or polyspiro cycloalkyl, preferably monospiro cycloalkyl and bispiro cycloalkyl, more preferably 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered monospiro cycloalkyl. Non-limiting examples of the spiro cycloalkyl include:

##STR00066##

[0213] The term “fused cycloalkyl” refers to a 5- to 20-membered carbon polycyclic group in which each ring shares a pair of adjacent carbon atoms with the other rings in the system, wherein one or more of the rings may contain one or more double bonds. The fused cycloalkyl is preferably 6- to 14-membered, more preferably 7- to 10-membered (e.g., 7, 8, 9 and 10-membered). According to the number of the formed rings, the fused cycloalkyl may be bicyclic, tricyclic, tetracyclic or polycyclic cycloalkyl, preferably bicyclic or tricyclic cycloalkyl, and more preferably 3-membered/4-membered, 3-membered/5-membered, 3-membered/6-membered, 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/4-membered, 5-membered/5-membered, 5-membered/6-membered, 6-membered/3-membered, 6-membered/4-membered, 6-membered/5-membered and 6-membered/6-membered bicyclic cycloalkyl. Non-limiting examples of the fused cycloalkyl include:

##STR00067##

[0214] The term “bridged cycloalkyl” refers to a 5- to 20-membered carbon polycyclic group in which any two rings share two carbon atoms that are not directly connected to each other, wherein the bridged cycloalkyl may contain one or more double bonds. The bridged cycloalkyl is preferably 6- to 14-membered, more preferably 7- to 10-membered (e.g., 7, 8, 9 and 10-membered). According to the number of the formed rings, the bridged cycloalkyl may be bicyclic, tricyclic, tetracyclic or polycyclic, preferably bicyclic, tricyclic or tetracyclic, and more preferably bicyclic or tricyclic. Non-limiting examples of the bridged cycloalkyl include:

##STR00068##

[0215] The cycloalkyl ring includes those in which the cycloalkyl described above (e.g., monocyclic, fused, spiro, and bridged cycloalkyl groups) is fused to an aryl, heteroaryl or heterocycloalkyl ring, wherein the ring connected to the parent structure is cycloalkyl. Non-limiting examples include indanyl, tetrahydronaphthyl, benzocycloheptanyl, and the like, preferably indanyl and tetrahydronaphthyl.

[0216] The cycloalkyl may be optionally substituted or unsubstituted, and when it is substituted, the substituent is preferably one or more of the following groups; it is substituted with one or more substituents independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio and oxo.

[0217] The term “heterocyclyl” refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent containing 3 to 20 ring atoms, wherein one or more of the ring atoms are heteroatoms selected from the group consisting of nitrogen, oxygen, S, S(O) and S(O).sub.2, excluding a cyclic portion of —O—O—, —O—S— or —S—S—, and the remaining ring atoms are carbon atoms. The heterocyclyl preferably contains 3 to 12 (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12) ring atoms, of which 1 to 4 (e.g., 1, 2, 3 and 4) are heteroatoms. The heterocyclyl preferably contains 3 to 8 ring atoms, of which 1 to 3 are heteroatoms. The heterocyclyl preferably contains 3 to 6 ring atoms, of which 1 to 3 are heteroatoms. Non-limiting examples of the monocyclic heterocyclyl include azetidinyl, pyrrolidinyl, imidazolidinyl, tetrahydrofuryl, tetrahydropyranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, and the like, preferably tetrahydropyranyl, piperidinyl and pyrrolidinyl. The polycyclic heterocyclyl includes spiro heterocyclyl, fused heterocyclyl, and bridged heterocyclyl.

[0218] The term “spiro heterocyclyl” refers to a 5- to 20-membered polycyclic heterocyclyl group in which monocyclic rings share one atom (referred to as the spiro atom), wherein one or more ring atoms are heteroatoms selected from the group consisting of nitrogen, oxygen, S, S(O) and S(O).sub.2, and the remaining ring atoms are carbon atoms. The spiro heterocyclyl may contain one or more double bonds. The spiro heterocyclyl is preferably 6- to 14-membered, more preferably 7- to 10-membered (e.g., 7, 8, 9 and 10-membered). According to the number of spiro atoms shared among the rings, the spiro heterocyclyl may be monospiro heterocyclyl, bispiro heterocyclyl or polyspiro heterocyclyl, preferably monospiro heterocyclyl and bispiro heterocyclyl, and more preferably 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered monospiro heterocyclyl. Non-limiting examples of the spiro heterocyclyl include:

##STR00069##

[0219] The term “fused heterocyclyl” refers to a 5- to 20-membered polycyclic heterocyclyl group in which each ring shares a pair of adjacent atoms with the other rings in the system, wherein one or more of the rings may contain one or more double bonds. In the fused heterocyclyl, one or more of the ring atoms are heteroatoms selected from the group consisting of nitrogen, oxygen, S, S(O) and S(O).sub.2, and the remaining ring atoms are carbon atoms. The fused heterocyclyl is preferably 6- to 14-membered, more preferably 7- to 10-membered (e.g., 7, 8, 9 and 10-membered). According to the number of the formed rings, the fused heterocyclyl may be bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclyl, preferably bicyclic or tricyclic fused heterocyclyl, and more preferably 3-membered/4-membered, 3-membered/5-membered, 3-membered/6-membered, 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/4-membered, 5-membered/5-membered, 5-membered/6-membered, 6-membered/3-membered, 6-membered/4-membered, 6-membered/5-membered and 6-membered/6-membered bicyclic fused heterocyclyl. Non-limiting examples of fused heterocyclyl include:

##STR00070##

[0220] The term “bridged heterocyclyl” refers to a 5- to 14-membered polycyclic heterocyclyl group in which any two rings share two atoms that are not directly connected to each other, wherein the bridged heterocyclyl may contain one or more double bonds. In the bridged heterocyclyl, one or more of the ring atoms are heteroatoms selected from the group consisting of nitrogen, oxygen, S, S(O) and S(O).sub.2, and the remaining ring atoms are carbon atoms. The bridged heterocyclyl is preferably 6- to 14-membered, more preferably 7- to 10-membered (e.g., 7, 8, 9 and 10-membered). According to the number of the formed rings, the bridged heterocyclyl may be bicyclic, tricyclic, tetracyclic or polycyclic, preferably bicyclic, tricyclic or tetracyclic, and more preferably bicyclic or tricyclic. Non-limiting examples of the bridged heterocyclyl include:

##STR00071##

[0221] The heterocyclyl ring includes those in which the heterocyclyl described above (e.g., monocyclic, fused, spiro and bridged heterocyclyl groups) is fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring connected to the parent structure is heterocyclyl. Non-limiting examples include:

##STR00072##

and the like.

[0222] The heterocyclyl may be optionally substituted or unsubstituted, and when it is substituted, the substituent is preferably one or more of the following groups; it is substituted with one or more substituents independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio and oxo.

[0223] The term “aryl” refers to a 6- to 20-membered, preferably 6- to 10-membered, and more preferably 6-membered carbon monocyclic or fused polycyclic (i.e., rings that share a pair of adjacent carbon atoms) group having a conjugated n-electron system such as phenyl and naphthyl. The aryl ring includes those in which the aryl described above is fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is an aryl ring. Non-limiting examples include:

##STR00073##

[0224] The aryl may be substituted or unsubstituted, and when it is substituted, the substituent is preferably one or more of the following groups; it is substituted with one or more substituents independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio and heterocycloalkylthio.

[0225] The term “heteroaryl” refers to a heteroaromatic system containing 1 to 4 (e.g., 1, 2, 3 and 4) heteroatoms and 5 to 20 ring atoms, wherein the heteroatoms are selected from the group consisting of oxygen, sulfur and nitrogen. The heteroaryl is preferably 5- to 10-membered (e.g. 5-, 6-, 7-, 8-, 9- and 10-membered) and contains 1 to 3 heteroatoms. The heteroaryl is more preferably 5- or 6-membered and contains 1 to 3 heteroatoms. Non-limiting examples are pyrazolyl, imidazolyl, furyl, thienyl, thiazolyl, oxazolyl, pyrrolyl, triazolyl, tetrazolyl, pyridyl, pyrimidinyl, thiadiazole, pyrazinyl, and the like. The heteroaryl ring may be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is heteroaryl. Non-limiting examples include:

##STR00074##

[0226] The heteroaryl may be optionally substituted or unsubstituted, and when it is substituted, the substituent is preferably one or more of the following groups; it is substituted with one or more substituents preferably independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio and heterocycloalkylthio.

[0227] The cycloalkyl, heterocyclyl, aryl and heteroaryl described above have 1 residue derived from the parent ring by removal of one hydrogen atom from a ring atom, or 2 residues derived from the parent ring by removal of two hydrogen atoms from the same ring atom or two different ring atoms.

[0228] The term “alkylthio” refers to —S-(alkyl) and —S-(unsubstituted cycloalkyl), wherein the alkyl is as defined above. Non-limiting examples of the alkylthio include: methylthio, ethylthio, propylthio, butylthio, cyclopropylthio, cyclobutylthio, cyclopentylthio and cyclohexylthio. The alkylthio may be optionally substituted or unsubstituted, and when it is substituted, the substituent is preferably one or more of the following groups; it is substituted with one or more substituents independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio and heterocycloalkylthio.

[0229] The term “cycloalkyloxy” refers to —O-cycloalkyl, wherein the cycloalkyl is as defined above.

[0230] The term “haloalkyl” refers to an alkyl group substituted with one or more halogens, wherein the alkyl group is as defined above.

[0231] The term “deuterated alkyl” refers to an alkyl group substituted with one or more deuterium atoms, wherein the alkyl group is as defined above. The term “haloalkoxy” refers to an alkoxy group substituted with a halogen, wherein the alkoxy group is as defined above.

[0232] The term “hydroxyalkyl” refers to an alkyl group substituted with one or more hydroxy groups, wherein the alkyl group is as defined above.

[0233] The term “halohydroxyalkyl” refers to a hydroxyalkyl group substituted with one or more halogens, wherein the hydroxyalkyl group is as defined above. The term “hydroxy” refers to —OH group.

[0234] The term “halogen” refers to fluorine, chlorine, bromine or iodine.

[0235] The term “amino” refers to —NH.sub.2.

[0236] The term “cyano” refers to —CN.

[0237] The term “nitro” refers to —NO.sub.2.

[0238] The term “aldehyde” refers to —C(O)H.

[0239] The term “carboxyl” refers to —C(O)OH.

[0240] The term “aldehyde” refers to —C(O)H.

[0241] The term “carboxylate” refers to —C(O)O(alkyl) or —C(O)O(cycloalkyl), wherein the alkyl and cycloalkyl are as defined above.

[0242] The term “hydroxy protective group” is a suitable group known in the art for protecting hydroxy. See the hydroxy protective groups in the literature (“Protective Groups in Organic Synthesis”, 5.sup.th Ed. T. W. Greene & P. G. M. Wuts). As an example, preferably, the hydroxy protective group may be (C.sub.1-10alkyl or aryl).sub.3silyl such as triethylsilyl, triisopropylsilyl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl, and the like; or may be C.sub.1-10 alkyl or substituted alkyl, preferably alkoxy or aryl-substituted alkyl, more preferably C.sub.1-6 alkoxy-substituted C.sub.1-6 alkyl or phenyl-substituted C.sub.1-6 alkyl, and most preferably C.sub.1-4 alkoxy-substituted C.sub.1-4 alkyl such as methyl, tert-butyl, allyl, benzyl, methoxymethyl (MOM), ethoxyethyl, 2-tetrahydropyranyl (THP), and the like; or may be (C.sub.1-10 alkyl or aryl)acyl such as formyl, acetyl, benzoyl, and the like; or may be (C.sub.1-6 alkyl or C.sub.6-10 aryl)sulfonyl; or may also be (C.sub.1-6 alkoxy or C.sub.6-10 aryloxy)carbonyl. The term “optional” or “optionally” means that the event or circumstance subsequently described may, but not necessarily, occur, and that the description includes instances where the event or circumstance occurs or does not occur. For example, “a heterocyclyl group optionally substituted with alkyl” means that alkyl may be, but not necessarily, present, and that the description includes instances where the heterocyclyl group is or is not substituted with alkyl.

[0243] “Substituted” means that one or more, preferably up to 5, more preferably 1-3 hydrogen atoms in the group are independently substituted with a corresponding number of substituents, wherein each of the substituents has an independent option (i.e., the substituents may be identical or different). It goes without saying that a substituent is only in its possible chemical position, and those skilled in the art will be able to determine (experimentally or theoretically) possible or impossible substitution without undue efforts. For example, it may be unstable when an amino or hydroxy group having a free hydrogen is bound to a carbon atom having an unsaturated (e.g., olefinic) bond.

[0244] The term “pharmaceutical composition” refers to a mixture containing one or more of the compounds described herein or a physiologically/pharmaceutically acceptable salt or pro-drug thereof, and other chemical components, and other components, for example physiologically/pharmaceutically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to promote the administration to an organism, which facilitates the absorption of the active ingredient, thereby exerting biological activities.

[0245] The term “pharmaceutically acceptable salt” refers to salts of the disclosed compounds which are safe and effective for use in the body of a mammal and possess the requisite biological activities.

[0246] The compounds disclosed herein include isotopic derivatives thereof. The term “isotopic derivative” refers to compounds that differ in structure only by having one or more enriched isotopic atoms. For example, compounds having the structure disclosed herein having “deuterium” or “tritium” in place of hydrogen, or .sup.18F-fluorine labeling (.sup.18F isotope) in place of fluorine, or .sup.11C-, .sup.13C- or .sup.14C-enriched carbon (.sup.11C-, .sup.13C- or .sup.14C-carbon labeling; .sup.13C- or .sup.14C-isotope) in place of a carbon atom are within the scope of the present disclosure. Such a compound can be used as an analytical tool or a probe in, for example, a biological assay, or may be used as a tracer for in vivo diagnostic imaging of disease, or as a tracer in a pharmacodynamic, pharmacokinetic or receptor study.

[0247] Various deuterated forms of the compound of general formula (I) of the present disclosure are those in which each of the available hydrogen atoms connected to the carbon atoms can be independently replaced by a deuterium atom. Unless otherwise stated, when a position is specifically designated to D or deuterium, that position is construed as deuterium having an abundance that is at least 3000 times greater than the natural abundance of deuterium (which is 0.015%) (i.e., at least 45% incorporation of deuterium). Those skilled in the art are able to synthesize the deuterated forms of the compound of general formula (I) with reference to the relevant literature. Commercially available deuterated starting materials can be used in preparing the deuterated forms of the compound of general formula (I), or they can be synthesized by using conventional techniques with deuterated reagents including, but not limited to, deuterated borane, tri-deuterated borane in tetrahydrofuran, deuterated lithium aluminum hydride, deuterated iodoethane, deuterated iodomethane, and the like.

[0248] For drugs and pharmacological active agents, the term “therapeutically effective amount” refers to an amount of a medicament or an agent that is sufficient to provide the desired effect but is non-toxic. The determination of the effective amount varies from person to person. It depends on the age and general condition of a subject, as well as the particular active substance used. The appropriate effective amount in a case may be determined by those skilled in the art in light of routine tests.

Synthesis of the Compounds of the Present Disclosure

[0249] In order to achieve the purpose of the present disclosure, the following technical schemes are adopted in the present disclosure:

##STR00075##

[0250] Provided is a method for preparing the compound of general formula (I) or the tautomer, mesomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof of the present disclosure, which comprises the following step:

[0251] subjecting a compound of general formula (IA) and a compound of general formula (IB) to a coupling reaction under alkaline conditions in the presence of a catalyst to give the compound of general formula (I), wherein:

[0252] X is Br;

[0253] ring A, Z, G.sup.1, G.sup.2, G.sup.3, G.sup.4, R.sup.1a, R.sup.1b, R.sup.2-R.sup.6, p, n and s are as defined in general formula (I).

[0254] Provided is a method for preparing the compound of general formula (II) or the tautomer, mesomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof of the present disclosure, which comprises the following step:

##STR00076##

[0255] subjecting a compound of general formula (IIA) and a compound of general formula (IIB) to a coupling reaction under alkaline conditions in the presence of a catalyst to give the compound of general formula (II),

[0256] wherein:

[0257] X is Br;

[0258] G.sup.1, Z, R.sup.1a, R.sup.1b, R.sup.2-R.sup.6, R.sup.8 r, q, t, n and s are as defined in general formula (II).

##STR00077##

[0259] Provided is a method for preparing the compound of general formula (III) or the tautomer, mesomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof of the present disclosure, which comprises the following step:

[0260] subjecting a compound of general formula (IIIA) and a compound of general formula (IIIB) to a coupling reaction under alkaline conditions in the presence of a catalyst to give the compound of general formula (III),

[0261] wherein:

[0262] X is Br;

[0263] G.sup.1, R.sup.1a, R.sup.1b, R.sup.2-R.sup.8, r and s are as defined in general formula (III).

##STR00078##

[0264] Provided is a method for preparing the compound of general formula (IIIa) or the tautomer, mesomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof of the present disclosure, which comprises the following step:

[0265] subjecting a compound of general formula (IIIaA) and a compound of general formula (IIIB) to a coupling reaction under alkaline conditions in the presence of a catalyst to give the compound of general formula (IIIa),

[0266] wherein:

[0267] X is Br;

[0268] G.sup.1, R.sup.1a, R.sup.1b, R.sup.2-R.sup.8, r and s are as defined in general formula (IIIa).

##STR00079##

[0269] Provided is a method for preparing the compound of general formula (IIG) or the tautomer, mesomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof of the present disclosure, which comprises the following step:

[0270] removing a hydroxy protective group from a compound of general formula (IIGA) in the presence of a phase transfer catalyst (preferably n-tetrabutylammonium fluoride) to give the compound of general formula (IIG),

[0271] wherein:

[0272] R.sup.w is a hydroxy protective group, and is preferably

##STR00080##

[0273] Z, G.sup.1, R.sup.1a, R.sup.1b, R.sup.2-R.sup.4, R.sup.6, R.sup.8, R.sup.11a, R.sup.11b, q, t, k, r, n and s are as defined in general formula (IIG).

##STR00081##

[0274] Provided is a method for preparing the compound of general formula (IIGa) or the tautomer, mesomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof of the present disclosure, which comprises the following step:

[0275] removing a hydroxy protective group from a compound of general formula (IIGaA) in the presence of a phase transfer reagent (preferably n-tetrabutylammonium fluoride) to give the compound of general formula (IIGa),

[0276] wherein:

[0277] R.sup.w is a hydroxy protective group, and is preferably

##STR00082##

[0278] Z, G.sup.1, R.sup.1a, R.sup.1b, R.sup.2-R.sup.4, R.sup.6, R.sup.8, R.sup.11a, R.sup.11b, q, t, k, r, n and s are as defined in general formula (IIGa).

[0279] The reagents that provide alkaline conditions in the above synthesis schemes 1-4 described above include organic bases including, but not limited to, triethylamine, N,N-diisopropylethylamine, n-butyllithium, lithium diisopropylamide, sodium acetate, potassium acetate, sodium tert-butoxide and potassium tert-butoxide; and inorganic bases including, but not limited to, sodium hydride, potassium phosphate, sodium carbonate, potassium carbonate or cesium carbonate, sodium hydroxide, lithium hydroxide and potassium hydroxide; and are preferably sodium tert-butoxide.

[0280] The catalysts in the above synthesis schemes 1-4 described above include, but are not limited to tetrakis(triphenylphosphine)palladium(0), palladium dichloride, palladium acetate, bis(dibenzylideneacetone)palladium, chlorine(2-dicyclohexylphosphino-2′,4′,6′-triisopropyl-1,1′-biphenyl) [2-(2′-amino-1,1′-biphenyl)] palladium, [1,1′-bis(diphenylphosphino)ferrocene] palladium(II) dichloride, [1,1′-bis(dibenzylphosphino)ferrocene]palladium dichloride or tris(dibenzylideneacetone)dipalladium(0);

[0281] The phase transfer reagents in the synthesis schemes 5 and 6 described above include, but are not limited to, benzyltriethylammonium chloride (TEBA), tetrabutylammonium bromide, tetrabutylammonium chloride, n-tetrabutylammonium fluoride, tetrabutylammonium hydrogen sulfate (TBAB), trioctylmethylammonium chloride, dodecyltrimethylammonium chloride and tetradecyltrimethylammonium chloride, and are preferably n-tetrabutylammonium fluoride;

[0282] The above reactions are preferably conducted in solvents including, but not limited to acetic acid, methanol, ethanol, n-butanol, toluene, acetonitrile, tetrahydrofuran, dichloromethane, petroleum ether, ethyl acetate, n-hexane, dimethyl sulfoxide, 1,4-dioxane, ethylene glycol dimethyl ether, water or N,N-dimethylformamide, and mixtures thereof.

DETAILED DESCRIPTION

[0283] The following examples further illustrate the present disclosure, but the present disclosure is not limited thereto.

EXAMPLES

[0284] The structure of the compound was determined by nuclear magnetic resonance (NMR) spectroscopy and/or mass spectrometry (MS). NMR shifts (δ) are given in units of 10.sup.−6 (ppm). NMR spectra were measured using a Bruker AVANCE-400 nuclear magnetic resonance instrument, with deuterated dimethyl sulfoxide (DMSO-d.sub.6), deuterated chloroform (CDCl.sub.3) and deuterated methanol (CD.sub.3OD) as determination solvents, with tetramethylsilane (TMS) as internal standard.

[0285] Mass spectra were measured using Agilent 1200/1290 DAD-6110/6120 Quadrupole MS liquid chromatography-mass spectrometry system (manufacturer: Agilent; MS model: 6110/6120 Quadrupole MS),

[0286] waters ACQuity UPLC-QD/SQD (manufacturer: waters; MS model: waters ACQuity Qda Detector/waters SQ Detector) and

[0287] THERMO Ultimate 3000-Q Exactive (manufacturer: THERMO; MS model: THERMO Q Exactive).

[0288] High performance liquid chromatography (HPLC) was performed using Agilent HPLC 1200DAD, Agilent HPLC 1200VWD and Waters HPLC e2695-2489 high pressure liquid chromatography.

[0289] Chiral HPLC was performed on Agilent 1260 DAD HPLC.

[0290] HPLC preparation was performed using Waters 2545-2767, Waters 2767-SQ Detecor2, Shimadzu LC-20AP and Gilson GX-281 preparative chromatographs.

[0291] Chiral preparation was performed on a Shimadzu LC-20AP preparative chromatograph.

[0292] A CombiFlash Rf200 (TELEDYNE ISCO) system was used for rapid preparation.

[0293] Huanghai HSGF254 or Qingdao GF254 silica gel plates of specifications 0.15 mm to 0.2 mm were adopted for thin layer chromatography (TLC) analysis and 0.4 mm to 0.5 mm for TLC separation and purification.

[0294] The silica gel column chromatography generally used 200 to 300-mesh silica gel (Huanghai, Yantai) as the carrier.

[0295] The mean kinase inhibition rates and IC50 values were measured using a NovoStar microplate reader (BMG, Germany).

[0296] Known starting materials described herein may be synthesized using or according to methods known in the art, or may be purchased from ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc., Chembee Chemicals, and other companies.

[0297] In the examples, the reactions can be performed in an argon atmosphere or a nitrogen atmosphere unless otherwise specified.

[0298] The argon atmosphere or nitrogen atmosphere means that the reaction flask is connected to a balloon containing about 1 L of argon or nitrogen.

[0299] A hydrogen atmosphere means that the reaction flask is connected to a balloon containing about 1 L of hydrogen.

[0300] Parr 3916EKX hydrogenator, Qinglan QL-500 hydrogenator or HC2-SS hydrogenator was used in the pressurized hydrogenation reactions.

[0301] The hydrogenation reactions usually involve 3 cycles of vacuumization and hydrogen purge.

[0302] A CEM Discover-S 908860 microwave reactor was used in the microwave reactions.

[0303] In the examples, a solution refers to an aqueous solution unless otherwise specified.

[0304] In the examples, the reaction temperature was room temperature, i.e., 20° C. to 30° C., unless otherwise specified.

[0305] The monitoring of the reaction progress in the examples was conducted by thin layer chromatography (TLC). The developing solvent for reactions, the eluent system for column chromatography purification and the developing solvent system for thin layer chromatography included: A: dichloromethane/methanol system, B: n-hexane/ethyl acetate system, and C: petroleum ether/ethyl acetate system. The volume ratio of the solvents was adjusted according to the polarity of the compound, or by adding a small amount of basic or acidic reagents such as triethylamine and acetic acid.

Example 1

(S)-1-(3-fluoropropyl)-N-(4-((5R,7R)-7-methyl-6-(2,2,2-trifluoroethyl)-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)phenyl)pyrrolidin-3-amine 1

[0306] ##STR00083##

##STR00084## ##STR00085## ##STR00086##

Step 1

tert-Butyl (S)-(1-(3-fluoropropyl)pyrrolidin-3-yl)carbamate 1b

[0307] tert-Butyl (S)-pyrrolidin-3-ylcarbamate 1a (1.86 g, 10 mmol, Accela) was dissolved in N,N-dimethylformamide (20 mL), and diisopropylethylamine (1.55 g, 12 mmol) was added, followed by the dropwise addition of 1-fluoro-3-iodopropane (207 mg, 11 mmol). The reaction mixture was stirred for 12 h. Water (50 mL) was added, followed by the extraction with ethyl acetate (50 mL×3) and washing with water (50 mL×2) and saturated sodium chloride solution (50 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 1b (1.92 g, 78% yield).

Step 2

(S)-1-(3-fluoropropyl)pyrrolidin-3-amine 1c

[0308] Compound 1b (1.23 g, 5 mmol) was dissolved in dichloromethane (10 mL), and a 5 M solution of hydrogen chloride in 1,4-dioxane (2 mL) was added dropwise in an ice bath. After addition, the reaction mixture was stirred at room temperature for 1.5 h and concentrated under reduced pressure. Saturated sodium bicarbonate solution (25 mL) was added, followed by the extraction with ethyl acetate (15 mL×3). The organic phases were combined, washed with saturated sodium chloride solution (15 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to give the title compound 1c (702 mg, 96% yield).

Step 3

(R)-1-(3,4-bis(benzyloxy)phenyl)-N-(2,2,2-trifluoroethyl)propan-2-amine 1e

[0309] (R)-1-(3,4-bis(benzyloxy)phenyl)propan-2-amine 1d (7.0 g, 20.1 mmol, prepared by using the well-known method in “European Journal of Medicinal Chemistry, 2014, 67(23), 35-36”) was dissolved in dioxane (100 mL), and diisopropylethylamine (7.8 g, 60.4 mmol), 2,2,2-trifluoroethyl trifluoromethanesulfonate (9.4 g, 40.3 mmol, prepared as disclosed in “Example 59 on page 69 of the specification of Patent Application US20140249162”) was added. The reaction mixture was stirred in an oil bath at 80° C. in an argon atmosphere for 20 h. The reaction mixture was cooled and concentrated under reduced pressure. Saturated sodium bicarbonate solution (50 mL) was added, followed by the extraction with ethyl acetate (100 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 1e (7.6 g, 88% yield).

[0310] MS m/z (ESI): 430.3 [M+1].

Step 4

(R)-4-(2-((2,2,2-trifluoroethyl)amino)propyl)-1,2-benzenediol 1f

[0311] Compound 1e (3.3 g, 7.7 mmol) was dissolved in methanol (10 mL), and palladium hydroxide on carbon (0.5 g, 3.8 mmol) was added in an argon atmosphere. The reaction mixture was stirred under hydrogen balloon for 3 h, and filtered. The filtrate was concentrated under reduced pressure to give the title compound 1f (1.8 g, 96% yield).

Step 5

(1R,3R)-1-(4-bromophenyl)-3-methyl-2-(2,2,2-threefluoroethyl)-1,2,3,4-tetrahydroisoquinoline-6,7-diol 1g

[0312] Compound 1f (3.0 g, 12.0 mmol) was dissolved in toluene (100 mL), and acetic acid (1.4 g, 23.3 mmol) and 4-bromo-benzaldehyde (4.4 g, 23.8 mmol) were added. The reaction mixture was stirred in an oil bath at 80° C. for 16 h, and the reaction was terminated. The reaction mixture was cooled and concentrated under reduced pressure. Water (100 mL) was added, and the aqueous phase was adjusted to about pH 8 by adding saturated sodium bicarbonate solution (200 mL) and extracted with ethyl acetate (100 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 1g (3.5 g, 70% yield).

[0313] MS m/z (ESI): 416.0 [M+1].

Step 6

(5R,7R)-5-(4-bromophenyl)-7-methyl-6-(2,2,2-trifluoroethyl)-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinoline 1h

[0314] Compound 1g (3.5 g, 8.4 mmol) was dissolved in N,N-dimethylformamide (100 mL), and dibromomethane (1.9 g, 10.9 mmol) and cesium carbonate (3.6 g, 11.0 mmol) were added. The reaction mixture was stirred in an oil bath at 70° C. for 16 h, and the reaction was terminated. The reaction mixture was cooled and concentrated under reduced pressure. Water (100 mL) was added, followed by the extraction with ethyl acetate (100 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 1h (2.4 g, 67% yield).

Step 7

(S)-1-(3-fluoropropyl)-N-(4-((5R,7R)-7-methyl-6-(2,2,2-trifluoroethyl)-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)phenyl)pyrrolidin-3-amine 1

[0315] Compound 1h (600 mg, 1.4 mmol) was dissolved in dioxane (10 mL), and compound 1c (225 mg, 1.5 mmol), 2-dicyclohexylphosphine-2′,6′-bis(N,N-dimethylamino)-1,1′-biphenyl (3 mg, 0.007 mmol), tris(dibenzylideneacetone)dipalladium(0) (20 mg, 0.02 mmol) and sodium tert-butoxide (471 mg, 4.9 mmol) were added. The reaction mixture was stirred in an oil bath at 105° C. in an argon atmosphere for 16 h, and the reaction was terminated. The reaction mixture was cooled and concentrated. Saturated sodium bicarbonate solution (20 mL) was added, followed by the extraction with ethyl acetate (50 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 1 (449 mg, 65% yield).

[0316] MS m/z (ESI): 494.2 [M+1].

[0317] 1H NMR (400 MHz, CD.sub.3OD) 6.84 (d, 2H), 6.49 (s, 1H), 6.43 (d, 2H), 6.20 (s, 1H), 5.75 (d, 2H), 4.64 (s, 1H), 4.45-4.42 (m, 1H), 4.33-4.30 (m, 1H), 3.91-3.87 (m, 1H), 3.18-3.15 (m, 2H), 2.83-2.80 (m, 2H), 2.66-2.40 (m, 8H), 1.85-1.60 (m, 3H), 0.92 (d, 3H).

Example 2

(S)—N-(3,5-difluoro-4-((5S,7R)-7-methyl-6-(2,2,2-trifluoroethyl)-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)phenyl)-1-(3-fluoropropyl)pyrrolidin-3-amine 2

[0318] ##STR00087##

##STR00088##

Step 1

(1S,3R)-1-(4-bromo-2,6-difluorophenyl)-3-methyl-2-(2,2,2-threefluoroethyl)-1,2,3,4-tetrahydroisoquinoline-6,7-diol 2a

[0319] Compound 1f (310 mg, 1.2 mmol) was dissolved in toluene (5 mL), and acetic acid (598 mg, 10.0 mmol) and 4-bromo-2,6-difluorobenzaldehyde (384 mg, 1.7 mmol) were added. The reaction mixture was stirred in an oil bath at 80° C. for 16 h, and the reaction was terminated. The reaction mixture was cooled and concentrated under reduced pressure. Water (10 mL) was added, and the aqueous phase was adjusted to about pH 8 by adding saturated sodium bicarbonate solution (20 mL) and extracted with ethyl acetate (10 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 2a (390 mg, 72% yield).

[0320] MS m/z (ESI): 451.9 [M+1].

Step 2

(5S,7R)-5-(4-bromo-2,6-difluorophenyl)-7-methyl-6-(2,2,2-trifluoroethyl)-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinoline 2b

[0321] Compound 2a (130 mg, 0.3 mmol) was dissolved in N,N-dimethylformamide (10 mL), and dibromomethane (60 mg, 0.3 mmol) and cesium carbonate (121 mg, 0.4 mmol) were added. The reaction mixture was stirred in an oil bath at 70° C. for 16 h, and the reaction was terminated. The reaction mixture was cooled and concentrated. Water (10 mL) was added, followed by the extraction with ethyl acetate (10 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 2b (85 mg, 61% yield).

[0322] MS m/z (ESI): 464.0 [M+1].

Step 3

(S)—N-(3,5-difluoro-4-((5S,7R)-7-methyl-6-(2,2,2-trifluoroethyl)-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)phenyl)-1-(3-fluoropropyl)pyrrolidin-3-amine 2

[0323] Compound 2b (70 mg, 0.2 mmol) was dissolved in dioxane (10 mL), and compound 1c (29 mg, 0.2 mmol), 2-dicyclohexylphosphine-2′,6′-bis(N,N-dimethylamino)-1,1′-biphenyl (2 mg, 0.005 mmol), tris(dibenzylideneacetone)dipalladium(0) (2 mg, 0.002 mmol) and sodium tert-butoxide (43 mg, 0.5 mmol) were added. The reaction mixture was stirred in an oil bath at 105° C. in an argon atmosphere for 16 h, and the reaction was terminated. The reaction mixture was cooled and concentrated under reduced pressure. Saturated sodium bicarbonate solution (20 mL) was added, followed by the extraction with ethyl acetate (50 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 2 (449 mg, 58% yield).

[0324] MS m/z (ESI): 530.1 [M+1].

[0325] 1H NMR (400 MHz, CD.sub.3OD) 6.45 (s, 1H), 6.07 (s, 1H), 6.03-6.00 (m, 2H), 5.73 (d, 2H), 4.94 (s, 1H), 4.46-4.43 (m, 1H), 4.34-4.31 (m, 1H), 3.87-3.83 (m, 1H), 3.38-3.35 (m, 1H), 3.13-3.02 (m, 2H), 2.86-2.42 (m, 9H), 1.87-1.63 (m, 3H), 0.92 (d, 3H).

Example 3

2,2-Difluoro-3-((5R,7R)-5-(4-(((S)-1-(3-fluoropropyl)pyrrolidin-3-yl)amino)phenyl)-7-methyl-7,8-dihydro-[1,3]dioxolo[4,5-g]isoquinolin-6(5H)-yl)propan-1-ol 3

[0326] ##STR00089##

##STR00090## ##STR00091## ##STR00092##

Step 1

(R)—N-(1-(3,4-bis(benzyloxy)phenyl)propan-2-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropan-1-amine 3a

[0327] Compound 1d (1.0 g, 3.0 mmol) was dissolved in dioxane (20 mL), and diisopropylethylamine (1.2 g, 9.0 mmol) and 3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyltrifluoromethanesulfonate (1.0 g, 2.0 mmol, prepared by using the well-known method in “Bioorganic &Medicinal Chemistry Letters, 2018, 28(14), 2528-2532”) were added. The reaction mixture was stirred in an oil bath at 80° C. in an argon atmosphere for 20 h. The reaction mixture was cooled and concentrated under reduced pressure. Saturated sodium bicarbonate solution (50 mL) was added, followed by the extraction with ethyl acetate (100 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 3a (1.7 g, 84% yield).

[0328] MS m/z (ESI): 680.2 [M+1].

Step 2

(R)-4-(2-((3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)amino)propyl)benzene-1,2-diol 3b

[0329] Compound 3a (1.4 g, 2.0 mmol) was dissolved in methanol (10 mL), and palladium hydroxide on carbon (0.2 g) was added in an argon atmosphere. The reaction mixture was stirred under hydrogen balloon for 3 h, and filtered. The filtrate was concentrated under reduced pressure to give the title compound 3b (0.9 g, 94% yield).

Step 3

(1R,3R)-1-(4-bromophenyl)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3-methyl-1,2,3,4-tetrahydroisoquinoline-6,7-diol 3c

[0330] Compound 3b (0.8 g, 1.6 mmol) was dissolved in toluene (10 mL), and acetic acid (0.2 g, 3.2 mmol) and 4-bromo-benzaldehyde (0.6 g, 3.2 mmol) were added. The reaction mixture was stirred in an oil bath at 80° C. for 16 h, and the reaction was terminated. The reaction mixture was cooled and concentrated under reduced pressure. Water (10 mL) was added, and the reaction mixture was adjusted to about pH 8 by slowly adding saturated sodium bicarbonate solution (20 mL) and extracted with ethyl acetate (10 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 3c (0.6 g, 56% yield).

[0331] MS m/z (ESI): 666.1 [M+1].

Step 4

(5R,7R)-5-(4-bromophenyl)-6-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-7-methyl-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinoline 3d

[0332] Compound 3c (0.2 g, 0.3 mmol) was dissolved in N,N-dimethylformamide (10 mL), and dibromomethane (0.07 g, 0.4 mmol) and cesium carbonate (0.1 g, 0.4 mmol) were added. The reaction mixture was stirred in an oil bath at 70° C. for 16 h, and the reaction was terminated. The reaction mixture was cooled and concentrated under reduced pressure. Water (10 mL) was added, followed by the extraction with ethyl acetate (10 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 3d (0.1 g, 47% yield).

Step 5

tert-Butyl (S)-3-((4-((5R,7R)-6-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-7-methyl-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)phenyl)amino)pyrrolidine-1-carboxylate 3e

[0333] Compound 3d (60 mg, 0.09 mmol) was dissolved in dioxane (10 mL), and tert-butyl (S)-3-aminopyrrolidine-1-carboxylate (19 mg, 0.1 mmol), 2,2′-bis-(diphenylphosphino)-1,1′-binaphthyl (12 mg, 0.02 mmol), tris(dibenzylideneacetone)dipalladium(0) (20 mg, 0.02 mmol) and sodium tert-butoxide (29 mg, 0.3 mmol) were added. The reaction mixture was stirred in an oil bath at 80° C. in an argon atmosphere for 16 h, and the reaction was terminated. The reaction mixture was cooled and concentrated. Saturated sodium bicarbonate solution (10 mL) was added, followed by the extraction with ethyl acetate (10 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 3e (39 mg, 55% yield).

[0334] MS m/z (ESI): 784.3 [M+1].

Step 6

(S)—N-(4-((5R,7R)-6-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-7-methyl-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)phenyl)pyrrolidin-3-amine 3f

[0335] Compound 3e (39 mg, 0.05 mmol) was dissolved in dichloromethane (5 mL), and a 5 M solution of hydrogen chloride in 1,4-dioxane (1 mL) was added dropwise in an ice bath. After addition, the reaction mixture was stirred at room temperature for 1.5 h and concentrated under reduced pressure. Saturated sodium bicarbonate solution (5 mL) was added, followed by the extraction with ethyl acetate (5 mL×3). The organic phases were combined, washed with saturated sodium chloride solution (5 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to give the title compound 3f (32 mg, 93% yield).

Step 7

(S)—N-(4-((5R,7R)-6-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-7-methyl-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)phenyl)-1-(3-fluoropropyl)pyrrolidin-3-amine 3g

[0336] Compound 3f (32 mg, 0.04 mmol) was dissolved in N,N-dimethylformamide (5 mL), and diisopropylethylamine (4 mg, 0.05 mmol) was added, followed by the dropwise addition of 1-fluoro-3-iodopropane (10 mg, 0.05 mmol). The reaction mixture was stirred for 12 h. Water (5 mL) was added, followed by the extraction with ethyl acetate (5 mL×3). The organic phases were combined, washed with water (5 mL×2) and saturated sodium chloride solution (5 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 3g (22 mg, 73% yield).

Step 8

2,2-Difluoro-3-((5R,7R)-5-(4-(((S)-1-(3-fluoropropyl)pyrrolidin-3-yl)amino)phenyl)-7-methyl-7,8-dihydro-[1,3]dioxolo[4,5-g]isoquinolin-6(5H)-yl)propan-1-ol 3

[0337] Compound 3g (20 mg, 0.03 mmol) was dissolved in dichloromethane (5 mL), and a 1 M solution of n-tetrabutylammonium fluoride in tetrahydrofuran (1 mL) was added dropwise in an ice bath. After addition, the reaction mixture was stirred at room temperature for 1.5 h and concentrated under reduced pressure. Saturated sodium bicarbonate solution (5 mL) was added, followed by the extraction with ethyl acetate (5 mL×3). The organic phases were combined, washed with saturated sodium chloride solution (5 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 3 (5 mg, 33% yield).

[0338] MS m/z (ESI): 506.3 [M+1].

[0339] 1H NMR (400 MHz, CD.sub.3OD) 6.84 (d, 2H), 6.49 (s, 1H), 6.45 (d, 2H), 6.17 (s, 1H), 5.75 (s, 2H), 4.67 (s, 1H), 4.45-4.42 (m, 1H), 4.34-4.31 (m, 1H), 3.96-3.93 (m, 1H), 3.72-3.69 (m, 1H), 3.57-3.54 (m, 1H), 2.99-2.89 (m, 2H), 2.70-2.61 (m, 6H), 2.43-2.40 (m, 2H), 1.89-1.67 (m, 5H), 0.91 (d, 3H).

Example 4

2,2-Difluoro-3-((5S,7R)-5-(5-(((S)-1-(3-fluoropropyl)pyrrolidin-3-yl)amino)pyridin-2-yl)-7-methyl-7,8-dihydro-[1,3]dioxolo[4,5-g]isoquinolin-6(5H)-yl)propan-1-ol 4

[0340] ##STR00093##

##STR00094## ##STR00095##

Step 1

(1S,3R)-1-(5-bromopyridin-2-yl)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3-methyl-1,2,3,4-tetrahydroisoquinoline-6,7-diol 4a

[0341] The title compound 4a (256 mg, 75% yield) was obtained by following the synthesis scheme of Example 3 with the starting material 4-bromo-benzaldehyde in step 3 replaced by 5-bromopyridylaldehyde.

[0342] MS m/z(ESI): 667.1 [M+1].

Step 2

(5S,7R)-5-(5-bromopyridin-2-yl)-6-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-7-methyl-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinoline 4b

[0343] The title compound 4b (163 mg, 65% yield) was obtained by following the synthesis scheme of Example 3 with the compound 3c in step 4 replaced by compound 4a.

Step 3

tert-Butyl (S)-3-((6-((5S,7R)-6-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-7-methyl-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)pyridin yl)amino)pyrrolidine-1-carboxylate 4c

[0344] The title compound 4c (40 mg, 68% yield) was obtained by following the synthesis scheme of Example 3 with the compound 3d in step 5 replaced by compound 4b.

[0345] MS m/z(ESI): 785.3 [M+1].

Step 4

6-((5S,7R)-6-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-7-methyl-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)-N—((S)-pyrrolidin-3-yl)pyridin-3-amine 4d

[0346] The title compound 4d (30 mg, 95% yield) was obtained by following the synthesis scheme of Example 3 with the compound 3e in step 6 replaced by compound 4c.

Step 5

6-((5S,7R)-6-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-7-methyl-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)-N—((S)-1-(3-fluoropropyl)pyrrolidin-3-yl)pyridin-3-amine 4e

[0347] The title compound 4e (25 mg, 75% yield) was obtained by following the synthesis scheme of Example 3 with the compound 3f in step 7 replaced by compound 4d.

Step 6

2,2-Difluoro-3-((5S,7R)-5-(5-(((S)-1-(3-fluoropropyl)pyrrolidin-3-yl)amino)pyridin-2-yl)-7-methyl-7,8-dihydro-[1,3]dioxolo[4,5-g]isoquinolin-6(5H)-yl)propan-1-ol 4

[0348] The title compound 4 (5 mg, 29% yield) was obtained by following the synthesis scheme of Example 3 with the compound 3g in step 8 replaced by compound 4e.

[0349] MS m/z (ESI): 507.2 [M+1].

[0350] 1H NMR (400 MHz, CD.sub.3OD) 7.84 (s, 1H), 7.05-6.96 (m, 2H), 6.62 (s, 1H), 6.21 (s, 1H), 5.86 (d, 2H), 4.77 (s, 1H), 4.58-4.55 (m, 1H), 4.44-4.43 (m, 1H), 3.74-3.71 (m, 1H), 3.62-3.58 (m, 1H), 3.26-3.24 (m, 2H), 2.97-2.95 (m, 2H), 2.76-2.52 (m, 6H), 1.72-1.64 (m, 4H), 1.45-1.43 (m, 2H), 0.92 (d, 3H).

Example 5

3-(5S,7R)-5-(2,6-difluoro-4-(((S)-1-(3-fluoropropyl)pyrrolidin-3-yl)amino)phenyl)-7-methyl-7,8-dihydro-[1,3]dioxolo[4,5-g]isoquinolin-6(5H)-yl)-2,2-difluoropropan-1-ol 5

[0351] ##STR00096##

##STR00097## ##STR00098##

Step 1

(1S,3R)-1-(4-bromo-2,6-difluorophenyl)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3-methyl-1,2,3,4-tetrahydroisoquinoline-6,7-diol 5a

[0352] The title compound 5a (3.0 g, 47% yield) was obtained by following the synthesis scheme of Example 3 with the starting material 4-bromo-benzaldehyde in step 3 replaced by 4-bromo-2,6-difluorobenzaldehyde.

[0353] MS m/z(ESI): 702.0 [M+1].

Step 2

(5S,7R)-5-(4-bromo-2,6-difluorophenyl)-6-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-7-methyl-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinoline 5b

[0354] The title compound 5b (1.6 g, 55% yield) was obtained by following the synthesis scheme of Example 3 with the compound 3c in step 4 replaced by compound 5a.

Step 3

tert-Butyl (S)-3-((4-((5S,7R)-6-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-7-methyl-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)-3,5-difluorophenyl)amino)pyrrolidine-1-carboxylate 5c

[0355] The title compound 5c (1.5 g, 65% yield) was obtained by following the synthesis scheme of Example 3 with the compound 3d in step 5 replaced by compound 5b.

[0356] MS m/z(ESI): 820.3 [M+1].

Step 4

(S)—N-(4-((5S,7R)-6-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-7-methyl-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)-3,5-difluorophenyl)pyrrolidin-3-amine 5d

[0357] The title compound 5d (1.3 g, 99% yield) was obtained by following the synthesis scheme of Example 3 with the compound 3e in step 6 replaced by compound 5c.

Step 5

(S)—N-(4-((5S,7R)-6-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-7-methyl-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)-3,5-difluorophenyl)-1-(3-fluoropropyl)pyrrolidin-3-amine 5e

[0358] The title compound 5e (1.2 g, 92% yield) was obtained by following the synthesis scheme of Example 3 with the compound 3f in step 7 replaced by compound 5d.

Step 6

3-((5S,7R)-5-(2,6-difluoro-4-(((S)-1-(3-fluoropropyl)pyrrolidin-3-yl)amino)phenyl)-7-methyl-7,8-dihydro-[1,3]dioxolo[4,5-g]isoquinolin-6(5H)-yl)-2,2-difluoro-1-propanol 5

[0359] The title compound 5 (300 mg, 36% yield) was obtained by following the synthesis scheme of Example 3 with the compound 3g in step 8 replaced by compound 5e.

[0360] MS m/z (ESI): 542.2 [M+1].

[0361] .sup.1H NMR (400 MHz, CD.sub.3OD) 6.56 (s, 1H), 6.18-6.16 (m, 3H), 5.86-5.84 (m, 2H), 4.99 (s, 1H), 4.59-4.56 (m, 1H), 4.49-4.47 (m, 1H), 4.05 (br, 1H), 3.84-3.71 (m, 2H), 3.55-3.53 (m, 2H), 3.15-3.07 (m, 3H), 2.88-2.68 (m, 4H), 2.53-2.38 (m, 2H), 2.03-1.96 (m, 2H), 1.84 (br, 1H), 1.40-1.31 (m, 1H), 1.06 (d, 3H).

Example 6

N—((S-1-(3-fluoropropyl)pyrrolidin-3-yl)-6-((5S,7R)-7-methyl-6-(2,2,2-trifluoroethyl)-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)pyridin-3-amine 6

[0362] ##STR00099##

##STR00100##

Step 1

(1S,3R)-1-(5-bromopyridin-2-yl)-3-methyl-2-(2,2,2-threefluoroethyl)-1,2,3,4-tetrahydroisoquinoline-6,7-diol6a

[0363] The title compound 6a (600 mg, 60% yield) was obtained by following the synthesis scheme of Example 2 with the starting material 4-bromo-2,6-difluorobenzaldehyde in step 1 replaced by 5-bromopyridylaldehyde.

[0364] MS m/z(ESI): 417.0 [M+1].

Step 2

(5S,7R)-5-(5-bromopyridin-2-yl)-7-methyl-6-(2,2,2-trifluoroethyl)-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinoline 6b

[0365] The title compound 6b (450 mg, 73% yield) was obtained by following the synthesis scheme of Example 2 with the compound 2a in step 2 replaced by compound 6a. MS m/z(ESI): 431.0 [M+1].

Step 3

N—((S)-1-(3-fluoropropyl)pyrrolidin-3-yl)-6-((5S,7R)-7-methyl-6-(2,2,2-trifluoroethyl)-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)pyridin-3-amine 6

[0366] The title compound 6 (200 mg, 44% yield) was obtained by following the synthesis scheme of Example 2 with the compound 2b in step 2 replaced by compound 6b.

[0367] MS m/z(ESI): 495.2 [M+1].

[0368] .sup.1H NMR (400 MHz, CD.sub.3OD) 7.81 (s, 1H), 7.12 (d, 1H), 7.00 (d, 1H), 6.60 (s, 1H), 6.21 (s, 1H), 5.86 (d, 2H), 4.76 (s, 1H), 4.56-4.54 (m, 1H), 4.43-4.41 (m, 1H), 4.01-4.00 (m, 1H), 3.47-3.31 (m, 2H), 3.14-3.11 (m, 1H), 2.94-2.61 (m, 8H), 2.37-2.35 (m, 1H), 1.96-1.90 (m, 2H), 1.88-1.74 (m, 1H), 1.08 (d, 3H).

Example 7

(S)—N-(4-((5S,7R)-2,2-difluoro-7-methyl-6-(2,2,2-trifluoroethyl)-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)-3,5-difluorophenyl)-1-(3-fluoropropyl)pyrrolidin amine 7

[0369] ##STR00101##

Example 8

6-((5S,7R)-2,2-difluoro-7-methyl-6-(2,2,2-trifluoroethyl)-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)-N—((S)-1-(3-fluoropropyl)pyrrolidin-3-yl)pyridin-3-amine 8

[0370] ##STR00102##

Example 9

5-Fluoro-N—((S)-1-(3-fluoropropyl)pyrrolidin-3-yl)-6-((5S,7R)-7-methyl-6-(2,2,2-trifluoroethyl)-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)pyridin-3-amine 9

[0371] ##STR00103##

##STR00104##

Step 1

(1S,3R)-1-(5-bromo-3-fluoropyridin-2-yl)-3-methyl-2-(2,2,2-trifluoroethyl)-1,2,3,4-tetrahydroisoquinoline-6,7-diol 9a

[0372] The title compound 9a (115 mg, 55% yield) was obtained by following the synthesis scheme of Example 2 with the starting material 4-bromo-2,6-difluorobenzaldehyde in step 1 replaced by 5-bromo-3-fluoropyridyl-2-aldehyde.

Step 2

(5S,7R)-5-(5-bromo-3-fluoropyridin-2-yl)-7-methyl-6-(2,2,2-trifluoroethyl)-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinoline 9b

[0373] The title compound 9b (58 mg, 58% yield) was obtained by following the synthesis scheme of Example 2 with the compound 2a in step 2 replaced by compound 9a.

Step 3

5-Fluoro-N—((S)-1-(3-fluoropropyl)pyrrolidin-3-yl)-6-((5S,7R)-7-methyl-6-(2,2,2-trifluoroethyl)-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)pyridin-3-amine 9

[0374] The title compound 9 (18 mg, 48% yield) was obtained by following the synthesis scheme of Example 2 with the compound 2b in step 3 replaced by compound 9b.

[0375] MS m/z (ESI): 513.2 [M+1].

[0376] .sup.1H NMR (400 MHz, CD.sub.3OD) 7.66-7.65 (m, 1H), 6.74-6.73 (m, 1H), 6.61 (s, 1H), 6.18 (s, 1H), 5.85 (d, 2H), 4.56-4.54 (m, 1H), 4.46-4.44 (m, 1H), 4.00-3.97 (m, 1H), 3.53-3.51 (m, 1H), 3.32-3.29 (m, 2H), 3.06-3.02 (m, 1H), 2.98-2.92 (m, 2H), 2.79-2.76 (m, 1H), 2.66-2.60 (m, 3H), 2.56-2.51 (m, 2H), 2.36-2.35 (m, 1H), 1.96-1.88 (m, 2H), 1.74-1.73 (m, 1H), 1.07 (d, 3H).

Example 10

3-((5S,7R)-5-(2,6-difluoro-4-(((S)-1-(3-fluoropropyl)pyrrolidin-3-yl)amino)phenyl)-2,2,7-trimethyl-7,8-dihydro-[1,3]dioxolo[4,5-g]isoquinolin-6(5H)-yl)-2,2-difluoropropan-1-ol 10

[0377] ##STR00105##

Example 11

2,2-Difluoro-3-((5S,7R)-5-(3-fluoro-5-(((S)-1-(3-fluoropropyl)pyrrolidin-3-yl)amino)pyridin-2-yl)-7-methyl-7,8-dihydro-[1,3]dioxolo[4,5-g]isoquinolin-6(5H)-yl)propan-1-ol 11

[0378] ##STR00106##

##STR00107## ##STR00108##

Step 1

(1S,3R)-1-(5-bromo-3-fluoropyridin-2-yl)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3-methyl-1,2,3,4-tetrahydroisoquinoline-6,7-diol 11a

[0379] The title compound 11a (356 mg, 71% yield) was obtained by following the synthesis scheme of Example 3 with the starting material 4-bromo-benzaldehyde in step 3 replaced by 5-bromo-3-fluoropyridylaldehyde.

[0380] MS m/z(ESI): 685.1 [M+1].

Step 2

(5S,7R)-5-(5-bromo-3-fluoropyridin-2-yl)-6-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-7-methyl-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinoline 11b

[0381] The title compound 11b (169 mg, 61% yield) was obtained by following the synthesis scheme of Example 3 with the compound 3c in step 4 replaced by compound 11a.

Step 3

tert-Butyl (S)-3-(((6-((5S,7R)-6-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-7-methyl-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)-5-fluoropyridin-3-yl)amino)pyrrolidine-1-carboxylate 11c

[0382] The title compound 11c (92 mg, 69% yield) was obtained by following the synthesis scheme of Example 3 with the compound 3d in step 5 replaced by compound 11b.

Step 4

6-((5S,7R)-6-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-7-methyl-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)-5-fluoro-N—((S)-pyrrolidin-3-yl)pyridin-3-amine 11d

[0383] The title compound 11d (62 mg, 95% yield) was obtained by following the synthesis scheme of Example 3 with the compound 3e in step 6 replaced by compound 11c.

Step 5

6-((5S,7R)-6-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-7-methyl-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)-5-fluoro-N—((S)-1-(3-fluoropropyl)pyrrolidin-3-yl)pyridin-3-amine 11e

[0384] The title compound 11e (35 mg, 71% yield) was obtained by following the synthesis scheme of Example 3 with the compound 3f in step 7 replaced by compound 11d.

Step 6

2,2-Difluoro-3-((5S,7R)-5-(3-fluoro-5-(((S)-1-(3-fluoropropyl)pyrrolidin-3-yl)amino)pyridin-2-yl)-7-methyl-7,8-dihydro-[1,3]dioxolo[4,5-g]isoquinolin-6(5H)-yl)propan-1-ol 11

[0385] The title compound 11 (15 mg, 39% yield) was obtained by following the synthesis scheme of Example 3 with the compound 3g in step 8 replaced by compound 11e.

[0386] MS m/z (ESI): 525.2 [M+1].

[0387] .sup.1H NMR (400 MHz, CD.sub.3OD) 7.68-7.67 (m, 1H), 6.76-6.73 (m, 1H), 6.60 (s, 1H), 6.15 (s, 1H), 5.85 (d, 2H), 4.56-4.54 (m, 1H), 4.47-4.44 (m, 1H), 4.01-3.98 (m, 1H), 3.75-3.70 (m, 1H), 3.60-3.52 (m, 2H), 3.13-2.95 (m, 4H), 2.80-2.51 (m, 7H), 2.38-2.34 (m, 1H), 1.97-1.88 (m, 2H), 1.76-1.72 (m, 1H), 1.05 (d, 3H).

Example 12

2,2-Difluoro-3-((5S,7R)-5-(5-(((S)-1-(3-fluoropropyl)pyrrolidin-3-yl)amino)pyridin-2-yl)-7-methyl-7,8-dihydro-[1,3]dioxolo[4,5-g]isoquinolin-6(5H)-yl-2,2-th)propan-1-ol 12

[0388] ##STR00109##

##STR00110##

Step 1

(5S,7R)-5-(5-bromopyridin-2-yl)-6-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-7-methyl-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinoline-2,2-th 12a

[0389] Compound 4a (350 mg, 0.5 mmol) was dissolved in N,N-dimethylformamide (10 mL), and dibromodideuteromethane (277 mg, 1.6 mmol) and cesium carbonate (512 mg, 1.6 mmol) were added. The reaction mixture was stirred in an oil bath at 70° C. for 16 h, and the reaction was terminated. The reaction mixture was cooled and concentrated under reduced pressure. Water (10 mL) was added, followed by the extraction with ethyl acetate (10 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 12a (170 mg, 48% yield).

[0390] MS m/z(ESI): 681.1 [M+1].

Step 2

6-((5S,7R)-6-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-7-methyl-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl-2,2-d.SUB.2.)—N—((S)-1-(3-fluoropropyl)pyrrolidin-3-yl)pyridin-3-amine 12b

[0391] Compound 12a (170 mg, 0.25 mmol) was dissolved in dioxane (10 mL), and compound 1c (44 mg, 0.3 mmol), 2,2′-bis-(diphenylphosphino)-1,1′-binaphthyl (12 mg, 0.02 mmol), tris(dibenzylideneacetone)dipalladium(0) (20 mg, 0.02 mmol) and sodium tert-butoxide (29 mg, 0.3 mmol) were added. The reaction mixture was stirred in an oil bath at 80° C. in an argon atmosphere for 16 h, and the reaction was terminated. The reaction mixture was cooled and concentrated. Saturated sodium bicarbonate solution (10 mL) was added, followed by the extraction with ethyl acetate (10 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 12b (103 mg, 55% yield).

Step 3

2,2-Difluoro-3-((5S,7R)-5-(5-(((S)-1-(3-fluoropropyl)pyrrolidin-3-yl)amino)pyridin-2-yl)-7-methyl-7,8-dihydro-[1,3]dioxolo[4,5-g]isoquinolin-6(5H)-yl-2,2-th)propan-1-ol 12

[0392] Compound 12b (103 mg, 0.14 mmol) was dissolved in dichloromethane (5 mL), and a 1 M solution of n-tetrabutylammonium fluoride in tetrahydrofuran (1 mL) was added dropwise in an ice bath. After addition, the reaction mixture was stirred at room temperature for 1.5 h and concentrated under reduced pressure. Saturated sodium bicarbonate solution (5 mL) was added, followed by the extraction with ethyl acetate (5 mL×3). The organic phases were combined, washed with saturated sodium chloride solution (5 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 12 (35 mg, 49% yield).

[0393] MS m/z (ESI): 509.1 [M+1].

[0394] .sup.1H NMR (400 MHz, CD.sub.3OD) 8.01 (dd, 1H), 7.68 (d, 1H), 7.63 (d, 1H), 6.74 (s, 1H), 6.47 (s, 1H), 5.19 (s, 1H), 4.64-4.62 (m, 1H), 4.54-4.52 (m, 1H), 4.42 (br, 1H), 3.83-3.71 (m, 5H), 3.45-3.41 (m, 5H), 3.09-3.05 (m, 1H), 2.91-2.82 (m, 1H), 2.67-2.62 (m, 1H), 2.21-2.12 (m, 4H), 1.14 (d, 3H).

Example 13

2,2-Difluoro-3-((5S,7R)-5-(5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-7-methyl-7,8-dihydro-[1,3]dioxolo[4,5-g]isoquinolin-6(5H)-yl-2,2-th)propan-1-ol 13

[0395] ##STR00111##

##STR00112##

Step 1

6-((5S,7R)-6-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-7-methyl-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl-2,2-d.SUB.2.)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine 13a

[0396] Compound 12a (86 mg, 0.13 mmol) was dissolved in dioxane (10 mL), and the compound 1-(3-fluoropropyl)azetidin-3-amine (44 mg, 0.3 mmol, prepared as disclosed in “Example 1 on page 50 of the specification of Patent Application WO2019228443”), 2,2′-bis-(diphenylphosphino)-1,1′-binaphthyl (12 mg, 0.02 mmol), tris(dibenzylideneacetone)dipalladium(0) (20 mg, 0.02 mmol) and sodium tert-butoxide (29 mg, 0.3 mmol) were added. The reaction mixture was stirred in an oil bath at 80° C. in an argon atmosphere for 16 h, and the reaction was terminated. The reaction mixture was cooled and concentrated. Saturated sodium bicarbonate solution (10 mL) was added, followed by the extraction with ethyl acetate (10 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 13a (47 mg, 51% yield).

Step 2

2,2-Difluoro-3-((5S,7R)-5-(5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-7-methyl-7,8-dihydro-[1,3]dioxolo[4,5-g]isoquinolin-6(5H)-yl-2,2-d.SUB.2.)propan-1-ol 13

[0397] Compound 13a (39 mg, 0.05 mmol) was dissolved in dichloromethane (5 mL), and a 1 M solution of n-tetrabutylammonium fluoride in tetrahydrofuran (1 mL) was added dropwise in an ice bath. After addition, the reaction mixture was stirred at room temperature for 1.5 h and concentrated under reduced pressure. Saturated sodium bicarbonate solution (5 mL) was added, followed by the extraction with ethyl acetate (5 mL×3). The organic phases were combined, washed with saturated sodium chloride solution (5 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 13 (11 mg, 42% yield).

[0398] MS m/z (ESI): 495.2 [M+1].

[0399] .sup.1H NMR (400 MHz, CD.sub.3OD) 7.80 (d, 1H), 7.04 (d, 1H), 6.94 (dd, 1H), 6.61 (s, 1H), 6.19 (s, 1H), 4.64 (br, 2H), 4.54-4.52 (m, 1H), 4.45-4.42 (m, 1H), 4.15-4.10 (m, 1H), 3.85-3.83 (m, 2H), 3.76-3.68 (m, 1H), 3.64-3.56 (m, 1H), 3.14-2.97 (m, 4H), 2.78-2.68 (m, 3H), 2.59-2.52 (m, 1H), 1.84-1.74 (m, 2H), 1.05 (d, 3H).

Example 14

2,2-Difluoro-3-((5S,7R)-5-(5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-7-methyl-7,8-dihydro-[1,3]dioxolo[4,5-g]isoquinolin-6(5H)-yl)propan-1-ol 14

[0400] ##STR00113##

##STR00114## ##STR00115##

Step 1

tert-Butyl 3-((6-((5S,7R)-6-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-7-methyl-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)pyridin-3-yl)amino)azetidine carboxylate 14a

[0401] Compound 4b (1.8 g, 2.65 mmol) was dissolved in toluene (50 mL), and tert-butyl 3-aminoazetidine-1-carboxylate (0.9 g, 5.21 mmol), 2,2′-bis-(diphenylphosphino)-1,1′-binaphthyl (165 mg, 0.26 mmol), tris(dibenzylideneacetone)dipalladium(0) (243 mg, 0.26 mmol) and sodium tert-butoxide (763 mg, 7.95 mmol) were added. The reaction mixture was stirred in an oil bath at 80° C. in an argon atmosphere for 16 h, and the reaction was terminated. The reaction mixture was cooled and concentrated. Saturated sodium bicarbonate solution (50 mL) was added, followed by the extraction with ethyl acetate (50 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 14a (1.2 g, 59% yield).

[0402] MS m/z(ESI): 771.2 [M+1].

Step 2

N-(azetidin-3-yl)-6-((5S,7R)-6-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-7-methyl-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)pyridin-3-amine 14b

[0403] Compound 14a (1.2 g, 1.56 mmol) was dissolved in dichloromethane (25 mL), and a 4 M solution of hydrogen chloride in 1,4-dioxane (2 mL) was added dropwise in an ice bath. After addition, the reaction mixture was stirred at room temperature for 1.5 h and concentrated under reduced pressure. Saturated sodium bicarbonate solution (15 mL) was added, followed by the extraction with ethyl acetate (15 mL×3). The organic phases were combined, washed with saturated sodium chloride solution (15 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to give the title compound 14b (0.85 g, 81% yield).

Step 3

6-((5S,7R)-6-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-7-methyl-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin amine 14c

[0404] Compound 14b (0.82 g, 1.22 mmol) was dissolved in N,N-dimethylformamide (10 mL), and diisopropylethylamine (0.48 mg, 3.68 mmol) was added, followed by the dropwise addition of 1-fluoro-3-iodopropane (0.35 mg, 1.84 mmol). The reaction mixture was stirred for 12 h. Water (15 mL) was added, followed by the extraction with ethyl acetate (15 mL×3). The organic phases were combined, washed with water (15 mL×2) and saturated sodium chloride solution (15 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 14c (0.7 g, 78% yield).

Step 4

2,2-Difluoro-3-((5S,7R)-5-(5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-7-methyl-7,8-dihydro-[1,3]dioxolo[4,5-g]isoquinolin-6(5H)-yl)propan-1-ol 14

[0405] Compound 14c (200 mg, 0.4 mmol) was dissolved in tetrahydrofuran (25 mL), and a 1 M solution of n-tetrabutylammonium fluoride in tetrahydrofuran (2 mL) was added dropwise in an ice bath. After addition, the reaction mixture was stirred at room temperature for 1.5 h and concentrated under reduced pressure. Saturated sodium bicarbonate solution (10 mL) was added, followed by the extraction with ethyl acetate (10 mL×3). The organic phases were combined, washed with saturated sodium chloride solution (10 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 14 (50 mg, 25% yield).

[0406] MS m/z (ESI): 493.2 [M+1].

[0407] .sup.1H NMR (400 MHz, CD.sub.3OD) 7.80 (d, 1H), 7.05 (d, 1H), 6.93 (dd, 1H), 6.61 (s, 1H), 6.20 (s, 1H), 5.85 (d, 2H), 4.77 (s, 1H), 4.54-4.52 (m, 1H), 4.45-4.43 (m, 1H), 4.15-4.12 (m, 1H), 3.88-3.85 (m, 2H), 3.76-3.57 (m, 3H), 3.14-3.05 (m, 4H), 2.75-2.69 (m, 3H), 2.57-2.53 (m, 1H), 1.84-1.76 (m, 2H), 1.06 (d, 3H).

Example 15

(5S,7R)-5-(2,6-difluoro-4-(((S)-1-(3-fluoropropyl)pyrrolidin-3-yl)oxy)phenyl)-7-methyl (2,2,2-trifluoroethyl)-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinoline 15

[0408] ##STR00116##

##STR00117## ##STR00118##

Step 1

(1S,3R)-1-(4-(benzyloxy)-2,6-difluorophenyl)-3-methyl-2-(2,2,2-trifluoroethyl)-1,2,3,4-tetrahydroisoquinoline-6,7-diol 15a

[0409] Compound 1f (167 mg, 0.7 mmol) was dissolved in toluene (100 mL), and trifluoroacetic acid (115 mg, 1.0 mmol) and 4-(benzyloxy)-2,6-difluorobenzaldehyde (250 mg, 1.0 mmol, prepared as disclosed in “Example 5 on page 61 of the specification of Patent Application WO2008080001”) were added. The reaction mixture was stirred in an oil bath at 80° C. for 16 h, and the reaction was terminated. The reaction mixture was cooled and concentrated under reduced pressure. Water (100 mL) was added, and the aqueous phase was adjusted to about pH 8 by adding saturated sodium bicarbonate solution (200 mL) and extracted with ethyl acetate (100 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 15a (235 mg, 73% yield).

[0410] MS m/z (ESI): 480.1 [M+1].

Step 2

(5S,7R)-5-(4-(benzyloxy)-2,6-difluorophenyl)-7-methyl-6-(2,2,2-trifluoroethyl)-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinoline 15b

[0411] Compound 15a (580 mg, 1.2 mmol) was dissolved in N,N-dimethylformamide (10 mL), and dibromomethane (421 g, 2.4 mmol) and cesium carbonate (1.2 g, 3.6 mmol) were added. The reaction mixture was stirred in an oil bath at 70° C. for 16 h, and the reaction was terminated. The reaction mixture was cooled and concentrated under reduced pressure. Water (10 mL) was added, followed by the extraction with ethyl acetate (100 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 15b (458 mg, 77% yield).

[0412] MS m/z (ESI): 492.1 [M+1].

Step 3

3,5-difluoro-4-((5S,7R)-7-methyl-6-(2,2,2-trifluoroethyl)-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)phenol 15c

[0413] Compound 15b (450 mg, 0.9 mmol) was dissolved in methanol (10 mL), and palladium hydroxide on carbon (0.1 g) was added in an argon atmosphere. The reaction mixture was stirred under hydrogen balloon for 3 h, and filtered. The filtrate was concentrated under reduced pressure to give the title compound 15c (0.3 g, 93% yield).

[0414] MS m/z (ESI): 402.1 [M+1].

Step 4

tert-Butyl (S)-3-(3,5-difluoro-4-((5S,7R)-7-methyl-6-(2,2,2-trifluoroethyl)-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)phenoxy)pyrrolidine-1-carboxylate 15d

[0415] tert-Butyl (R)-3-hydroxypyrrolidine-1-carboxylate (82 mg, 0.4 mmol) was dissolved in a dry tetrahydrofuran (5 mL) solution, and tri-n-butylphosphine (240 mg, 1.2 mmol) and azodicarboxylic acid dipiperidide compound (300 mg, 1.2 mmol) were successively added in an ice bath. After stirring for 1 h, compound 15c (159 mg, 0.4 mmol) was added. The reaction mixture was stirred in an argon atmosphere for another 3 h and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 15d (153 mg, 67% yield).

[0416] MS m/z (ESI): 571.0 [M+1].

Step 5

(5S,7R)-5-(2,6-difluoro-4-(((S)-pyrrolidin-3-yl)oxy)phenyl)-7-methyl-6-(2,2,2-trifluoroethyl)-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinoline 15e

[0417] Compound 15d (240 mg, 0.4 mmol) was dissolved in dichloromethane (5 mL), and a 5 M solution of hydrogen chloride in 1,4-dioxane (1 mL) was added dropwise in an ice bath. After addition, the reaction mixture was stirred at room temperature for 1.5 h and concentrated under reduced pressure. Saturated sodium bicarbonate solution (5 mL) was added, followed by the extraction with ethyl acetate (5 mL×3). The organic phases were combined, washed with saturated sodium chloride solution (5 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to give the title compound 15e (220 mg, 90% yield).

Step 6

(5S,7R)-5-(2,6-difluoro-4-(((S)-1-(3-fluoropropyl)pyrrolidin-3-yl)oxy)phenyl)-7-methyl-6-(2,2,2-trifluoroethyl)-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinoline 15

[0418] Compound 15e (200 mg, 0.4 mmol) was dissolved in N,N-dimethylformamide (5 mL), and diisopropylethylamine (44 mg, 0.4 mmol) was added, followed by the dropwise addition of 1-fluoro-3-iodopropane (160 mg, 0.9 mmol). The reaction mixture was stirred for 12 h. Water (5 mL) was added, followed by the extraction with ethyl acetate (5 mL×3). The organic phases were combined, washed with water (5 mL×2) and saturated sodium chloride solution (5 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 15 (50 mg, 22% yield).

[0419] MS m/z (ESI): 531.1 [M+1].

[0420] .sup.1H NMR (400 MHz, CD.sub.3OD) 6.59 (s, 1H), 6.52-6.49 (m, 2H), 6.18 (s, 1H), 5.85 (d, 2H), 5.15 (s, 1H), 4.91-4.89 (m, 1H), 4.56-4.53 (m, 1H), 4.46-4.44 (m, 1H), 3.49-3.47 (m, 1H), 3.20-3.16 (m, 1H), 2.93-2.88 (m, 4H), 2.67-2.63 (m, 2H), 2.56-2.51 (m, 2H), 2.40-2.36 (m, 1H), 1.98-1.89 (m, 4H), 1.08 (d, 3H).

Example 16

1-(3-Fluoropropyl)-N-(4-((5R,7R)-7-methyl-6-(2,2,2-trifluoroethyl)-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)phenyl)azetidin-3-amine 16

[0421] ##STR00119##

[0422] The title compound 16 (27 mg, 41% yield) was obtained by following the synthesis scheme of Example 1 with the compound 1c in step 7 replaced by 1-(3-fluoropropyl)azetidin-3-amine (prepared as disclosed in “Example 1 on page 50 of the specification of Patent Application WO2019228443”).

[0423] MS m/z (ESI): 480.1 [M+1].

[0424] .sup.1H NMR (400 MHz, CD.sub.3OD) 7.04 (d, 2H), 6.64 (s, 1H), 6.61-6.52 (m, 2H), 6.31 (s, 1H), 5.87 (d, 2H), 4.69-4.61 (m, 2H), 4.53-4.39 (m, 3H), 4.18-4.16 (m, 1H), 3.94-3.90 (m, 1H), 3.48-3.37 (m, 4H), 3.10-3.05 (m, 1H), 2.87-2.82 (m, 1H), 2.61-2.56 (m, 1H), 2.07-1.96 (m, 3H), 1.09 (d, 3H).

Example 17

N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((5S,7R)-7-methyl-6-(2,2,2-trifluoroethyl)-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl)pyridin-3-amine 17

[0425] ##STR00120##

##STR00121##

[0426] Compound 6b (100 mg, 0.2 mmol) was dissolved in dioxane (5 mL), and the compound 1-(3-fluoropropyl)azetidin-3-amine (47 mg, 0.4 mmol, prepared as disclosed in “Example 1 on page 50 of the specification of Patent Application WO2019228443”), 2-dicyclohexylphosphine-2′,6′-bis(N,N-dimethylamino)-1,1′-biphenyl (3 mg, 0.007 mmol), tris(dibenzylideneacetone)dipalladium(0) (20 mg, 0.02 mmol) and sodium tert-butoxide (48 mg, 0.5 mmol) were added. The reaction mixture was stirred in an oil bath at 105° C. in an argon atmosphere for 16 h, and the reaction was terminated. The reaction mixture was cooled and concentrated. Saturated sodium bicarbonate solution (20 mL) was added, followed by the extraction with ethyl acetate (50 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 17 (25 mg, 22% yield).

[0427] MS m/z (ESI): 481.2 [M+1].

[0428] .sup.1H NMR (400 MHz, CD.sub.3OD) 7.78 (s, 1H), 7.14 (d, 1H), 6.98 (d, 1H), 6.60 (s, 1H), 6.20 (s, 1H), 5.83 (d, 2H), 4.76 (s, 1H), 4.54-4.52 (m, 1H), 4.44-4.42 (m, 1H), 4.13-4.09 (m, 1H), 3.84-3.81 (m, 2H), 3.47-3.45 (m, 1H), 3.30-2.90 (m, 4H), 2.71-2.53 (m, 4H), 1.82-1.74 (m, 2H), 1.08 (d, 3H).

Example 18

2,2-Difluoro-3-((5R,7R)-5-(4-((1-(3-fluoropropyl)azetidin-3-yl)amino)phenyl)-7-methyl-7,8-dihydro-[1,3]dioxolo[4,5-g]isoquinolin-6(5H)-yl)propan-1-ol 18

[0429] ##STR00122##

##STR00123##

Step 1

3-((5R,7R)-5-(4-bromophenyl)-7-methyl-7,8-dihydro-[1,3]dioxolo[4,5-g]isoquinolin-6(5H)-yl)-2,2-difluoropropan-1-ol 18a

[0430] Compound 3d (300 mg, 0.4 mmol) was dissolved in dichloromethane (10 mL), and a 1 M solution of n-tetrabutylammonium fluoride in tetrahydrofuran (2 mL) was added dropwise in an ice bath. After addition, the reaction mixture was stirred at room temperature for 1.5 h and concentrated under reduced pressure. Saturated sodium bicarbonate solution (10 mL) was added, followed by the extraction with ethyl acetate (10 mL×3). The organic phases were combined, washed with saturated sodium chloride solution (10 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 18a (84 mg, 43% yield).

Step 2

2,2-Difluoro-3-((5R,7R)-5-(4-((1-(3-fluoropropyl)azetidin-3-yl)amino)phenyl)-7-methyl-7,8-dihydro-[1,3]dioxolo[4,5-g]isoquinolin-6(5H)-yl)propan-1-ol 18

[0431] Compound 18a (150 mg, 0.34 mmol) was dissolved in dioxane (5 mL), and the compound 1-(3-fluoropropyl)azetidin-3-amine (90 mg, 0.68 mmol, prepared as disclosed in “Example 1 on page 50 of the specification of Patent Application WO2019228443”), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (20 mg, 0.03 mmol), palladium acetate (4 mg, 0.02 mmol) and cesium carbonate (222 mg, 0.68 mmol) were added. The reaction mixture was stirred in an oil bath at 105° C. in an argon atmosphere for 16 h, and the reaction was terminated. The reaction mixture was cooled and concentrated. Saturated sodium bicarbonate solution (20 mL) was added, followed by the extraction with ethyl acetate (50 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 18 (25 mg, 15% yield).

[0432] MS m/z (ESI): 492.2 [M+1].

[0433] .sup.1H NMR (400 MHz, CD.sub.3OD) 6.95 (d, 2H), 6.61 (s, 1H), 6.50 (d, 2H), 6.28 (s, 1H), 5.87 (d, 2H), 4.78 (s, 1H), 4.52-4.51 (m, 1H), 4.44-4.42 (m, 1H), 4.10-4.09 (m, 1H), 3.86-3.78 (m, 3H), 3.71-3.66 (m, 1H), 3.29-3.28 (m, 1H), 3.11-2.98 (m, 3H), 2.75-2.67 (m, 4H), 2.54-2.49 (m, 1H), 1.82-1.74 (m, 2H), 1.04 (d, 3H).

Example 19

2,2-Difluoro-3-((5R,7R)-5-(4-(((S)-1-(3-fluoropropyl)pyrrolidin-3-yl)amino)phenyl)-7-methyl-7,8-dihydro-[1,3]dioxolo[4,5-g]isoquinolin-6(5H)-yl-2,2-th)propan-1-ol 19

[0434] ##STR00124##

##STR00125##

Step 1: (5S,7R)-5-(4-bromophenyl)-6-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-7-methyl-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinoline-2,2-th 19a

[0435] Compound 3c (2.6 g, 4 mmol) was dissolved in N,N-dimethylformamide (10 mL), and dibromodideuteromethane (2.8 g, 16 mmol) and cesium carbonate (5.1 g, 16 mmol) were added. The reaction mixture was stirred in an oil bath at 70° C. for 16 h, and the reaction was terminated. The reaction mixture was cooled and concentrated under reduced pressure. Water (100 mL) was added, followed by the extraction with ethyl acetate (100 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 19a (1.3 g, 49% yield).

Step 2

(S)—N-(4-((5R,7R)-6-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-7-methyl-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]isoquinolin-5-yl-2,2-th)phenyl)-1-(3-fluoropropyl)pyrrolidin-3-amine 19b

[0436] Compound 19a (1.0 g, 1.5 mmol) was dissolved in dioxane (20 mL), and compound 1c (0.26 g, 1.8 mmol), 2,2′-bis-(diphenylphosphino)-1,1′-binaphthyl (0.1 g, 0.2 mmol), tris(dibenzylideneacetone)dipalladium(0) (0.1 g, 0.1 mmol) and sodium tert-butoxide (0.3 g, 3.0 mmol) were added. The reaction mixture was stirred in an oil bath at 80° C. in an argon atmosphere for 16 h, and the reaction was terminated. The reaction mixture was cooled and concentrated. Saturated sodium bicarbonate solution (20 mL) was added, followed by the extraction with ethyl acetate (50 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 19b (0.6 g, 52% yield).

Step 3

2,2-Difluoro-3-((5R,7R)-5-(4-(((S)-1-(3-fluoropropyl)pyrrolidin-3-yl)amino)phenyl)-7-methyl-7,8-dihydro-[1,3]dioxolo[4,5-g]isoquinolin-6(5H)-yl-2,2-th)propan-1-ol 19

[0437] Compound 19b (0.3 g, 0.4 mmol) was dissolved in dichloromethane (5 mL), and a 1 M solution of n-tetrabutylammonium fluoride in tetrahydrofuran (2 mL) was added dropwise in an ice bath. After addition, the reaction mixture was stirred at room temperature for 1.5 h and concentrated under reduced pressure. Saturated sodium bicarbonate solution (10 mL) was added, followed by the extraction with ethyl acetate (10 mL×3). The organic phases were combined, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 19 (63 mg, 31% yield).

[0438] MS m/z (ESI): 508.0 [M+1].

[0439] .sup.1H NMR (400 MHz, CD.sub.3OD) 6.95 (d, 2H), 6.61 (s, 1H), 6.56 (d, 2H), 6.29 (s, 1H), 4.78 (s, 1H), 4.56-4.54 (m, 1H), 4.47-4.44 (m, 1H), 4.05-4.01 (m, 1H), 3.86-3.64 (m, 3H), 3.11-2.96 (m, 2H), 2.84-2.53 (m, 8H), 2.37-2.32 (m, 1H), 1.99-1.90 (m, 2H), 1.77-1.72 (m, 1H), 1.04 (d, 3H).

Example 20 (Comparative Example)

(7R,9S)-8-(2,2-difluoro-3-hydroxypropyl)-9-(5-((1-(3-fluoropropyl)azetidin yl)amino)pyridin-2-yl)-7-methyl-6,7,8,9-tetrahydro-1H-pyrrolo[3,4-h]isoquinoline-1,3(2H)-dione 20

[0440] ##STR00126##

##STR00127## ##STR00128##

Step 1

(1S,3R)-1-(5-bromopyridin-2-yl)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3-methyl-1,2,3,4-tetrahydroisoquinoline-7,8-diol 20a

[0441] Compound 3b (0.8 g, 1.6 mmol) was dissolved in toluene (10 mL), and acetic acid (0.2 g, 3.2 mmol) and 5-bromopyridylaldehyde (0.6 g, 3.2 mmol) were added. The reaction mixture was stirred in an oil bath at 80° C. for 16 h, and the reaction was terminated. The reaction mixture was cooled and concentrated under reduced pressure. Water (20 mL) was added, and the reaction mixture was adjusted to about pH 8 by slowly adding saturated sodium bicarbonate solution (20 mL) and extracted with ethyl acetate (50 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 20a (0.3 g, 28% yield).

Step 2

(1S,3R)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-1-(5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-3-methyl-1,2,3,4-tetrahydroisoquinoline-7,8-diol 20b

[0442] Compound 20a (300 mg, 0.28 mmol) was dissolved in dioxane (10 mL), and 1-(3-fluoropropyl)azetidin-3-amine (111 mg, 0.84 mmol), 2-dicyclohexylphosphine-2′,6′-bis(N,N-dimethylamino)-1,1′-biphenyl (15 mg, 0.04 mmol), tris(dibenzylideneacetone)dipalladium(0) (56 mg, 0.06 mmol) and sodium tert-butoxide (269 mg, 2.8 mmol) were added. The reaction mixture was stirred in an oil bath at 105° C. in an argon atmosphere for 16 h, and the reaction was terminated. The reaction mixture was cooled and concentrated under reduced pressure. Saturated sodium bicarbonate solution (20 mL) was added, followed by the extraction with ethyl acetate (50 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 20b (109 mg, 54% yield).

Step 3

(1S,3R)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-1-(5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-3-methyl-1,2,3,4-tetrahydroisoquinoline-7,8-diyl bis(trifluoromethylsulfonate) 20c

[0443] Compound 20b (400 mg, 0.56 mmol) was dissolved in dichloromethane (25 mL), and triethylamine (170 mg, 1.68 mmol), N,N-dimethylpyridin-4-amine (7 mg, 0.06 mmol) and N-phenylbis(trifluoromethylsulfonimide) (500 mg, 1.40 mmol) were added. The reaction mixture was allowed to react at room temperature for 3 h, and the reaction was terminated. The reaction mixture was cooled and concentrated under reduced pressure. Saturated sodium bicarbonate solution (20 mL) was added, followed by the extraction with ethyl acetate (50 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 20c (450 mg, 82% yield).

Step 4

(1S,3R)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-1-(5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-3-methyl-1,2,3,4-tetrahydroisoquinoline-7,8-dinitrile 20d

[0444] Compound 20c (30 mg, 0.03 mmol) was dissolved in N,N-dimethylformamide (5 mL), and zinc cyanide (11 mg, 0.1 mmol), tris(dibenzylideneacetone)dipalladium(0) (3 mg, 0.003 mmol) and 1,1′-bis(diphenylphosphino)ferrocene (2 mg, 0.004 mmol) were added. The reaction mixture was stirred in an oil bath at 100° C. in an argon atmosphere for 3 h, and the reaction was terminated. The reaction mixture was cooled and concentrated under reduced pressure. Saturated sodium bicarbonate solution (20 mL) was added, followed by the extraction with ethyl acetate (50 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 20d (16 mg, 72% yield).

[0445] MS m/z (ESI): 737.3 [M+1].

Step 5

(7R,9S)-8-(2,2-difluoro-3-hydroxypropyl)-9-(5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-7-methyl-6,7,8,9-tetrahydro-1H-pyrrolo[3,4-h]isoquinoline-1,3(2H)-dione 20

[0446] Compound 20d (16 mg, 0.02 mmol) was dissolved in sulfuric acid (30 mg, 0.3 mmol). The solution was stirred in an oil bath at 90° C. for 2 h, and the reaction was terminated. The reaction mixture was cooled and concentrated under reduced pressure. Saturated sodium bicarbonate solution (10 mL) was added, followed by the extraction with ethyl acetate (10 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 20 (3 mg, 29% yield).

[0447] MS m/z (ESI): 518.2 [M+1].

[0448] .sup.1H NMR (400 MHz, CD.sub.3OD) 7.67 (d, 1H), 7.62 (d, 1H), 7.57 (d, 1H), 7.28 (d, 1H), 6.96 (dd, 1H), 5.93 (s, 1H), 4.53-4.51 (m, 1H), 4.43-4.41 (m, 1H), 4.10-4.05 (m, 1H), 3.94-3.88 (m, 2H), 3.81-3.78 (m, 2H), 3.30-3.26 (m, 2H), 2.98-2.95 (m, 2H), 2.81-2.63 (m, 5H), 1.83-1.77 (m, 2H), 1.13 (d, 3H).

Example 21 (Comparative Example)

(8R,10S)-9-(2,2-difluoro-3-hydroxypropyl)-10-(5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-8-methyl-2,3,7,8,9,10-hexahydropyrrolophthalazine-1,4-dione 21

[0449] ##STR00129##

[0450] Compound 20 (6 mg, 0.01 mmol) was dissolved in ethanol (5 mL), and hydrazine hydrate (5 mg, 0.1 mmol) was added. The reaction mixture was stirred in an oil bath at 80° C. for 24 h, and the reaction was terminated. The reaction mixture was cooled and concentrated under reduced pressure. Saturated sodium bicarbonate solution (10 mL) was added, followed by the extraction with ethyl acetate (10 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by column chromatography with developing solvent system B to give the title compound 21 (3 mg, 49% yield).

[0451] MS m/z (ESI): 533.2 [M+1].

[0452] .sup.1H NMR (400 MHz, CD.sub.3OD) 8.08 (d, 1H), 7.71 (d, 1H), 7.55 (d, 1H), 7.27 (d, 1H), 6.97 (dd, 1H), 6.65 (s, 1H), 4.55-4.53 (m, 1H), 4.45-4.43 (m, 1H), 4.18-4.11 (m, 1H), 3.97-3.91 (m, 4H), 3.29-3.15 (m, 4H), 2.87-2.81 (m, 4H), 2.71-2.63 (m, 1H), 1.86-1.78 (m, 2H), 1.11 (d, 3H).

Biological Evaluation

[0453] The present disclosure is further described and explained below with reference to test examples, but these examples are not intended to limit the scope of the present disclosure.

Test Example 1. Inhibition of Binding of E to ER by Compounds Disclosed Herein

[0454] The compounds of the present disclosure can inhibit the binding of E (estrogen) to ER (estrogen receptor), thereby blocking the binding of the complex of E and ER to ERE (estrogen response element), and consequently the expression of downstream luciferase proteins. The inhibitory effects of the compounds of the present disclosure on the binding of E to ER was tested by using the following method.

1. Objective

[0455] This experiment is intended to test the inhibitory effects of the compounds on the binding of E to ER, and to evaluate the in vitro activity of the compounds according to the IC.sub.50 values.

2. Method

[0456] An ERE was cloned upstream of the luciferase gene, and MCF-7/ERE-luciferase monoclonal cells were selected by transfection of MCF-7 (TCHu74, National Collection of Authenticated Cell Cultures). The MCF-7/ERE-luciferase cells were seeded into a 96-well plate with an MEM (hyclone, SH30024.01B) medium containing 10% charcoal stripped FBS (Moregate, FBSF), 1% sodium pyruvate (sigma, S8636), 1% non-essential amino acids (sigma, M7145) and 500 μg/mL G418 at a density of 30,000 cells/well and cultured at 37° C. with 5% CO.sub.2. 20 mM stock solutions of the drugs are prepared, serially diluted 10-fold with 100% DMSO, and then diluted 20-fold with the medium. After the cells were cultured for 24 h, the medium was removed. 0.1 nM estradiol (sigma, E2758) and 10 μL of a medium-diluted drug were added to each well, and DMSO was added to the control group. The mixtures were well mixed by gentle shaking. The cells were cultured in an incubator at 37° C. with 5% CO.sub.2 for 24 h, and then the cell culture liquid was removed, followed by the addition of 50 μL of a luciferase substrate (Promega, E6110) to each well. The plate was let stand at room temperature in the dark for 10-15 min, and the chemiluminescence signal values were determined.

3. Results

[0457] The inhibitory effects of the compounds of the present disclosure on the binding of E to ER were tested through the above experiment. The chemiluminescence signal value was plotted against the compound concentration (in log form) using Graphpad Prism, and the IC.sub.50 values of the compounds were obtained. The results are shown in Table 1.

TABLE-US-00003 TABLE 1 Inhibitory effects of the compounds disclosed herein on the binding of E to ER Example IC.sub.50 (nM) 1 0.14 2 0.10 3 0.44 4 0.37 5 0.05 6 0.27 9 0.46 11 0.35 12 1.13 13 0.69 14 0.5 15 0.19 16 0.27 17 0.43 18 0.07 19 0.07

[0458] Conclusion: The compounds claimed by the present disclosure had significant inhibitory effects on the binding of E to ER.

Test Example 2. Inhibitory Effects of Compounds Disclosed Herein on Proliferation of MCF-7 Cells

1. Objective

[0459] This experiment was intended to test the inhibitory effects of the compounds disclosed herein on the proliferation of MCF-7 Cells by using the ATP method, and to evaluate the in vitro activity of the compounds according to the IC.sub.50 values.

2. Method

[0460] MCF-7 cells (TCHu74, National Collection of Authenticated Cell Cultures) were seeded into a 96-well plate with an MEM (hyclone, SH30024.01B) medium containing 10% FBS (Gibco, 10099-141), 1% sodium pyruvate (sigma, 58636) and 1% non-essential amino acid (sigma, M7145) at a density of 4,000 cells/well and cultured at 37° C. with 5% CO.sub.2. 20 mM stock solutions of the compounds are prepared, serially diluted with 100% DMSO to a final concentration of 1000×, and then diluted 20-fold with a medium containing 2% FBS. After the cells were cultured for 24 h, the medium was removed. 90 μL of the medium containing 2% FBS and 10 μL of a drug were added to each well, and 10 μL of DMSO was added to the control group. The mixtures were well mixed by gentle shaking. The blank group contained only 100 μL of the medium containing 2% FBS. The cells were cultured in an incubator at 37° C. with 5% CO.sub.2 for 72 h, and then 50 μL of mixed Cell Titer-Glo (Promega, G7571) was added to each well. The mixtures were well mixed by shaking and let stand at room temperature for 10 min, and the chemiluminescence signal values were determined.

3. Data Analysis

[0461] The chemiluminescence signal value was plotted against the compound concentration (in log form) using Graphpad Prism, and the IC.sub.50 values of the compounds were obtained. The results are shown in Table 2.

TABLE-US-00004 TABLE 2 Inhibitory effects of the compounds disclosed herein on proliferation of MCF-7 cells Example IC.sub.50 (nM) 1 0.31 2 0.17 3 0.15 4 0.28 5 0.03 6 0.34 9 0.35 11 0.23 12 1.12 13 0.29 14 0.36 15 0.3 17 0.44 18 0.07 19 0.07 20 >10000 21 >10000

[0462] Note: Examples 20 and 21 are comparative examples.

[0463] Conclusion: The compounds claimed by the present disclosure had significant inhibitory effects on the proliferation of MCF-7 cells, and Comparative Examples 20 and 21 showed no activity.

Test Example 3. Biological Evaluation of Inhibition of Proliferation of ERα Mutant-Expressing MCF7 Cells by Compounds Disclosed Herein

1. Objective

[0464] This experiment was intended to test the inhibitory activity of the compounds disclosed herein against the proliferation of ERα mutant-expressing MCF7 cells.

2. Method

Site-Directed Mutagenesis and Cell Line Construction

[0465] The mutant estrogen receptor α (ERα) Y537S of human ERα protein was obtained by site-directed mutagenesis in a two-primer PCR manner using the cDNA of wild-type ESR1 gene (Accession No. NM000125) as a template. The sequences of the primers used in the mutation are as follows (the underlined nucleotides are the sites of mutation): Y537S: F-AAG AAC GTG GTG CCC CTC TCT GAC CTG CTG CTG GAG ATG (SEQ ID NO: 1); R-CAT CTC CAG CAG CAG GTC AGA GAG GGG CAC CAC GTT CTT (SEQ ID NO: 2). The cDNA of the mutant ESR1 was cloned into the lentiviral vector of interest pCDH-CMV-MCS-EF1-Puro. The lentiviral plasmid bearing the mutant ESR1 gene sequence and the lentiviral packaging plasmid were then transfected into HEK-293T cells (ATCC, CRL-3216) by using Lipofectamine 3000 Transfection Reagent (ThermoFisher Scientific, Cat # L3000075). Forty-eight hours after transfection, the virus-bearing medium supernatant was filtered and ultracentrifuged to obtain a virus precipitate, which was then resuspended in an appropriate amount of medium. The suspension was added to MCF7 cells (ATCC, HTB-22), followed by the addition of polybrene at a final concentration of 8 μg/mL. The mixture was incubated overnight. Two days after transfection, 1 μg/mL puromycin was added to the cell culture liquid for resistance screening. About two weeks later, an MCF7 cell line capable of stably expressing the ERαY537S mutant was obtained.

Cell Proliferation Inhibition Assay

[0466] The ERα mutant-expressing MCF7 cells were cultured in an MEM (GE Healthcare, SH30024.01) complete medium containing 10% fetal bovine serum. On the first day of the experiment, the cells were seeded into a 96-well plate with complete medium at a density of 3,000 cells/well to form 100 μL of cell suspension per well. The plate was incubated overnight in a cell incubator at 37° C. with 5% CO.sub.2. The following day the medium was removed, and 135 μL of an MEM incomplete medium containing 2% fetal bovine serum, and 15 μL of a test compound prepared at different concentrations with the incomplete medium were added to each well. The final concentrations of the compound were 9 concentration points obtained by 4-fold serial dilution from 100 nM. A blank control containing 0.5% DMSO was set. The plate was incubated in a cell incubator at 37° C. with 5% CO.sub.2 for 144 h. On day 8, the 96-well cell culture plate was taken out. 150 μL of CellTiter-Glo® Luminescent Cell Viability Assay (Promega, G7573) was added to each well. The plate was let stand at room temperature for 10 min, and the luminescence signal values was read using a multilabel microplate reader (PerkinElmer, VICTOR 3). The IC.sub.50 values for the inhibitory activity of the compounds were calculated according to the compound concentrations and the luminescence signal values and are shown in Table 3.

3. Results

[0467]

TABLE-US-00005 TABLE 3 IC.sub.50 values for the inhibitory effects of the compounds disclosed herein on the proliferation of ERα mutant-expressing MCF7 cells Example IC.sub.50 (nM) 1 1.38 2 0.78 3 0.43 4 0.89 5 0.07 6 0.82 9 1.35 11 0.71 12 3.49 13 1.36 14 1.27 15 0.68 17 2.19 18 0.25 19 0.43

[0468] Conclusion: The compounds claimed by the present disclosure had significant inhibitory effects on the proliferation of ERα mutant-expressing MCF7 cells.

Test Example 4. Degradation of ERα by Compounds Disclosed Herein

1. Objective

[0469] This experiment was intended to test the degradation of ERα by the compounds disclosed herein. This method was used to test the degradation of ERα by the compounds disclosed herein.

2. Method

[0470] ERα-positive breast cancer cell line MCF-7 cells were cultured in a DMEM/F12 medium (HyClone, SH30023.01) containing 10% fetal bovine serum (Corning, 35-010-CV). On the first day of the experiment, after being digested, the cells were washed once with a phenol red-free DMEM/F12 medium (ThermoFisher, 11039-021) containing 5% charcoal stripped fetal bovine serum (BIOSUN, BS-0004-500), then resuspended, and counted, and the cell density was adjusted to 1.79×10.sup.5 cells/mL. 280 μL of the cell suspension was added to each well of a 48-well plate (Corning, 3548), and the cells were cultured overnight in an incubator at 37° C. with 5% CO.sub.2. The following day the compounds were serially diluted with DMSO and further diluted with the phenol red-free DMEM/F12 medium containing 5% charcoal-stripped fetal bovine serum. 20 μL of a diluted compound was added to each well of the 48-well plate at final concentrations of 3000, 300, 30, 3, 0.3, 0.03 and 0.003 nM. The 48-well plate was placed in the incubator for 16 to 18 h. A 96-well plate was coated with the capture antibody from the human ERα/NR3A1 total protein assay kit (R&D, DYC5715-5). 1 μg/mL capture antibody was prepared in PBS, and 100 μL of the antibody was added to each well of the 96-well plate (Corning, 3590). The plate was placed in an incubator at 26° C. overnight. On day 3, the antibody-coated 96-well plate was washed once with PBS, and 200 μL of PBS containing 1% BSA was added to each well. The plate was incubated in an incubator at 37° C. for 1.5 h to be blocked. The cell culture medium supernatant was discarded. The cells were washed once with PBS, and 60 μL of a cell lysis buffer was added to each well. The cell lysis buffer was PBS containing 6 M urea, 1 mM EDTA, 0.5% TritonX-100, 1 mM PMSF and a protease inhibitor (Roche, 04693159001). The cells were lysed on ice for 15 min, and 300 μL of PBS containing 1 mM EDTA and 0.5% TritonX-100 was added to each well to dilute the urea to 1 M. The blocking buffer in the blocked 96-well plate was discarded, and 100 μL of diluted cell lysis buffer was added to each well. The plate was incubated in an incubator at 37° C. for 2 h and washed 5 times with PBS. A biotinylated assay antibody was diluted to 0.4 μg/mL with PBS containing 1% BSA, and then 100 μL of the assay antibody was added to each well. The plate was incubated in an incubator at 37° C. for 1 h. The plate was then washed five more times, and 100 μL of avidin-HRP diluted 200-fold with PBS containing 1% BSA was added to each well. The plate was incubated at 37° C. for 30 min. The plate was washed five more times, and 100 μL of TMB substrate was added to each well. The plate was incubated at room temperature until blue color appeared, and 100 μL of stop solution was added to each well. OD450 signal values were read using a PHERAstar multi-mode microplate reader. The IC.sub.50 values for the inhibitory activity of the compounds were calculated using Graphpad Prism software. The maximum degradation rates of the compounds are the ratios of the level of ERα remaining in the cells after the cells were treated with 3000 nM compounds to the level of ERα remaining in cells after the cells were treated with 3000 nM Fulvestrant.

3. Results

[0471] The EC.sub.50 values determined for the compounds disclosed herein in the degradation of ERα are shown in Table 4.

TABLE-US-00006 TABLE 4 Degradation of ERα by the compounds disclosed herein Example EC.sub.50 (nM) Emax degradation (%) Fulvestrant 0.06 100 1 0.32 94 2 0.24 104 3 0.19 129 4 0.21 115 5 0.04 108 6 0.17 104 9 1.04 107 11 0.45 91 12 0.15 101 13 0.38 107 14 0.5 92 15 0.15 95 17 1.24 104 18 0.1 106 19 0.13 100 20 >10000 — 21 >10000 —

[0472] Note: Examples 20 and 21 are comparative examples.

[0473] Conclusion: The compounds claimed by the present disclosure significantly degraded ERα, and Comparative Examples 20 and 21 showed no activity.

Test Example 5. Inhibition of Enzymatic Activity of Site of Metabolism of Midazolam in Human Liver Microsome CYP3A4 by Compounds Disclosed Herein

[0474] The inhibition of the enzymatic activity of the site of metabolism of midazolam in human liver microsome CYP3A4 by the compounds disclosed herein was tested by using the following method.

I. Materials and Instruments

[0475] 1. Phosphate buffer (20×PBS, purchased from Sangon);

[0476] 2. NADPH (ACROS, A2646-71-1);

[0477] 3. Human liver microsome (Corning Gentest, Cat No, 452161, Lot No. 9050002, Donor, 36);

[0478] 4. ABI QTrap 4000 LC-MS System (AB Sciex);

[0479] 5. ZORBAX Extend-C18, 3×50 mm, 3.5 μm (Agilent, USA);

[0480] 6. CYP probe substrate (midazolam, TRC, M343000/3 μM) and positive control inhibitor (ketoconazole, SIGMA, Cat No. K1003-100MG).

II. Procedures

[0481] A 100 mM PBS buffer was prepared. A 0.25 mg/mL microsome solution, a 7.5 mM MgCl.sub.2 and a 5 mM NADPH solution were prepared using the buffer. A 30 mM stock solution was diluted with DMSO to obtain 30 mM, 10 mM, 3 mM, 1 mM, 0.3 mM, 0.03 mM, 0.003 mM and 0 mM serial solutions I, which were further diluted 200-fold with phosphate buffer (PBS) to obtain serial test solutions II (150, 50, 15, 5, 1.5, 0.15, 0.015 and 0 μM). A midazolam working solution was diluted with PBS to 15 μM.

[0482] 40 μL of a 0.25 mg/mL microsome solution prepared in 7.5 mM MgCl.sub.2 and 20 μL of each of the 15 μM midazolam working solution and the compound working solutions (150, 50, 15, 5, 1.5, 0.15, 0.015 and 0 μM) were measured out and well mixed. Ketoconazole at the same concentration was used in place of the compounds as a positive control group. The mixtures were pre-incubated with 5 mM NADPH solution at 37° C. for 5 min at the same time. After 5 minutes, 20 μL of NADPH was added to each well to start reactions. The mixtures were incubated for 30 min. Duplicate samples were set for all of the incubated samples. After 30 minutes, 250 μL of internal standard-containing acetonitrile was added to all the samples. The mixtures were well mixed, shaken at 800 rpm for 10 min, and then centrifuged at 3700 rpm for 10 min. 100 μL of the supernatant and 80 μL of ultrapure water were well mixed and subjected to LC-MS/MS analysis.

[0483] The IC.sub.50 values of the drugs for the site of metabolism of midazolam in CYP3A4 were calculated using Graphpad Prism and are shown in Table 5.

TABLE-US-00007 TABLE 5 IC.sub.50 values of the compounds disclosed herein for the site of metabolism of midazolam in human liver microsome CYP3A4 Example IC.sub.50 (μM) 1 15.76 2 >30 3 10 12 8.18 13 15.38

[0484] Conclusion: The compounds claimed by the present disclosure had weak inhibitory effects on the site of metabolism of midazolam in human liver microsome CYP3A4, showing better safety, suggesting that metabolic drug interaction based on the site of metabolism of midazolam in CYP3A4 does not occur.

Test Example 6. Inhibition of Enzymatic Activity of Site of Metabolism of Testosterone in Human Liver Microsome CYP3A4 by Compounds Disclosed Herein

[0485] The inhibition of the enzymatic activity of the site of metabolism of testosterone in human liver microsome CYP3A4 by the compounds disclosed herein was tested by using the following method.

I. Materials and Instruments

[0486] 1. Phosphate buffer (20×PBS, purchased from Sangon);

[0487] 2. NADPH (ACROS, A2646-71-1);

[0488] 3. Human liver microsome (Corning Gentest, Cat No, 452161, Lot No. 905002, Donor36);

[0489] 4. ABI QTrap 4000 LC-MS System (AB Sciex);

[0490] 5. ZORBAX Extend-C18, 3×50 mm, 3.5 μm (Agilent, USA);

[0491] 6. CYP probe substrate (testosterone, Vocko, CAS No. [58-22-0]/75 μM) and positive control inhibitor (ketoconazole, SIGMA, Cat No. K1003-100MG).

II. Procedures

[0492] A 100 mM PBS buffer was prepared. A 100 mM PBS buffer was prepared. A 0.25 mg/mL microsome solution, a 7.5 mM MgCl.sub.2 and a 5 mM NADPH solution were prepared using the buffer. A 30 mM stock solution was diluted with DMSO to obtain 30 mM, 10 mM, 3 mM, 1 mM, 0.3 mM, 0.03 mM, 0.003 mM and 0 mM serial solutions I, which were further diluted 200-fold with phosphate buffer (PBS) to obtain serial test solutions II (150, 50, 15, 5, 1.5, 0.15, 0.015 and 0 μM). A testosterone working solution was diluted with PBS to 375

[0493] 40 μL of a 0.25 mg/mL microsome solution prepared in 7.5 mM MgCl.sub.2 and 20 μL of each of the 375 μM testosterone working solution and the compound working solutions (150, 50, 15, 5, 1.5, 0.15, 0.015 and 0 μM) were measured out and well mixed. Ketoconazole at the same concentration was used in place of the compounds as a positive control group. The mixtures were pre-incubated with 5 mM NADPH solution at 37° C. for 5 min at the same time. After 5 minutes, 20 μL of NADPH was added to each well to start reactions. The mixtures were incubated for 30 min. After 30 minutes, 250 μL of internal standard-containing acetonitrile was added to all the samples. The mixtures were well mixed, shaken at 800 rpm for 10 min, and then centrifuged at 3700 rpm for 10 min. 100 μL of the supernatant and 80 μL of ultrapure water were well mixed and subjected to LC-MS/MS analysis.

[0494] The IC.sub.50 values of the drugs for the site of metabolism of testosterone in CYP3A4 were calculated using Graphpad Prism and are shown in Table 6.

TABLE-US-00008 TABLE 6 IC.sub.50 values of the compounds disclosed herein for the site of metabolism of testosterone in human liver microsome CYP3A4 Example IC.sub.50 (μM) 1 7.31 2 >30 3 14.57 4 15.50 12 23.77 13 >30

[0495] Conclusion: The compounds claimed by the present disclosure had weak inhibitory effects on the site of metabolism of testosterone in human liver microsome CYP3A4, showing better safety.

Test Example 7. Time-Dependent Inhibition of Enzymatic Activity of Site of Metabolism of Midazolam in Human Liver Microsome CYP3A4 by Compounds Disclosed Herein

[0496] The time-dependent inhibition of the CYP enzymatic activity of the site of metabolism of midazolam in human liver microsome CYP3A4 by the compounds disclosed herein was tested by using the following method.

I. Materials and Instruments

[0497] 1. Phosphate buffer (20×PBS, purchased from Sangon);

[0498] 2. NADPH (ACROS, A2646-71-1);

[0499] 3. Human liver microsome (Corning Gentest, Cat No, 452161, Lot No. 9050002, Donor, 36);

[0500] 4. ABI QTrap 4000 LC-MS System (AB Sciex);

[0501] 5. ZORBAX Extend-C18, 3×50 mm, 3.5 μm (Agilent, USA);

[0502] 6. CYP probe substrate (midazolam, TRC, M343000/3 μM) and positive control inhibitor (verapamil, Adamas Reagent Co, Ltd, Cat No. 25904A).

II. Procedures

[0503] A 100 mM PBS buffer was prepared. A 0.25 mg/mL microsome solution, a 7.5 mM MgCl.sub.2 and a 5 mM NADPH solution were prepared using the buffer. A 30 mM stock solution was diluted with DMSO to obtain 30 mM, 10 mM, 3 mM, 1 mM, 0.3 mM, 0.1 mM, 0.03 mM and 0 mM serial solutions I, which were further diluted 200-fold with phosphate buffer (PBS) to obtain serial test solutions II (150, 50, 15, 5, 1.5, 0.5, 0.15 and 0 μM). A midazolam working solution was diluted with PBS to 15 μM.

[0504] The serial test solutions prepared above were well mixed by shaking and aliquoted in 20 μL portions into corresponding reaction plates (+NADPH, T0 and −NADPH groups were set). Three parallel tests were set. 40 μL of the liver microsome working solution was added to each 96-well plate. 20 μL of a corresponding substrate solution was added to the T0 plate. 20 μL of NADPH was added to the TO plate and +NADPH group. A timer was started, and the plates were incubated in a water bath at 37° C. After 30 min of incubation, the TO plate was taken out, and the reaction was terminated with 250 μL of an internal standard-containing ACN solution. The +NADPH group was supplemented with 20 μL of the corresponding substrate solution, and the −NADPH group was supplemented with 20 μL of the corresponding substrate solution and 20 μL of NADPH. A timer was started, and the plates were incubated in a water bath at 37° C. After 30 min of incubation, the plates were taken out, and the reactions were terminated with 250 μL of an internal standard-containing ACN solution. Then the plates were shaken on a shaker at 800 rpm for 10 min and centrifuged at 4000 rpm for 15 min. 100 μL of the supernatant and 80 μL of ultrapure water were well mixed and subjected to LC-MS/MS analysis.

[0505] The IC.sub.50 values of the drugs for the site of metabolism of midazolam in CYP3A4 were calculated using Graphpad Prism and are shown in Table 7.

TABLE-US-00009 TABLE 7 IC.sub.50 values and IC.sub.50 shifts of the compounds disclosed herein for the site of metabolism of midazolam in human liver microsome CYP3A4 (+)NADPH (−)NADPH Example IC.sub.50 (μM) IC.sub.50 (μM) IC.sub.50 Shift 4 2.43 2.47 1.0 12 9.16 10.17 0.9 13 13.11 19.59 0.7

[0506] Conclusion: The compounds claimed by the present disclosure had weak inhibitory effects on the site of metabolism of midazolam in human liver microsome CYP3A4, showing better safety, suggesting that metabolic drug interaction based on the site of metabolism of midazolam in CYP3A4 does not occur.

Test Example 8. Time-Dependent Inhibition of Enzymatic Activity of Site of Metabolism of Testosterone in Human Liver Microsome CYP3A4 by Compounds Disclosed Herein

[0507] The time-dependent inhibition of the enzymatic activity of the site of metabolism of testosterone in human liver microsome CYP3A4 by the compounds disclosed herein was tested by using the following method.

I. Materials and Instruments

[0508] 1. Phosphate buffer (20×PBS, purchased from Sangon);

[0509] 2. NADPH (ACROS, A2646-71-1);

[0510] 3. Human liver microsome (Corning Gentest, Cat No, 452161, Lot No. 905002, Donor36);

[0511] 4. ABI QTrap 4000 LC-MS System (AB Sciex);

[0512] 5. ZORBAX Extend-C18, 3×50 mm, 3.5 μm (Agilent, USA);

[0513] 6. CYP probe substrate (testosterone, Vocko, CAS No. [58-22-0]/75 μM) and positive control inhibitor (verapamil, Adamas Reagent Co Ltd, Cat No. 25904A).

II. Procedures

[0514] A 100 mM PBS buffer was prepared. A 0.25 mg/mL microsome solution, a 7.5 mM MgCl.sub.2 and a 5 mM NADPH solution were prepared using the buffer. A 30 mM stock solution was diluted with DMSO to obtain 30 mM, 10 mM, 3 mM, 1 mM, 0.3 mM, 0.1 mM, 0.03 mM and 0 mM serial solutions I, which were further diluted 200-fold with phosphate buffer (PBS) to obtain serial test solutions II (150, 50, 15, 5, 1.5, 0.5, 0.15 and 0 μM). A midazolam working solution was diluted with PBS to 15 μM.

[0515] The serial test solutions prepared above were well mixed by shaking and aliquoted in 20 μL portions into corresponding reaction plates (+NADPH, T0 and −NADPH groups were set). Three parallel tests were set. 40 μL of the liver microsome working solution was added to each 96-well plate. 20 μL of a corresponding substrate solution was added to the T0 plate. 20 μL of NADPH was added to the TO plate and +NADPH group. A timer was started, and the plates were incubated in a water bath at 37° C. After 30 min of incubation, the TO plate was taken out, and the reaction was terminated with 250 μL of an internal standard-containing ACN solution. The +NADPH group was supplemented with 20 μL of the corresponding substrate solution, and the −NADPH group was supplemented with 20 μL of the corresponding substrate solution and 20 μL of NADPH. A timer was started, and the plates were incubated in a water bath at 37° C. After 30 min of incubation, the plates were taken out, and the reactions were terminated with 250 μL of an internal standard-containing ACN solution. Then the plates were shaken on a shaker at 800 rpm for 10 min and centrifuged at 4000 rpm for 15 min. 100 μL of the supernatant and 80 μL of ultrapure water were well mixed and subjected to LC-MS/MS analysis.

[0516] The IC.sub.50 values of the drugs for the site of metabolism of testosterone in CYP3A4 were calculated using Graphpad Prism and are shown in Table 8.

TABLE-US-00010 TABLE 8 IC.sub.50 values and IC.sub.50 shifts of the compounds disclosed herein for the site of metabolism of testosterone in human liver microsome CYP3A4 (+)NADPH (−)NADPH Example IC.sub.50 (μM) IC.sub.50 (μM) IC.sub.50 Shift 4 2.4 2.5 1.0 12 11.6 25.2 0.5 13 >30 >30 No TDI

[0517] Conclusion: The compounds claimed by the present disclosure had weak inhibitory effects on the site of metabolism of testosterone in human liver microsome CYP3A4, showing better safety, suggesting that metabolic drug interaction based on CYP3A4 does not occur.

Test Example 9

1. Objective

[0518] The blocking of hERG potassium currents by the compounds disclosed herein was tested in a stable cell strain transfected with hERG potassium channels using automated patch clamp.

2. Method

2.1. Materials and Instruments

2.1.1. Materials:

[0519]

TABLE-US-00011 Reagent Supplier Cat. No. FBS GIBCO 10099 Sodium pyruvate solution sigma S8636-100ML MEM non-essential amino sigma M7145-100ML acid solution (100×) G418 sulfate Enzo ALX-380-013-G005 MEM Hyclone SH30024.01B hERG cDNA Synthesized by Gene sequence GENEWIZ NM_000238.4-

2.1.2. Instruments:

[0520]

TABLE-US-00012 Instrument Supplier Model Patchliner 4 channel nanion 2-03-03100-002 Patchliner cleaning station nanion 2-02-03201-005 Patchliner cell bank nanion 2-02-03105-000 Elektrodenchloridierer nanion 3-02-03533-000 Patchliner HEAK EPC10 patch clamp nanion 1-01-10012-000 amplifier Osmometer Gonoter Gonoter 030 pH meter Mettle Toledo FE20

2.2. Procedures of Automated Patch Clamp

[0521] An HEK293-hERG stable cell strain was subcultured in an MEM/EBSS medium (10% FBS, 400 μg/mL G418, 1% MEM non-essential amino acid solution (100×), 1% sodium pyruvate solution) at a density of 1:4, and an automated patch clamp experiment was conducted between hour 48 and hour 72 after the start of the culture. On the day of the experiment, the cells were digested with 0.25% trypsin, then centrifuged, collected, and resuspended in an extracellular fluid (140 mM NaCl, 4 mM KCl, 1 mM MgCl.sub.2, 2 mM CaCl.sub.2), 5 mM D-glucose monohydrate, 10 mM Hepes, pH 7.4, 298 mOsm) to form a cell suspension. The cell suspension was placed on the cell bank of the Patchliner instrument, which then applied the cells to a chip (NPC-16) by means of a negative pressure controller. The negative pressure drew individual cells to the wells of the chip. When a whole-cell configuration was formed, the instrument was given hERG currents according to a set hERG current-voltage program and then automatically performed compound perfusion from low concentrations to high concentrations. Data were recorded and analyzed using the HEAK Patchmaster, HEAK EPC10 patch clamp amplifier (Nanion), Pathliner software and Pathcontrol HT software, and the currents of the compounds at each concentration and the current of the blank control were analyzed.

2.3. Results

[0522] The blocking of hERG potassium currents by the compounds disclosed herein was tested through the above assay, and the IC.sub.50 values determined are shown in Table 9.

TABLE-US-00013 TABLE 9 IC.sub.50 of the compounds disclosed herein for the blocking of hERG potassium currents Example IC.sub.50 (μM) 11 >30 12 >30 13 >30

[0523] Note: IC.sub.50≥30 μM indicates no inhibitory activity; 30>IC.sub.50≥10 μM indicates weak inhibitory activity; 10>IC.sub.50≥1 μM indicates moderate inhibitory activity; IC.sub.50<1 μM indicates strong inhibitory activity.

[0524] Conclusion: The compounds claimed by the present disclosure have no inhibitory activity against hERG, showing better safety.