Certain chemical compositions and their methods of use

Abstract

This disclosure provides CDK9 inhibitors. Methods of treating a disease or condition are also provided, comprising administering one of the CDK9 inhibitors disclosed herein to a subject requiring treatment. In some embodiments, the disease or condition to be treated is cancer. In some embodiments, the disease or condition is liver cancer.

Description

[0001] Cross-references

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 025,474, filed May 15, 2020, which is incorporated herein by reference. Background Technology

[0003] Despite progress, cancer treatment remains relatively challenging. Systemic therapies such as chemotherapy can be toxic and have negative side effects on patients. While CDK9 inhibitors have shown promise as small-molecule cancer therapeutics, their potential utility is limited by poor targeting of cancerous tissue and the resulting peripheral exposure. Therefore, there is a need to develop CDK9 inhibitors as small-molecule cancer therapeutics with improved targeting. Summary of the Invention

[0004] On the one hand, this paper provides compounds of formula (I):

[0005]

[0006] Or its pharmaceutically acceptable salt, wherein:

[0007] Ring A is selected from C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C 6-10 Aryl and 6- to 10-membered heteroaryl groups;

[0008] X 1 Selected from N and CR 11 ;

[0009] X 2 Selected from N and CR 12 ;

[0010] X 3 Selected from N and CR 13 ;

[0011] X 4 Selected from N and CR 14 ;

[0012] R 1 Selected from H, C 1-6 Alkyl, C 1-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C 6-10 Aryl, 6-membered to 10-membered heteroaryl; wherein each alkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl group is independently and optionally substituted by one or more substituents selected from oxo, halogen, -OR 18 C 1-4 Alkyl, C1-4 Alkoxy, C 1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups, -CN and -NR 20 R 21 ;

[0013] R 2 Selected from halogen, -CN, -OR 18 -SOR 15 -SO2R 15 -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)R 18 -C(O)OR 18 -NR 19 C(O)R 18 -NR 19 C(O)NR 16 R 17 -NR 19 SO2R 15 -NR 19 SO2NR 16 R 17 C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C 6-10 Aryl and 6- to 10-membered heteroaryl groups; wherein each alkyl, alkenyl, ynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl group is independently and optionally substituted by one or more substituents selected from oxo, halogen, -OR 18 C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups, -CN and -NR 20 R 21 ;

[0014] R 3 Selected from H, halogen, -CN, -OR 18 -SOR 15 -SO2R 15 -NR 16 R 17 -C(O)NR 16 R 17-SO2NR 16 R 17 -C(O)R 18 -C(O)OR 18 -NR 19 C(O)R 18 -NR 19 C(O)NR 16 R 17 -NR 19 SO2R 15 -NR 19 SO2NR 16 R 17 C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C 6-10 Aryl and 6- to 10-membered heteroaryl groups; wherein each alkyl, alkenyl, ynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl group is independently and optionally substituted by one or more substituents selected from oxo, halogen, -OR 18 C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups, -CN and -NR 20 R 21 ;

[0015] R 4 Selected from C 1-6 Alkyl, C 2-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C 6-10 Aryl, 6- to 10-member heteroaryl, -O(C 0-4 Alkyl)C 3-6 cycloalkyl, -O(C) 0-4 Alkyl groups (3- to 10-membered heterocyclic alkyl groups), -O(C 0-4 Alkyl)C 6-10 Aryl, -O(C 0-4 Alkyl groups (6 to 10 aryl groups), -O(C) 0-4 Alkyl)C(O)OR 18 -O(C 0-4 alkyl)C(O)NR 19 SO2R 15 -O(C 0-4 Alkyl)SO2 NR 19C(O)R 18 -O(C 3-6 cycloalkyl)C 3-6 cycloalkyl, -O(C) 3-6 Cycloalkyl (3- to 10-membered heterocycloalkyl), -O (C 3-6 cycloalkyl)C 6-10 Aryl, -O(C 3-6 cycloalkyl (6- to 10-membered heteroaryl), -O (C 3-6 Cycloalkyl)C(O)OR 18 -(C 1-4 Alkyl)C 3-6 cycloalkyl, -(C 1-4 Alkyl groups (3- to 10-membered heterocyclic alkyl groups), -(C 1-4 Alkyl)C 6-10 Aryl, -(C 1-4 alkyl (6- to 10-membered heteroaryl) and -(C 1-4 Alkyl)C(O)OR 18 Each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl group is independently and optionally substituted by one or more substituents selected from C10. 1-4 Alkyl, oxo, halogen, -OR 18 -CN, -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)R 18 -C(O)OR 18 -(C 1-4 Alkyl)OC(O)(C 1-4 alkyl), -(C 1-4 Alkyl)OC(O)OR 18 -NR 19 C(O)R 18 -NR 19 C(O)NR 16 R 17 -NR 19 SO2R 15 and -NR 19 SO2NR 16 R 17 ;and

[0016] R 4’ and R 4” Each is independently selected from H and C. 1-6 Alkyl, C 2-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6Cycloalkyl, 3- to 10-membered heterocycloalkyl, C 6-10 Aryl, 6- to 10-member heteroaryl, -O(C 0-4 Alkyl)C 3-6 cycloalkyl, -O(C) 0-4 Alkyl groups (3- to 10-membered heterocyclic alkyl groups), -O(C 0-4 Alkyl)C 6-10 Aryl, -O(C 0-4 Alkyl groups (6 to 10 aryl groups), -O(C) 0-4 Alkyl)C(O)OR 18 -O(C 0-4 alkyl)C(O)NR 19 SO2R 15 -O(C 0-4 Alkyl)SO2NR 19 C(O)R 18 -O(C 3-6 cycloalkyl)C 3-6 cycloalkyl, -O(C) 3-6 Cycloalkyl (3- to 10-membered heterocycloalkyl), -O (C 3-6 cycloalkyl)C 6-10 Aryl, -O(C 3-6 cycloalkyl (6- to 10-membered heteroaryl), -O (C 3-6 Cycloalkyl)C(O)OR 18 -(C 1-4 Alkyl)C 3-6 cycloalkyl, -(C 1-4 Alkyl groups (3- to 10-membered heterocyclic alkyl groups), -(C 1-4 Alkyl)C 6-10 Aryl and -(C 1-4 Alkyl group (6- to 10-membered heteroaryl group); wherein each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl group is independently and optionally substituted by one or more substituents selected from oxo, halogen, -OR 18 -CN, -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)R 18 -C(O)OR 18 -(C 1-4 Alkyl)OC(O)(C 1-4 alkyl), -(C 1-4 Alkyl)OC(O)OR 18 -NR 19 C(O)R 18 -NR19 C(O)NR 16 R 17 -NR 19 SO2R 15 and -NR 19 SO2NR 16 R 17 ;or

[0017] R 3 It is H; and

[0018] R 4 R 4’ and R 4” Together with the carbon atoms they are attached to, they form -C(O)R 18 Or 6 to 10 yuan for heteroaryl compounds;

[0019] R 5 Selected from H, C 1-6 Alkyl, C 1-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C 6-10 Aryl and 6- to 10-membered heteroaryl groups; wherein each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl group is independently and optionally substituted by one or more substituents selected from oxo, halogen, -OR 18 C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups, -CN and -NR 20 R 21 ;

[0020] R 6 and R 7 Each is independently selected from H and C. 1-6 Alkyl, C 1-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C 6-10 Aryl, 6- to 10-membered heteroaryl, -(C 1-4 Alkyl)C 3-6 cycloalkyl, -(C 1-4 Alkyl groups (3- to 10-membered heterocyclic alkyl groups), -(C 1-4 Alkyl)C 6-10 Aryl and -(C 1-4 Alkyl group (6- to 10-membered heteroaryl group); wherein each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl group is independently and optionally substituted by one or more substituents selected from oxo, halogen, -OR 18 C1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups, -CN and -NR 20 R 21 ;or

[0021] R 6 and R 7 Together with the nitrogen atoms to which they are attached, they form 3- to 10-membered heterocyclic alkyl groups optionally substituted with one or more substituents selected from oxo, halogen, and -OR groups. 18 C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups, -CN and -NR 20 R 21 ;

[0022] R 8 and R 9 Each is independently selected from H, halogen, -CN, -OR 18 -SOR 15 -SO2R 15 -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)R 18 -C(O)OR 18 -NR 19 C(O)R 18 -NR 19 C(O)NR 16 R 17 -NR 19 SO2R 15 -NR 19 SO2NR 16 R 17 C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Heteroalkyl and C 1-6 Haloalkyl; wherein each alkyl, alkenyl, and ynyl group is independently and optionally substituted by one or more substituents selected from oxo, halogen, -OR 18 C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups, -CN and -NR20 R 21 ;

[0023] Each R 10 Independently selected from halogen, -CN, -OR 18 -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)R 18 -C(O)OR 18 -NR 19 C(O)R 18 C 1-6 Alkyl, C 1-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl; wherein each alkyl, cycloalkyl, and heterocycloalkyl group is independently and optionally substituted by one or more substituents selected from oxo, halogen, -OR 18 C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups, -CN and -NR 20 R 21 ;

[0024] R 11 R 12 R 13 and R 14 Each is independently selected from H, halogen, -CN, -OR 18 -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)R 18 -C(O)OR 18 -NR 19 C(O)R 18 C 1-6 Alkyl, C 1-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl; wherein each alkyl, cycloalkyl, and heterocycloalkyl group is independently and optionally substituted by one or more substituents selected from oxo, halogen, -OR 18 C 1-4 Alkyl, C 1-4 Alkoxy, C1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups, -CN and -NR 20 R 21 ;

[0025] Each R 15 Selected independently from C 1-4 Alkyl, C 1-4 Haloalkyl, C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C 6-10 Aryl and 6- to 10-membered heteroaryl groups;

[0026] Each R 16 and R 17 Independently selected from H and C 1-4 Alkyl, C 1-4 Heteroalkyl, C 1-4 Haloalkyl, C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C 6-10 Aryl, 6- to 10-membered heteroaryl; or

[0027] R 16 and R 17 They can form 3- to 10-membered heterocyclic alkyl groups together with the nitrogen atoms they are attached to; each R 18 Independently selected from H and C 1-4 Alkyl, C 1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups and C 3-6 cycloalkyl;

[0028] Each R 19 Independently selected from H and C 1-4 Alkyl, C 1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups and C 3-6 cycloalkyl;

[0029] Each R 20 and R 21 Independently selected from H and C 1-4 Alkyl, C 1-4 Heteroalkyl, C 1-4 Haloalkyl, C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C 6-10 Aryl, 6- to 10-membered heteroaryl; or

[0030] R 20 and R 21 They can form 3- to 10-membered heterocyclic alkyl groups together with the nitrogen atoms they are attached to; and

[0031] n is 0, 1, 2, 3 or 4.

[0032] In some implementations, ring A is selected from C. 3-6 Cycloalkyl and 3- to 10-membered heterocyclic alkyl groups. In some embodiments, ring A is C. 3-6 Cycloalkyl groups. In some embodiments, cycloA is selected from:

[0033]

[0034] In some implementations, ring A is selected from:

[0035]

[0036]

[0037] In some implementations, ring A is selected from:

[0038]

[0039] In some implementations, ring A is selected from:

[0040]

[0041] In some implementation schemes, X 1 X 2 X 3 and X 4 One of them is N. In some implementations, X 1 X 2 X 3 and X 4 None of them are N.

[0042] In some implementation schemes, R 11 R 12 R 13 and R 14 Each is independently selected from H, halogen, -CN, -OR 18 -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)R 18 -C(O)OR 18 -NR 19 C(O)R 18 C 1-6 Alkyl and C 1-6 Halogenated alkyl groups. In some embodiments, R 11 R 12 R 13 and R 14Each is independently selected from H, halogen, -CN, -OR 18 -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)R 18 -C(O)OR 18 and -NR 19 C(O)R 18 In some implementations, R 11 R 12 R 13 and R 14 Each is independently selected from H, halogen, -CN, -OR 18 and -NR 16 R 17 In some implementations, R 11 It is chlorine, and R 12 R 13 and R 14 Each is H.

[0043] In some implementation schemes, R 1 Selected from H, C 1-6 Alkyl, C 1-6 Halogenated alkyl groups and C 3-6 Cycloalkyl. In some embodiments, R 1 It is Me. In some implementations, R 1 It is H.

[0044] In some implementation schemes, R 2 Selected from halogen, -CN, -OR 18 -SOR 15 -SO2R 15 -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)R 18 -C(O)OR 18 -NR 19 C(O)R 18 -NR 19 C(O)NR 16 R 17 -NR 19 SO2R 15 -NR 19 SO2NR 16 R 17 C1-6 Alkyl, C 1-6 Halogenated alkyl groups and C 3-6 Cycloalkyl. In some embodiments, R 2 Selected from halogens, -CN, -OH, -OMe, -OEt, -NH2, -NHMe, -NMe2, Me, Et, n-Pr, i-Pr, -CF3, and cyclopropyl. In some embodiments, R 2 It's me.

[0045] In some implementation schemes, R 3 Selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl and 3- to 10-membered heterocyclic alkyl groups. In some embodiments, R 3 Selected from H, Me, Et, -CF3, and cyclopropyl. In some embodiments, R 3 It is H.

[0046] In some implementation schemes, R 2 It is Me and R 3 It is H.

[0047] In some implementation schemes,

[0048] R 4 Selected from C 6-10 Aryl, 6- to 10-member heteroaryl, -O(C 0-4 Alkyl)C 3-6 cycloalkyl, -O(C) 0-4 Alkyl groups (3- to 10-membered heterocyclic alkyl groups), -O(C 0-4 Alkyl)C 6-10 Aryl, -O(C 0-4 Alkyl groups (6 to 10 aryl groups), -O(C) 0-4 Alkyl)C(O)OR 18 -O(C 0-4 alkyl)C(O)NR 19 SO2R 15 -O(C 0-4 Alkyl)SO2 NR 19 C(O)R 18 -O(C 3-6 cycloalkyl)C 3-6 cycloalkyl, -O(C) 3-6 Cycloalkyl (3- to 10-membered heterocycloalkyl), -O (C 3-6 cycloalkyl)C 6-10 Aryl, -O(C 3-6 cycloalkyl (6- to 10-membered heteroaryl), -O (C 3-6 Cycloalkyl)C(O)OR18 -(C 1-4 Alkyl)C 3-6 cycloalkyl, -(C 1-4 Alkyl groups (3- to 10-membered heterocyclic alkyl groups), -(C 1-4 Alkyl)C 6-10 Aryl, -(C 1-4 alkyl (6- to 10-membered heteroaryl) and -(C 1-4 Alkyl)C(O)OR 18 Each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl group is independently and optionally substituted by one or more substituents selected from C10. 1-4 Alkyl, oxo, halogen, -OR 18 -CN, -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)R 18 -C(O)OR 18 -(C 1-4 Alkyl)OC(O)(C 1-4 alkyl), -(C 1-4 Alkyl)OC(O)OR 18 -NR 19 C(O)R 18 -NR 19 C(O)NR 16 R 17 -NR 19 SO2R 15 and -NR 19 SO2NR 16 R 17 ;and

[0049] R 4’ and R 4” Each is independently selected from H and C. 1-6 Alkyl, C 1-6 Halogenated alkyl groups and C 3-6 Cycloalkyl; wherein each alkyl group and cycloalkyl group is independently and optionally substituted with one or more substituents selected from halogens, -OR 18 -CN and -NR 16 R 17 ;or

[0050] R 3 It is H; and

[0051] R 4 R 4’ and R 4”Together with the carbon atoms they are attached to, they form -C(O)R 18 Or 6 to 10 yuan for mixed aromatic compounds.

[0052] In some implementation schemes,

[0053] R 4 Selected from 6- to 10-member heteroaryl groups, -O(C 0-4 Alkyl groups (3- to 10-membered heterocyclic alkyl groups), -O(C 0-4 Alkyl)C 6-10 Aryl, -O(C 0-4 Alkyl groups (6 to 10 aryl groups), -O(C) 0-4 Alkyl)C(O)OR 18 -O(C 0-4 alkyl)C(O)NR 19 SO2R 15 -O(C 0-4 Alkyl)SO2NR 19 C(O)R 18 -O(C 3-6 cycloalkyl (6- to 10-membered heteroaryl), -O (C 3-6 Cycloalkyl)C(O)OR 18 -(C 1-4 alkyl (6- to 10-membered heteroaryl) and -(C 1-4 Alkyl)C(O)OR 18 Each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl group is independently and optionally substituted by one or more substituents selected from C10. 1-4 Alkyl, oxo, halogen, -OR 18 -CN, -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)OR 18 -(C 1-4 Alkyl)OC(O)(C 1-4 alkyl), -(C 1-4 Alkyl)OC(O)OR 18 -NR 19 C(O)R 18 -NR 19 C(O)NR 16 R 17 -NR 19 SO2R 15 and -NR 19 SO2NR 16 R 17 ;and

[0054] R 4’ and R 4” Both are H; or

[0055] R 3 It is H; and

[0056] R 4 R 4’ and R 4” Together with the carbon atoms they are attached to, they form -C(O)R 18 Or 6 to 10 yuan for mixed aromatic compounds.

[0057] In some implementation schemes,

[0058] R 4 Selected from 6- to 10-member heteroaryl groups, -O(C 0-4 Alkyl groups (3- to 10-membered heterocyclic alkyl groups), -O(C 0-4 Alkyl)C 6-10 Aryl, -O(C 0-4 Alkyl groups (6 to 10 aryl groups), -O(C) 0-4 Alkyl)C(O)OR 18 -O(C 0-4 alkyl)C(O)NR 19 SO2R 15 -O(C 0-4 Alkyl)SO2NR 19 C(O)R 18 -O(C 3-6 cycloalkyl (6- to 10-membered heteroaryl), -O (C 3-6 Cycloalkyl)C(O)OR 18 -(C 1-4 alkyl (6- to 10-membered heteroaryl) and -(C 1-4 Alkyl)C(O)OR 18 Each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl group is independently and optionally substituted by one or more substituents selected from C10. 1-4 Alkyl, oxo, halogen, -OR 18 -CN, -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)OR 18 -(C 1-4 Alkyl)OC(O)(C 1-4 alkyl), -(C 1-4 Alkyl)OC(O)OR 18-NR 19 C(O)R 18 -NR 19 C(O)NR 16 R 17 -NR 19 SO2R 15 and -NR 19 SO2NR 16 R 17 ;and

[0059] R 4’ and R 4” Both are H.

[0060] In some implementation schemes,

[0061] R 3 It is H; and

[0062] R 4 R 4’ and R 4” Together with the carbon atoms they are attached to, they form -C(O)R 18 Or 6 to 10 yuan for mixed aromatic compounds.

[0063] In some implementation schemes, R 5 Selected from H, C 1-6 Alkyl, C 1-6 Halogenated alkyl groups and C 3-6 Cycloalkyl. In some embodiments, R 5 It is Me. In some implementations, R 5 It is H.

[0064] In some implementation schemes,

[0065] R 6 and R 7 Each is independently selected from H, -(C 1-4 Alkyl)C 3-6 cycloalkyl, -(C 1-4 Alkyl groups (3- to 10-membered heterocyclic alkyl groups), -(C 1-4 Alkyl)C 6-10 Aryl and -(C 1-4 Alkyl group (6- to 10-membered heteroaryl group); wherein each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl group is independently and optionally substituted by one or more substituents selected from oxo, halogen, -OR 18 C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups, -CN and -NR 20 R 21 ;or

[0066] R 6 and R 7 Together with the nitrogen atoms to which they are attached, they form 3- to 10-membered heterocyclic alkyl groups optionally substituted with one or more substituents selected from oxo, halogen, and -OR groups. 18 C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups, -CN and -NR 20 R 21 .

[0067] In some implementation schemes, R 6 and R 7 Each is independently selected from H and -(C 1-4 Alkyl group (3- to 10-membered heterocyclic alkyl group); wherein each alkyl group and heterocyclic alkyl group is independently and optionally substituted by one or more substituents selected from halogens, -OR 18 -CN and -NR 20 R 21 .

[0068] In some implementation schemes, R 6 and R 7 One is H and the other is

[0069] In some implementation schemes, R 8 and R 9 Each is independently selected from H, halogen, -CN, -OR 18 -SO2R 15 -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)OR 18 -NR 19 C(O)R 18 -NR 19 SO2R 15 In some implementations, R 8 and R 9 Each is independently selected from H, halogen, -CN, -OR 18 and -NR 16 R 17 In some implementations, R 8 and R 9 Both are H.

[0070] In some implementations, n is 0, 1, or 2. In some implementations, n is 0.

[0071] In some embodiments, the compound of formula (I) is represented by formula (IA):

[0072]

[0073] In some embodiments, the compound of formula (I) is represented by formula (IB):

[0074]

[0075] In some embodiments, the compound of formula (I) is represented by formula (IC), formula (ID), formula (IE), or formula (IF):

[0076]

[0077]

[0078] In some implementation schemes, the compound is selected from:

[0079]

[0080]

[0081]

[0082]

[0083]

[0084]

[0085]

[0086]

[0087]

[0088]

[0089]

[0090] On the other hand, this article provides a pharmaceutical composition comprising the compound described herein or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0091] On the other hand, this article provides methods for treating a disease or condition in a patient in need, comprising administering to a subject a therapeutically effective amount of the compound described herein or a pharmaceutically acceptable salt thereof or a pharmaceutical composition described herein.

[0092] In some implementations, the disease or symptom is cancer. In some implementations, the cancer is selected from leukemia, breast cancer, prostate cancer, ovarian cancer, colon cancer, cervical cancer, lung cancer, lymphoma, and liver cancer. In some implementations, the cancer is liver cancer.

[0093] Incorporation

[0094] All publications, patents and patent applications mentioned in this specification are incorporated herein by reference to the extent that each individual publication, patent or patent application is specifically and individually cited and incorporated herein by reference. Attached Figure Description

[0095] The novel features of the invention are particularly set forth in the appended claims. A better understanding of the features and advantages of the invention will be gained by referring to the following detailed description of illustrative embodiments utilizing the principles of the invention, and the accompanying drawings, in which:

[0096] Figure 1 The ratio of compound concentration in the liver to the concentration in the blood of CD-1 mice after a single oral administration of a 5 mg / kg suspension of the compound is shown.

[0097] Figure 2 The ratio of compound concentration in blood collected from the jugular vein to the portal vein of Sprague-Dawley (SD) rats after a single oral administration of a 5 mg / kg suspension of the compound is shown.

[0098] Figure 3 The mean weight changes (relative to day 1) of BALB / c nude mice treated with the medium and compound are shown. Detailed Implementation

[0099] definition

[0100] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains.

[0101] As used herein, unless the context clearly indicates otherwise, the singular forms “a,” “one,” and “the” include plural indicators.

[0102] When used in conjunction with chemical moieties such as alkyl, alkenyl, or ynyl groups, the term "C" x-y"This refers to groups containing x to y carbons in the chain. For example, the term "C" 1-6 "Alkyl" refers to a substituted or unsubstituted saturated hydrocarbon group, including straight-chain alkyl and branched-chain alkyl groups containing 1 to 6 carbons. Term -C x-y Alkylene – refers to a substituted or unsubstituted alkylene chain having x to y carbons in the alkylene chain. For example, -C 1-6 The alkylene group can be selected from methylene, ethylene, propylene, butylene, pentylene, and hexylene, any of which may be optionally substituted.

[0103] "Alkyl" refers to a substituted or unsubstituted saturated hydrocarbon group, including straight-chain alkyl and branched-chain alkyl groups. Alkyl groups can contain 1 to 12 carbon atoms (e.g., C12, C23, C12, C23, C23, C24 ... 1-12 Alkyl groups, such as those with 1 to 8 carbon atoms (C 1-8 Alkyl groups or 1 to 6 carbon atoms (C 1-6 Alkyl groups. Exemplary alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, hexyl, heptyl, octyl, nonyl, and decyl. The alkyl group is attached to the remainder of the molecule by a single bond. Unless otherwise specified in the specification, the alkyl group may optionally be substituted with one or more substituents (such as those described herein).

[0104] "Halogenated alkyl" refers to an alkyl group that is substituted with one or more halogens. Exemplary halogenated alkyl groups include trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, and 1,2-dibromoethyl.

[0105] "Alkenyl" refers to a substituted or unsubstituted hydrocarbon group, including straight-chain or branched alkenyl groups containing at least one double bond. Alkenyl groups can contain 2 to 12 carbon atoms (e.g., C64, C12, C23 ... 2-12 Alkenyl groups include vinyl (i.e., vinyl), prop-1-enyl, but-1-enyl, pent-1-enyl, pent-1,4-dienyl, etc. Unless otherwise specified in the specification, alkenyl groups may optionally be substituted with one or more substituents (such as those described herein).

[0106] "Alynyl" refers to a substituted or unsubstituted hydrocarbon group, including straight-chain or branched alkynyl groups containing at least one triple bond. Alynyl groups can contain 2 to 12 carbon atoms (e.g., C64, C12, C23 ... 2-12 (Alynyl group). Exemplary alkynyl groups include ethynyl, propynyl, butynyl, pentylyl, hexynyl, etc. Unless otherwise specified in the specification, the alkynyl group may optionally be substituted by one or more substituents (such as those described herein).

[0107] “Heteroalkyl,” “heteroalkenyl,” and “heteroyneyl” refer to substituted or unsubstituted alkyl, alkenyl, and ynyl groups, each having one or more skeletal chain atoms selected from atoms other than carbon. Exemplary skeletal chain atoms selected from atoms other than carbon include, for example, O, N, P, Si, S, or combinations thereof, wherein nitrogen, phosphorus, and sulfur atoms may optionally be oxidized, and nitrogen heteroatoms may optionally be quaternized. If given, numerical ranges refer to the total chain length. For example, 3- to 8-membered heteroalkyl groups have chain lengths of 3 to 8 atoms. Connection to the remainder of the molecule can be achieved by heteroatoms or carbon atoms in the heteroalkyl, heteroalkenyl, or heteroyneyl chain. Unless otherwise specified in the specification, the heteroalkyl, heteroalkenyl, or heteroyneyl group may optionally be substituted with one or more substituents (such as those described herein).

[0108] “Aryl” refers to an aromatic ring in which each atom forming the ring is a carbon atom. The aryl group may be optionally substituted. Examples of aryl groups include, but are not limited to, phenyl and naphthyl. In some embodiments, the aryl group is phenyl. Depending on the structure, the aryl group may be a monovalent or divalent group (i.e., arylene). Unless otherwise specified in the specification, the term “aryl” or the prefix “aromatic” (such as in “arylene”) means including an optionally substituted aryl group.

[0109] "Heteroaryl" refers to a 3- to 12-membered aromatic ring containing at least one heteroatom, wherein each heteroatom may be independently selected from N, O, and S. As used herein, the heteroaryl ring may be selected from monocyclic or bicyclic rings and fused or bridged ring systems, wherein at least one ring in the system is an aromatic ring, i.e., according to Hückel theory, it contains a cyclic, delocalized (4n+2)π-electron system. The heteroatom in the heteroaryl may optionally be oxidized. One or more nitrogen atoms (if present) may optionally be quaternized. Wherever the valence allows, the heteroaryl may be attached to the rest of the molecule by any atom in the heteroaryl (such as a carbon or nitrogen atom in the heteroaryl). Examples of heteroaryl include, but are not limited to, azirrolyl, acridinel, benzimidazolyl, benzoindolyl, 1,3-benzodioxonyl, benzofuranyl, benzoxazolyl, benzo[d]thiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxane Benz[b][1,4]oxazinyl, 1,4-benzodioxane, benzonaphthuronyl, benzooxazolyl, benzodioxoctenyl, benzodioxinyl, benzopyranyl, benzopyranoneyl, benzofuranyl, benzofuranoneyl, benzothiopheneyl (benzothiocenyl), benzothiophene[3,2-d]pyrimidinyl, benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridyl, carbazoleyl, cyclopentano[d]pyrimidinyl, 6,7-dihydro-5H-cyclopentano[4,5]thiophene[2,3-d]pyrimidinyl, 5,6-dihydrobenzo[h]quinazolinyl, 5, 6-Dihydrobenzo[h]pyrazinyl, 6,7-Dihydro-5H-benzo[6,7]cyclopento[1,2-c]pyridazinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanoneyl, furano[3,2-c]pyridinyl, 5,6,7,8,9,10-hexahydrocyclooctano[d]pyrimidinyl, 5,6,7,8,9,10-hexahydrocyclooctano[d]pyridazinyl, 5,6,7,8,9,10-hexahydrocyclooctano[d]pyridinyl, isothiazolyl, imidazolyl, indazole, indole, indazole, isoindole, indolinyl, isoindolinyl, isoquinolinyl, inazinyl Isoxazolyl, 5,8-methylbridged-5,6,7,8-tetrahydroquinazolinyl, naphridinyl, 1,6-naphridinoneyl, oxadiazolyl, 2-oxozazolyl, oxazolyl, ethylene oxide, 5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purineyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyridinyl, pyrido[3,2-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrazinyl, pyridinyl, pyrrolyl Quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, 5,6,7,8-tetrahydroquinazolinyl, 5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl, 6,7,8,9-tetrahydro-5H-cyclopentano[4,5]thieno[2,3-d]pyrimidinyl, 5,6,7,8-tetrahydropyridino[4,5-c]pyridazinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl, thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pyridinyl, and phenylthioyl (i.e., thienoyl). Unless otherwise specified in the specification, the heteroaryl group may optionally be substituted by one or more substituents (such as those described herein).

[0110] The term "cycloalkyl" refers to a monocyclic or polycyclic non-aromatic group in which each atom forming the ring (i.e., the skeleton atom) is a carbon atom. In some embodiments, the cycloalkyl group is saturated or partially unsaturated. In some embodiments, the cycloalkyl group is a spirocyclic or bridged compound. In some embodiments, the cycloalkyl group is fused to an aromatic ring (in which case, the cycloalkyl group is bonded through the carbon atoms of the non-aromatic ring). Cycloalkyl groups include groups having 3 to 10 ring atoms. Representative cycloalkyl groups include, but are not limited to, cycloalkyl groups having 3 to 10 carbon atoms, 3 to 8 carbon atoms, 3 to 6 carbon atoms, or 3 to 5 carbon atoms. Monocyclic cycloalkyl groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic groups include, for example, adamantyl, 1,2-dihydronaphthyl, 1,4-dihydronaphthyl, tetrahydronaphthyl, decahydronaphthyl, 3,4-dihydronaphthyl-1(2H)-one, spiro[2.2]pentyl, norbornyl, and bicyclo[1.1.1]pentyl. Unless otherwise specified in the specification, cycloalkyl groups may be optionally substituted.

[0111] The term "heterocyclic alkyl" refers to a cycloalkyl group containing at least one heteroatom selected from nitrogen, oxygen, and sulfur. Unless otherwise specified in the specification, the heterocyclic alkyl group can be a monocyclic or bicyclic system, which can include fused ring systems (where the heterocyclic alkyl group is bonded through non-aromatic ring atoms when fused with an aryl or heteroaryl ring) or bridged ring systems. The nitrogen, carbon, or sulfur atom in the heterocyclic group may optionally be oxidized. The nitrogen atom may optionally be quaternized. The heterocyclic alkyl group may be partially or fully saturated. Examples of heterocyclic alkyl groups include, but are not limited to, dioxacyclopentyl, thienyl[1,3]dithiaalkyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, decahydroisoquinolinyl, imidazolinyl, imidazoalkyl, isothiazolinyl, isoxazolinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopiperidinyl, oxazolinyl, piperidinyl, piperazinyl, 4-piperidinoneyl, pyrrolyl, pyrazolyl, quininecycloyl, thiazoalkyl, tetrahydrofuranyl, trithiaalkyl, tetrahydropyranyl, thiomorpholinyl, thiomorpholinyl, 1-oxo-thiomorpholinyl, 1,1-dioxo-thiomorpholinyl. The term heterocyclic alkyl also includes all cyclic forms of carbohydrates, including but not limited to monosaccharides, disaccharides, and oligosaccharides. Unless otherwise stated, heterocyclic alkyl groups have 2 to 12 carbons in the ring. It should be understood that when referring to the number of carbon atoms in a heterocyclic alkyl group, the number of carbon atoms in the heterocyclic alkyl group is different from the total number of atoms (including heteroatoms) constituting the heterocyclic alkyl group (i.e., the skeletal atoms of the heterocyclic alkyl ring). Unless otherwise specified in the specification, the heterocyclic alkyl group may be optionally substituted.

[0112] The term "substituted" refers to a portion of a structure having a substituent for hydrogen on one or more carbon or heteroatoms. It should be understood that "substituted" or "substituted" includes the implicit condition that such substitution meets the permissible valence of the substituted atom and the substituent, and that the substitution produces a stable compound, for example, one that does not spontaneously undergo transformation (such as by rearrangement, cyclization, elimination, etc.). As used herein, the term "substituted" means including all permissible substituents in organic compounds. In a broad sense, permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents in organic compounds. For suitable organic compounds, permissible substituents can be one or more and can be the same or different. For the purposes of this disclosure, heteroatoms (such as nitrogen) can have hydrogen substituents and / or any permissible substituent in organic compounds that meet the heteroatom valence as described herein. Substituents may include any substituents described herein, such as halogens, hydroxyl groups, carbonyl groups (such as carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl groups (such as thioesters, thioacetates, or thioformates), alkoxy groups, phosphoryl groups, phosphate esters, phosphonates, phosphonites, amino groups, amide groups, amidine, imine, cyano, nitro, azide, mercapto, alkylthio, sulfate esters, sulfonates, aminosulfonyl, sulfonamide, sulfonyl, heterocyclic, aralkyl, carbocyclic, heterocyclic, cycloalkyl, heterocyclic, aromatic, and heteroaromatic moieties.

[0113] Those skilled in the art will understand that the substituted element itself can be substituted, if appropriate. Unless specifically stated as “unsubstituted,” references to chemical parts herein should be understood to include substituted variants. For example, references to “heteroaryl” groups or parts implicitly include both substituted and unsubstituted variants.

[0114] When substituents are specified by their conventional chemical formula (written from left to right), they contain chemically identical substituents as produced by a structure written from right to left; for example, -CH2O- is equivalent to -OCH2-.

[0115] "Optional" or "optionally" means that the event described below may or may not occur, and the description includes instances where the event or situation occurs and instances where it does not occur. For example, "optionally substituted aryl" means that the aryl group may or may not be substituted, and the description includes substituted aryl groups and aryl groups without substituents.

[0116] The compounds disclosed herein also include crystalline and amorphous forms of those compounds, pharmaceutically acceptable salts, and active metabolites of these compounds having the same type of activity, including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, nonsolventized polymorphs (including anhydrous forms), conformational polymorphs, and amorphous forms of the compounds, as well as mixtures thereof.

[0117] The compounds described herein may exhibit their natural isotopic abundances, or one or more atoms may be artificially enriched in specific isotopes having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number predominantly found in nature. All isotopic variants of the compounds of the present disclosure, whether radioactive or not, are included within the scope of the present disclosure. For example, hydrogen has three naturally occurring isotopes, designated as 1 H (protium), 2 H (deuterium), and 3 H (tritium). Protium is the most abundant hydrogen isotope in nature. Enrichment of deuterium can provide certain therapeutic advantages, such as increased in vivo half-life and / or exposure, or can provide compounds that can be used to study the in vivo pathways of drug elimination and metabolism. Isotopically enriched compounds can be prepared by conventional techniques well known to those skilled in the art.

[0118] "Isomers" are different compounds having the same molecular formula. "Stereoisomers" are isomers that differ only in the way the atoms are arranged in space. "Enantiomers" are a pair of stereoisomers that are non-superimposable mirror images of each other. A 1:1 mixture of a pair of enantiomers is a "racemic" mixture. In appropriate cases, the term "(±)" is used to designate a racemic mixture. "Diastereomers" or "diastereoisomers" are stereoisomers that have at least two asymmetric atoms but are not mirror images of each other. Absolute stereochemistry is specified according to the Cahn Ingold Prelog R-S system. When a compound is a pure enantiomer, the stereochemistry of each chiral carbon can be specified as R or S. A resolved compound of unknown absolute configuration can be designated as (+) or (-) according to the direction (right or left) in which it rotates plane-polarized light at the sodium D line wavelength. Some of the compounds described herein contain one or more asymmetric centers and thus can give rise to enantiomers, diastereomers, and other stereoisomeric forms, and their asymmetric centers can be defined as (R)- or (S)- according to absolute stereochemistry. The chemical entities, pharmaceutical compositions, and methods of the present invention are intended to include all such possible stereoisomers, including racemic mixtures, optically pure forms, diastereomeric mixtures, and intermediate mixtures. Optically active (R)- and (S)-isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. The optical activity of a compound can be analyzed by any suitable method, including but not limited to chiral chromatography and polarimetry, and the predominance of one stereoisomer over another can be determined.

[0119] A chemical entity having a carbon-carbon double bond or a carbon-nitrogen double bond can exist in the Z- or E-form (or cis or trans form). In addition, some chemical entities may exist in various tautomeric forms. Unless otherwise specified, the chemical entities described herein are also intended to include all Z-, E-, and tautomeric forms.

[0120] If necessary, the separation and purification of the chemical entities and intermediates described herein can be performed by any suitable separation or purification procedure, such as filtration, extraction, crystallization, column chromatography, thin-layer chromatography, or thick-layer chromatography, or a combination of these procedures. Specific instructions for suitable separation and isolation procedures can be obtained by referring to the examples below. However, other equivalent separation or isolation procedures may also be used.

[0121] When no stereochemistry is specified, certain small molecules described herein include, but are not limited to (where possible) their isomers, such as enantiomers and diastereomers, mixtures of enantiomers (including racemates), mixtures of diastereomers, and other mixtures thereof, provided they can be prepared by those skilled in the art through routine experiments. In these cases, a single enantiomer or diastereomer (i.e., the optically active form) can be obtained by asymmetric synthesis or by resolution of a mixture of racemates or diastereomers. If possible, resolution of a mixture of racemates or diastereomers can be accomplished, for example, by conventional methods, such as crystallization in the presence of a resolving agent, or by chromatographic analysis using, for example, a chiral high-performance liquid chromatography (HPLC) column. Furthermore, a mixture of two enantiomers enriched in one of the two enantiomers can be purified by recrystallization and / or grinding to provide a further optically enriched form of the major enantiomer. Additionally, such small molecules include the Z- and E-forms (or cis and trans) of certain small molecules having carbon-carbon or carbon-nitrogen double bonds. When certain small molecules described herein exist in various tautomer forms, the term "certain small molecules" is intended to include all tautomer forms of certain small molecules.

[0122] The term "salt" or "pharmaceutically acceptable salt" refers to a salt derived from a variety of organic and inorganic counterions well known in the art. Pharmaceutically acceptable acid addition salts can be formed from inorganic and organic acids. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid. Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, and salicylic acid. Pharmaceutically acceptable base addition salts can be formed from inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum. Organic bases that can be derivatized include, for example, primary, secondary, and tertiary amines, substituted amines (including naturally occurring substituted amines), cyclic amines, basic ion exchange resins, and particularly isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In some embodiments, pharmaceutically acceptable base addition salts are selected from ammonium, potassium, sodium, calcium, and magnesium salts.

[0123] As used herein, the phrase “pharmaceutically acceptable excipient” or “pharmaceutically acceptable carrier” refers to a pharmaceutically acceptable material, composition, or medium, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each carrier must be “acceptable,” meaning it is compatible with other components of the formulation and will not cause harm to the patient. Some examples of materials that can be used as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) tragacanth gum powder; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository wax; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, and corn oil. (10) Soybean oil; (11) Diols, such as propylene glycol; (12) Polyols, such as glycerol, sorbitol, mannitol and polyethylene glycol; (13) Esters, such as ethyl oleate and ethyl laurate; (14) Agar; (15) Buffers, such as magnesium hydroxide and aluminum hydroxide; (16) Alginate; (17) Atherless water; (18) Isotonic saline; (19) Ringer's solution; (20) Ethanol; (21) Phosphate buffer solution; and (22) Other non-toxic and compatible substances used in pharmaceutical preparations.

[0124] The term "effective amount" or "therapeutic effective amount" refers to the amount of a compound described herein sufficient to affect the intended application (including, but not limited to, the treatment of a disease as defined below). Therapeutic effective amounts can vary depending on the intended therapeutic application (in vivo) or the subject and the condition of the disease being treated (e.g., the subject's weight and age, the severity of the disease, the route of administration, etc.), which can be readily determined by those skilled in the art. The term also applies to doses that will induce a specific response in target cells (e.g., a reduction in platelet adhesion and / or cell migration). Specific doses will vary depending on the particular compound selected, the dosing regimen followed, whether it is administered in combination with other compounds, the timing of administration, the tissue to which it is administered, and the physical delivery system carrying it.

[0125] As used herein, "treatment" refers to a method of obtaining a beneficial or desired outcome with respect to a disease, condition, or medical condition, including but not limited to therapeutic and / or preventive benefits. Therapeutic benefits may include, for example, eradicating or improving an underlying condition that is being treated. Furthermore, therapeutic benefits may include, for example, eradicating or improving one or more physiological symptoms associated with an underlying condition, such that improvement is observed in a subject, even though the subject may still have the underlying condition. In some embodiments, for preventive benefits, the composition is applied to a subject at risk of developing a specific disease, or to a subject who reports one or more physiological symptoms of a disease, even if the disease may not yet have been diagnosed.

[0126] As used herein, the term "therapeutic effect" includes therapeutic benefits and / or preventive benefits as described above. Preventive effects include delaying or eliminating the onset of a disease or condition, delaying or eliminating the onset of symptoms of a disease or condition, slowing, stopping, or reversing the progression of a disease or condition, or any combination thereof.

[0127] As used herein, the terms “co-administration,” “combination administration,” and their grammatical equivalents include the administration of two or more agents to an animal (including a human) such that the two agents and / or their metabolites are simultaneously present in the subject. Co-administration includes simultaneous administration of a single composition, administration of a single composition at different times, or administration of a composition in the presence of two agents.

[0128] The terms “antagonist” and “inhibitor” are used interchangeably, and they refer to compounds that have the ability to inhibit the biological function (e.g., activity, expression, binding, protein-protein interactions) of a target protein or enzyme. Therefore, the terms “antagonist” and “inhibitor” are defined in the context of the biological function of the target protein. While preferred antagonists herein interact specifically with the target (e.g., bind to the target), compounds that inhibit the biological activity of a target protein by interacting with other members of the signal transduction pathway in which the target protein is a member are also specifically included in this definition. Preferred biological activity inhibited by antagonists is associated with tumor development, growth, or spread.

[0129] compound

[0130] On the one hand, this paper provides compounds of formula (I):

[0131]

[0132] Or its pharmaceutically acceptable salt, wherein:

[0133] Ring A is selected from C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C 6-10 Aryl and 6- to 10-membered heteroaryl groups;

[0134] X 1 Selected from N and CR 11 ;

[0135] X 2 Selected from N and CR 12 ;

[0136] X 3 Selected from N and CR 13 ;

[0137] X 4 Selected from N and CR 14 ;

[0138] R1 Selected from H, C 1-6 Alkyl, C 1-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C 6-10 Aryl, 6-membered to 10-membered heteroaryl; wherein each alkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl group is independently and optionally substituted by one or more substituents selected from oxo, halogen, -OR 18 C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups, -CN and -NR 20 R 21 ;

[0139] R 2 Selected from halogen, -CN, -OR 18 -SOR 15 -SO2R 15 -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)R 18 -C(O)OR 18 -NR 19 C(O)R 18 -NR 19 C(O)NR 16 R 17 -NR 19 SO2R 15 -NR 19 SO2NR 16 R 17 C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C 6-10 Aryl and 6- to 10-membered heteroaryl groups; wherein each alkyl, alkenyl, ynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl group is independently and optionally substituted by one or more substituents selected from oxo, halogen, -OR 18 C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Heteroalkyl, C 1-4Halogenated alkyl groups, -CN and -NR 20 R 21 ;

[0140] R 3 Selected from H, halogen, -CN, -OR 18 -SOR 15 -SO2R 15 -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)R 18 -C(O)OR 18 -NR 19 C(O)R 18 -NR 19 C(O)NR 16 R 17 -NR 19 SO2R 15 -NR 19 SO2NR 16 R 17 C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C 6-10 Aryl and 6- to 10-membered heteroaryl groups; wherein each alkyl, alkenyl, ynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl group is independently and optionally substituted by one or more substituents selected from oxo, halogen, -OR 18 C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups, -CN and -NR 20 R 21 ;

[0141] R 4 Selected from C 1-6 Alkyl, C 2-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C 6-10 Aryl, 6- to 10-member heteroaryl, -O(C 0-4 Alkyl)C 3-6 cycloalkyl, -O(C) 0-4Alkyl groups (3- to 10-membered heterocyclic alkyl groups), -O(C 0-4 Alkyl)C 6-10 Aryl, -O(C 0-4 Alkyl groups (6 to 10 aryl groups), -O(C) 0-4 Alkyl)C(O)OR 18 -O(C 0-4 alkyl)C(O)NR 19 SO2R 15 -O(C 0-4 Alkyl)SO2 NR 19 C(O)R 18 -O(C 3-6 cycloalkyl)C 3-6 cycloalkyl, -O(C) 3-6 Cycloalkyl (3- to 10-membered heterocycloalkyl), -O (C 3-6 cycloalkyl)C 6-10 Aryl, -O(C 3-6 cycloalkyl (6- to 10-membered heteroaryl), -O (C 3-6 Cycloalkyl)C(O)OR 18 -(C 1-4 Alkyl)C 3-6 cycloalkyl, -(C 1-4 Alkyl groups (3- to 10-membered heterocyclic alkyl groups), -(C 1-4 Alkyl)C 6-10 Aryl, -(C 1-4 alkyl (6- to 10-membered heteroaryl) and -(C 1-4 Alkyl)C(O)OR 18 Each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl group is independently and optionally substituted by one or more substituents selected from C10. 1-4 Alkyl, oxo, halogen, -OR 18 -CN, -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)R 18 -C(O)OR 18 -(C 1-4 Alkyl)OC(O)(C 1-4 alkyl), -(C 1-4 Alkyl)OC(O)OR 18 -NR 19 C(O)R 18 -NR 19 C(O)NR 16 R 17-NR 19 SO2R 15 and -NR 19 SO2NR 16 R 17 ;and

[0142] R 4’ and R 4” Each is independently selected from H and C. 1-6 Alkyl, C 2-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C 6-10 Aryl, 6- to 10-member heteroaryl, -O(C 0-4 Alkyl)C 3-6 cycloalkyl, -O(C) 0-4 Alkyl groups (3- to 10-membered heterocyclic alkyl groups), -O(C 0-4 Alkyl)C 6-10 Aryl, -O(C 0-4 Alkyl groups (6 to 10 aryl groups), -O(C) 0-4 Alkyl)C(O)OR 18 -O(C 0-4 alkyl)C(O)NR 19 SO2R 15 -O(C 0-4 Alkyl)SO2NR 19 C(O)R 18 -O(C 3-6 cycloalkyl)C 3-6 cycloalkyl, -O(C) 3-6 Cycloalkyl (3- to 10-membered heterocycloalkyl), -O (C 3-6 cycloalkyl)C 6-10 Aryl, -O(C 3-6 cycloalkyl (6- to 10-membered heteroaryl), -O (C 3-6 Cycloalkyl)C(O)OR 18 -(C 1-4 Alkyl)C 3-6 cycloalkyl, -(C 1-4 Alkyl groups (3- to 10-membered heterocyclic alkyl groups), -(C 1-4 Alkyl)C 6-10 Aryl and -(C 1-4 Alkyl group (6- to 10-membered heteroaryl group); wherein each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl group is independently and optionally substituted by one or more substituents selected from oxo, halogen, -OR 18 -CN, -NR 16 R 17 -C(O)NR 16 R17 -SO2NR 16 R 17 -C(O)R 18 -C(O)OR 18 -(C 1-4 Alkyl)OC(O)(C 1-4 alkyl), -(C 1-4 Alkyl)OC(O)OR 18 -NR 19 C(O)R 18 -NR 19 C(O)NR 16 R 17 -NR 19 SO2R 15 and -NR 19 SO2NR 16 R 17 ;or

[0143] R 3 It is H; and

[0144] R 4 R 4’ and R 4” Together with the carbon atoms they are attached to, they form -C(O)R 18 Or 6 to 10 yuan for heteroaryl compounds;

[0145] R 5 Selected from H, C 1-6 Alkyl, C 1-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C 6-10 Aryl and 6- to 10-membered heteroaryl groups; wherein each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl group is independently and optionally substituted by one or more substituents selected from oxo, halogen, -OR 18 C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups, -CN and -NR 20 R 21 ;

[0146] R 6 and R 7 Each is independently selected from H and C. 1-6 Alkyl, C 1-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C 6-10Aryl, 6- to 10-membered heteroaryl, -(C 1-4 Alkyl)C 3-6 cycloalkyl, -(C 1-4 Alkyl groups (3- to 10-membered heterocyclic alkyl groups), -(C 1-4 Alkyl)C 6-10 Aryl and -(C 1-4 Alkyl group (6- to 10-membered heteroaryl group); wherein each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl group is independently and optionally substituted by one or more substituents selected from oxo, halogen, -OR 18 C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups, -CN and -NR 20 R 21 ;or

[0147] R 6 and R 7 Together with the nitrogen atoms to which they are attached, they form 3- to 10-membered heterocyclic alkyl groups optionally substituted with one or more substituents selected from oxo, halogen, and -OR groups. 18 C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups, -CN and -NR 20 R 21 ;

[0148] R 8 and R 9 Each is independently selected from H, halogen, -CN, -OR 18 -SOR 15 -SO2R 15 -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)R 18 -C(O)OR 18 -NR 19 C(O)R 18 -NR 19 C(O)NR 16 R 17 -NR 19 SO2R 15 -NR 19 SO2NR 16 R 17 C1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Heteroalkyl and C 1-6 Haloalkyl; wherein each alkyl, alkenyl, and ynyl group is independently and optionally substituted by one or more substituents selected from oxo, halogen, -OR 18 C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups, -CN and -NR 20 R 21 ;

[0149] Each R 10 Independently selected from halogen, -CN, -OR 18 -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)R 18 -C(O)OR 18 -NR 19 C(O)R 18 C 1-6 Alkyl, C 1-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl; wherein each alkyl, cycloalkyl, and heterocycloalkyl group is independently and optionally substituted by one or more substituents selected from oxo, halogen, -OR 18 C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups, -CN and -NR 20 R 21 ;

[0150] R 11 R 12 R 13 and R 14 Each is independently selected from H, halogen, -CN, -OR 18 -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)R 18 -C(O)OR18 -NR 19 C(O)R 18 C 1-6 Alkyl, C 1-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl; wherein each alkyl, cycloalkyl, and heterocycloalkyl group is independently and optionally substituted by one or more substituents selected from oxo, halogen, -OR 18 C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups, -CN and -NR 20 R 21 ;

[0151] Each R 15 Selected independently from C 1-4 Alkyl, C 1-4 Haloalkyl, C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C 6-10 Aryl and 6- to 10-membered heteroaryl groups;

[0152] Each R 16 and R 17 Independently selected from H and C 1-4 Alkyl, C 1-4 Heteroalkyl, C 1-4 Haloalkyl, C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C 6-10 Aryl, 6- to 10-membered heteroaryl; or

[0153] R 16 and R 17 They can form 3- to 10-membered heterocyclic alkyl groups together with the nitrogen atoms they are attached to; each R 18 Independently selected from H and C 1-4 Alkyl, C 1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups and C 3-6 cycloalkyl;

[0154] Each R 19 Independently selected from H and C 1-4 Alkyl, C 1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups and C 3-6 cycloalkyl;

[0155] Each R 20 and R 21 Independently selected from H and C 1-4 Alkyl, C1-4 Heteroalkyl, C 1-4 Haloalkyl, C 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C 6-10 Aryl, 6- to 10-membered heteroaryl; or

[0156] R 20 and R 21 They can form 3- to 10-membered heterocyclic alkyl groups together with the nitrogen atoms they are attached to; and

[0157] n is 0, 1, 2, 3 or 4.

[0158] In some implementations, ring A is selected from C. 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C 6-10 Aryl and 6- to 10-membered heteroaryl groups. In some embodiments, ring A is selected from C. 3-6 Cycloalkyl, 3- to 10-membered heterocycloalkyl and C 6-10 Aryl. In some embodiments, ring A is selected from C. 3-6 Cycloalkyl and 3- to 10-membered heterocyclic alkyl groups. In some embodiments, ring A is a 3- to 10-membered heterocyclic alkyl group. In some embodiments, ring A is C 6-10 Aryl. In some embodiments, ring A is a 6- to 10-membered heteroaryl group. In some embodiments, ring A is C. 3-6 Cycloalkyl groups. In some embodiments, cycloA is selected from:

[0159]

[0160] In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is

[0161] In some implementations, ring A is selected from:

[0162]

[0163] In some implementations, ring A is selected from:

[0164]

[0165] In some implementations, ring A is selected from:

[0166]

[0167] In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is In some implementations, ring A is

[0168] In some implementation schemes, X 1 X 2 X 3 and X 4 None of them are N. In some implementations, X 1 X 2X 3 and X 4 One of them is N. In some implementations, X 1 X 2 X 3 and X 4 The two in it are N. In some implementations, X 1 X 2 X 3 and X 4 The three in the list are N. In some implementations, X 1 X 2 X 3 and X 4 Both are N. In some implementations, X 1 It is N. In some implementations, X 2 It is N. In some implementations, X 3 It is N. In some implementations, X 4 It is N. In some implementations, X 1 and X 2 It is N. In some implementations, X 1 and X 3 It is N. In some implementations, X 1 and X 4 It is N. In some implementations, X 2 and X 3 It is N. In some implementations, X 2 and X 4 It is N. In some implementations, X 3 and X 4 It is N. In some implementations, X 1 X 2 and X 3 It is N. In some implementations, X 1 X 2 and X 4 It is N. In some implementations, X 1 X 3 and X 4 It is N. In some implementations, X 2 X 3 and X 4 It is N. In some implementations, X 1 X 2 X 3 and X 4 It is N.

[0169] In some implementation schemes, R 11 R 12 R 13 and R 14Each is independently selected from H, halogen, -CN, -OR 18 -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)R 18 -C(O)OR 18 -NR 19 C(O)R 18 C 1-6 Alkyl and C 1-6 Halogenated alkyl groups. In some embodiments, R 11 R 12 R 13 and R 14 Each is independently selected from H, halogen, -CN, -OR 18 -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)R 18 -C(O)OR 18 and -NR 19 C(O)R 18 In some implementations, R 11 R 12 R 13 and R 14 Each is independently selected from H, halogen, -CN, -OR 18 and -NR 16 R 17 In some implementations, R 11 R 12 R 13 and R 14 One of them is a halogen and the rest is H. In some implementations, R 11 It is chlorine, and R 12 R 13 and R 14 Each is H.

[0170] In some implementation schemes, R 1 Selected from H, C 1-6 Alkyl, C 1-6 Halogenated alkyl groups and C 3-6 Cycloalkyl. In some embodiments, R 1 It is C 1-6 Alkyl group. In some embodiments, R 1 It is C 1-6Halogenated alkyl groups. In some embodiments, R 1 It is C 3-6 Cycloalkyl. In some embodiments, R 1 Selected from H, Me, Et, n-Pr, i-Pr, -CF3, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. In some embodiments, R 1 Selected from H, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. In some embodiments, R 1 Selected from Me, Et, n-Pr, and i-Pr. In some implementations, R 1 It is Me. In some implementations, R 1 It is H.

[0171] In some implementation schemes, R 2 Selected from halogen, -CN, -OR 18 -SOR 15 -SO2R 15 -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)R 18 -C(O)OR 18 -NR 19 C(O)R 18 -NR 19 C(O)NR 16 R 17 -NR 19 SO2R 15 -NR 19 SO2NR 16 R 17 C 1-6 Alkyl, C 1-6 Halogenated alkyl groups and C 3-6 Cycloalkyl. In some embodiments, R 2 Selected from halogens, -CN, -OH, -OMe, -OEt, -NH2, -NHMe, -NMe2, Me, Et, n-Pr, i-Pr, -CF3, and cyclopropyl. In some embodiments, R 2 Selected from Me, Et, n-Pr, and i-Pr. In some implementations, R 2 It is Me. In some implementations, R 2 It is -CF3. In some implementations, R 2 It is cyclopropyl.

[0172] In some implementation schemes, R 3 Selected from H, C 1-6Alkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl and 3- to 10-membered heterocyclic alkyl groups. In some embodiments, R 3 It is C 1-6 Alkyl group. In some embodiments, R 3 It is C 1-6 Halogenated alkyl groups. In some embodiments, R 3 It is C 3-6 Cycloalkyl. In some embodiments, R 3 It is a 3- to 10-membered heterocyclic alkyl group. In some embodiments, R 3 Selected from H, Me, Et, -CF3, and cyclopropyl. In some embodiments, R 3 It is H. In some implementations, R 3 It is Me. In some implementations, R 3 It is -CF3.

[0173] In some implementation schemes, R 2 It is Me and R 3 It is H.

[0174] In some implementation schemes,

[0175] R 4 Selected from C 6-10 Aryl, 6- to 10-member heteroaryl, -O(C 0-4 Alkyl)C 3-6 cycloalkyl, -O(C) 0-4 Alkyl groups (3- to 10-membered heterocyclic alkyl groups), -O(C 0-4 Alkyl)C 6-10 Aryl, -O(C 0-4 Alkyl groups (6 to 10 aryl groups), -O(C) 0-4 Alkyl)C(O)OR 18 -O(C 0-4 alkyl)C(O)NR 19 SO2R 15 -O(C 0-4 Alkyl)SO2 NR 19 C(O)R 18 -O(C 3-6 cycloalkyl)C 3-6 cycloalkyl, -O(C) 3-6 Cycloalkyl (3- to 10-membered heterocycloalkyl), -O (C 3-6 cycloalkyl)C 6-10 Aryl, -O(C 3-6 cycloalkyl (6- to 10-membered heteroaryl), -O (C 3-6 Cycloalkyl)C(O)OR 18 -(C1-4 Alkyl)C 3-6 cycloalkyl, -(C 1-4 Alkyl groups (3- to 10-membered heterocyclic alkyl groups), -(C 1-4 Alkyl)C 6-10 Aryl, -(C 1-4 alkyl (6- to 10-membered heteroaryl) and -(C 1-4 Alkyl)C(O)OR 18 Each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl group is independently and optionally substituted by one or more substituents selected from C10. 1-4 Alkyl, oxo, halogen, -OR 18 -CN, -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)R 18 -C(O)OR 18 -(C 1-4 Alkyl)OC(O)(C 1-4 alkyl), -(C 1-4 Alkyl)OC(O)OR 18 -NR 19 C(O)R 18 -NR 19 C(O)NR 16 R 17 -NR 19 SO2R 15 and -NR 19 SO2NR 16 R 17 ;and

[0176] R 4’ and R 4” Each is independently selected from H and C. 1-6 Alkyl, C 1-6 Halogenated alkyl groups and C 3-6 Cycloalkyl; wherein each alkyl group and cycloalkyl group is independently and optionally substituted with one or more substituents selected from halogens, -OR 18 -CN and -NR 16 R 17 ;or

[0177] R 3 It is H; and

[0178] R 4 R 4’ and R 4” Together with the carbon atoms they are attached to, they form -C(O)R18 Or 6 to 10 yuan for mixed aromatic compounds.

[0179] In some implementation schemes,

[0180] R 4 Selected from 6- to 10-member heteroaryl groups, -O(C 0-4 Alkyl groups (3- to 10-membered heterocyclic alkyl groups), -O(C 0-4 Alkyl)C 6-10 Aryl, -O(C 0-4 Alkyl groups (6 to 10 aryl groups), -O(C) 0-4 Alkyl)C(O)OR 18 -O(C 0-4 alkyl)C(O)NR 19 SO2R 15 -O(C 0-4 Alkyl)SO2NR 19 C(O)R 18 -O(C 3-6 cycloalkyl (6- to 10-membered heteroaryl), -O (C 3-6 Cycloalkyl)C(O)OR 18 -(C 1-4 alkyl (6- to 10-membered heteroaryl) and -(C 1-4 Alkyl)C(O)OR 18 Each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl group is independently and optionally substituted by one or more substituents selected from C10. 1-4 Alkyl, oxo, halogen, -OR 18 -CN, -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)OR 18 -(C 1-4 Alkyl)OC(O)(C 1-4 alkyl), -(C 1-4 Alkyl)OC(O)OR 18 -NR 19 C(O)R 18 -NR 19 C(O)NR 16 R 17 -NR 19 SO2R 15 and -NR 19 SO2NR 16 R 17 ;and

[0181] R4’ and R 4” Both are H; or

[0182] R 3 It is H; and

[0183] R 4 R 4’ and R 4” Together with the carbon atoms they are attached to, they form -C(O)R 18 Or 6 to 10 yuan for mixed aromatic compounds.

[0184] In some implementation schemes,

[0185] R 4 Selected from 6- to 10-member heteroaryl groups, -O(C 0-4 Alkyl groups (3- to 10-membered heterocyclic alkyl groups), -O(C 0-4 Alkyl)C 6-10 Aryl, -O(C 0-4 Alkyl groups (6 to 10 aryl groups), -O(C) 0-4 Alkyl)C(O)OR 18 -O(C 0-4 alkyl)C(O)NR 19 SO2R 15 -O(C 0-4 Alkyl)SO2NR 19 C(O)R 18 -O(C 3-6 cycloalkyl (6- to 10-membered heteroaryl), -O (C 3-6 Cycloalkyl)C(O)OR 18 -(C 1-4 alkyl (6- to 10-membered heteroaryl) and -(C 1-4 Alkyl)C(O)OR 18 Each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl group is independently and optionally substituted by one or more substituents selected from C10. 1-4 Alkyl, oxo, halogen, -OR 18 -CN, -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)OR 18 -(C 1-4 Alkyl)OC(O)(C 1-4 alkyl), -(C 1-4 Alkyl)OC(O)OR 18 -NR 19 C(O)R18 -NR 19 C(O)NR 16 R 17 -NR 19 SO2R 15 and -NR 19 SO2NR 16 R 17 ;and

[0186] R 4’ and R 4” Both are H.

[0187] In some implementation schemes,

[0188] R 3 It is H; and

[0189] R 4 R 4’ and R 4” Together with the carbon atoms they are attached to, they form -C(O)R 18 Or 6 to 10 yuan for mixed aromatic compounds.

[0190] In some implementation schemes,

[0191] R 3 It is H; and

[0192] R 4 R 4’ and R 4” Together with the carbon atoms they are attached to, they form -C(O)R 18 Or 6 to 10 yuan for mixed aromatic compounds.

[0193] In some implementation schemes,

[0194] R 3 It is H; and

[0195] R 4 R 4’ and R 4” Together with the carbon atoms they are attached to, they form 6- to 10-membered heteroaryl groups.

[0196] In some implementation schemes, R 5 Selected from H, C 1-6 Alkyl, C 1-6 Halogenated alkyl groups and C 3-6 Cycloalkyl. In some embodiments, R 5 It is C 1-6 Halogenated alkyl groups. In some embodiments, R 5 It is C 3-6 Cycloalkyl. In some embodiments, R 5 It is C 1-6Alkyl group. In some embodiments, R 5 It is Me, Et, n-Pr, or i-Pr. In some implementations, R 5 It is Me. In some implementations, R 5 It is H.

[0197] In some implementation schemes,

[0198] R 6 and R 7 Each is independently selected from H, -(C 1-4 Alkyl)C 3-6 cycloalkyl, -(C 1-4 Alkyl groups (3- to 10-membered heterocyclic alkyl groups), -(C 1-4 Alkyl)C 6-10 Aryl and -(C 1-4 Alkyl group (6- to 10-membered heteroaryl group); wherein each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl group is independently and optionally substituted by one or more substituents selected from oxo, halogen, -OR 18 C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups, -CN and -NR 20 R 21 ;or

[0199] R 6 and R 7 Together with the nitrogen atoms to which they are attached, they form 3- to 10-membered heterocyclic alkyl groups optionally substituted with one or more substituents selected from oxo, halogen, and -OR groups. 18 C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Heteroalkyl, C 1-4 Halogenated alkyl groups, -CN and -NR 20 R 21 .

[0200] In some implementation schemes, R 6 and R 7 Each is independently selected from H and -(C 1-4 Alkyl group (3- to 10-membered heterocyclic alkyl group); wherein each alkyl group and heterocyclic alkyl group is independently and optionally substituted by one or more substituents selected from halogens, -OR 18 -CN and -NR 20 R 21 .

[0201] In some implementation schemes, R 6 and R 7One is H and the other is In some implementation schemes, R 6 It is H and R 7 yes In some implementation schemes, R 7 It is H and R 6 yes

[0202] In some implementation schemes, R 8 and R 9 Each is independently selected from H, halogen, -CN, -OR 18 -SO2R 15 -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 -C(O)OR 18 -NR 19 C(O)R 18 -NR 19 SO2R 15 In some implementations, R 8 and R 9 Each is independently selected from H, halogen, -CN, -OR 18 and -NR 16 R 17 In some implementations, R 8 and R 9 Both are H.

[0203] In some implementations, n is 0, 1, 2, 3, or 4. In some implementations, n is 0, 1, 2, or 3. In some implementations, n is 0, 1, or 2. In some implementations, n is 0 or 1. In some implementations, n is 0. In some implementations, n is 1. In some implementations, n is 2. In some implementations, n is 3. In some implementations, n is 4.

[0204] In some embodiments, the compound of formula (I) is represented by formula (IA):

[0205]

[0206] In some embodiments, the compound of formula (I) is represented by formula (IB):

[0207]

[0208] In some embodiments, the compound of formula (I) is represented by formula (IC), formula (ID), formula (IE), or formula (IF):

[0209]

[0210]

[0211] In some embodiments, the compound of formula (I) is represented by formula (IC):

[0212]

[0213] In some implementations, the compound of formula (I) is represented by formula (ID):

[0214]

[0215] In some embodiments, the compound of formula (I) is represented by formula (IE):

[0216]

[0217] In some embodiments, the compound of formula (I) is represented by formula (IF):

[0218]

[0219] In some implementation schemes, the compound is selected from:

[0220]

[0221]

[0222]

[0223]

[0224]

[0225]

[0226]

[0227]

[0228]

[0229]

[0230]

[0231] In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is In some implementation schemes, the compound is

[0232] On the other hand, this article provides a pharmaceutical composition comprising the compound described herein or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0233] On the other hand, this article provides methods for treating a disease or condition in a patient in need, comprising administering to a subject a therapeutically effective amount of the compound described herein or a pharmaceutically acceptable salt thereof or a pharmaceutical composition described herein.

[0234] In some implementations, the disease or symptom is cancer. In some implementations, the cancer is selected from leukemia, breast cancer, prostate cancer, ovarian cancer, colon cancer, cervical cancer, lung cancer, lymphoma, and liver cancer. In some implementations, the cancer is leukemia. In some implementations, the cancer is breast cancer. In some implementations, the cancer is prostate cancer. In some implementations, the cancer is ovarian cancer. In some implementations, the cancer is colon cancer. In some implementations, the cancer is cervical cancer. In some implementations, the cancer is lung cancer. In some implementations, the cancer is lymphoma. In some implementations, the cancer is liver cancer.

[0235] How to use

[0236] In one aspect, the present invention provides a method for treating a proliferative condition in a subject of need, comprising administering to the subject a compound having formula (I) as further disclosed herein. In some embodiments, the method for treating a proliferative condition comprises administering a CDK9 inhibitor to the subject. In some embodiments, the compound having formula (I) is a CDK9 inhibitor. In some embodiments, the proliferative condition is a cancerous condition. In some further embodiments, the cancerous condition is selected from leukemia, breast cancer, prostate cancer, ovarian cancer, colon cancer, cervical cancer, lung cancer, lymphoma, and liver cancer. In some embodiments, the cancerous condition is liver cancer.

[0237] In some embodiments, the CDK9 inhibitors disclosed herein highly target the liver. In some embodiments, the CDK9 inhibitors disclosed herein exhibit better liver targeting compared to known CDK9 inhibitors. In some embodiments, the CDK9 inhibitors disclosed herein accumulate in the liver while avoiding peripheral exposure to nearby tissues. In some embodiments, the CDK9 inhibitors disclosed herein have reduced peripheral exposure to nearby tissues compared to known CDK9 inhibitors. In some embodiments, the CDK9 inhibitors disclosed herein have reduced toxicity compared to known CDK9 inhibitors.

[0238] In a further embodiment, the present invention provides a method for treating cancer conditions, wherein a compound having formula (I) (e.g., a CDK9 inhibitor) is effective in one or more methods of inhibiting cancer cell proliferation, inhibiting cancer cell metastasis, reducing the severity or incidence of symptoms associated with the presence of cancer cells, and promoting an immune response against tumor cells. In some embodiments, the method includes administering a therapeutically effective amount of a compound having formula (I) to cancer cells. In some embodiments, the compound having formula (I) is a CDK9 inhibitor. In some embodiments, the administration is performed in vitro. In other embodiments, the administration is performed in vivo.

[0239] As used herein, a therapeutically effective dose of a CDK9 inhibitor is defined as an amount sufficient to achieve the intended application (including, but not limited to, disease treatment as defined herein). Subtherapeutic doses of CDK9 inhibitors for the treatment of the intended disease condition are also considered in this approach.

[0240] The amount of CDK9 inhibitor administered may vary depending on the intended therapeutic application (in vitro or in vivo) or the subject and the condition of the disease being treated (e.g., the subject's weight and age, the severity of the disease, the method of administration, etc.), which can be readily determined by those skilled in the art.

[0241] Measuring the inhibition of biological effects by CDK9 may involve performing an assay on a biological sample, such as a sample from a subject. Depending on the assay, any of a variety of samples may be selected. Examples of samples include, but are not limited to, blood samples (e.g., plasma or serum), exhaled respiratory condensate samples, bronchoalveolar lavage fluid, sputum samples, urine samples, and tissue samples.

[0242] Subjects treated with CDK9 inhibitors can be monitored to determine the effectiveness of the treatment, and the treatment regimen can be adjusted based on the subject's physiological response to the treatment. For example, if CDK9 degradation inhibits the biological effect above or below a threshold, the dosage or frequency can be reduced or increased, respectively. If the treatment is determined to be effective, the method may also include continuing treatment. If the treatment is determined to be effective, the method may include maintaining, gradually reducing, decreasing, or discontinuing the administration of the compound used for treatment. If the compound used for treatment is determined to be ineffective, the method may include increasing its administration. Alternatively, if the treatment is determined to be ineffective, the method may include discontinuing treatment. In some embodiments, treatment with the CDK9 inhibitor is discontinued if the inhibition of the biological effect is above or below a threshold (such as lack of response or adverse reaction). The biological effect can be a change in any of a variety of physiological indicators.

[0243] Generally speaking, CDK9 inhibitors are compounds that inhibit one or more biological effects of CDK9. Such biological effects can be inhibited by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or more.

[0244] In some other embodiments, this method can be used to treat CDK9-related disease conditions. Any disease condition directly or indirectly caused by abnormal activity or expression levels of CDK9 can be the intended disease condition. In some embodiments, the disease condition is a proliferative disease, such as, but not limited to, cancer, as described herein. In some embodiments, the disease condition is cancer. The role of CDK9 in tumorigenesis and tumor progression is associated with many human cancers. Therefore, agents targeting CDK9 have therapeutic value.

[0245] The data presented in the examples below demonstrate the anticancer effects of CDK9 inhibitors. Therefore, this method is particularly useful for treating proliferative conditions, such as tumors.

[0246] In some implementations, the methods of administering CDK9 inhibitors described herein are used to treat hematologic malignancies, breast cancer, prostate cancer, ovarian cancer, colon cancer, cervical cancer, lung cancer, lymph node cancer, liver cancer, or any combination thereof.

[0247] Treatment effect

[0248] In some implementations, treatment efficacy is measured based on its effectiveness in treating proliferative conditions such as cancer. Generally, the therapeutic efficacy of the methods and compositions of this invention for treating proliferative conditions (e.g., cancer, whether benign or malignant) can be measured by the degree to which the methods and compositions promote: inhibition of tumor cell proliferation, inhibition of tumor angiogenesis, eradication of tumor cells, reduction of tumor growth rate, and / or reduction of at least one tumor size. Several parameters that need to be considered in determining treatment efficacy are discussed herein. Clinicians can establish appropriate combinations of parameters for specific situations. The progress of the methods of this invention in treating cancer (e.g., reducing tumor size or eradicating cancer cells) can be determined using any suitable method, such as those currently used in clinical practice to track tumor size and cancer progression. The primary efficacy parameter for evaluating the methods and compositions of this invention in treating cancer is preferably a reduction in tumor size. Tumor size can be calculated using any suitable technique, such as measuring dimensions, or using available computer software to estimate tumor volume, such as FreeFlight software developed at Wake Forest University, which is capable of accurately estimating tumor volume. Tumor size can be determined by tumor visualization using, for example, CT, ultrasound, SPECT, spiral CT, MRI, radiography, etc. In the implementation plan for surgical resection of the tumor after the treatment period, the presence and size of the tumor tissue can be determined by total analysis of the tissue to be resected and / or by pathological analysis of the resected tissue.

[0249] In some desired embodiments, due to the methods and compositions of the present invention, tumor growth is stabilized (i.e., the size of one or more tumors increases by no more than 1%, 5%, 10%, 15%, or 20%, and / or there is no metastasis). In some embodiments, the tumor is stable for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or more weeks. In some embodiments, the tumor is stable for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or more months. In some embodiments, the tumor is stable for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more years. Preferably, the method of the present invention reduces the size of the tumor by at least about 5% (e.g., at least about 10%, 15%, 20%, or 25%). More preferably, the tumor size is reduced by at least about 30% (e.g., at least about 35%, 40%, 45%, 50%, 55%, 60%, or 65%). More preferably, the tumor size is reduced by at least about 70% (e.g., at least about 75%, 80%, 85%, 90%, or 95%). Most preferably, the tumor is completely eliminated or reduced below the detectable level. In some embodiments, the subject remains tumor-free for at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or more weeks after treatment (e.g., remission). In some embodiments, the subject remains tumor-free for at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or more months after treatment. In some embodiments, the subject remains tumor-free for at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more years after treatment.

[0250] In some embodiments, the efficacy of the method of the present invention in reducing tumor size can be determined by measuring the percentage of necrotic (i.e., dead) tissue of the surgically removed tumor at the end of the treatment period. In some further embodiments, the treatment is considered effective if the percentage of necrosis in the removed tissue is greater than about 20% (e.g., at least about 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%), more preferably about 90% or greater (e.g., about 90%, 95%, or 100%). Most preferably, the percentage of necrosis in the removed tissue is 100%, i.e., tumor tissue is absent or undetectable.

[0251] The efficacy of the method of the present invention can be determined by a number of secondary parameters. Examples of secondary parameters include, but are not limited to, detection of new tumors, detection of tumor antigens or biomarkers (e.g., CEA, PSA, or CA-125), biopsy, surgical downgrading (i.e., the surgical stage of a tumor changes from unresectable to resectable), PET scans, survival, progression-free survival, time to disease progression, quality of life assessments (such as Clinical Benefit Response Assessment), etc., all of which can indicate the overall progression (or regression) of human cancer. Biopsy is particularly useful in detecting the eradication of cancer cells within tissue. Radioimmunoassay (RAID) is used to locate and stage tumors using serum levels of tumor-derived and / or tumor-associated markers (antigens) (“tumor markers” or “tumor-associated antigens”), and can be used as a diagnostic criterion before treatment, a diagnostic indicator of recurrence after treatment, and an indicator of treatment efficacy after treatment. Examples of tumor markers or tumor-associated antigens that can be evaluated as indicators of treatment efficacy include, but are not limited to, carcinoembryonic antigen (CEA), prostate-specific antigen (PSA), CA-125, CA19-9, ganglioside molecules (e.g., GM2, GD2, and GD3), MART-1, heat shock proteins (e.g., gp96), sialic acid Tn (STn), tyrosinase, MUC-1, HER-2 / neu, c-erb-B2, KSA, PSMA, p53, RAS, EGF-R, VEGF, MAGE, and gp100. Other tumor-associated antigens are known in the art. The combination of RAID technology with endoscopic detection systems can also effectively differentiate small tumors from surrounding tissues (see, for example, U.S. Patent No. 4,932,412).

[0252] In other desired embodiments, treatment of a human patient with cancer according to the method of the invention is demonstrated by one or more of the following results: (a) complete disappearance of the tumor (i.e., complete remission); (b) a reduction in tumor size of about 25% to about 50% within at least four weeks after the end of the treatment period compared to the pre-treatment tumor size; (c) a reduction in tumor size of about 50% within at least four weeks after the end of the treatment period compared to the pre-treatment tumor size; and (d) a reduction in a specific tumor-associated antigen level of at least 2% (e.g., a reduction of about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%) within about 4-12 weeks after the end of the treatment period compared to the pre-treatment level. While a reduction of at least 2% in the tumor-associated antigen level is preferred, any reduction in the tumor-associated antigen level is evidence of treatment of the patient's cancer by the method of the invention. For example, for unresectable locally advanced pancreatic cancer, a reduction in CA19-9 tumor-associated antigen level of at least 10% within 4-12 weeks after the end of the treatment period compared to the pre-treatment CA19-9 level can demonstrate treatment. Similarly, for locally advanced rectal cancer, a reduction of at least 10% in CEA tumor-associated antigen levels within 4–12 weeks after the end of treatment, compared to pre-treatment CEA levels, can be considered evidence of treatment.

[0253] Regarding quality of life assessments, such as clinical benefit response criteria, the therapeutic benefits of the treatment according to the present invention can be demonstrated based on pain intensity, analgesic consumption, and / or Karnofsky Performance Scale scores. Alternatively or additionally, treatment of cancer in human patients is demonstrated by: (a) a reduction of at least 50% (e.g., at least 60%, 70%, 80%, 90%, or 100%) in pain intensity reported by the patient prior to treatment, such as during any consecutive four-week period within 12 weeks after the end of treatment; (b) a reduction of at least 50% (e.g., at least 60%, 70%, 80%, 90%, or 100%) in analgesic consumption reported by the patient prior to treatment, such as during any consecutive four-week period within 12 weeks after the end of treatment; and / or (c) an increase of at least 20 points (e.g., at least 30, 50, 70, or 90 points) in the patient's reported Kanovsky Sexual Energy Scale score prior to treatment, such as during any consecutive four-week period within 12 weeks after the end of treatment.

[0254] Treatment of proliferative disorders (e.g., cancer, whether benign or malignant) in human patients is ideally demonstrated by one or more of the aforementioned results (in any combination), but alternative or additional results from reference tests and / or other tests may also demonstrate the effectiveness of the treatment.

[0255] In some embodiments, due to the method of the present invention, tumor size is reduced, preferably with no significant adverse events in the subjects. Adverse events are classified or “graded” by the Cancer Treatment Evaluation Program (CTEP) of the National Cancer Institute (NCI), with grade 0 representing the least severe adverse side effect and grade 4 representing the most severe adverse event. Ideally, the method of the present invention is associated with the least severe adverse events, such as grade 0, 1, or 2 adverse events in the CTEP / NCI classification. However, as discussed herein, while reduction in tumor size is preferred, it is not necessary, because even if tumor cells are eradicated, the actual size of the tumor may not shrink. Eradication of cancer cells is sufficient to achieve a therapeutic effect. Similarly, any reduction in tumor size is sufficient to achieve a therapeutic effect.

[0256] The detection, monitoring, and rating of various human cancers are further described in Cancer Facts and Figures 2001, American Cancer Society, New York, NY, and international patent application WO 01 / 24684. Therefore, clinicians can use standard tests to determine the efficacy of various embodiments of the methods of this invention in treating cancer. However, in addition to tumor size and spread, clinicians may also consider the patient's quality of life and survival when assessing treatment effectiveness.

[0257] In some embodiments, administration of a CDK9 inhibitor provides improved therapeutic efficacy. Improved efficacy can be measured using any method known in the art, including but not limited to the methods described herein. In some embodiments, using appropriate measures (e.g., tumor size reduction, duration of tumor size stability, duration of metastasis-free events, duration of disease-free survival), improved therapeutic efficacy is defined as an improvement of at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, 95%, 100%, 110%, 120%, 150%, 200%, 300%, 400%, 500%, 600%, 700%, 1000%, or more. Using appropriate measures (e.g., tumor size reduction, duration of tumor size stabilization, duration of metastasis-free period, duration of disease-free survival), the improved efficacy can also be expressed as an improvement factor, such as at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 1000, 10000 or more.

[0258] Pharmaceutical Composition

[0259] The compositions of this disclosure can be formulated into any suitable pharmaceutical formulation. The pharmaceutical compositions of this disclosure typically comprise an active ingredient (e.g., a compound of formula (I) or a pharmaceutically acceptable salt and / or coordination complex thereof) and one or more pharmaceutically acceptable excipients or carriers, including but not limited to: inert solid diluents and fillers, diluents, sterile aqueous solutions and various organic solvents, penetration enhancers, solubilizers and adjuvants. The compositions of this disclosure can be formulated into any suitable pharmaceutical formulation. In some embodiments, the pharmaceutically acceptable carrier or excipient is selected from water, ethanol, glycerol, chitosan, alginate, chondroitin, vitamin E, mineral oil and dimethyl sulfoxide (DMSO).

[0260] Pharmaceutical formulations can be provided in any suitable form, depending on the route of administration. In some embodiments, the pharmaceutical compositions disclosed herein can be formulated into dosage forms for administration to a subject. In some embodiments, the pharmaceutical compositions are formulated for oral, intravenous, intra-arterial, aerosol, parenteral, oral, topical, transdermal, rectal, intramuscular, subcutaneous, intraosseous, intranasal, intrapulmonary, transmucosal, inhalation, and / or intraperitoneal administration. In some embodiments, the dosage form is formulated for oral administration. For example, the pharmaceutical compositions can be formulated into pills, tablets, capsules, inhalers, liquid suspensions, liquid emulsions, gels, or powders. In some embodiments, the pharmaceutical compositions can be formulated into unit doses in liquid, gel, semi-liquid, semi-solid, or solid forms.

[0261] The amount of each compound administered depends on the mammal being treated, the severity of the condition or illness, the rate of administration, the disposal of the compound, and the prescribing physician's considerations. However, the effective dose can range from about 0.001 to about 100 mg / kg body weight / day, either as a single dose or in fractions. In some cases, dose levels below the lower limit of the aforementioned range may be sufficient, while in others, larger doses may be used without causing any harmful side effects, for example, by dividing such a larger dose into several smaller doses for administration throughout the day. In some embodiments, the effective dose may be provided in a pulsed administration manner (i.e., administering the compound for several consecutive days, followed by several consecutive days without administration).

[0262] In some embodiments, the composition is provided in one or more unit doses. For example, the composition may be administered in doses of 1, 2, 3, 4, 5, 6, 7, 14, 30, 60 or more. Such amounts may be administered daily, for example, as a single dose administered once, twice, or three or more times daily. However, the daily doses described herein should not be construed as requiring daily administration of a single dose. For example, if one of the agents is provided in a suitable sustained-release form, two or more daily doses may be administered at a lower frequency, for example, as a pre-injectable or pre-oral medication, every two days to monthly or longer. Most typically and conveniently for the subject, a CDK9 inhibitor may be administered once daily, for example, in the morning, evening, or during the day.

[0263] The unit doses can be administered simultaneously or sequentially. The composition can be administered over an extended period of treatment. Illustratively, the period of treatment can be at least about one month, for example at least about three months, at least about six months, or at least about one year. In some cases, administration can continue for the remainder of the subject's life.

[0264] In some implementations, the CDK9 inhibitor may be administered as part of a treatment regimen that includes the simultaneous or sequential administration of one or more second agents (e.g., 1, 2, 3, 4, 5, or more second agents). When administered sequentially, the CDK9 inhibitor may be administered before or after one or more second agents. When administered simultaneously, the CDK9 inhibitor and one or more second agents may be administered via the same route (e.g., injection at the same site; simultaneous oral tablet administration), via different routes (e.g., oral tablet administration concurrent with intravenous infusion), or as part of the same combination (e.g., a solution containing the CDK9 inhibitor and one or more second agents).

[0265] The combination therapy according to the invention can be effective over a wide range of dosages. For example, in the treatment of adults, examples of dosages that can be used are 0.01 to 1000 mg, 0.5 to 100 mg, 1 to 50 mg / day, and 5 to 40 mg / day. The exact dosage will depend on the chosen agent, route of administration, form of the compound, the patient being treated, the patient's weight, and the preferences and experience of the attending physician.

[0266] Pharmaceutical compositions for oral administration

[0267] In some embodiments, this disclosure provides pharmaceutical compositions for oral administration comprising at least one compound of this disclosure and a pharmaceutical excipient suitable for oral administration. The composition may be in solid, liquid, gel, semi-liquid, or semi-solid form. In some embodiments, the composition further comprises a second pharmaceutical agent.

[0268] In some embodiments, the present invention provides a solid pharmaceutical composition for oral administration comprising: (i) a CDK9 inhibitor; and (ii) a pharmaceutical excipient suitable for oral administration. In some embodiments, the composition further comprises: (iii) a third or even a fourth agent. In some embodiments, each compound or agent is present in a therapeutically effective amount. In other embodiments, one or more compounds or agents are present in a subtherapeutic amount, and the compounds and agents work synergistically to provide a therapeutically effective pharmaceutical composition.

[0269] The pharmaceutical compositions of this disclosure suitable for oral administration can be presented in discrete dosage forms, such as hard capsules or soft capsules, flat capsules, troche, lozenges, tablets, or liquids or aerosol sprays, each containing a predetermined amount of an active ingredient as a powder or granules, a solution or suspension in an aqueous or non-aqueous liquid, an oil-in-water emulsion or a water-in-oil liquid emulsion, or a dispersible powder or granules, syrup, or elixir. Such dosage forms can be prepared by any pharmaceutical method, which generally includes the step of binding the active ingredient to a carrier. Generally, the composition is prepared by uniformly and closely mixing the active ingredient with a liquid carrier or a finely segmented solid carrier, or both, and then, if desired, shaping the product into the desired presentation form. For example, tablets can be prepared by compression or molding, optionally having one or more excipients. Compressed tablets can be prepared by compressing the active ingredient in a free-flowing form (such as powder or granules) in a suitable machine, optionally mixed with excipients, such as, but not limited to, binders, lubricants, inert diluents, and / or surfactants or dispersants. Molded tablets can be prepared by molding a mixture of powdered compounds wetted with an inert liquid diluent in a suitable machine.

[0270] This disclosure also includes anhydrous pharmaceutical compositions and dosage forms containing active ingredients, as water can promote the degradation of some compounds. For example, in the pharmaceutical field, water (e.g., 5%) can be added as a means of simulating long-term storage to determine properties such as shelf life or the stability of the formulation over time. The anhydrous pharmaceutical compositions and dosage forms of this disclosure can be prepared using anhydrous or low-water-content ingredients and under low-moisture or low-humidity conditions. The lactose-containing pharmaceutical compositions and dosage forms of this disclosure can be prepared as anhydrous if substantial contact with moisture and / or humidity is anticipated during manufacturing, packaging, and / or storage. Anhydrous pharmaceutical compositions can be prepared and stored to maintain their anhydrous properties. Therefore, anhydrous compositions can be packaged using known materials that prevent exposure to water, allowing them to be contained in suitable formulation kits. Examples of suitable packaging include, but are not limited to, sealing foil, plastics, unit-dose containers, blister packs, and strip packs.

[0271] According to conventional pharmaceutical compounding techniques, active ingredients can be combined with drug carriers in a closely mixed form. The carrier can take various forms depending on the desired formulation for administration. In preparing compositions for oral dosage forms, in the case of oral liquid formulations (such as suspensions, solutions, or elixirs) or aerosols, any commonly used pharmaceutical medium can be used as the carrier, such as water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents, etc.; or, in some embodiments where lactose is not used, in the case of oral solid dosage forms, carriers such as starch, sugar, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, and disintegrants can be used. For example, suitable carriers for solid oral dosage forms include powders, capsules, and tablets. If desired, tablets can be coated using standard aqueous or non-aqueous techniques.

[0272] Binders suitable for pharmaceutical compositions and dosage forms include, but are not limited to, corn starch, potato starch or other starches, gelatin, natural and synthetic gums such as gum arabic, sodium alginate, alginic acid, other alginates, tragacanth gum powder, guar gum, cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate, calcium carboxymethyl cellulose, sodium carboxymethyl cellulose), polyvinylpyrrolidone, methyl cellulose, pregelatinized starch, hydroxypropyl methyl cellulose, microcrystalline cellulose and mixtures thereof.

[0273] Examples of suitable fillers for the pharmaceutical compositions and dosage forms disclosed herein include, but are not limited to, talc, calcium carbonate (e.g., granules or powder), microcrystalline cellulose, powdered cellulose, glucose binders, kaolin, mannitol, silicic acid, sorbitol, starch, pregelatinized starch, and mixtures thereof.

[0274] Disintegrants may be used in the compositions of this disclosure to provide tablets that disintegrate upon exposure to an aqueous environment. Excessive disintegrant may result in tablets that may disintegrate in the bottle. Insufficient disintegrant may be insufficient to cause disintegration and may alter the rate and extent of release of the active ingredient from the dosage form. A sufficient amount of disintegrant, neither too little nor too much, may be used to form dosage forms of the compounds disclosed herein so as not to adversely alter the release of the active ingredient. The amount of disintegrant used may vary depending on the type of formulation and the administration method, and may be readily discerned by one of ordinary skill in the art. About 0.5 to about 15% by weight of disintegrant or about 1 to about 5% by weight of disintegrant may be used in the pharmaceutical composition. Disintegrants that can be used to form pharmaceutical compositions and dosage forms disclosed herein include, but are not limited to, agar, alginate, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, croscarmellose, polycrylene potassium, sodium glycolate starch, potato or cassava starch, other starches, pregelatinized starches, other corn starches, clay, other alginates, other celluloses, gums, or mixtures thereof.

[0275] Lubricants that can be used to form the pharmaceutical compositions and dosage forms disclosed herein include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oils (e.g., peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar, or mixtures thereof. Additional lubricants include, for example, syloid silica gel, coagulated aerosols of synthetic silica, or mixtures thereof. Lubricants may optionally be added in an amount less than about 1% by weight of the pharmaceutical composition.

[0276] When oral administration requires an aqueous suspension and / or elixir, the active ingredient may be combined with various sweeteners or flavorings, colorants or dyes, and, if desired, with emulsifiers and / or suspending agents, as well as diluents such as water, ethanol, propylene glycol, glycerin, and various combinations thereof.

[0277] Tablets may be uncoated or coated using known techniques to delay disintegration and absorption in the gastrointestinal tract, thereby providing sustained action over a longer period. For example, delaying materials such as glyceryl monostearate or glyceryl distearate may be used. Formulations for oral administration may also be presented as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate, or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with an aqueous or oily medium (e.g., peanut oil, liquid paraffin, or olive oil).

[0278] Surfactants that can be used to form the pharmaceutical compositions and dosage forms disclosed herein include, but are not limited to, hydrophilic surfactants, lipophilic surfactants, and mixtures thereof. That is, mixtures of hydrophilic surfactants, mixtures of lipophilic surfactants, or mixtures of at least one hydrophilic surfactant and at least one lipophilic surfactant may be used.

[0279] Suitable hydrophilic surfactants typically have an HLB value of at least 10, while suitable lipophilic surfactants typically have an HLB value of about 10. The empirical parameter used to characterize the relative hydrophilicity and hydrophobicity of nonionic amphiphilic compounds is the hydrophilic-lipophilic balance (“HLB” value). Surfactants with lower HLB values ​​are more lipophilic or hydrophobic and have greater solubility in oils, while surfactants with higher HLB values ​​are more hydrophilic and have greater solubility in aqueous solutions. Hydrophilic surfactants are generally considered to be those compounds with HLB values ​​greater than about 10, as well as anionic, cationic, or zwitterionic compounds for which the HLB scale is not typically applicable. Similarly, lipophilic (i.e., hydrophobic) surfactants are compounds with HLB values ​​equal to or less than about 10. However, the HLB value of a surfactant is only a rough guideline commonly used to formulate industrial, pharmaceutical, and cosmetic emulsions.

[0280] Hydrophilic surfactants can be ionic or nonionic. Suitable ionic surfactants include, but are not limited to, alkylammonium salts; fusidates; fatty acid derivatives of amino acids, oligopeptides, and polypeptides; glycerol ester derivatives of amino acids, oligopeptides, and polypeptides; lecithin and hydrogenated lecithin; lysophosphatidylcholine and hydrogenated lysophosphatidylcholine; phospholipids and their derivatives; lysophosphatidylcholine and its derivatives; carnitine fatty acid ester salts; alkyl sulfates; fatty acid salts; sodium docusate; acyl lactates; monoacetylated and diacetylated tartrate esters of monoglycerides and diglycerides; succinylated monoglycerides and diglycerides; citrate esters of monoglycerides and diglycerides; and mixtures thereof.

[0281] In the above group, ionic surfactants include, for example: lecithin, lysophosphatidylcholine, phospholipids, lysophosphatidylcholine and their derivatives; carnitine fatty acid ester salts; alkyl sulfates; fatty acid salts; sodium docusate; acyl lactates; monoacetylated and diacetylated tartrate esters of monoglycerides and diglycerides; succinylated monoglycerides and diglycerides; citrate esters of monoglycerides and diglycerides; and mixtures thereof.

[0282] Ionic surfactants can be lecithin, lysophosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidic acid, phosphatidylserine, lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylserine, PEG-phosphatidylethanolamine, PVP-phosphatidylethanolamine, lactyl esters of fatty acids, stearoyl-2-lactic acid ester, stearoyl lactate, succinylated monoglycerides, monoacetylated and diacetylated tartrate esters of monoglycerides / diglycerides, citrate esters of monoglycerides / diglycerides, cholycylsarcosine, hexanoate, caprylate, caprate, laurate, myristate, palmitate, oleate, castor oil ester, linoleate, linolenic acid ester, stearate, dodecyl sulfate, tetradecyl sulfate, sodium docusate, lauroylcarnitine, palmitoylcarnitine, myristoylcarnitine, and their salts and mixtures in ionized form.

[0283] Hydrophilic nonionic surfactants may include, but are not limited to, alkyl glucosides; alkyl maltodextrins; alkyl thioglucosides; lauryl polyethylene glycol glycerides; polyoxyalkylene alkyl ethers, such as polyethylene glycol alkyl ethers; polyoxyalkylene alkylphenols, such as polyethylene glycol alkylphenols; polyoxyalkylene alkylphenol fatty acid esters, such as polyethylene glycol fatty acid monoesters and polyethylene glycol fatty acid diesters; polyethylene glycol glycerol fatty acid esters; polyglycerol fatty acid esters; polyoxyalkylene sorbitan fatty acid esters, such as polyethylene glycol sorbitan fatty acid esters; hydrophilic transesterification products of polyols with at least one member of glycerides, vegetable oils, hydrogenated vegetable oils, fatty acids, and sterols; polyoxyethylene sterols, their derivatives, and analogs; polyoxyethyleneized vitamins and their derivatives; polyoxyethylene-polyoxypropylene block copolymers; and mixtures thereof; polyethylene glycol sorbitan fatty acid esters and hydrophilic transesterification products of polyols with at least one member of triglycerides, vegetable oils, and hydrogenated vegetable oils. Polyols can be glycerol, ethylene glycol, polyethylene glycol, sorbitol, propylene glycol, pentaerythritol, or sugars.

[0284] Other hydrophilic nonionic surfactants include, but are not limited to, PEG-10 laurate, PEG-12 laurate, PEG-20 laurate, PEG-32 laurate, PEG-32 dilaurate, PEG-12 oleate, PEG-15 oleate, PEG-20 oleate, PEG-20 dioleate, PEG-32 oleate, PEG-200 oleate, PEG-400 oleate, PEG-15 stearate, PEG-32 distearate, PEG-40 stearate, PEG-100 stearate, PEG-20 dilaurate, PEG-25 glyceryl trioleate, PEG-32 dioleate, PEG-20 glyceryl laurate, PEG-30 glyceryl laurate, PEG-20 glyceryl stearate, PEG-20 glyceryl oleate, PEG-30 glyceryl oleate, PEG-30 glyceryl laurate, PEG-40 glyceryl laurate, and PEG-40 palm kernel oleate. Oils, PEG-50 hydrogenated castor oil, PEG-40 castor oil, PEG-35 castor oil, PEG-60 castor oil, PEG-40 hydrogenated castor oil, PEG-60 hydrogenated castor oil, PEG-60 corn oil, PEG-6 glyceryl caprylate / caprylate, PEG-8 glyceryl caprylate / caprylate, polyglycerol-10 laurate, PEG-30 cholesterol, PEG-25 phytosterols, PEG-30 soybean sterols, PEG-20 trioleate, PEG PEG-40 Sorbitan Oleate, PEG-80 Sorbitan Laurate, Polysorbate 20, Polysorbate 80, POE-9 Lauryl Ether, POE-23 Lauryl Ether, POE-10 Oil-based Ether, POE-20 Oil-based Ether, POE-20 Stearyl Ether, Tocopherol PEG-100 Succinate, PEG-24 Cholesterol, Polyglycerol-10 Oleate, Tween 40, Tween 60, Sucrose Monostearate, Sucrose Monolaurate, Sucrose Monopalmitate, PEG 10-100 Nonylphenol Series, PEG 15-100 Octylphenol Series, and Poloxamer.

[0285] Suitable lipophilic surfactants, by way of example only, include: fatty alcohols; glycerol fatty acid esters; acetylated glycerol fatty acid esters; lower alcohol fatty acid esters; propylene glycol fatty acid esters; sorbitan fatty acid esters; polyethylene glycol sorbitan fatty acid esters; sterols and sterol derivatives; polyoxyethylene sterols and sterol derivatives; polyethylene glycol alkyl ethers; sugar esters; sugar ethers; lactic acid derivatives of monoglycerides and diglycerides; hydrophobic transesterification products of polyols with at least one member of glycerides, vegetable oils, hydrogenated vegetable oils, fatty acids, and sterols; oil-soluble vitamin / vitamin derivatives; and mixtures thereof. Within this group, preferred lipophilic surfactants include glycerol fatty acid esters, propylene glycol fatty acid esters, and mixtures thereof, or hydrophobic transesterification products of polyols with at least one member of vegetable oils, hydrogenated vegetable oils, and triglycerides.

[0286] In one embodiment, the composition may include a solubilizer to ensure good solubilization and / or dissolution of the compounds of this disclosure and to minimize precipitation of the compounds of this disclosure. This is particularly important for compositions intended for non-oral use (e.g., compositions for injection). Solubilizers may also be added to increase the solubility of hydrophilic drugs and / or other components (such as surfactants), or to maintain the composition as a stable or homogeneous solution or dispersion.

[0287] Examples of suitable solubilizers include, but are not limited to, the following: alcohols and polyols, such as ethanol, isopropanol, butanol, benzyl alcohol, ethylene glycol, propylene glycol, butanediol and its isomers, glycerol, pentaerythritol, sorbitol, mannitol, diethylene glycol monoethyl ether, dimethyl isosorbide, polyethylene glycol, polypropylene glycol, polyvinyl alcohol, hydroxypropyl methylcellulose and other cellulose derivatives, cyclodextrins and cyclodextrin derivatives; ethers of polyethylene glycol with an average molecular weight of about 200 to about 6000, such as tetrahydrofurfuryl PEG ether (glycofurol) or methoxy PEG; amides and other nitrogen-containing compounds, such as 2-pyrrolidone, 2-piperidinone, ε-caprolactone Amines, N-alkylpyrrolidones, N-hydroxyalkylpyrrolidones, N-alkylpiperidones, N-alkylcaprolactams, dimethylacetamide, and polyvinylpyrrolidone; esters, such as ethyl propionate, tributyl citrate, acetyl triethyl citrate, acetyl tributyl citrate, triethyl citrate, ethyl oleate, ethyl octanoate, ethyl butyrate, triacetyl triacetate, propylene glycol monoacetate, propylene glycol diacetate, ε-caprolactone and its isomers, δ-valerolactone and its isomers, β-butyrolactone and its isomers; and other solubilizers known in the art, such as dimethylacetamide, dimethyl isosorbide, N-methylpyrrolidone, monooctyl ether, diethylene glycol monoethyl ether, and water.

[0288] Mixtures of solubilizers may also be used. Examples include, but are not limited to, glyceryl triacetate, triethyl citrate, ethyl oleate, ethyl octanoate, dimethylacetamide, N-methylpyrrolidone, N-hydroxyethylpyrrolidone, polyvinylpyrrolidone, hydroxypropyl methylcellulose, hydroxypropyl cyclodextrin, ethanol, polyethylene glycol 200-100, poly(ethylene glycol) tetrahydrofurfuryl ether (glycofurol), diethylene glycol monoethyl ether, propylene glycol, and dimethyl isosorbide diester. Particularly preferred solubilizers include sorbitol, glycerol, glyceryl triacetate, ethanol, PEG-400, poly(ethylene glycol) tetrahydrofurfuryl ether, and propylene glycol.

[0289] There are no particular limitations on the amount of solubilizer that may be included. The amount of a given solubilizer may be limited to a biologically acceptable amount, which can be readily determined by those skilled in the art. In some cases, including an amount of solubilizer far exceeding a biologically acceptable amount may be advantageous, for example, to maximize the concentration of the drug and to remove excess solubilizer before serving the composition to the patient using conventional techniques such as distillation or evaporation. If present, the weight percentage of the solubilizer may be 10%, 25%, 50%, 100%, or up to about 200% based on the combined weight of the drug and other excipients. Very small amounts of solubilizer, such as 5%, 2%, 1%, or less, may also be used if desired. Typically, the solubilizer may be present in amounts from about 1% to about 100% by weight, and more typically from about 5% to about 25% by weight.

[0290] The composition may also include one or more pharmaceutically acceptable additives and excipients. Such additives and excipients include, but are not limited to, anti-sticking agents, defoamers, buffers, polymers, antioxidants, preservatives, chelating agents, viscosity modifiers, tonicifiers, flavoring agents, coloring agents, taste enhancers, opacifiers, suspensions, binders, fillers, plasticizers, lubricants, and mixtures thereof.

[0291] In addition, acids or bases may be incorporated into the composition to facilitate processing, enhance stability, or for other reasons. Examples of pharmaceutically acceptable bases include amino acids, amino acid esters, ammonium hydroxide, potassium hydroxide, sodium hydroxide, sodium bicarbonate, aluminum hydroxide, calcium carbonate, magnesium hydroxide, magnesium aluminum silicate, synthetic aluminum silicate, synthetic hydrocalcite, magnesium aluminum hydroxide, diisopropylethylamine, ethanolamine, ethylenediamine, triethanolamine, triethylamine, triisopropanolamine, trimethylamine, tri(hydroxymethyl)aminomethane (TRIS), etc. Bases that are salts of pharmaceutically acceptable acids are also suitable. These pharmaceutically acceptable acids include acetic acid, acrylic acid, adipic acid, alginic acid, alkanolamine, amino acids, ascorbic acid, benzoic acid, boric acid, butyric acid, carbonic acid, citric acid, fatty acids, formic acid, fumaric acid, gluconic acid, hydroquinone sulfonic acid, isoascorbic acid, lactic acid, maleic acid, oxalic acid, p-bromobenzenesulfonic acid, propionic acid, p-toluenesulfonic acid, salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid, mercaptoacetic acid, toluenesulfonic acid, uric acid, etc. Salts of polybasic acids, such as sodium phosphate, disodium hydrogen phosphate, and sodium dihydrogen phosphate, can also be used. When the base is a salt, the cation can be any convenient and pharmaceutically acceptable cation, such as ammonium, alkali metals, alkaline earth metals, etc. Examples can include, but are not limited to, sodium, potassium, lithium, magnesium, calcium, and ammonium.

[0292] Suitable acids are pharmaceutically acceptable organic or inorganic acids. Examples of suitable inorganic acids include hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, boric acid, and phosphoric acid. Examples of suitable organic acids include acetic acid, acrylic acid, adipic acid, alginic acid, alkanesulosic acid, amino acids, ascorbic acid, benzoic acid, boric acid, butyric acid, carbonic acid, citric acid, fatty acids, formic acid, fumaric acid, gluconic acid, hydroquinone sulfonic acid, isoascorbic acid, lactic acid, maleic acid, methanesulfonic acid, oxalic acid, p-bromobenzenesulfonic acid, propionic acid, p-toluenesulfonic acid, salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid, thioglycolic acid, toluenesulfonic acid, and uric acid.

[0293] Example

[0294] The following abbreviations are used in the examples: ATP = adenosine triphosphate, DBU = 1,8-diazabicyclo[5.4.0]undec-7-ene, DCE = 1,2-dichloroethane, DCM = dichloromethane, DEA = diethylamine, DHP = dihydropyran, DIPEA = N,N-diisopropylethylamine, DME = ethylene glycol dimethyl ether, DMF = dimethylformamide, DMSO = dimethyl sulfoxide, EtOH = ethanol, DTT = dithiothreitol, HPLC = high performance liquid chromatography, PMB = p-methoxybenzyl, PPTS = pyridinium p-toluenesulfonate, SFC = supercritical fluid chromatography, TBME = tert-butylmethyl ether, TEA = triethylamine, TFA = trifluoroacetic acid, THF = tetrahydrofuran, THP = tetrahydropyran.

[0295] All chemicals, reagents and solvents were purchased from commercial sources (if available) and used without further purification.

[0296] Example 1: Synthesis of 4-(((2'-(((1R,4R)-4-aminocyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (intermediate 1)

[0297]

[0298] Step 1: Preparation of tetrahydro-4H-pyran-4,4-dicarboxynitrile (INT-2)

[0299]

[0300] DBU (26.26 g, 172.48 mmol) was added to a DMF (30 mL) solution of 1-bromo-2-(2-bromoethoxy)ethane (INT-1, 20 g, 86.24 mmol) and malononitrile (6.27 g, 94.86 mmol). The reaction mixture was stirred at 85 °C for 3 h, cooled to ambient temperature, diluted with water (100 mL), and extracted with ethyl acetate (100 mL × 3). The combined organic phases were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to provide tetrahydro-4H-pyran-4,4-dicarboxynitrile (INT-2, 12.61 g, crude product) as a brown solid. The crude product was used directly in the next step without further purification. 1 HNMR (400MHz, methanol-d4) δ=3.86-3.77(m,4H), 2.32-2.21(m,4H).

[0301] Step 2: Preparation of 4-(aminomethyl)tetrahydro-2H-pyran-4-carboxynitrile (INT-3)

[0302]

[0303] NaBH4 (7.50 g, 198.31 mmol) was added fractionally to a solution of tetrahydro-4H-pyran-4,4-dicarboxynitrile (INT-2, 9.0 g, 66.10 mmol) in EtOH (270 mL). The reaction mixture was stirred at 20 °C for 4 h, quenched with water (200 mL), and extracted with ethyl acetate (200 mL × 3). The combined organic phases were washed with brine (150 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to provide 4-(aminomethyl)tetrahydro-2H-pyran-4-carboxynitrile (INT-3, 7.23 g, 78% yield) as a brown oil. The crude product was used directly in the next step without further purification.

[0304] Step 3: Preparation of 4-(((6-bromopyridin-2-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (INT-4)

[0305]

[0306] TEA (13.05 g, 128.94 mmol) was added to a DMSO (80 mL) solution of 4-(aminomethyl)tetrahydro-2H-pyran-4-carboxynitrile (INT-3, 7.23 g, 51.58 mmol) and 2-bromo-6-fluoro-pyridine (7.72 g, 43.84 mmol). The reaction mixture was stirred at 130 °C for 18 h, cooled to ambient temperature, diluted with ethyl acetate (100 mL), washed with saturated NaHCO3 solution and brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The resulting residue was purified by silica gel chromatography (Biotage 20 g silica gel rapid column; 0-25% petroleum ether in ethyl acetate, @40 mL / min) to provide 4-(((6-bromopyridin-2-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (INT-4, 3.8 g, 18.35% yield) as a pale green solid. MS(ESI)m / z = 296.1[M+H] + .

[0307] Step 4: Preparation of 4-(((5'-chloro-2'-fluoro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (INT-5)

[0308]

[0309] Na₂CO₃ (2M, 16.04 mL) was added to a DME (40 mL) solution of 4-(((6-bromopyridin-2-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (INT-4, 3.8 g, 12.83 mmol), (5-chloro-2-fluoro-4-pyridinyl)boronic acid (3.37 g, 19.25 mmol), and Pd(dppf)Cl₂ (938.84 mg, 1.28 mmol). The reaction mixture was sealed and stirred at 110 °C for 4 hours under N₂. After cooling to ambient temperature, the mixture was diluted with water (40 mL) and extracted with ethyl acetate (70 mL × 3). The combined organic phases were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The resulting residue was purified by silica gel chromatography (Biotage 40g silica gel rapid column; 20-26% petroleum ether in ethyl acetate, @80mL / min) to provide 4-(((5'-chloro-2'-fluoro-[2,4'-bipyridin]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (INT-5, 3.6g, 73% yield) as a yellow oil. MS (ESI) m / z = 347.1 [M+H] + .

[0310] Step 5: Preparation of 4-(((2'-(((1R,4R)-4-aminocyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (intermediate 1)

[0311]

[0312] TEA (2.53 g, 24.97 mmol) was added to a DMSO (50 mL) solution of 4-(((5'-chloro-2'-fluoro-[2,4'-bipyridin]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (INT-5, 4.33 g, 12.49 mmol) and cyclohexane-1,4-diamine (2.14 g, 18.73 mmol). The reaction mixture was stirred at 110 °C for 16 h, diluted with water (40 mL), and extracted with ethyl acetate (100 mL × 3). The combined organic phases were washed with brine (60 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The resulting residue was dissolved in ethyl acetate (100 mL), added dropwise to HCl / dioxane (50 mL), filtered, and washed with ethyl acetate. The resulting solid was dissolved in water (150 mL), alkalized to pH 9 with NaHCO3, and extracted with ethyl acetate (100 mL × 3). The combined organic phases were washed with brine (60 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to provide 4-(((2'-(((1R,4R)-4-aminocyclohexyl)amino)-5'-chloro-[2,4'-bipyridin]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (intermediate 1, 4.1 g, 61.11% yield) as a brown solid. MS (ESI) m / z = 441.3 [M+H] + .

[0313] Example 2: Synthesis of 1-((2-(tetrahydro-2H-pyran-2-yl)-2H-tetrazol-5-yl)methoxy)prop-2-one (intermediate 2)

[0314]

[0315] Step 1: Preparation of 5-(chloromethyl)-2H-tetrazole (INT-7)

[0316]

[0317] Add NaN3 (4.87 g, 74.97 mmol) to a solution of AlCl3 (3.67 g, 27.55 mmol) in THF (50 mL). Stir the reaction mixture at 60 °C for 2 hours and cool to 20 °C. Add 2-chloroacetonitrile (INT-6, 2.0 g, 26.49 mmol), and heat the reaction mixture to 70 °C and stir for 24 hours. Concentrate the reaction mixture and acidify the resulting residue to pH 2 with an aqueous HCl solution (37%) and extract with EtOAc (100 mL × 3). Dry the combined organic layers with Na2SO4, filter, and concentrate to provide 5-(chloromethyl)-2H-tetrazole (INT-7, 3.0 g, crude product) as a white solid. The crude product was used directly in the next step without further purification. 1 H NMR (400MHz, DMSO-d6): δ = 5.08 (s, 2H).

[0318] Step 2: Preparation of 5-(chloromethyl)-2-(tetrahydro-2H-pyran-2-yl)-2H-tetrazole (INT-8)

[0319]

[0320] PPTS (203.54 mg, 0.809 mmol) was added to a solution of 5-(chloromethyl)-2H-tetrazole (INT-7, 3.0 g, 25.31 mmol) and DHP (4.26 g, 50.62 mmol) in acetone (100 mL). The reaction mixture was stirred at 45 °C for 3 hours and concentrated. The residue was purified by silica gel chromatography (Biotage 20 g silica gel rapid column; 0-10% petroleum ether in ethyl acetate, @40 mL / min) to provide 5-(chloromethyl)-2-(tetrahydro-2H-pyran-2-yl)-2H-tetrazole (INT-8, 4.6 g, 89% yield) as a colorless oil. 1 H NMR (400MHz, DMSO-d6): δ = 6.18 (dd, J = 8.0, 2.8Hz, 1H), 5.06 (s, 2H), 3.81-3.76 (m, 2H), 2.51 -2.26(m,1H),2.25-2.13(m,1H),2.12-1.98(m,1H),1.64-1.62(m,1H),1.61-1.601(m,2H).

[0321] Step 3: Preparation of 5-(((2-methylallyl)oxy)methyl)-2-(tetrahydro-2H-pyran-2-yl)-2H-tetrazole (INT-9)

[0322]

[0323] To a solution of 2-methylprop-2-en-1-ol (1.07 g, 14.80 mmol, 1.25 mL) in DMF (2 mL), NaH (789 mg, 19.74 mmol, 60% purity) was added, and the reaction mixture was stirred at 0 °C for 0.5 h. 5-(chloromethyl)-2-(tetrahydro-2H-pyran-2-yl)-2H-tetraazole (INT-8, 2.0 g, 9.87 mmol) was added, and the reaction mixture was stirred at 25 °C for 2 h, quenched with brine (50 mL), and extracted with EtOAc (50 mL × 3). The combined organic layers were washed with brine (50 mL × 7), dried over Na₂SO₄, filtered, and concentrated to provide 5-(((2-methylallyl)oxy)methyl)-2-(tetrahydro-2H-pyran-2-yl)-2H-tetraazole (INT-9, 2.3 g, 97% yield) as a white solid. The crude product can be used directly in the next step without further purification. 1 H NMR (400MHz, DMSO-d6): δ = 6.15 (dd, J = 8.0, 3.2Hz, 1H), 4.95 (s, 1H), 4.89 (s, 1H), 4.71 (s, 2H), 3.95 (s, 2H), 3.82-3.7 5(m,2H),2.26-2.25(m,1H),2.25-2.24(m,1H),2.16-2.10(m,1H),1.67-1.66(m,1H),1.64(s,3H),1.62-1.58(m,2H).

[0324] Step 4: Preparation of 1-((2-(tetrahydro-2H-pyran-2-yl)-2H-tetrazol-5-yl)methoxy)prop-2-one (intermediate 2)

[0325]

[0326] A solution of NaIO4 (2.06 g, 9.65 mmol) in H2O (8 mL) was added dropwise to a solution of 5-(((2-methylallyl)oxy)methyl)-2-(tetrahydro-2H-pyran-2-yl)-2H-tetrazole (INT-9, 1.0 g, 4.20 mmol) and K2OsO4·2H2O (15.46 mg, 0.042 mmol) in H2O (3.6 mL) and THF (4 mL). The reaction mixture was stirred at 25 °C for 12 h, filtered, and extracted with EtOAc (30 mL × 5). The combined organic layers were dried over Na2SO4, filtered, and concentrated to provide 1-((2-(tetrahydro-2H-pyran-2-yl)-2H-tetrazole-5-yl)methoxy)prop-2-one (intermediate 2, 800 mg, 79% yield) as a yellow oil. 1H NMR (400MHz, DMSO-d6): δ = 6.17 (dd, J = 8.0, 3.6Hz, 1H), 5.06 (s, 2H), 3.84-3.73 (m, 3H), 2.2 4-2.15(m,1H),2.15-2.13(m,1H),1.99-1.75(m,1H),1.74-1.72(m,4H),1.64-1.59(m,3H). MS(ESI)m / z=263.3[M+Na] + .

[0327] Example 3: Synthesis of 2-(2-oxopropoxy)ethyl acetate (intermediate 3)

[0328]

[0329] Step 1: Preparation of 2-((2-methylallyl)oxy)ethyl acetate (INT-11)

[0330]

[0331] NaH (610 mg, 15.26 mmol) was added to a DMF (12 mL) solution of 2-methylprop-2-en-1-ol (1.0 g, 13.87 mmol). The reaction mixture was stirred at 0 °C for 10 min, and ethyl 2-bromoacetate (INT-10, 2.32 g, 13.87 mmol) was added. The reaction mixture was stirred at 0 °C for 1 h, quenched with water (5 mL) at 0 °C, and extracted with ethyl acetate (20 mL × 3). The combined organic phases were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The resulting residue was purified by silica gel rapid column chromatography (0-20% ethyl acetate in petroleum ether) to provide ethyl 2-((2-methylallyl)oxy)acetate as a colorless liquid (INT-11, 780 mg, 36% yield). 1 H NMR (400MHz, DMSO-d6) δ = 4.89-4.86 (m, 1H), 4.57-4.45 (m, 1H), 4.11 (s, 2H), 4.09-4.06(m,2H),3.92-3.90(m,2H),1.68-1.67(m,3H),1.23-1.16(m,3H). MS(ESI)m / z=159.3[M+H] + .

[0332] Step 2: Preparation of 2-(2-oxopropoxy)ethyl acetate (intermediate 3)

[0333]

[0334] A solution of K₂O₄·2H₂O (34.94 mg, 0.1 mmol) in a mixture of H₂O (4 mL) and THF (9 mL) and a solution of NaIO₄ (4.66 g, 21.81 mmol) in H₂O (12 mL) were added to a solution of ethyl 2-((2-methylallyl)oxy)ethyl acetate (INT-11, 1.5 g, 9.48 mmol) in THF (8 mL). The reaction mixture was stirred at 20 °C for 11 h and concentrated under vacuum. The resulting residue was purified by silica gel chromatography (Biotage 20 g silica gel rapid column; 0-10% petroleum ether in ethyl acetate, @40 mL / min) to provide ethyl 2-(2-oxopropoxy)ethyl acetate as a colorless liquid (intermediate 3, 850 mg, 56% yield). 1 H NMR (400MHz, CDCl3) δ = 4.24-4.14 (m, 6H), 2.17 (s, 3H), 1.27 (t, J = 7.2Hz, 3H).

[0335] Example 4: Synthesis of 1-(1-(4-methoxybenzyl)-1H-tetrazol-5-yl)prop-2-one (intermediate 4)

[0336]

[0337] Step 1: Preparation of 4-hydrazino-6-methylpyrimidine (INT-13)

[0338]

[0339] To a solution of 4-chloro-6-methylpyrimidine (INT-12, 2.5 g, 19.45 mmol) in dioxane (30 mL), NH₂NH₂·H₂O (1.67 g, 33.45 mmol) and K₂CO₃ (2.74 g, 19.84 mmol) were added. The reaction mixture was stirred at 100 °C for 6 h, diluted with H₂O (20 mL), and extracted with EtOAc (50 mL × 3). The combined organic layers were washed with brine (50 mL), dried over Na₂SO₄, filtered, and concentrated under vacuum to provide 4-hydrazino-6-methylpyrimidine (INT-13, 2 g, 16.11 mmol, 82.85% yield) as a yellow solid. The crude product was used directly in the next step without further purification.

[0340] Step 2: Preparation of (Z)-N-(1-(1H-tetrazol-5-yl)prop-1-en-2-yl)formamide (INT-14)

[0341]

[0342] HCl (2M, 3.62 mL) and NaNO₂ (500.19 mg, 7.25 mmol) were added to a solution of 4-hydrazino-6-methylpyrimidine (INT-13, 0.9 g, 7.25 mmol) in H₂O (2 mL). The reaction mixture was stirred at 0 °C for 8 hours and filtered. The filter cake was concentrated under reduced pressure to provide (Z)-N-(1-(1H-tetrazol-5-yl)prop-1-en-2-yl)carboxamide (INT-14, 0.6 g, 3.92 mmol, 54.04% yield) as a brown solid. The crude product was used directly in the next step without further purification. MS (ESI) m / z = 296.1 [M+H] + .

[0343] Step 3: Preparation of 5-(2-oxopropyl)-1H-tetrazole-1-onium chloride (INT-15)

[0344]

[0345] HCl (1 mL) was added to a solution of (Z)-N-(1-(1H-tetrazol-5-yl)prop-1-en-2-yl)formamide (INT-14, 0.8 g, 5.22 mmol) in H₂O (4 mL). The reaction mixture was stirred at 80 °C for 12 h and concentrated under reduced pressure to provide 5-(2-oxopropyl)-1H-tetrazol-1-onium chloride as a yellow solid (INT-15, 0.7 g, 4.31 mmol, 82.42% yield). The crude product was used directly in the next step without further purification.

[0346] Step 5: Preparation of 1-(1-(4-methoxybenzyl)-1H-tetrazol-5-yl)prop-2-one (intermediate 4)

[0347]

[0348] DIPEA (3.07 g, 23.79 mmol, 4.14 mL, 5 equivalents) was added to a THF (15 mL) solution of 5-(2-oxopropyl)-1H-tetrazole-1-onium chloride (INT-15, 600 mg, 4.76 mmol, 1 equivalent) and 1-(chloromethyl)-4-methoxybenzene (1.12 g, 7.14 mmol, 971.83 μL, 1.5 equivalents). The reaction mixture was stirred at 45 °C for 16 h, diluted with water (20 mL), and extracted with DCM (30 mL × 3). The combined organic layers were washed with brine (40 mL), dried over Na₂SO₄, filtered, and concentrated. The resulting residue was subjected to rapid silica gel chromatography (…). 20g Purification was performed using a silica gel rapid column (0-6% methanol / dichloromethane, @35 mL / min) to provide 1-(1-(4-methoxybenzyl)-1H-tetrazol-5-yl)prop-2-one as a pale yellow oil (intermediate 4, 440 mg, 1.14 mmol, 24.04% yield, 64% purity). MS (ESI) m / z = 247.2 [M+H] + .

[0349] Example 5: Synthesis of 4-(((2'-(((1R,4R)-4-(((R)-1-((2H-tetrazol-5-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (Compound 2) and 4-(((2'-(((1S,4R)-4-(((S)-1-((2H-tetrazol-5-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (Compound 3)

[0350]

[0351] NaBH(OAc)3 (336 mg, 1.59 mmol) was added to a solution of 4-(((2'-(((1R,4R)-4-aminocyclohexyl)amino)-5'-chloro-[2,4'-bipyridin]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (intermediate 1, 500 mg, 1.13 mmol) and 1-((2-(tetrahydro-2H-pyran-2-yl)-2H-tetrazol-5-yl)methoxy)prop-2-one (intermediate 2, 354 mg, 1.47 mmol) in AcOH (204 mg, 3.40 mmol) and DCE (15 mL) at 0 °C. The mixture was stirred at 25 °C for 12 h, quenched with MeOH (5 mL), and concentrated. The resulting residue was purified by preparative HPLC (Phenomenex Luna C18 75×30 mm (3 μm particle size); 0–35% acetonitrile / water (0.225% FA); 35 min; 25 mL / min) to provide 4-(((2'-((4-((1-((2H-tetrazol-5-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)5'-chloro-[2,4'-bipyridin]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (compound 1, 140 mg, 21% yield) as a white solid. MS (ESI) m / z = 581.5 [M+H] + .

[0352] 4-(((2'-((4-(((1-((2H-tetrazol-5-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (compound 1) was subjected to chiral SFC (Chiralpak OD-3 100×4.6mm (3μm particle size); 5-40% EtOH (0.05%) DEA / CO2; 8 min; 2.8 mL / min) separation to provide 4-(((2'-(((1R,4R)-4-(((R)-1-((2H-tetrazol-5-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (compound 2, 35 mg, 35% yield) , 97% purity, >99% ee) and 4-(((2'-(((1S,4R)-4-(((S)-1-((2H-tetrazol-5-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (compound 3, 35 mg, 35% yield, 99% purity, 97% ee).

[0353] Compound 2: 1 H NMR (400MHz, CDCl3): δ = 8.04 (s, 1H), 7.48 (t, J = 7.6Hz, 1H), 6.91 (d, J = 7.6Hz, 1H), 6.54 (s, 1H), 6.49 (d, J = 8.0Hz, 2H), 5.09-5.05 (m, 1H), 4.87-4.83(m,2H),4.45(d,J=8.0Hz,1H),3.97-3.95(m,2H),3.87-3.85(m,1H),3.74-3.63(m,6H),3.57-3.55(m,2H),3.22-3.21(m,1H). 2.41-2.40(m,2H),2.30-2.10(m,2H),1.91-1.77(m,4H),1.76-1.70(m,2H),1.50-1.48(m,3H),1.29-1.26(m,2H). MS(ESI)m / z=581.5[M+H] + .

[0354] Compound 3: 1H NMR (400MHz, CDCl3): δ = 8.05 (s, 1H), 7.48 (t, J = 7.6Hz, 1H), 6.91 (d, J = 7.6Hz, 1H), 6.54 (s, 1H), 6.49 (d, J = 8.0Hz, 2H), 5.08-5.05 (m, 1H), 4.89-4.83(m,2H),4.45(d,J=8.0Hz,1H),3.97-3.95(m,2H),3.87-3.85(m,1H),3.74-3.63(m,6H),3.57-3.55(m,2H),3.22-3.21(m,1H). 2.41-2.40(m,2H),2.30-2.10(m,2H),1.91-1.77(m,4H),1.76-1.70(m,2H),1.50-1.48(m,3H),1.29-1.26(m,2H). MS(ESI)m / z=581.5[M+H] + .

[0355] Example 6: Synthesis of 2-((R)-2-(((1R,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4]-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)ethyl acetate (compound 5) and 2-((S)-2-(((1R,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4]-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)ethyl acetate (compound 6)

[0356]

[0357] NaBH(OAc)3 (134 mg, 0.63 mmol) was added to a DCE (2 mL) solution of 4-(((2'-(((1R,4R)-4-aminocyclohexyl)amino)-5'-chloro-[2,4'-bipyridin]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (intermediate 1, 200 mg, 0.45 mmol), ethyl acetate 2-(2-oxopropoxy)acetate (intermediate 3, 80 mg, 0.5 mmol), and AcOH (82 mg, 1.36 mmol). The reaction mixture was stirred at 20 °C for 20 h, quenched with water (10 mL) at 0 °C, and extracted with ethyl acetate (30 mL × 3). The combined organic phases were washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The resulting residue was purified by preparative HPLC (Phenomenex Luna C18 100×30mm (5μm particle size); 10-40% acetonitrile / water (0.225% FA); 10 min; 25 mL / min) to provide ethyl acetate (compound 4) as a white solid.

[0358] 2-(2-((4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)ethyl acetate (compound 4) was subjected to chiral SFC (Chiralpak AD-3100 × 4.6 mm (3 μm particle size); 5-40% EtOH (0.05%) DEA / CO2; 8 min; 2.8 mL / min) further separation yielded 2-((R)-2-(((1R,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4]-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)ethyl acetate (compound 5, 18.8 mg, 7% yield, >99% purity, >99% ee) and 2-((S)-2-(((1R,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4]-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)ethyl acetate (compound 6, 21.8 mg, 8% yield, 99% purity, 90% ee) as white solids).

[0359] Compound 5: 11H NMR (400 MHz, CDCl3) δ = 8.11 (s, 1H), 7.53 (t, J = 8.0 Hz, 1H), 6.97 (d, J = 8.0 Hz, 1H), 6.55 (s, 1H), 6.51 (d, J = 8.0 Hz, 1H), 4.78 (t, J = 4.0 Hz, 1H), 4.43 (d, J = 8.0 Hz, 1H), 4.30 - 4.17 (m, 2H), 4.11 (s, 2H), 4.05 - 3.97 (m, 2H), 3.81 - 3.76 (m, 2H), 3.74 - 3.66 (m, 2H), 3.61 - 3.49 (m, 2H), 3.47 - 3.34 (m, 1H), 3.19 - 3.01 (m, 1H), 2.82 - 2.47 (m, 1H), 2.22 - 2.13 (m, 2H), 2.11 - 2.00 (m, 2H), 1.97 - 1.90 (m, 2H), 1.80 - 1.72 (m, 2H), 1.32 - 1.20 (m, 7H), 1.15 - 1.06 (m, 3H). MS (ESI) m / z = 585.4 [M+H] + .

[0360] Compound 6: 1 1H NMR (400 MHz, CDCl3) δ = 8.44 (s, 1H), 8.10 (s, 1H), 7.52 (t, J = 8.0 Hz, 1H), 6.97 (d, J = 4.0 Hz, 1H), 6.57 (s, 1H), 6.52 (d, J = 8.0 Hz, 1H), 4.82 (t, J = 8.0 Hz, 1H), 4.28 - 4.20 (m, 2H), 4.17 - 4.11 (m, 2H), 4.03 - 3.97 (m, 2H), 3.81 - 3.64 (m, 7H), 3.31 - 3.27 (m, 1H), 3.02 - 2.92 (m, 1H), 2.28 - 2.11 (m, 4H), 1.97 - 1.89 (m, 2H), 1.81 - 1.72 (m, 2H), 1.67 - 1.46 (m, 2H), 1.33 - 1.25 (m, 8H). MS (ESI) m / z = 585.4 [M+H] + .

[0361] Example 7: Synthesis of 2-((R)-2-(((1R,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)acetic acid (compound 8) and 2-((S)-2-(((1R,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)acetic acid (compound 9)

[0362]

[0363] To a solution of ethyl acetate (compound 4, 210 mg, 0.35 mmol) in THF (3 mL) and H₂O (1 mL), NaOH (143 mg, 3.59 mmol) was added. The reaction mixture was stirred at 25 °C for 2 hours. The aqueous phase was acidified to pH 4 with aqueous HCl (1 N) and concentrated under vacuum. The resulting residue was dissolved in DMF, filtered, concentrated under vacuum, and purified by preparative HPLC (Phenomenex Luna C18 100×40 mm (3 μm particle size); 15-45% acetonitrile / water (0.225% FA); 10 min; 25 mL / min) to provide 2-(2-((4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)acetic acid (compound 7).

[0364] 2-(2-((4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)acetic acid (compound 7) was further purified by chiral SFC (Chiralpak AD-3 100×4.6mm (3μm particle size); 5-40% EtOH (0.05% DEA) / CO2; 8 min; 2.8 mL / min) to provide 2-((R)-2-(((1R,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)acetic acid (compound 8), 32.5 mg, 1 6% yield, 98% purity, >99% ee) and 2-((S)-2-(((1R,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)acetic acid (compound 9, 34.8 mg, 17% yield, 99% purity, >99% ee).

[0365] Compound 8: 1 H NMR (400MHz, DMSO-d6) δ = 8.16 (s, 1H), 8.03 (s, 1H), 7.54-7.45 (m, 1H), 7.12-7. 05(m,1H),6.79-6.70(m,2H),6.68-6.60(m,2H),3.92-3.86(m,2H),3.71-3.62( m,7H),3.49-3.46(m,2H),3.04-2.92(m,2H),2.13-2.00(m,4H),1.87-1.80(m,2 H),1.71-1.62(m,2H),1.54-1.44(m,2H),1.30-1.21(m,2H),1.17-1.11(m,3H). MS(ESI)m / z = 557.3 [M+H] + .

[0366] Compound 9: 1H NMR(400MHz, DMSO-d6)δ=8.17(s,1H),8.03(s,1H),7.54-7.45(m,1H),7.13-7.04(m ,1H),6.78-6.71(m,2H),6.69-6.64(m,1H),6.63-6.60(m,1H),3.92-3.86(m,2H),3. 70-3.61(m,7H),3.50-3.46(m,2H),3.04-2.90(m,2H),2.14-2.02(m,4H),1.88-1.80 (m,2H),1.73-1.62(m,2H),1.54-1.43(m,2H),1.33-1.21(m,2H),1.18-1.10(m,3H). MS(ESI)m / z=557.3[M+H] + .

[0367] Example 8: Synthesis of 4-(((2'-(((1R,4R)-4-(((R)-1-(1H-tetrazol-5-yl)prop-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (Compound 12) and 4-(((2'-(((1S,4R)-4-(((S)-1-(1H-tetrazol-5-yl)prop-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (Compound 13)

[0368]

[0369] Step 1: Preparation of 4-(((5'-chloro-2'-(((1R,4R)-4-((1-(1-(4-methoxybenzyl)-1H-tetrazol-5-yl)prop-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (compound 10)

[0370]

[0371] Add AcOH (137.34 mg, 2.29 mmol, 130.80 μL, 2 equivalents) and NaBH(OAc)3 (727.05 mg, 3.43 mmol, 3 equivalents) to a DCE (20 mL) solution of 4-(((2'-(((1R,4R)-4-aminocyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (intermediate 1, 504.24 mg, 1.14 mmol, 1 equivalent) and 1-(1-(4-methoxybenzyl)-1H-tetrazol-5-yl)prop-2-one (intermediate 4, 440 mg, 1.14 mmol, 64% purity, 1 equivalent) to a solution of 4-(((2'-(((1R,4R)-4-aminocyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (intermediate 1, 504.24 mg, 1.14 mmol, 1 equivalent) and NaBH(OAc)3 (727.05 mg, 3.43 mmol, 3 equivalents). The reaction mixture was stirred at 20°C for 16 hours, diluted with water (40 mL), alkalized to pH 7-8 with saturated NaHCO3, and extracted with DCM (60 mL × 3). The combined organic layers were washed with brine (80 mL), dried over Na2SO4, filtered, and concentrated. The resulting residue was subjected to rapid silica gel chromatography (20 g) Purification was performed using a silica gel rapid column chromatography (0-8% methanol / dichloromethane, 35 mL / min) to provide 4-(((5'-chloro-2'-(((1R,4R)-4-((1-(1-(4-methoxybenzyl)-1H-tetrazol-5-yl)prop-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (compound 10, 150 mg, 207.83 μmol, 18.17% yield, 93% purity). MS (ESI) m / z = 671.2 [M+H] + .

[0372] Step 2: Preparation of 4-(((2'-(((1R,4R)-4-(((R)-1-(1H-tetrazol-5-yl)prop-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 12) and 4-(((2'-(((1S,4R)-4-(((S)-1-(1H-tetrazol-5-yl)prop-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 13)

[0373]

[0374] A solution of 4-(((5'-chloro-2'-(((1R,4R)-4-((1-(1-(4-methoxybenzyl)-1H-tetrazol-5-yl)propyl-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (compound 10, 140 mg, 208.57 μmol, 1 equivalent) in TFA (38.50 g, 337.65 mmol, 25.00 mL, 1618.88 equivalents) was stirred at 40 °C for 12 hours, concentrated under vacuum, diluted with water (20 mL), and extracted with TBME (20 mL × 2). The aqueous layer was alkalized to pH 8 with saturated NaHCO3 and extracted with DCM (40 mL × 5). The combined DCM layers were washed with brine (50 mL), dried over Na₂SO₄, filtered, and concentrated to provide a yellow oil of 4-(((2'-(((1R,4R)-4-((1-(1H-tetrazol-5-yl)propyl-2-yl)amino)cyclohexyl)amino)5'-chloro-[2,4'-bipyridin]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (110 mg, crude product) (compound 11). MS (ESI) m / z = 551.4 [M+H] + .

[0375] 4-(((2'-(((1R,4R)-4-((1-(1H-tetrazol-5-yl)propyl-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (compound 11) was subjected to chiral SFC (Chiralpak IC 250×30mm (10μm particle size); 60% MeOH (0.1%) Further purification of NH3 / H2O / CO2 yielded 4-(((2'-(((1R,4R)-4-(((R)-1-(1H-tetrazol-5-yl)prop-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (Compound 12, 30 mg, 76.7% purity) and 4-(((2'-(((1S,4R)-4-(((S)-1-(1H-tetrazol-5-yl)prop-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (Compound 13, 50 mg, 93.6% purity). Compound 12 was further purified by preparative HPLC (Phenomenex Luna C18 80×40 mm (3 μm particle size); 0-30% acetonitrile / water (0.05% HCl); 11 min) to provide a yellow solid hydrochloride of compound 12 (21 mg, 35.74 μmol, 17.91% yield, >99% purity). Compound 13 was further purified by preparative HPLC (Phenomenex Luna C18 80×40 mm (3 μm particle size); 0-30% acetonitrile / water (0.05% HCl); 11 min) to provide a yellow solid hydrochloride of compound 13 (26 mg, 44.25 μmol, 22.17% yield, >99% purity).

[0376] Compound 12: 1¹H NMR (400MHz, methanol-d⁴) δ=8.15(s, 1H), 7.93(t, J=8.1Hz, 1H), 7.30(s, 1H), 7.22(d, J=8.8Hz, 1H), 7.09(d, J=7.2Hz, 1H), 3.99(dd, J=3.2, 11.2Hz, 3H), 3.88-3.78(m, 3H). 3.70-3.61(m,2H),3.57-3.45(m,2H),3.40-3.33(m,1H),2.29(t,J=14.4Hz,4H),1 .98(d,J=13.6Hz,2H),1.83-1.71(m,4H),1.68-1.54(m,2H),1.39(d,J=6.4Hz,3H). MS(ESI)m / z=551.2[M+H] + .

[0377] Compound 13: 1 H NMR (400MHz, methanol-d4) δ = 8.17 (s, 1H), 8.03 (t, J = 8.0Hz, 1H), 7.35 (s, 2H), 7.13 (d, J = 7.2Hz, 1H), 4.00 (dd, J = 3.2, 11.6Hz,3H),3.89(s,2H),3.87-3.79(m,1H),3.70-3.61(m,2H),3.57-3.47(m,2H),3.41-3.33(m,1H),2.30(br t,J=14.4Hz,4H),2.01(br d,J=13.2Hz,2H),1.85-1.73(m,4H),1.69-1.56(m,2H),1.40(d,J=6.4Hz,3H). MS(ESI)m / z=551.2[M+H] + .

[0378] Example 9: Synthesis of 2-(2-oxopropoxy)isopropyl acetate (intermediate 5)

[0379]

[0380] Step 1: Preparation of 2-((2-methylallyl)oxy)isopropyl acetate (INT-17)

[0381]

[0382] NaH (1.66 g, 41.43 mmol, 60% purity) was added to a DMF (50 mL) solution of 2-methylprop-2-en-1-ol (1.99 g, 27.62 mmol) at 0 °C over 15 minutes, followed by the slow addition of isopropyl 2-bromoacetate (INT-16, 5 g, 27.62 mmol). The mixture was stirred at 0 °C for 1 hour. The mixture was quenched with water (25 mL) at 0 °C and then extracted with ethyl acetate (30 mL × 3). The combined organic phases were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated under vacuum, and then purified by silica gel rapid column chromatography (ethyl acetate in petroleum ether 0% to 3%) to obtain isopropyl 2-(2-methylallyloxy)acetate as a colorless liquid (INT-17, 2.22 g, 46% yield). 1 H NMR (400MHz, DMSO-d6): δ=5.05-4.8(m,3H),4.03(s,2H),4.0-3.85(m,2H),1.68(s,3H),1.21(s,3H),1.19(s,3H).

[0383] Step 2: Preparation of 2-(2-oxopropoxy)ethyl acetate (intermediate 5)

[0384]

[0385] To a solution of 2-(2-methylallyloxy)acetic acid isopropyl ester (INT-17, 2.22 g, 12.89 mmol) in THF (25 mL) and H2O (24 mL), K2O·2H2O (48 mg, 0.129 mmol) and NaIO4 (6.07 g, 28.36 mmol) were added. The mixture was stirred at 20 °C for 12 hours. The mixture was diluted with H2O (10 mL) and extracted with DCM (20 mL × 3). The organic phases were combined, washed with brine (50 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum to give a residue. The residue was purified by silica gel chromatography (10–15% ethyl acetate in petroleum ether) to give 2-acetoneoxyacetic acid isopropyl ester as a pale yellow liquid (intermediate 5, 1.06 g, 47% yield). 1 H NMR (400MHz, DMSO-d6): δ = 5.13-5.04 (m, 1H), 4.19 (s, 2H), 4.13 (s, 2H), 2.17 (s, 3H), 1.26 (s, 3H), 1.24 (s, 3H).

[0386] Example 10: Synthesis of 2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)isopropyl acetate (compound 15) and 2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)isopropyl acetate (compound 16)

[0387]

[0388] NaBH(OAc)3 (67 mg, 0.317 mmol) was added to a DCE (2 mL) solution of 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (intermediate 1, 100 mg, 0.227 mmol), 2-propenyloxyacetic acid isopropyl ester (intermediate 5, 43 mg, 0.249 mmol), and AcOH (41 mg, 0.680 mmol). The mixture was stirred at 20 °C for 20 hours. The mixture was quenched with H2O (5 mL). The resulting mixture was extracted with EtOAc (20 mL × 3). The organic phases were combined, washed with brine (50 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The mixture was then purified by silica gel chromatography (15–20% MeOH in DCM) to give 2-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]isopropyl acetate (compound 14, 50 mg, 12% yield) as a dark brown solid.

[0389] Racemic 2-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]isopropyl acetate (compound 14, 50 mg, 0.083 mmol) was subjected to chiral SFC (Chiralpak AD 250 x 30 mm (10 μm particle size); 70% Isopropyl acetate (compound 15, 3.9 mg, 8% yield) was separated by EtOH / CO2 and further purified by preparative HPLC (HCOOH conditions) to obtain 2-[(2R)-2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]acetate as a white solid (compound 15, 3.9 mg, 8% yield) and 2-[(2S)-2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]acetate as a brown solid (compound 16, 11.2 mg, 21% yield).

[0390] Compound 15: 1 H NMR (400MHz, CDCl3): δ = 8.49 (s, 1H), 8.09 (s, 1H), 7.52 (t, J = 7.6Hz, 1H), 6.97 (d, J = 7.2Hz, 1H), 6.6-6.5 (m, 2H), 5.15-5.0 (m, 1H) ,4.86(t,J=6.8Hz,1H),4.56(s,1H),4.15-4.0(m,2H),4.0-3.9(m,2H),3.78(d,J=6.8Hz,2H),3.75-3.7(m,2H),3.7-3.5(m,3H). 3.4-3.25(m,1H),3.05-2.9(m,1H),2.3-2.1(m,4H),1.95-1.85(m,2H),1.8-1.7(m,2H),1.65-1.45(m,2H),1.3-1.2(m,11H).

[0391] Compound 16: 1H NMR (400MHz, CDCl3): δ = 8.1 (s, 1H), 7.51-7.5 (m, 1H), 6.97 (d, J = 7.2Hz, 1H), 6.51-6.5 (m, 2H), 5.2-5.0 (m, 1H) ,4.82(t,J=7.6Hz,1H),4.45(d,J=8.4Hz,1H),4.05(d,J=0.8Hz,2H),4.05-3.95(m,2H),3.77(d,J=7.6Hz,2H), 3.75-3.65(m,2H),3.6-3.45(m,2H),3.4-3.3(m,1H),3.15-3.0(m,1H),2.65-2.5(m,1H),2.2-2.1(m,2H),2.1 -1.95(m,2H),1.95-1.85(m,2H),1.8-1.75(m,2H),1.3-1.25(m,8H),1.25-1.15(m,2H),1.06(d,J=6.4Hz,3H).

[0392] Example 11: Synthesis of 2-(2-oxopropoxy)tert-butyl acetate (intermediate 6)

[0393]

[0394] Step 1: Preparation of 2-((2-methylallyl)oxy)tert-butyl acetate (INT-19)

[0395]

[0396] Add 18 mL of 50% NaOH aqueous solution to a toluene (18 mL) solution of 2-methylprop-2-en-1-ol (296 mg, 4.10 mmol), then add NBu4HSO4 (1.13 g, 3.33 mmol), and stir the mixture at 15–20 °C for 30 min. Then slowly add tert-butyl 2-bromoacetate (INT-18, 500 mg, 2.56 mmol), and stir the mixture at 15–20 °C for 1.5 h. Extract the mixture with ethyl acetate (300 mL × 3). Wash the combined organic phases with brine (200 mL), dry with anhydrous sodium sulfate, filter, and concentrate under vacuum. Then purify the mixture by silica gel chromatography (3% ethyl acetate in petroleum ether) to give tert-butyl 2-(2-methylallyloxy)acetate (INT-19, 0.35 g, 72% yield) as a colorless liquid. 1 H NMR (400MHz, CDCl3): δ = 4.98 (s, 1H), 4.92 (s, 1H), 3.98 (s, 2H), 3.94 (s, 2H), 1.75 (s, 3H), 1.48 (s, 9H).

[0397] Step 2: Preparation of 2-(2-oxopropoxy)tert-butyl acetate (intermediate 6)

[0398]

[0399] To a solution of 2-(2-methylallyloxy)tert-butyl acetate (INT-19, 4.5 g, 24.16 mmol) in THF (50 mL) and H₂O (50 mL), K₂O₄·2H₂O (89 mg, 0.242 mmol) and NaIO₄ (11.37 g, 53.16 mmol) were added. The mixture was stirred at 20 °C for 12 hours. The mixture was diluted with H₂O (100 mL) and extracted with DCM (200 mL × 3). The organic phases were combined, washed with brine (50 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under vacuum. The purified solution was purified by silica gel chromatography (6-10% ethyl acetate in petroleum ether) to give 2-acetone-oxyethyl tert-butyl acetate (intermediate 6, 2.99 g, 66% yield) as a pale yellow liquid. 1 H NMR (400MHz, CDCl3): δ = 4.17 (s, 2H), 4.05 (s, 2H), 2.20 (s, 3H), 1.46 (s, 9H).

[0400] Example 12: Synthesis of 2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)tert-butyl acetate (compound 18) and 2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)tert-butyl acetate (compound 19)

[0401]

[0402] NaBH(OAc)3 (intermediate 6, 135 mg, 0.635 mmol) was added to a DCE (2 mL) solution of 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (intermediate 1, 200 mg, 0.454 mmol), 2-propenyloxyacetic acid tert-butyl ester (94 mg, 0.499 mmol), and AcOH (82 mg, 1.36 mmol). The mixture was stirred at 20 °C for 20 hours. The mixture was quenched with H2O (5 mL). The resulting mixture was extracted with EtOAc (20 mL × 3). The organic phases were combined, washed with brine (50 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The mixture was then purified by silica gel chromatography (15–20% MeOH in DCM) to give 2-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]tert-butyl acetate (compound 17, 110 mg, 25% yield) as a dark brown solid.

[0403] Racemic 2-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]tert-butyl acetate (compound 17, 92 mg) was subjected to chiral SFC (Chiralpak AD 250x30 mm (10 μm particle size); 75%) 2-[(2R)-2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]tert-butyl acetate (compound 18, 20.3 mg, 22% yield) and 2-[(2S)-2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]amino]cyclohexyl]amino]propoxy]tert-butyl acetate (compound 19, 26.2 mg, 28% yield) were separated by EtOH / CO2 and further purified by preparative HPLC (HCOOH conditions) to obtain tert-butyl acetate as a white solid.

[0404] Compound 18: 1H NMR (400MHz, CDCl3): δ = 8.47 (s, 1H), 8.09 (s, 1H), 7.52 (t, J = 7.2Hz, 1H), 6.97 (d, J = 7.2Hz, 1H), 6.6-6.5 (m, 2H),5.0-4.8(m,1H),4.58(s,1H),4.1-3.9(m,4H),3.8-3.75(m,2H),3.75-3.65(m,4H),3.65-3.55(m,1H). 3.4-3.3(m,1H),3.05-2.95(m,1H),2.3-2.1(m,4H),1.97-1.87(m 2H),1.8-1.7(m,2H),1.7-1.5(m,2H),1.49(s,9H),1.32-1.22(m,5H).

[0405] Compound 19: 1 H NMR (400MHz, CDCl3): δ = 8.1 (s, 1H), 7.51 (t, J = 8.0Hz, 1H), 6.97 (d, J = 7.2Hz, 1H), 6.6-6.4 (m, 2H), 4.81 ( t,J=6.4Hz,1H),4.44(d,J=7.6Hz,1H),4.1-4.0(m,4H),3.77(d,J=7.2Hz,2H),3.7(t,J=11.2Hz,2H),3.6 -3.45(m,2H),3.4-3.3(m,1H),3.15-3.0(m,1H),2.7-2.45(m,1H),2.2-2.1(m,2H),2.1-1.95(m,2H),1. 95-1.9(m,2H),1.8-1.75(m,2H),1.48(s,9H),1.3-1.25(m,2H),1.25-1.2(m,2H),1.06(d,J=6.4Hz,3H).

[0406] Example 13: Synthesis of (1-(5-(((2-methylallyl)oxy)methyl)-2H-tetrazol-2-yl)ethyl)carbonate (INT-21) and (1-(5-(((2-methylallyl)oxy)methyl)-1H-tetrazol-1-yl)ethyl)carbonate (INT-22)

[0407]

[0408] Step 1: Preparation of 5-(((2-methylallyl)oxy)methyl)-2H-tetrazole (INT-20)

[0409]

[0410] Add 4-methylbenzenesulfonic acid (3.68 g, 21.36 mmol) to a MeOH (50 mL) solution of 5-(2-methylallyloxymethyl)-2-tetrahydropyran-2-yl-tetraazole (INT-9, 5.09 g, 21.36 mmol). Stir the mixture at 20 °C for 12 hours. Concentrate the mixture under reduced pressure. Purify the residue by silica gel chromatography (0-50% ethyl acetate in petroleum ether) to obtain 5-(2-methylallyloxymethyl)-2H-tetraazole (INT-20, 600 mg, 18% yield) as a white solid. 1 H NMR (400MHz, CDCl3) δ = 5.02 (d, J = 6.0Hz, 2H), 4.91 (s, 2H), 4.07 (s, 2H), 1.77 (s, 3H).

[0411] Step 2: Preparation of (1-(5-(((2-methylallyl)oxy)methyl)-2H-tetrazol-2-yl)ethyl)carbonate (INT-21) and (1-(5-(((2-methylallyl)oxy)methyl)-1H-tetrazol-1-yl)ethyl)carbonate (INT-22)

[0412]

[0413] Add DIEA (5.03 g, 38.92 mmol) and 1-chloroethyl ethyl carbonate (5.94 g, 38.92 mmol) to a DMF (6 mL) solution of 5-(2-methylallyloxymethyl)-2H-tetrazole (INT-20, 600 mg, 3.89 mmol). Stir the mixture at 70 °C for 16 hours. Dilute the mixture with ethyl acetate (100 mL) and wash it with brine (100 mL × 3). The organic layer was dried with anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the residue, which was purified by rapid column chromatography (silica gel, 100-200 mesh, 0-10% ethyl acetate in petroleum ether) to obtain 1-[5-(2-methylallyloxymethyl)tetrazol-1-yl]ethyl carbonate in yellow oil (INT-21, 326 mg, 31% yield) and 1-[5-(2-methylallyloxymethyl)tetrazol-2-yl]ethyl carbonate in yellow oil (INT-22, 464 mg, 44% yield).

[0414] INT-21: 1H NMR (400MHz, CDCl3): δ = 7.00 (q, J = 6.4Hz, 1H), 5.06-4.88 (m, 4H), 4.254.14 (m, 2H ), 3.96 (q, J = 12.4Hz, 2H), 1.98 (d, J = 6.0Hz, 3H), 1.74 (s, 3H), 1.31-1.27 (m, 3H).

[0415] INT-22: 1 H NMR (400MHz, CDCl3) δ = 7.19 (q, J = 6.4Hz, 1H), 5.00 (d, J = 27.2Hz, 2H), 4.76 (s, 2H), 4.29-4.19(m,2H),4.05(s,2H),2.04-2.01(m,3H),1.77(s,3H),1.31-1.28(m,3H).

[0416] Example 14: Synthesis of (R)-(1-(5-((2-oxopropoxy)methyl)-2H-tetrazol-2-yl)ethyl)carbonate (intermediate 7) and (S)-(1-(5-((2-oxopropoxy)methyl)-2H-tetrazol-2-yl)ethyl)carbonate (intermediate 8)

[0417]

[0418] To a solution of 1-[5-(2-methylallyloxymethyl)tetrazol-2-yl]ethyl carbonate (INT-21, 464 mg, 1.72 mmol) in THF (10 mL) and H₂O (10 mL), K₂O₄·2H₂O (32 mg, 0.086 mmol) and NaIO₄ (845 mg, 3.95 mmol) were added. The mixture was stirred at 15 °C for 4 hours. The mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL × 3). The combined organic phases were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the residue, which was purified by rapid column chromatography (silica gel, 100-200 mesh, 0-20% ethyl acetate in petroleum ether) to obtain 1-[5-(acetoneyloxymethyl)tetrazol-2-yl]ethyl carbonate (INT-23, 400 mg, 86% yield) as a yellow oil.

[0419] 1-[5-(acetoneoxymethyl)tetrazol-2-yl]ethyl carbonate (INT-23, 400 mg, 1.47 mmol) was separated by chiral SFC to obtain [(1R)-1-[5-(acetoneoxymethyl)tetrazol-2-yl]ethyl]carbonate (intermediate 7, 143 mg, 36% yield, 90% purity) and [(1S)-1-[5-(acetoneoxymethyl)tetrazol-2-yl]ethyl]carbonate (intermediate 8, 290 mg, 73% yield) in yellow oil.

[0420] Intermediate 7: 1 H NMR (400MHz, MeOD): δ = 7.23 (q, J = 6.0Hz, 1H), 4.87 (s, 2H), 4.29 (s, 2H), 4.26-4.18 (m, 2H), 2.12 (s, 3H), 1.96 (d, J = 6.0Hz, 3H), 1.27 (t, J = 7.2Hz, 3H).

[0421] Intermediate 8: 1 H NMR (400MHz, MeOD): δ = 7.23 (q, J = 6.0Hz, 1H), 4.87 (s, 2H), 4.30 (s, 2H), 4.25-4.17 (m, 2H), 2.12 (s, 3H), 1.96 (d, J = 6.4Hz, 3H), 1.27 (t, J = 6.8Hz, 3H).

[0422] Example 15: Synthesis of (R)-(1-(5-((2-oxopropoxy)methyl)-1H-tetrazol-1-yl)ethyl)carbonate (intermediate 9) and (S)-(1-(5-((2-oxopropoxy)methyl)-1H-tetrazol-1-yl)ethyl)carbonate (intermediate 10)

[0423]

[0424] Add K₂O₄·2H₂O (22 mg, 0.060 mmol) and NaIO₄ (593 mg, 2.77 mmol) to a solution of 1-[5-(2-methylallyloxymethyl)tetrazol-1-yl]ethyl ethyl carbonate (INT-22, 326 mg, 1.21 mmol) in THF (7 mL) and H₂O (7 mL). Stir the mixture at 15 °C for 4 hours. Dilute the reaction mixture with water (50 mL) and extract it with ethyl acetate (50 mL × 3). The combined organic phases were washed with brine (100 mL), dried with anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the residue, which was purified by rapid column chromatography (silica gel, 100-200 mesh, 0-25% ethyl acetate in petroleum ether) and then purified by chiral SFC to obtain yellow oil [(1R)-1-[5-(acetoneoxymethyl)tetrazol-1-yl]ethyl]carbonate (intermediate 9, 142 mg, 43% yield) and yellow oil [(1S)-1-[5-(acetoneoxymethyl)tetrazol-1-yl]ethyl]carbonate (intermediate 10, 195 mg, 59% yield).

[0425] Intermediate 9: 1 H NMR (400MHz, MeOD): δ = 7.16 (q, J = 6.0Hz, 1H), 5.16-4.97 (m, 2H), 4.35 (d, J = 2.8Hz, 1H), 4.22-4.14 (m, 2H), 2.09 (s, 3H), 1.95 (d, J = 6.0Hz, 3H), 1.25 (t, J = 6.8Hz, 3H).

[0426] Intermediate 10: 1 H NMR (400MHz, MeOD): δ = 7.16 (q, J = 6.0Hz, 1H), 5.17-4.96 (m, 2H), 4.35 (d, J = 2.8Hz, 2H), 4.23-4.12 (m, 2H), 2.09 (s, 3H), 1.95 (d, J = 6.0Hz, 3H), 1.25 (t, J = 7.2Hz, 3H).

[0427] Example 16: Synthesis of (R)-1-(5-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-2H-tetrazole-2-yl)ethyl carbonate (compound 21) and (R)-1-(5-(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-2H-tetrazole-2-yl)ethyl carbonate (compound 22)

[0428]

[0429] HOAc (68 mg, 1.13 mmol) and NaBH(OAc)3 (112 mg, 0.53 mmol) were added to a DCE (2 mL) solution of [(1R)-1-[5-(acetoneoxymethyl)tetrazol-2-yl]ethyl]carbonate (intermediate 7, 123 mg, 0.45 mmol) and 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (intermediate 1, 166 mg, 0.38 mmol) (intermediate 1). The mixture was stirred at 20 °C for 16 h. The mixture was quenched with water (50 mL) at 0 °C and extracted with DCM (50 mL × 3). The combined organic phases were washed with saturated brine (50 mL), dried with anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the residue, which was purified by RP-HPLC (16 to 46% acetonitrile in water and 0.225% formic acid) to obtain ethyl carbonate [(1R)-1-[5-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxymethyl]tetrazole-2-yl]ethyl]carbonate (compound 20, 168 mg, 63% yield).

[0430] [(1R)-1-[5-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxymethyl]tetrazol-2-yl]ethyl]carbonate (compound 20, 100 mg, 0.14 mmol) was separated by chiral SFC to obtain a white solid of [(1R)-1-[5-[[(2R)-2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-yl]ethyl]carbonate (compound 20, 100 mg, 0.14 mmol)). [-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]methyl]tetrazol-2-yl]ethyl]ethyl]carbonate (compound 21, 11.6 mg, 11% yield) and [(1R)-1-[5-[[(2S)-2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]methyl]tetrazol-2-yl]ethyl]carbonate (compound 22, 22.4 mg, 20% yield) as white solids).

[0431] Compound 21: 1 H NMR (400MHz, MeOD): δ = 8.54 (s, 1H), 7.97 (s, 1H), 7.53-7.40 (m, 1H), 7.25 (q, J = 6.0Hz, 1H ),6.84(d,J=7.2Hz,1H),6.70(s,1H),6.63(d,J=8.4Hz,1H),4.98-4.92(m,2H),4.28-4.1 3(m,2H),4.00-3.83(m,3H),3.75(s,2H),3.73-3.55(m,6H),2.25-2.11(m,4H),1.96(d,J =6.0Hz,3H),1.93-1.85(m,2H),1.82-1.71(m,2H),1.60-1.46(m,2H),1.38-1.21(m,8H).

[0432] Compound 22: 1H NMR (400MHz, MeOD): δ = 8.55 (s, 1H), 7.97 (s, 1H), 7.53-7.45 (m, 1H), 7.25 (q, J = 6.0Hz, 1H), 6.8 4(d,J=7.2Hz,1H),6.70(s,1H),6.62(d,J=8.0Hz,1H),4.87(s,2H),4.26-4.15(m,2H),3.99-3 .92(m,2H),3.82-3.74(m,3H),3.69-3.59(m,4H),3.51-3.42(m,1H),3.15-2.99(m,1H),2.21- 2.07(m,4H),1.96(d,J=6.4Hz,3H),1.93-1.86(m,2H),1.82-1.72(m,2H),1.50-1.23(m,10H).

[0433] Example 17: Synthesis of (S)-1-(5-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-2H-tetrazole-2-yl)ethyl carbonate (compound 24) and (S)-1-(5-(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-2H-tetrazole-2-yl)ethyl carbonate (compound 25)

[0434]

[0435] HOAc (149 mg, 2.48 mmol) and NaBH(OAc)3 (245 mg, 1.16 mmol) were added to a DCE (3 mL) solution of [(1S)-1-[5-(acetoneoxymethyl)tetrazol-2-yl]ethyl]carbonate (intermediate 8, 270 mg, 0.99 mmol) and 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (intermediate 1, 364 mg, 0.83 mmol) (intermediate) at 0 °C. The mixture was stirred at 20 °C for 16 h. The mixture was quenched with water (50 mL) at 0 °C and extracted with DCM (50 mL × 3). The combined organic phases were washed with saturated brine (50 mL), dried with anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the residue, which was purified by RP-HPLC (16 to 46% acetonitrile in water and 0.225% formic acid) to obtain ethyl carbonate [(1S)-1-[5-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxymethyl]tetrazole-2-yl]ethyl]carbonate (compound 23, 260 mg, 44% yield).

[0436] [(1S)-1-[5-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxymethyl]tetrazol-2-yl]ethyl]carbonate (compound 23, 100 mg, 0.14 mmol) was separated by chiral SFC to obtain a white solid of [(1S)-1-[5-[[(2R)-2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-]ethyl]carbonate (compound 23, 100 mg, 0.14 mmol)). [-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]methyl]tetrazol-2-yl]ethyl]ethyl]carbonate (compound 24, 17.3 mg, 16% yield) and [(1S)-1-[5-[[(2S)-2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]methyl]tetrazol-2-yl]ethyl]carbonate (compound 25, 28.5 mg, 26% yield) as white solids).

[0437] Compound 24: 1H NMR (400MHz, MeOD): δ = 8.53 (s, 1H), 7.98 (s, 1H), 7.53-7.45 (m, 1H), 7.25 (q, J = 6.0Hz, 1H), 6.83 (d ,J=7.2Hz,1H),6.70(s,1H),6.63(d,J=8.4Hz,1H),4.92(s,2H),4.26-4.14(m,2H),3.99-3.86(m, 3H),3.75(s,2H),3.72-3.60(m,5H),3.28-3.20(m,1H),2.25-2.12(m,4H),1.97(d,J=6.0Hz,3H), 1.93-1.86(m,2H),1.83-1.72(m,2H),1.61-1.47(m,2H),1.40-1.31(m,5H),1.26(t,J=7.2Hz,3H).

[0438] Compound 25: 1 H NMR (400MHz, MeOD): δ = 8.55 (s, 1H), 7.97 (s, 1H), 7.53-7.44 (m, 1H), 7.25 (q, J = 6.4Hz, 1H), 6.8 4(d,J=6.8Hz,1H),6.69(s,1H),6.62(d,J=8.4Hz,1H),4.89(s,2H),4.26-4.14(m,2H),3.99-3 .92(m,2H),3.82-3.74(m,3H),3.70-3.59(m,4H),3.52-3.42(m,1H),3.12-3.00(m,1H),2.20- 2.07(m,4H),1.96(d,J=6.4Hz,3H),1.93-1.86(m,2H),1.82-1.71(m,2H),1.54-1.25(m,10H).

[0439] Example 18: Synthesis of (R)-1-(5-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-1H-tetrazol-1-yl)ethyl ethyl carbonate (compound 27) and (R)-1-(5-(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-1H-tetrazol-1-yl)ethyl ethyl carbonate (compound 28)

[0440]

[0441] HOAc (67 mg, 1.12 mmol) and NaBH(OAc)3 (111 mg, 0.52 mmol) were added to a DCE (2 mL) solution of [(1R)-1-[5-(acetoneoxymethyl)tetrazol-1-yl]ethyl]carbonate (intermediate 9, 122 mg, 0.45 mmol) and 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (intermediate 1, 165 mg, 0.37 mmol) at 0 °C. The mixture was stirred at 20 °C for 16 h under N2. The mixture was quenched with water (50 mL) at 0 °C and extracted with DCM (50 mL × 3). The combined organic phases were washed with saturated brine (50 mL), dried with anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the residue, which was purified by preparative HPLC (45 to 75% acetonitrile in water and 0.225% formic acid) to obtain ethyl carbonate [(1R)-1-[5-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxymethyl]tetrazole-1-yl]ethyl]carbonate (compound 26, 73 mg, 27% yield).

[0442] Ethyl [(1R)-1-[5-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxymethyl]tetrazol-1-yl]ethyl]carbonate (compound 26, 73 mg, 0.11 mmol) was separated by chiral SFC to obtain a white solid of [(1R)-1-[5-[[(2R)-2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-]ethyl]carbonate. [-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]methyl]tetrazol-1-yl]ethyl]ethyl]carbonate (compound 27, 14.5 mg, 19% yield) and [(1R)-1-[5-[[(2S)-2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]methyl]tetrazol-1-yl]ethyl]carbonate (compound 28, 17.1 mg, 21% yield) as white solids).

[0443] Compound 27: 11H NMR (400 MHz, MeOD): δ = 7.95 (s, 1H), 7.52 - 7.46 (m, 1H), 7.02 (q, J = 6.0 Hz, 1H), 6.85 (d, J = 6.8 Hz, 1H), 6.69 (s, 1H), 6.62 (d, J = 8.4 Hz, 1H), 5.12 - 5.00 (m, 2H), 4.26 - 4.14 (m, 2H), 3.99 - 3.92 (m, 2H), 3.75 (s, 2H), 3.69 - 3.59 (m, 4H), 3.51 - 3.45 (m, 1H), 3.29 - 3.27 (m, 1H), 3.18 - 3.12 (m, 1H), 2.16 - 1.99 (m, 4H), 1.95 (d, J = 6.0 Hz, 3H), 1.92 - 1.90 (m, 1H), 1.89 - 1.85 (m, 1H), 1.82 - 1.72 (m, 2H), 1.35 - 1.24 (m, 7H), 1.16 - 1.06 (m, 3H).

[0444] Compound 28: 1 1H NMR (400 MHz, MeOD): δ = 8.54 (s, 1H), 7.97 (s, 1H), 7.52 - 7.46 (m, 1H), 6.93 (q, J = 6.0 Hz, 1H), 6.84 (d, J = 7.2 Hz, 1H), 6.70 (s, 1H), 6.62 (d, J = 8.4 Hz, 1H), 5.18 - 5.06 (m, 2H), 4.25 - 4.13 (m, 2H), 3.99 - 3.91 (m, 2H), 3.86 - 3.77 (m, 1H), 3.75 (s, 2H), 3.73 - 3.46 (m, 5H), 3.21 - 3.10 (m, 1H), 2.25 - 2.08 (m, 4H), 1.96 (d, J = 6.0 Hz, 3H), 1.92 - 1.86 (m, 2H), 1.82 - 1.71 (m, 2H), 1.60 - 1.42 (m, 2H), 1.38 - 1.24 (m, 8H).

[0445] Example 19: Synthesis of (S)-1-(5-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-1H-tetrazol-1-yl)ethyl ethyl carbonate (compound 30) and (S)-1-(5-(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-1H-tetrazol-1-yl)ethyl ethyl carbonate (compound 31)

[0446]

[0447] HOAc (97 mg, 1.61 mmol) and NaBH(OAc)3 (159 mg, 0.75 mmol) were added to a DCE (2 mL) solution of [(1S)-1-[5-(acetoneoxymethyl)tetrazol-1-yl]ethyl]carbonate (intermediate 10, 175 mg, 0.64 mmol) and 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (intermediate 1, 236 mg, 0.54 mmol) at 0 °C. The mixture was stirred at 20 °C for 16 hours under N2. The mixture was quenched with water (50 mL) at 0 °C and extracted with DCM (50 mL × 3). The combined organic phases were washed with saturated brine (50 mL), dried with anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the residue, which was purified by RP-HPLC (15 to 45% acetonitrile in water and 0.225% formic acid) to obtain ethyl carbonate [(1S)-1-[5-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxymethyl]tetrazole-1-yl]ethyl]carbonate (compound 29, 204 mg, 54% yield).

[0448] [(1S)-1-[5-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxymethyl]tetrazol-1-yl]ethyl]carbonate (compound 29, 100 mg, 0.14 mmol) was separated by chiral SFC to obtain a white solid of [(1S)-1-[5-[[(2R)-2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-]ethyl]carbonate (compound 29, 100 mg, 0.14 mmol)). [-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]methyl]tetrazol-1-yl]ethyl]ethyl]carbonate (compound 30, 25.4 mg, 25% yield) and [(1S)-1-[5-[[(2S)-2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]methyl]tetrazol-1-yl]ethyl]carbonate (compound 31, 27.1 mg, 24% yield) as white solids).

[0449] Compound 30: 1 H NMR (400MHz, MeOD): δ = 7.95 (s, 1H), 7.52-7.45 (m, 1H), 7.06-6.98 (m, 1H), 6.89-6.81 (m, 1H),6.69(s,1H),6.62(d,J=8.4Hz,1H),5.11-4.95(m,2H),4.26-4.14(m,2H),3.99-3.92 (m,2H),3.75(s,2H),3.70-3.44(m,6H),3.16-3.10(m,1H),2.14-1.98(m,4H),1.95(d,J =6.4Hz,3H),1.92-1.85(m,2H),1.81-1.72(m,2H),1.34-1.21(m,7H),1.15-1.04(m,3H).

[0450] Compound 31: 1H NMR (400MHz, MeOD): δ = 8.54 (s, 1H), 7.97 (s, 1H), 7.52-7.45 (m, 1H), 6.94 (q, J = 6.4Hz, 1H), 6.84 (d ,J=6.8Hz,1H),6.69(s,1H),6.63(d,J=8.0Hz,1H),5.21-5.06(m,2H),4.28-4.12(m,2H),3.99-3.9 2(m,2H),3.88-3.83(m,1H),3.75(s,2H),3.70-3.52(m,5H),3.26-3.12(m,1H),2.23-2.10(m,4H), 1.96(d,J=6.0Hz,3H),1.92-1.85(m,2H),1.81-1.72(m,2H),1.59-1.44(m,2H),1.43-1.23(m,8H).

[0451] Example 20: Synthesis of 2-methyl-5-(((2-methylallyl)oxy)methyl)-2H-tetrazole (INT-26) and 1-methyl-5-(((2-methylallyl)oxy)methyl)-1H-tetrazole (INT-27)

[0452]

[0453] Step 1: Preparation of 1-((2H-tetrazol-5-yl)methoxy)prop-2-one (INT-25)

[0454]

[0455] Add 4-methylbenzenesulfonic acid (2.2 g, 12.59 mmol) to a MeOH solution of 5-(2-methylallyloxymethyl)-2-tetrahydropyran-2-yl-tetraazole (intermediate 2, 3.0 g, 12.59 mmol) in 1 mL. Stir the mixture at 20 °C for 12 h. Concentrate the mixture under reduced pressure. Purify the residue by rapid silica gel chromatography (silica gel, 100-200 mesh, 0-40% ethyl acetate in petroleum ether) to obtain 5-(2-methylallyloxymethyl)-2H-tetraazole as a colorless oil (INT-25, 2.1 g, 13.62 mmol, 54% yield). 1 H NMR (400MHz, CDCl3): δ5.00-4.85 (m, 4H), 3.95 (s, 2H), 1.62 (s, 3H).

[0456] Step 2: Preparation of 2-methyl-5-(((2-methylallyl)oxy)methyl)-2H-tetrazole (INT-26) and 1-methyl-5-(((2-methylallyl)oxy)methyl)-1H-tetrazole (INT-27)

[0457]

[0458] Diazomethyl(trimethyl)silane (2M, 9.73 mL) was added to a THF (10 mL) solution of 5-(2-methylallyloxymethyl)-2H-tetrazole (INT-25, 1.0 g, 6.49 mmol). The mixture was stirred at 20 °C for 2 hours. The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by rapid column chromatography (silica gel, 100-200 mesh, 10-30% ethyl acetate in petroleum ether) to obtain 2-methyl-5-(2-methylallyloxymethyl)tetrazole (INT-26, 470 mg, 2.79 mmol, 43% yield) and 1-methyl-5-(2-methylallyloxymethyl)tetrazole (INT-27, 330 mg, 1.96 mmol, 30% yield) as colorless oil.

[0459] INT-26: 1 H NMR (400MHz, CDCl3) δ5.00 (d, J = 24.0Hz, 2H), 4.74 (s, 2H), 4.36 (s, 3H), 4.05 (s, 2H), 1.77 (s, 3H).

[0460] INT-27: 1 H NMR (400MHz, CDCl3) δ4.99 (d, J = 6.0Hz, 2H), 4.83 (s, 2H), 4.14 (s, 3H), 3.95 (s, 2H), 1.75 (s, 3H).

[0461] Example 21: Synthesis of 1-((2-methyl-2H-tetrazol-5-yl)methoxy)prop-2-one (intermediate 11)

[0462]

[0463] Add K₂O₅sO₄·2H₂O (49 mg, 0.13 mmol) and NaIO₄ (1.32 g, 6.15 mmol) to a solution of 2-methyl-5-(2-methylallyloxymethyl)tetrazole (INT-26, 450 mg, 2.68 mmol) in THF (4 mL) and H₂O (3 mL). Stir the mixture at 20 °C for 4 hours. Dilute the mixture with ethyl acetate (100 mL) and wash it with brine (100 mL). Dry the organic extract with anhydrous sodium sulfate and filter. Concentrate the filtrate under reduced pressure. Purify the residue by rapid column chromatography (silica gel, 100-200 mesh, 0-5% methanol in dichloromethane) to obtain 1-[(2-methyltetrazole-5-yl)methoxy]prop-2-one as a colorless oil (intermediate 11, 310 mg, 1.82 mmol, 68% yield). 1 H NMR (400MHz, CDCl3): δ4.87(s,2H), 4.37(s,3H), 4.22(s,2H), 2.18(s,2H).

[0464] Example 22: Synthesis of 1-((1-methyl-1H-tetrazol-5-yl)methoxy)prop-2-one (intermediate 12)

[0465]

[0466] Add K₂O₄·2H₂O (34 mg, 0.09 mmol) and NaIO₄ (907 mg, 4.24 mmol) to a solution of 1-methyl-5-(2-methylallyloxymethyl)tetrazole (INT-27, 310 mg, 1.84 mmol) in THF (3 mL) and H₂O (2 mL). Stir the mixture at 20 °C for 4 hours. Dilute the mixture with ethyl acetate (100 mL) and wash it with brine (100 mL). Dry the organic extract with anhydrous sodium sulfate and filter. Concentrate the filtrate under reduced pressure. Purify the residue by rapid column chromatography (silica gel, 100-200 mesh, 0-80% methanol in dichloromethane) to obtain 1-[(1-methyltetrazole-5-yl)methoxy]prop-2-one as a black oil (intermediate 12, 210 mg, 1.23 mmol, 67% yield). 1 H NMR (400MHz, CDCl3) δ4.86 (s, 2H), 4.16 (s, 3H), 4.15 (s, 2H), 2.05 (s, 2H).

[0467] Example 23: Synthesis of 4-(((5'-chloro-2'-(((1R,4r)-4-(((R)-1-((2-methyl-2H-tetrazol-5-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (compound 33) and 4-(((5'-chloro-2'-(((1S,4r)-4-(((S)-1-((2-methyl-2H-tetrazol-5-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (compound 34)

[0468]

[0469] To a DCE (5 mL) solution of 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (intermediate 1, 600 mg, 1.36 mmol) and 1-[(2-methyltetrazol-5-yl)methoxy]prop-2-one (intermediate 11, 278 mg, 1.63 mmol), HOAc (245 mg, 4.08 mmol) and NaBH(OAc)3 (404 mg, 1.90 mmol) were added. The mixture was stirred at 20 °C for 36 h. The mixture was quenched with water (50 mL) at 0 °C and extracted with DCM (50 mL × 3). The combined organic phases were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to dryness. The residue was purified by preparative HPLC (column: Ultimate C18 150*40mm*10um; mobile phase: [water (0.225% FA)-ACN]; B%: 10%-40%, 10min) to obtain 4-[[[6-[5-chloro-2-[[4-[[1-methyl-2-[(2-methyltetrazol-5-yl)methoxy]ethyl]amino]cyclohexyl]amino]-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (compound 32, 600 mg, 736 μmol, 54% yield).

[0470] 4-(((5'-chloro-2'-(((1r,4r)-4-((1-((2-methyl-2H-tetrazol-5-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (compound 32, 100 mg, 0.17 mmol) was separated by chiral SFC to obtain 4-(((5'-chloro-2'-(((1S,4r)-4-(((S)-1-((2-methyl-2H-tetrazol-5-yl)methoxy)propyl-2-yl)amino) Cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 33, 11.2 mg, 10.5% yield, 94% purity) and 4-(((5'-chloro-2'-(((1R,4r)-4-(((R)-1-((2-methyl-2H-tetrazol-5-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 34, 22 mg, 21.6% yield, 98% purity).

[0471] Compound 33: 1 H NMR (400MHz, CDCl3): δ = 7.95 (s, 1H), 7.50-7.46 (m, 1H), 6.85 (d, J = 7.2Hz, 1H), 6.69 (s, 1H), 6.62 (d, J = 8.4Hz, 1H), 4.76 (s, 2 H),4.37(s,3H),3.97-3.93(m,2H),3.75(s,2H),3.66-3.55(m,4H),3.45-3.41(m,1H),3.12-3.08(m,1H),2.64-2.63(m,1H). 2.11-1.97(m,4H),1.90-1.87(m,2H),1.80-1.72(m,2H),1.32-1.22(m,4H),1.05(d,J=6.4Hz,3H).

[0472] Compound 34: 1H NMR (400MHz, CDCl3): δ = 8.54 (s, 1H), 7.97 (s, 1H), 7.51-7.47 (m, 1H), 6.83 (d, J = 6.8Hz, 1H ),6.69(s,1H),6.62(d,J=8.4Hz,1H),4.89(s,2H),4.39(s,3H),3.97-3.94(m,2H),3.90-3 .80(m,1H),3.75(s,2H),3.67-3.61(m,5H),3.23-3.17(m,1H),2.21-2.12(m,5H),1.91-1. 87(m,2H),1.80-1.73(m,2H),1.54-1.51(m,2H),1.39-1.36(m,1H),1.32(d,J=6.4Hz,3H).

[0473] Example 24: Synthesis of 4-(((5'-chloro-2'-(((1R,4r)-4-(((R)-1-((1-methyl-1H-tetrazol-5-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (compound 36) and 4-(((5'-chloro-2'-(((1S,4r)-4-(((S)-1-((1-methyl-1H-tetrazol-5-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (compound 37)

[0474]

[0475] To a DCE (5 mL) solution of 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (intermediate 1, 410 mg, 0.93 mmol) and 1-[(1-methyltetrazol-5-yl)methoxy]prop-2-one (intermediate 12, 190 mg, 1.12 mmol), HOAc (168 mg, 2.79 mmol) and NaBH(OAc)3 (276 mg, 1.30 mmol) were added. The mixture was stirred at 20 °C for 36 h. The mixture was quenched with water (50 mL) at 0 °C and extracted with DCM (50 mL × 3). The combined organic phases were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Ultimate C18 150*40mm*10um; mobile phase: [water (0.225% FA)-ACN]; B%: 10%-40%, 10min) to obtain 4-[[[6-[5-chloro-2-[[4-[[1-methyl-2-[(1-methyltetrazol-5-yl)methoxy]ethyl]amino]cyclohexyl]amino]-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (compound 35, 300 mg, 367.98 μmol, 40% yield).

[0476] 4-[[[6-[5-chloro-2-[[4-[[[1-methyl-2-[(1-methyltetrazol-5-yl)methoxy]ethyl]amino]cyclohexyl]amino]-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxylonitrile (compound 35, 100 mg, 0.17 mmol) was separated by chiral SFC to obtain a white solid of 4-[[[6-[5-chloro-2-[[4-[[(1S)-1-methyl-2-[(1-methyltetrazol-5-yl)methoxy]ethyl]amino]cyclohexyl]amino] [Hexyl]amino]-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxylon (compound 36, 18.7 mg, 18.2% yield, 97% purity) and 4-[[[6-[5-chloro-2-[[4-[[(1R)-1-methyl-2-[(1-methyltetrazol-5-yl)methoxy]ethyl]amino]cyclohexyl]amino]-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxylon (compound 37, 21.6 mg, 21.6% yield, 98% purity).

[0477] Compound 36: 1H NMR (400MHz, MeOD): δ = 7.95 (s, 1H), 7.50-7.46 (m, 1H), 6.85 (d, J = 7.2Hz, 1H), 6.69 (s, 1H), 6.62 (d, J = 8.4Hz, 1H), 4.90 (s, 2H) ),4.13(s,3H),3.97-3.93(m,2H),3.75(s,2H),3.66-3.61(m,3H),3.56-3.52(m,1H),3.48-3.44(m,1H),3.17-3.13(m,1H). 2.66(s,1H),2.11-2.09(m,2H),2.03-1.98(m,2H),1.90-1.87(m,2H),1.80-1.73(m,2H),1.31-1.24(m,4H),1.10(d,J=6.4Hz,3H).

[0478] Compound 37: 1 H NMR (400MHz, MeOD): δ = 7.96 (s, 1H), 7.51-7.47 (m, 1H), 6.84 (d, J = 7.2Hz, 1H), 6.6 9(s,1H),6.62(d,J=8.0Hz,1H),4.93(s,2H),4.13(s,3H),3.97-3.93(m,2H),3.75 (s,2H),3.67-3.61(m,4H),3.56-3.52(m,1H),2.87-2.84(m,1H),2.15-2.01(m,5H ),1.90-1.87(m,2H),1.80-1.72(m,2H),1.39-1.29(m,4H),1.19(d,J=6.4Hz,3H).

[0479] Example 25: Synthesis of tert-butyl (2-(2-oxopropoxy)ethyl)carbamate (intermediate 13)

[0480]

[0481] Step 1: Preparation of tert-butyl (2-((2-methylallyl)oxy)ethyl)carbamate (INT-29)

[0482]

[0483] To a solution of N-(2-hydroxyethyl)carbamate tert-butyl ester (INT-28, 1.0 g, 6.20 mmol) in DCM (12 mL), 3-bromo-2-methyl-prop-1-ene (921 mg, 6.82 mmol), TBAB (1.40 g, 4.34 mmol), and NaOH aqueous solution (10 M, 6 mL) were added. The mixture was stirred at 25 °C for 12 hours. The mixture was diluted with water (10 mL) and extracted with DCM (20 mL × 2). The combined organic layers were dried over Na₂SO₄, filtered, and concentrated under vacuum to obtain the residue. The residue was purified by silica gel chromatography (Biotage 4 g silica gel fast column; gradient eluent of 0-10% petroleum ether in ethyl acetate, @40 mL / min) to obtain N-[2-(2-methylallyloxy)ethyl]carbamate tert-butyl ester (INT-29, 880 mg, 66% yield) as a colorless oil. 1 H NMR (400MHz, CDCl3): δ = 4.96 (s, 1H), 4.91 (s, 1H), 3.89 (s, 2H), 3.49-3.46 (m, 2H), 3.35-3.32 (m, 2H), 1.74 (s, 3H), 1.45 (s, 9H).

[0484] Step 2: Preparation of (2-(2-oxopropoxy)ethyl)carbamate tert-butyl ester (intermediate 13)

[0485]

[0486] Add K₂O₄·2H₂O (17 mg, 0.05 mmol) and NaIO₄ (457 mg, 2.14 mmol) to a solution of N-[2-(2-methylallyloxy)ethyl]carbamate tert-butyl ester (200 mg, 0.93 mmol) in THF (3 mL) and H₂O (3 mL). Stir the mixture at 15 °C for 4 hours. Dilute the mixture with water (20 mL) and extract with ethyl acetate (25 mL × 2). The combined organic phases were washed with 20 mL of Na2S2O3 aqueous solution and 20 mL of brine, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to obtain the residue. The residue was purified by silica gel chromatography (Biotage 4 g silica gel fast column; gradient eluent of 0-50% petroleum ether in ethyl acetate, @40 mL / min) to obtain N-(2-propenyloxyethyl)carbamate tert-butyl ester (intermediate 13, 200 mg, 99% yield) as a pale yellow oil. 1 H NMR (400MHz, CDCl3): δ = 4.10 (s, 2H), 3.58-3.56 (m, 2H), 3.37-3.35 (m, 2H), 2.15 (s, 3H), 1.46 (s, 9H).

[0487] Example 26: Preparation of N-(2-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)ethyl)-1,1,1-trifluoromethanesulfonamide (compound 40)

[0488]

[0489] Step 1: Preparation of (2-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)ethyl)tert-butyl carbamate (compound 38)

[0490]

[0491] To a DCE (5 mL) solution of 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (intermediate 1, 300 mg, 0.68 mmol), N-(2-acetoneoxyethyl)carbamate tert-butyl ester (intermediate 13, 163 mg, 0.75 mmol), HOAc (123 mg, 2.04 mmol), and NaBH(OAc)3 (202 mg, 0.95 mmol) were added. The mixture was stirred at 20 °C under nitrogen for 4 hours. The mixture was quenched with water (5 mL) at 0 °C and then extracted with DCM (50 mL × 3). The combined organic phases were washed with saturated Na2CO3 aqueous solution (50 mL) and brine (50 mL), dried with anhydrous sodium sulfate, filtered and concentrated, and purified by silica gel chromatography (Biotage 4 g silica gel rapid column; gradient elution of 0-15% methanol in dichloromethane, @40 mL / min) to obtain N-[2-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]ethyl]tert-butyl carbamate (compound 38, 250 mg, 58% yield) as a yellow oil.

[0492] Step 2: Preparation of 4-(((2'-(((1r,4r)-4-((1-(2-aminoethoxy)prop-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 39)

[0493]

[0494] TFA (710.17 mg, 6.23 mmol) was added to a DCM (5 mL) solution of N-[2-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]ethyl]carbamate (compound 38, 200 mg, 0.31 mmol). The mixture was stirred at 20 °C for 2 hours. The mixture was concentrated and then extracted with DCM (30 mL × 3). The combined organic phases were washed with saturated Na2CO3 aqueous solution (30 mL) and brine (30 mL), dried with anhydrous sodium sulfate, filtered and concentrated to give a yellow solid 4-[[[6-[2-[[4-[[2-(2-aminoethoxy)-1-methyl-ethyl]amino]cyclohexyl]amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (compound 39, 210 mg, crude product).

[0495] Step 3: Preparation of N-(2-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)ethyl)-1,1,1-trifluoromethanesulfonamide (compound 40)

[0496]

[0497] TEA (77 mg, 0.76 mmol) and Tf₂O (94 mg, 0.33 mmol) were added to a DCM (5 mL) solution of 4-[[[6-[2-[[4-[[2-(2-aminoethoxy)-1-methyl-ethyl]amino]cyclohexyl]amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (compound 39, 165 mg, 0.30 mmol) at -78 °C. The mixture was stirred at 20 °C for 3 h. The mixture was quenched with water (5 mL) and then extracted with DCM (30 mL × 3). The combined organic phases were washed with saturated Na₂CO₃ aqueous solution (30 mL) and brine (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The reaction was concentrated and analyzed by preparative TLC (dichloromethane:methanol = 10:1, R f=0.24) to obtain the crude product, which was further purified by preparative HPLC (instrument: preparative HPLC-WI column: Phenomenex Luna C18 100*30mm*3um; mobile phase: water (0.225% FA)-ACN; start B: 6%, end B: 56%, gradient time (8min)) to obtain N-[2-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]ethyl]-1,1,1-trifluoro-methanesulfonamide (compound 40, 83 mg, 60% yield, FA salt) as a pale yellow solid. 1 H NMR (400MHz, CDCl3): δ = 8.37 (s, 1H), 8.04 (s, 1H), 7.51 (t, J = 7.6Hz, 1H), 6.95 (d, J=7.2Hz,1H),6.59(s,1H),6.52(d,J=8.4Hz,1H),5.01-4.97(m,1H),4.00-3.96(m ,2H),3.77-3.55(m,9H),3.46(brs,3H),3.08-3.02(m,1H),2.28-2.13(m,4H),1. 90(d,J=13.6Hz,2H),1.80-1.63(m,4H),1.40(d,J=6.4Hz,3H),1.35-1.26(m,2H).

[0498] Example 27: Synthesis of N-(2-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)ethyl)-1,1-difluoromethanesulfonamide (compound 41)

[0499]

[0500] TEA (90 mg, 0.89 mmol) and difluoromethanesulfonyl chloride (67 mg, 0.44 mmol) were added to a DCM (3 mL) solution of 4-[[[6-[2-[[4-[[2-(2-aminoethoxy)-1-methyl-ethyl]amino]cyclohexyl]amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (compound 39, 240 mg, 0.44 mmol) at -78 °C. The mixture was stirred at 20 °C for 1 hour. The mixture was quenched with water (8 mL) and then extracted with DCM (50 mL × 3). The combined organic phases were washed with saturated Na2CO3 (50 mL) and brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The reaction was concentrated and analyzed by preparative TLC (dichloromethane:methanol = 10:1, R f =0.13) to obtain the crude product, which was further purified by preparative HPLC (instrument: preparative HPLC-WI column: Phenomenex Luna C18100*30mm*3um; mobile phase: water (0.225% FA)-ACN; start B: 6%, end B: 56%, gradient time (8min)) to obtain N-[2-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]ethyl]-1,1-difluoro-methanesulfonamide (compound 41, 50.5 mg, 62% yield, FA salt) as a pale yellow solid. 1 H NMR (400MHz, CDCl3): δ=8.37(s,1H),8.06(s,1H),7.53(t,J=7.2Hz,1H),6.97(d,J=7.6 Hz,1H),6.60(s,1H),6.52(d,J=8.0Hz,1H),6.20(t,J=54.0Hz,1H),4.92-4.90(m,1H),4 .00-3.96(m,2H),3.77-3.61(m,10H),3.46(brs,3H),3.08-3.02(m,1H),2.29-2.21(m,4 H), 1.90 (d, J = 13.6Hz, 2H), 1.80-1.63 (m, 4H), 1.41 (d, J = 6.8Hz, 3H), 1.35-1.26 (m, 2H).

[0501] Example 28: Synthesis of 1-(2-oxopropoxy)-N-(2,2,2-trifluoroethyl)methanesulfonamide (intermediate 14)

[0502]

[0503] In five synthetic steps, chloromethanesulfonyl chloride (INT-30) was converted into 1-(2-oxopropoxy)-N-(2,2,2-trifluoroethyl)methanesulfonamide (intermediate 14).

[0504] Example 29: Synthesis of 1-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-N-(2,2,2-trifluoroethyl)methanesulfonamide (compound 43) and 1-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-N-(2,2,2-trifluoroethyl)methanesulfonamide (compound 44)

[0505]

[0506] HOAc and NaBH(OAc)3 were added to a DCE solution of 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (intermediate 1) and 1-(2-oxopropoxy)-N-(2,2,2-trifluoroethyl)methanesulfonamide (intermediate 14). The mixture was stirred at 20 °C for 36 h. The mixture was quenched with water at 0 °C and extracted with DCM. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain 1-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-N-(2,2,2-trifluoroethyl)methanesulfonamide (compound 42).

[0507] 1-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-N-(2,2,2-trifluoroethyl)methanesulfonamide (compound 42) was separated by chiral SFC to obtain 1-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl) (Amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-N-(2,2,2-trifluoroethyl)methanesulfonamide (compound 43) and 1-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-N-(2,2,2-trifluoroethyl)methanesulfonamide (compound 44).

[0508] Example 30: Synthesis of ethyl 1-(2-oxopropoxy)cyclopropane-1-carboxylate (intermediate 15)

[0509]

[0510] Step 1: Preparation of ethyl 1-((2-methylallyl)oxy)cyclopropane-1-carboxylate (INT-32)

[0511]

[0512] NaH was added to a DMF solution of ethyl 1-hydroxycyclopropane-1-carboxylate (INT-31), and the reaction mixture was stirred at 0°C for 0.5 h. 1-Bromo-2-methyl-2-propene was added, and the reaction mixture was stirred at 25°C for 2 h, quenched with brine, and extracted with EtOAc. The combined organic layers were washed with brine, dried over Na₂SO₄, filtered, and concentrated to provide ethyl 1-((2-methylallyl)oxy)cyclopropane-1-carboxylate (INT-32). The crude product was used directly in the next step without further purification.

[0513] Step 2: Preparation of ethyl 1-(2-oxopropoxy)cyclopropane-1-carboxylate (intermediate 15)

[0514]

[0515] K₂O₄·2H₂O and NaIO₄ were added to a solution of 1-((2-methylallyl)oxy)cyclopropane-1-carboxylate (INT-32) in THF and H₂O. The mixture was stirred at 15°C for 4 hours. The mixture was diluted with water and extracted with ethyl acetate. The combined organic phases were washed with an aqueous solution of Na₂S₂O₃ and brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the residue. The residue was purified by silica gel chromatography to obtain 1-(2-oxopropoxy)cyclopropane-1-carboxylate (intermediate 15).

[0516] Example 31: Synthesis of ethyl 1-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylate (compound 46) and ethyl 1-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylate (compound 47)

[0517]

[0518] HOAc and NaBH(OAc)3 were added to a DCE solution of 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (intermediate 1) and ethyl 1-(2-oxopropoxy)cyclopropane-1-carboxylate (intermediate 15). The mixture was stirred at 20 °C for 36 h. The mixture was quenched with water at 0 °C and extracted with DCM. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain ethyl 1-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylate (compound 45).

[0519] 1-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylic acid ethyl ester (compound 45) was obtained by chiral SFC separation to obtain 1-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl) ethyl cyclopropane-1-carboxylate (compound 46) and ethyl cyclopropane-1-carboxylate (compound 47).

[0520] Example 32: Synthesis of ethyl 2-methyl-2-(2-oxopropoxy)propionate (intermediate 16)

[0521]

[0522] Step 1: Preparation of ethyl 2-methyl-2-((2-methylallyl)oxy)propionate (INT-34)

[0523]

[0524] NaH was added to a DMF solution of ethyl 2-hydroxy-2-methylpropionate (INT-33), and the reaction mixture was stirred at 0°C for 0.5 h. 1-Bromo-2-methyl-2-propene was added, and the reaction mixture was stirred at 25°C for 2 h, quenched with brine, and extracted with EtOAc. The combined organic layers were washed with brine, dried over Na₂SO₄, filtered, and concentrated to provide ethyl 2-methyl-2-((2-methylallyl)oxy)propionate (INT-34). The crude product was used directly in the next step without further purification.

[0525] Step 2: Preparation of ethyl 2-methyl-2-(2-oxopropoxy)propionate (intermediate 16)

[0526]

[0527] K₂O₄·2H₂O and NaIO₄ were added to a solution of ethyl 2-methyl-2-((2-methylallyl)oxy)propionate (INT-34) in THF and H₂O. The mixture was stirred at 15°C for 4 hours. The mixture was diluted with water and extracted with ethyl acetate. The combined organic phases were washed with an aqueous solution of Na₂S₂O₃ and brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the residue. The residue was purified by silica gel chromatography to obtain ethyl 2-methyl-2-(2-oxopropoxy)propionate (intermediate 16).

[0528] Example 33: Synthesis of ethyl 2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropionate (compound 49) and ethyl 2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropionate (compound 50)

[0529]

[0530] HOAc and NaBH(OAc)3 were added to a DCE solution of 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (intermediate 1) and ethyl 2-methyl-2-(2-oxopropoxy)propionate (intermediate 16). The mixture was stirred at 20°C for 36 hours. The mixture was quenched with water at 0°C and extracted with DCM. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain ethyl 2-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropionate (compound 48).

[0531] Ethyl 2-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropionate (compound 48) was obtained by chiral SFC separation to obtain 2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl) Ethyl 2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropionate (compound 50).

[0532] Example 34: Synthesis of (R)-2-(2-oxopropoxy)propionate ethyl ester (intermediate 17)

[0533]

[0534] Step 1: Preparation of (R)-2-((2-methylallyl)oxy)propionate ethyl ester (INT-36)

[0535]

[0536] NaH was added to a DMF solution of ethyl (R)-2-hydroxypropionate (INT-35), and the reaction mixture was stirred at 0°C for 0.5 h. 1-Bromo-2-methyl-2-propene was added, and the reaction mixture was stirred at 25°C for 2 h, quenched with brine, and extracted with EtOAc. The combined organic layers were washed with brine, dried over Na₂SO₄, filtered, and concentrated to provide ethyl (R)-2-((2-methylallyl)oxy)propionate (INT-36). The crude product was used directly in the next step without further purification.

[0537] Step 2: Preparation of (R)-2-(2-oxopropoxy)propionate ethyl ester (intermediate 17)

[0538]

[0539] K₂O₄·2H₂O and NaIO₄ were added to a solution of (R)-2-((2-methylallyl)oxy)propionate (INT-36) in THF and H₂O. The mixture was stirred at 15 °C for 4 hours. The mixture was diluted with water and extracted with ethyl acetate. The combined organic phases were washed with an aqueous solution of Na₂S₂O₃ and brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the residue. The residue was purified by silica gel chromatography to obtain (R)-2-(2-oxopropoxy)propionate (intermediate 17).

[0540] Example 35: Synthesis of (R)-2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)ethyl propionate (compound 52) and (R)-2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)ethyl propionate (compound 53)

[0541]

[0542] HOAc and NaBH(OAc)3 were added to a DCE solution of 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (intermediate 1) and (R)-2-(2-oxopropoxy)propionate ethyl ester (intermediate 17). The mixture was stirred at 20 °C for 36 h. The mixture was quenched with water at 0 °C and extracted with DCM. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain ethyl (2R)-2-(2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propionate (compound 51).

[0543] Ethyl propionate (compound 51) was obtained by chiral SFC separation of (2R)-2-(2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)acetate (compound 51). Ethyl pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propionate (compound 52) and (R)-2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propionate (compound 53).

[0544] Example 36: Synthesis of ethyl (S)-2-(2-oxopropoxy)propionate (intermediate 18)

[0545]

[0546] Step 1: Preparation of ethyl (S)-2-((2-methylallyl)oxy)propionate (INT-38)

[0547]

[0548] NaH was added to a DMF solution of ethyl (S)-2-hydroxypropionate (INT-37), and the reaction mixture was stirred at 0°C for 0.5 h. 1-Bromo-2-methyl-2-propene was added, and the reaction mixture was stirred at 25°C for 2 h, quenched with brine, and extracted with EtOAc. The combined organic layers were washed with brine, dried over Na₂SO₄, filtered, and concentrated to provide ethyl (S)-2-((2-methylallyl)oxy)propionate (INT-38). The crude product was used directly in the next step without further purification.

[0549] Step 2: Preparation of (S)-2-(2-oxopropoxy)propionate ethyl ester (intermediate 18)

[0550]

[0551] K₂O₄·2H₂O and NaIO₄ were added to a solution of (S)-2-((2-methylallyl)oxy)propionate (INT-38) in THF and H₂O. The mixture was stirred at 15 °C for 4 hours. The mixture was diluted with water and extracted with ethyl acetate. The combined organic phases were washed with an aqueous solution of Na₂S₂O₃ and brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the residue. The residue was purified by silica gel chromatography to obtain (S)-2-(2-oxopropoxy)propionate (intermediate 18).

[0552] Example 37: Synthesis of (S)-2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)ethyl propionate (compound 55) and (S)-2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)ethyl propionate (compound 56)

[0553]

[0554] HOAc and NaBH(OAc)3 were added to a DCE solution of 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (intermediate 1) and ethyl (S)-2-(2-oxopropoxy)propionate (intermediate 18). The mixture was stirred at 20 °C for 36 h. The mixture was quenched with water at 0 °C and extracted with DCM. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain ethyl (2S)-2-(2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propionate (compound 54).

[0555] Ethyl propionate (compound 54) was obtained by chiral SFC separation of (2S)-2-(2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)propionate (compound 54). Ethyl pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propionate (compound 55) and (S)-2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propionate (compound 56).

[0556] Example 38: Synthesis of 1-((6-nitropyridin-2-yl)methoxy)prop-2-one (intermediate 19)

[0557]

[0558] Step 1: Preparation of 2-(((2-methylallyl)oxy)methyl)-6-nitropyridine (INT-40)

[0559]

[0560] NaH was added to a DMF solution of (6-nitropyridin-2-yl)methanol (INT-39), and the reaction mixture was stirred at 0°C for 0.5 h. 1-Bromo-2-methyl-2-propene was added, and the reaction mixture was stirred at 25°C for 2 h, quenched with brine, and extracted with EtOAc. The combined organic layers were washed with brine, dried over Na₂SO₄, filtered, and concentrated to provide 2-(((2-methylallyl)oxy)methyl)-6-nitropyridinium (INT-40). The crude product was used directly in the next step without further purification.

[0561] Step 2: Preparation of ethyl (S)-2-(2-oxopropoxy)propionate (intermediate 19)

[0562]

[0563] Add K₂O₄·2H₂O and NaIO₄ to a solution of 2-(((2-methylallyl)oxy)methyl)-6-nitropyridine (INT-40) in THF and H₂O. Stir the mixture at 15°C for 4 hours. Dilute the mixture with water and extract with ethyl acetate. Wash the combined organic phases with an aqueous solution of Na₂S₂O₃ and brine, dry with anhydrous sodium sulfate, filter and concentrate under vacuum to obtain the residue. Purify the residue by silica gel chromatography to obtain ethyl (S)-2-(2-oxopropoxy)propionate (intermediate 19).

[0564] Example 39: Synthesis of N-(6-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)pyridin-2-yl)methanesulfonamide (compound 58) and N-(6-(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)pyridin-2-yl)methanesulfonamide (compound 59)

[0565]

[0566] HOAc and NaBH(OAc)3 were added to a DCE solution of 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (intermediate 1) and ethyl (S)-2-(2-oxopropoxy)propionate (intermediate 19). The mixture was stirred at 20°C for 36 hours. The mixture was quenched with water at 0°C and extracted with DCM. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC, reduced with iron and acetic acid, and then acylated with methanesulfonyl chloride and triethylamine to obtain N-(6-((2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)pyridin-2-yl)methanesulfonamide (compound 57).

[0567] N-(6-((2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)pyridin-2-yl)methanesulfonamide (compound 57) was obtained by chiral SFC separation. Compound 58 and N-(6-(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)pyridine-2-yl)methanesulfonamide (compound 59).

[0568] Example 40: Synthesis of (R)-3-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)ethyl)butyrate (compound 61) and (S)-3-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)ethyl)butyrate (compound 62)

[0569]

[0570] 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxylon (intermediate 1) was treated with ethyl 2-butenoate to obtain ethyl 3-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)butyrate (compound 60).

[0571] Ethyl 3-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butyrate (compound 60) was obtained by chiral SFC separation to obtain (R)-3-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran)) (Compound 61) and (S)-3-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)ethyl butyrate (Compound 62).

[0572] Example 41: Synthesis of (R)-3-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)tert-butyl butyrate (compound 64) and (S)-3-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)tert-butyl butyrate (compound 65)

[0573]

[0574] 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxylon (intermediate 1) was treated with tert-butyl 2-butenoate to obtain tert-butyl 3-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)tert-butyl butyrate (compound 63).

[0575] 3-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)tert-butyl butyrate (compound 63) was obtained by chiral SFC separation to obtain (R)-3-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran)) -4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)tert-butyl butyrate (compound 64) and (S)-3-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)tert-butyl butyrate (compound 65).

[0576] Example 42: Synthesis of (R)-3-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butyric acid (compound 66)

[0577]

[0578] (R)-3-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)tert-butyl butyrate (compound 64) was treated with trifluoroacetic acid to obtain (R)-3-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butyrate (compound 66).

[0579] Example 43: Synthesis of (S)-3-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butyric acid (compound 67)

[0580]

[0581] (S)-3-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)tert-butyl butyrate (compound 65) was treated with trifluoroacetic acid to obtain (S)-3-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butyrate (compound 67).

[0582] Example 44: (R)-1-(5-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-2H-tetrazole-2-yl)-2-methylpropylneopentate (compound 68), ( S)-1-(5-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-2H-tetrazole-2-yl)-2-methylpropylneovalerate (compound 69), 4-(((5'- Chloro-2'-(((1R,4r)-4-(((R)-1-((1-((R)-1-((3,3-dimethylbut-1-en-2-yl)oxy)-2-methylpropyl)-1H-tetrazol-5-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (compound) Synthesis of (compound 70) and (S)-1-(5-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-1H-tetrazol-1-yl)-2-methylpropylneopentate (compound 71)

[0583]

[0584] Compound 2 was obtained by treating 4-(((2'-(((1R,4r)-4-(((R)-1-((2H-tetrazol-5-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (compound 2) with (S)-1-chloro-2-methylpropylneovalerate and Ag2O and then separating by chiral SFC to obtain (R)-1-(5-(((R)-2-(((1r, 4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-2H-tetrazole-2-yl)-2-methylpropyl neopentanoate (compound 68), (S)-1-(5-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)- [2,4'-Bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-2H-tetrazol-2-yl)-2-methylpropylneopentate (compound 69), 4-(((5'-chloro-2'-(((1R,4r)-4-(((R)-1-((1-(((R)-1-((3,3-dimethylbut-1-en-2-yl)oxy)-2-methylpropyl)-1H-tetrazol-5-yl)methoxy)prop-2-yl)amino)cyclohexyl)amino)- [2,4'-Bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 70) and (S)-1-(5-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-Bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-1H-tetrazole-1-yl)-2-methylpropylneovalerate (compound 71).

[0585] Example 45: (R)-1-(5-(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-2H-tetrazole-2-yl)-2-methylpropyl neopentanoate (compound 72), ( S)-1-(5-(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-2H-tetrazole-2-yl)-2-methylpropylneopentate (compound 73), 4-(((5'- Chloro-2'-(((1S,4r)-4-(((S)-1-((1-((R)-1-((3,3-dimethylbut-1-en-2-yl)oxy)-2-methylpropyl)-1H-tetrazol-5-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (compound) Synthesis of (Compound 74) and (S)-1-(5-(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-1H-tetrazol-1-yl)-2-methylpropylneopentate (Compound 75)

[0586]

[0587] Compound 3 was prepared by treating 4-(((2'-(((1S,4r)-4-(((S)-1-((2H-tetrazol-5-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridin]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (compound 3) with (S)-1-chloro-2-methylpropylneovalerate and Ag2O and then separated by chiral SFC to obtain (R)-1-(5-(((S)-2-(((1r, 4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-2H-tetrazole-2-yl)-2-methylpropyl neopentanoate (compound 72), (S)-1-(5-(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)- [2,4'-Bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-2H-tetrazol-2-yl)-2-methylpropylneovalerate (compound 73), 4-(((5'-chloro-2'-(((1S,4r)-4-(((S)-1-((1-((R)-1-((3,3-dimethylbut-1-en-2-yl)oxy)-2-methylpropyl)-1H-tetrazol-5-yl)methoxy)prop-2-yl)amino)cyclohexyl)amino)- [2,4'-Bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (compound 74) and (S)-1-(5-(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-Bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-1H-tetrazole-1-yl)-2-methylpropylneovalerate (compound 75).

[0588] Example 46: Synthesis of 1-((4-methoxypyrimidin-2-yl)methoxy)prop-2-one (Intermediate 20)

[0589]

[0590] Step 1: Preparation of 4-methoxy-2-(((2-methylallyl)oxy)methyl)pyrimidine (INT-42)

[0591]

[0592] NaH was added to a DMF solution of (4-methoxypyrimidin-2-yl)methanol (INT-41), and the reaction mixture was stirred at 0°C for 0.5 h. 1-Bromo-2-methyl-2-propene was added, and the reaction mixture was stirred at 25°C for 2 h, quenched with brine, and extracted with EtOAc. The combined organic layers were washed with brine, dried over Na₂SO₄, filtered, and concentrated to provide 4-methoxy-2-(((2-methylallyl)oxy)methyl)pyrimidin (INT-42). The crude product was used directly in the next step without further purification.

[0593] Step 2: Preparation of 1-((4-methoxypyrimidin-2-yl)methoxy)prop-2-one (intermediate 20)

[0594]

[0595] Add K₂O₄·2H₂O and NaIO₄ to a solution of 4-methoxy-2-(((2-methylallyl)oxy)methyl)pyrimidine (INT-42) in THF and H₂O. Stir the mixture at 15°C for 4 hours. Dilute the mixture with water and extract with ethyl acetate. Wash the combined organic phases with an aqueous solution of Na₂S₂O₃ and brine, dry with anhydrous sodium sulfate, filter and concentrate under vacuum to obtain the residue. Purify the residue by silica gel chromatography to obtain 1-((4-methoxypyrimidin-2-yl)methoxy)prop-2-one (intermediate 20).

[0596] Example 47: Synthesis of 4-(((5'-chloro-2'-(((1R,4r)-4-(((R)-1-((5-fluoro-6-oxo-1,6-dihydropyrimidin-2-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (Compound 77) and 4-(((5'-chloro-2'-(((1S,4r)-4-(((S)-1-((5-fluoro-6-oxo-1,6-dihydropyrimidin-2-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (Compound 78)

[0597]

[0598] HOAc and NaBH(OAc)3 were added to a DCE solution of 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (intermediate 1) and 1-((4-methoxypyrimidin-2-yl)methoxy)prop-2-one (intermediate 20). The mixture was stirred at 20 °C for 36 hours. The mixture was quenched with water at 0 °C and extracted with DCM. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC, silanized with TMSI, and fluorinated with SelectFluor to obtain 4-(((5'-chloro-2'-(((1r,4r)-4-((1-((5-fluoro-6-oxo-1,6-dihydropyrimidin-2-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (compound 76).

[0599] Compound 76 was obtained by chiral SFC separation of 4-(((5'-chloro-2'-(((1r,4r)-4-((1-((5-fluoro-6-oxo-1,6-dihydropyrimidin-2-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (compound 76). )methoxy)prop-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylon (compound 77) and 4-(((5'-chloro-2'-(((1S,4r)-4-(((S)-1-((5-fluoro-6-oxo-1,6-dihydropyrimidin-2-yl)methoxy)prop-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylon (compound 78).

[0600] Example 48: Synthesis of tert-butyl 1-(2-oxopropoxy)cyclopropane-1-carboxylate (intermediate 21)

[0601]

[0602] Step 1: Preparation of tert-butyl 1-((2-methylallyl)oxy)cyclopropane-1-carboxylate (INT-44)

[0603]

[0604] NaH was added to a DMF solution of 1-hydroxycyclopropane-1-carboxylate tert-butyl ester (INT-43), and the reaction mixture was stirred at 0 °C for 0.5 h. 1-Bromo-2-methyl-2-propene was added, and the reaction mixture was stirred at 25 °C for 2 h, quenched with brine, and extracted with EtOAc. The combined organic layers were washed with brine, dried over Na₂SO₄, filtered, and concentrated to provide 1-((2-methylallyl)oxy)cyclopropane-1-carboxylate tert-butyl ester (INT-44). The crude product was used directly in the next step without further purification.

[0605] Step 2: Preparation of tert-butyl 1-(2-oxopropoxy)cyclopropane-1-carboxylate (intermediate 21)

[0606]

[0607] Add K₂O₅·2H₂O and NaIO₄ to a solution of 1-((2-methylallyl)oxy)cyclopropane-1-carboxylic acid tert-butyl ester (INT-44) in THF and H₂O. Stir the mixture at 15°C for 4 hours. Dilute the mixture with water and extract with ethyl acetate. Wash the combined organic phases with aqueous Na₂S₂O₃ solution and brine, dry with anhydrous sodium sulfate, filter and concentrate under vacuum to obtain the residue. Purify the residue by silica gel chromatography to obtain 1-(2-oxopropoxy)cyclopropane-1-carboxylic acid tert-butyl ester (intermediate 21).

[0608] Example 49: Synthesis of 1-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)tert-butyl cyclopropane-1-carboxylate (compound 80) and 1-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)tert-butyl cyclopropane-1-carboxylate (compound 81)

[0609]

[0610] HOAc and NaBH(OAc)3 were added to a DCE solution of 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (intermediate 1) and tert-butyl 1-(2-oxopropoxy)cyclopropane-1-carboxylate (intermediate 21). The mixture was stirred at 20 °C for 36 h. The mixture was quenched with water at 0 °C and extracted with DCM. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain tert-butyl 1-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylic acid (compound 79).

[0611] 1-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-tert-butyl ester (compound 79) was obtained by chiral SFC separation to obtain 1-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl) )methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylic acid tert-butyl ester (compound 80) and 1-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylic acid tert-butyl ester (compound 81).

[0612] Example 50: Synthesis of (R)-3-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butyric acid (compound 66)

[0613]

[0614] 1-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylic acid tert-butyl ester (compound 80) was treated with trifluoroacetic acid to obtain 1-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylic acid (compound 82).

[0615] Example 51: Synthesis of (S)-3-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butyric acid (compound 67)

[0616]

[0617] 1-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylic acid tert-butyl ester (compound 81) was treated with trifluoroacetic acid to obtain 1-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylic acid (compound 83).

[0618] Example 52: Synthesis of tert-butyl 2-methyl-2-(2-oxopropoxy)propionate (intermediate 22)

[0619]

[0620] Step 1: Preparation of tert-butyl 2-methyl-2-((2-methylallyl)oxy)propionate (INT-46)

[0621]

[0622] NaH was added to a DMF solution of tert-butyl 2-hydroxy-2-methylpropionate (INT-45), and the reaction mixture was stirred at 0°C for 0.5 h. 1-Bromo-2-methyl-2-propene was added, and the reaction mixture was stirred at 25°C for 2 h, quenched with brine, and extracted with EtOAc. The combined organic layers were washed with brine, dried over Na₂SO₄, filtered, and concentrated to provide tert-butyl 2-methyl-2-((2-methylallyl)oxy)propionate (INT-46). The crude product was used directly in the next step without further purification.

[0623] Step 2: Preparation of tert-butyl 2-methyl-2-(2-oxopropoxy)propionate (intermediate 22)

[0624]

[0625] To a solution of tert-butyl 2-methyl-2-((2-methylallyl)oxy)propionate (INT-46) in THF and H2O, K2O·2H2O and NaIO4 were added. The mixture was stirred at 15°C for 4 hours. The mixture was diluted with water and extracted with ethyl acetate. The combined organic phases were washed with an aqueous solution of Na2S2O3 and brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the residue. The residue was purified by silica gel chromatography to obtain tert-butyl 2-methyl-2-(2-oxopropoxy)propionate (intermediate 22).

[0626] Example 53: Synthesis of tert-butyl 2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropionate (compound 85) and tert-butyl 2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropionate (compound 86)

[0627]

[0628] HOAc and NaBH(OAc)3 were added to a DCE solution of 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (intermediate 1) and tert-butyl 2-methyl-2-(2-oxopropoxy)propionate (intermediate 22). The mixture was stirred at 20 °C for 36 hours. The mixture was quenched with water at 0 °C and extracted with DCM. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain tert-butyl 2-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropionate (compound 84).

[0629] 2-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropionate tert-butyl ester (compound 84) was obtained by chiral SFC separation to obtain 2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl) 2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropionate tert-butyl ester (compound 86).

[0630] Example 54: Synthesis of tert-butyl-2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropionic acid (compound 87)

[0631]

[0632] 2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropionic acid tert-butyl ester (compound 85) was treated with trifluoroacetic acid to obtain 2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropionic acid (compound 87).

[0633] Example 55: Synthesis of tert-butyl-2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropionic acid (compound 88)

[0634]

[0635] 2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropionic acid tert-butyl ester (compound 86) was treated with trifluoroacetic acid to obtain 2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropionic acid (compound 88).

[0636] Example 56: Synthesis of (R)-2-(2-oxopropoxy)propionate tert-butyl ester (intermediate 23)

[0637]

[0638] Step 1: Preparation of (R)-2-((2-methylallyl)oxy)tert-butyl propionate (INT-48)

[0639]

[0640] NaH was added to a DMF solution of (R)-2-hydroxypropionate tert-butyl (INT-47), and the reaction mixture was stirred at 0 °C for 0.5 h. 1-Bromo-2-methyl-2-propene was added, and the reaction mixture was stirred at 25 °C for 2 h, quenched with brine, and extracted with EtOAc. The combined organic layers were washed with brine, dried over Na₂SO₄, filtered, and concentrated to provide (R)-2-((2-methylallyl)oxy)propionate tert-butyl (INT-48). The crude product was used directly in the next step without further purification.

[0641] Step 2: Preparation of (R)-2-(2-oxopropoxy)propionate tert-butyl ester (intermediate 23)

[0642]

[0643] Add K₂O₄·2H₂O and NaIO₄ to a solution of (R)-2-((2-methylallyl)oxy)propionate tert-butyl ester (INT-48) in THF and H₂O. Stir the mixture at 15°C for 4 hours. Dilute the mixture with water and extract with ethyl acetate. Wash the combined organic phases with aqueous Na₂S₂O₃ solution and brine, dry with anhydrous sodium sulfate, filter and concentrate under vacuum to obtain the residue. Purify the residue by silica gel chromatography to obtain (R)-2-(2-oxopropoxy)propionate tert-butyl ester (intermediate 23).

[0644] Example 57: Synthesis of (R)-2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)tert-butyl propionate (compound 90) and (R)-2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)tert-butyl propionate (compound 91)

[0645]

[0646] HOAc and NaBH(OAc)3 were added to a DCE solution of 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (intermediate 1) and (R)-2-(2-oxopropoxy)propionate tert-butyl ester (intermediate 23). The mixture was stirred at 20 °C for 36 h. The mixture was quenched with water at 0 °C and extracted with DCM. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain (2R)-2-(2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)tert-butyl propionate (compound 89).

[0647] (2R)-2-(2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)tert-butyl propionate (compound 89) was obtained by chiral SFC separation to obtain (R)-2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-yl)) (Compound 90) and (R)-2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)tert-butyl propionate (Compound 91).

[0648] Example 58: Synthesis of (R)-2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propionic acid (compound 92)

[0649]

[0650] (R)-2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)tert-butyl propionate (compound 90) was treated with trifluoroacetic acid to obtain (R)-2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propionate (compound 92).

[0651] Example 59: Synthesis of (R)-2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propionic acid (compound 93)

[0652]

[0653] (R)-2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)tert-butyl propionate (compound 91) was treated with trifluoroacetic acid to obtain (R)-2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propionic acid (compound 93).

[0654] Example 60: Synthesis of (S)-2-(2-oxopropoxy)propionate tert-butyl ester (intermediate 24)

[0655]

[0656] Step 1: Preparation of (S)-2-((2-methylallyl)oxy)tert-butyl propionate (INT-50)

[0657]

[0658] NaH was added to a DMF solution of (S)-2-hydroxypropionate tert-butyl (INT-49), and the reaction mixture was stirred at 0 °C for 0.5 h. 1-Bromo-2-methyl-2-propene was added, and the reaction mixture was stirred at 25 °C for 2 h, quenched with brine, and extracted with EtOAc. The combined organic layers were washed with brine, dried over Na₂SO₄, filtered, and concentrated to provide (S)-2-((2-methylallyl)oxy)propionate tert-butyl (INT-50). The crude product was used directly in the next step without further purification.

[0659] Step 2: Preparation of (S)-2-(2-oxopropoxy)propionate tert-butyl ester (intermediate 24)

[0660]

[0661] Add K₂O₄·2H₂O and NaIO₄ to a solution of (S)-2-((2-methylallyl)oxy)propionate tert-butyl ester (INT-50) in THF and H₂O. Stir the mixture at 15°C for 4 hours. Dilute the mixture with water and extract with ethyl acetate. Wash the combined organic phases with an aqueous solution of Na₂S₂O₃ and brine, dry with anhydrous sodium sulfate, filter and concentrate under vacuum to obtain the residue. Purify the residue by silica gel chromatography to obtain (S)-2-(2-oxopropoxy)propionate tert-butyl ester (intermediate 24).

[0662] Example 61: Synthesis of (S)-2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)tert-butyl propionate (compound 95) and (S)-2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)tert-butyl propionate (compound 96)

[0663]

[0664] HOAc and NaBH(OAc)3 were added to a DCE solution of 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (intermediate 1) and (S)-2-(2-oxopropoxy)tert-butyl propionate (intermediate 24). The mixture was stirred at 20 °C for 36 h. The mixture was quenched with water at 0 °C and extracted with DCM. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain (2S)-2-(2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)tert-butyl propionate (compound 94).

[0665] (2S)-2-(2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridin]-2'-yl)amino)cyclohexyl)amino)propoxy)tert-butyl propionate (compound 94) was obtained by chiral SFC separation to obtain (S)-2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-) tert-butyl propionate (compound 95) and (S)-2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)tert-butyl propionate (compound 96).

[0666] Example 62: Synthesis of (S)-2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propionic acid (compound 97)

[0667]

[0668] (S)-2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)tert-butyl propionate (compound 95) was treated with trifluoroacetic acid to obtain (S)-2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propionic acid (compound 97).

[0669] Example 63: Synthesis of (S)-2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propionic acid (compound 98)

[0670]

[0671] (S)-2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)tert-butyl propionate (compound 96) was treated with trifluoroacetic acid to obtain (S)-2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propionic acid (compound 98).

[0672] Example 64: Synthesis of 1-(1-(2-triphenylmethyl-2H-tetrazol-5-yl)cyclopropoxy)prop-2-one (intermediate 25)

[0673]

[0674] Step 1: Preparation of 5-(1-((2-methylallyl)oxy)cyclopropyl)-2-triphenylmethyl-2H-tetrazole (INT-52)

[0675]

[0676] NaH was added to a DMF solution of 1-(2-triphenylmethyl-2H-tetrazole-5-yl)cycloprop-1-ol (INT-51), and the reaction mixture was stirred at 0°C for 0.5 h. 1-Bromo-2-methyl-2-propene was added, and the reaction mixture was stirred at 25°C for 2 h, quenched with brine, and extracted with EtOAc. The combined organic layers were washed with brine, dried over Na₂SO₄, filtered, and concentrated to provide 5-(1-((2-methylallyl)oxy)cyclopropyl)-2-triphenylmethyl-2H-tetrazole (INT-52). The crude product was used directly in the next step without further purification.

[0677] Step 2: Preparation of 1-(1-(2-triphenylmethyl-2H-tetrazol-5-yl)cyclopropoxy)prop-2-one (intermediate 25)

[0678]

[0679] Add K₂O₄·2H₂O and NaIO₄ to a solution of 5-(1-((2-methylallyl)oxy)cyclopropyl)-2-triphenylmethyl-2H-tetrazole (INT-52) in THF and H₂O. Stir the mixture at 15°C for 4 hours. Dilute the mixture with water and extract with ethyl acetate. Wash the combined organic phases with aqueous Na₂S₂O₃ solution and brine, dry with anhydrous sodium sulfate, filter and concentrate under vacuum to obtain the residue. Purify the residue by silica gel chromatography to obtain 1-(1-(2-triphenylmethyl-2H-tetrazole-5-yl)cyclopropoxy)prop-2-one (intermediate 25).

[0680] Example 65: Synthesis of 4-(((2'-(((1R,4r)-4-(((R)-1-(1-(2H-tetrazol-5-yl)cyclopropoxy)propyl-2-yl)amino)cyclohexyl)amino)5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (Compound 100) and 4-(((2'-(((1S,4r)-4-(((S)-1-(1-(2H-tetrazol-5-yl)cyclopropoxy)propyl-2-yl)amino)cyclohexyl)amino)5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (Compound 101)

[0681]

[0682] HOAc and NaBH(OAc)3 were added to a DCE solution of 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (intermediate 1) and 1-(1-(2-triphenylmethyl-2H-tetrazol-5-yl)cyclopropoxy)prop-2-one (intermediate 25). The mixture was stirred at 20 °C for 36 hours. The mixture was quenched with water at 0 °C and extracted with DCM. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC and deprotected with HCl to obtain 4-(((2'-(((1r,4r)-4-((1-(1-(2H-tetrazol-5-yl)cyclopropoxy)propyl-2-yl)amino)cyclohexyl)amino)5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 99).

[0683] 4-(((2'-(((1r,4r)-4-((1-(1-(2H-tetrazol-5-yl)cyclopropoxy)propyl-2-yl)amino)cyclohexyl)amino)5'-chloro-[2,4'-bipyridin]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (compound 99) was separated by chiral SFC to obtain 4-(((2'-(((1R,4r)-4-(((R)-1-(1-(2H-tetrazol-5-yl)cyclopropoxy)propyl-2-yl) )amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 100) and 4-(((2'-(((1S,4r)-4-(((S)-1-(1-(2H-tetrazol-5-yl)cyclopropoxy)prop-2-yl)amino)cyclohexyl)amino)5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 101).

[0684] Example 66: Synthesis of 1-((2-(2-triphenylmethyl-2H-tetrazol-5-yl)prop-2-yl)oxy)prop-2-one (intermediate 26)

[0685]

[0686] Step 1: Preparation of 5-(2-((2-methylallyl)oxy)propyl-2-yl)-2-triphenylmethyl-2H-tetrazole (INT-54)

[0687]

[0688] NaH was added to a DMF solution of 2-(2-triphenylmethyl-2H-tetrazol-5-yl)prop-2-ol (INT-53), and the reaction mixture was stirred at 0°C for 0.5 h. 1-Bromo-2-methyl-2-propene was added, and the reaction mixture was stirred at 25°C for 2 h, quenched with brine, and extracted with EtOAc. The combined organic layers were washed with brine, dried over Na₂SO₄, filtered, and concentrated to provide 5-(2-((2-methylallyl)oxy)prop-2-yl)-2-triphenylmethyl-2H-tetrazol (INT-54). The crude product was used directly in the next step without further purification.

[0689] Step 2: Preparation of 1-((2-(2-triphenylmethyl-2H-tetrazol-5-yl)prop-2-yl)oxy)prop-2-one (intermediate 26)

[0690]

[0691] Add K₂O₄·2H₂O and NaIO₄ to a solution of 5-(2-((2-methylallyl)oxy)prop-2-yl)-2-triphenylmethyl-2H-tetrazole (INT-54) in THF and H₂O. Stir the mixture at 15°C for 4 hours. Dilute the mixture with water and extract with ethyl acetate. Wash the combined organic phases with aqueous Na₂S₂O₃ solution and brine, dry with anhydrous sodium sulfate, filter and concentrate under vacuum to obtain the residue. Purify the residue by silica gel chromatography to obtain 1-((2-(2-triphenylmethyl-2H-tetrazole-5-yl)prop-2-yl)oxy)prop-2-one (intermediate 26).

[0692] Example 67: Synthesis of 4-(((2'-(((1R,4r)-4-(((R)-1-((2-(2H-tetrazol-5-yl)prop-2-yl)oxy)prop-2-yl)amino)cyclohexyl)amino)5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 103) and 4-(((2'-(((1S,4r)-4-(((S)-1-((2-(2H-tetrazol-5-yl)prop-2-yl)oxy)prop-2-yl)amino)cyclohexyl)amino)5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 104)

[0693]

[0694] HOAc and NaBH(OAc)3 were added to a DCE solution of 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (intermediate 1) and 1-((2-(2-triphenylmethyl-2H-tetrazol-5-yl)prop-2-yl)oxy)prop-2-one (intermediate 26). The mixture was stirred at 20 °C for 36 hours. The mixture was quenched with water at 0 °C and extracted with DCM. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC and deprotected with HCl to obtain 4-(((2'-(((1r,4r)-4-((1-((2-(2H-tetraazol-5-yl)prop-2-yl)oxy)prop-2-yl)amino)cyclohexyl)amino)5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 102).

[0695] 4-(((2'-(((1r,4r)-4-((1-((2-(2H-tetrazol-5-yl)propyl-2-yl)oxy)propyl-2-yl)amino)cyclohexyl)amino)5'-chloro-[2,4'-bipyridin]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 102) was separated by chiral SFC to obtain 4-(((2'-(((1R,4r)-4-(((R)-1-((2-(2H-tetrazol-5-yl)propyl-2-yl)oxy)propyl- 2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 103) and 4-(((2'-(((1S,4r)-4-(((S)-1-((2-(2H-tetrazol-5-yl)prop-2-yl)oxy)prop-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 104).

[0696] Example 68: Synthesis of 1-(1-(2-triphenylmethyl-2H-tetrazol-5-yl)ethoxy)prop-2-one (intermediate 27)

[0697]

[0698] Step 1: Preparation of 5-(1-((2-methylallyl)oxy)ethyl)-2-triphenylmethyl-2H-tetrazole (INT-56)

[0699]

[0700] NaH was added to a DMF solution of 1-(2-triphenylmethyl-2H-tetrazole-5-yl)ethanol-1-ol (INT-55), and the reaction mixture was stirred at 0°C for 0.5 h. 1-Bromo-2-methyl-2-propene was added, and the reaction mixture was stirred at 25°C for 2 h, quenched with brine, and extracted with EtOAc. The combined organic layers were washed with brine, dried over Na₂SO₄, filtered, and concentrated to provide 5-(1-((2-methylallyl)oxy)ethyl)-2-triphenylmethyl-2H-tetrazole (INT-56). The crude product was used directly in the next step without further purification.

[0701] Step 2: Preparation of 1-(1-(2-triphenylmethyl-2H-tetrazol-5-yl)ethoxy)prop-2-one (intermediate 27)

[0702]

[0703] Add K₂O₄·2H₂O and NaIO₄ to a solution of 5-(1-((2-methylallyl)oxy)ethyl)-2-triphenylmethyl-2H-tetrazole (INT-56) in THF and H₂O. Stir the mixture at 15°C for 4 hours. Dilute the mixture with water and extract with ethyl acetate. Wash the combined organic phases with aqueous Na₂S₂O₃ solution and brine, dry with anhydrous sodium sulfate, filter and concentrate under vacuum to obtain the residue. Purify the residue by silica gel chromatography to obtain 1-(1-(2-triphenylmethyl-2H-tetrazole-5-yl)ethoxy)prop-2-one (intermediate 27).

[0704] Example 69: 4-(((2'-(((1R,4r)-4-(((2R,3S)-3-((2H-tetrazol-5-yl)methoxy)but-2-yl)amino)cyclohexyl)amino)5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (Compound 106), 4-(((2'-(((1S,4r)-4-(((S)-1-((R)-1-(2H-tetrazol-5-yl)ethoxy)propyl-2-yl)amino)cyclohexyl)amino)5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (Compound 107) Synthesis of 4-(((2'-(((1R,4r)-4-(((2R,3R)-3-((2H-tetrazol-5-yl)methoxy)but-2-yl)amino)cyclohexyl)amino)5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 108) and 4-(((2'-(((1S,4r)-4-(((S)-1-((S)-1-(2H-tetrazol-5-yl)ethoxy)prop-2-yl)amino)cyclohexyl)amino)5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 109)

[0705]

[0706] HOAc and NaBH(OAc)3 were added to a DCE solution of 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (intermediate 1) and 1-(1-(2-triphenylmethyl-2H-tetrazol-5-yl)ethoxy)prop-2-one (intermediate 27). The mixture was stirred at 20°C for 36 hours. The mixture was quenched with water at 0°C and extracted with DCM. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC and deprotected with HCl to obtain 4-(((2'-(((1r,4r)-4-((1-((2-(2H-tetraazol-5-yl)prop-2-yl)oxy)prop-2-yl)amino)cyclohexyl)amino)5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 102).

[0707] 4-(((2'-(((1r,4r)-4-((1-((2-(2H-tetrazol-5-yl)prop-2-yl)oxy)prop-2-yl)amino)cyclohexyl)amino)5'-chloro-[2,4'-bipyridin]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 102) was separated by chiral SFC to obtain 4-(((2'-(((1R,4r)-4- (((2R,3S)-3-((2H-tetrazol-5-yl)methoxy)but-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 106), 4-(((2'-(((1S,4r)-4-(((S)-1-((R)-1-(2H-tetrazol-5-yl)ethoxy)prop- 2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 107), 4-(((2'-(((1R,4r)-4-(((2R,3R)-3-((2H-tetrazol-5-yl)methoxy)but-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6 -yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 108) and 4-(((2'-(((1S,4r)-4-(((S)-1-((S)-1-(2H-tetrazol-5-yl)ethoxy)prop-2-yl)amino)cyclohexyl)amino)5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 109).

[0708] Example 70: Synthesis of 5-((2-oxopropoxy)methyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (intermediate 28)

[0709]

[0710] Step 1: Preparation of 5-(hydroxymethyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (INT-58)

[0711]

[0712] In three synthetic steps, hydrazine hydrochloride (INT-57) was converted into 5-(hydroxymethyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (INT-58).

[0713] Step 2: Preparation of 5-((2-oxopropoxy)methyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (intermediate 28)

[0714]

[0715] In four synthetic steps, 5-hydroxymethyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (INT-58) was converted into 5-((2-oxopropoxy)methyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (intermediate 28).

[0716] Example 71: Synthesis of 4-(((5'-chloro-2'-(((1R,4r)-4-(((R)-1-((5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 111) and 4-(((5'-chloro-2'-(((1S,4r)-4-(((S)-1-((5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylonitrile (compound 112)

[0717]

[0718] HOAc and NaBH(OAc)3 were added to a DCE solution of 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carboxynitrile (intermediate 1) and 5-((2-oxopropoxy)methyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (intermediate 28). The mixture was stirred at 20 °C for 36 hours. The mixture was quenched with water at 0 °C and extracted with DCM. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain 4-(((5'-chloro-2'-(((1r,4r)-4-((1-((5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (compound 110).

[0719] 4-(((5'-chloro-2'-(((1r,4r)-4-((1-((5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxynitrile (compound 110) was obtained by chiral SFC separation to obtain 4-(((5'-chloro-2'-(((1R,4r)-4-(((R)-1-((5-oxo-4,5-dihydro-1H-1,2,4-triazol- 3-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylon (compound 111) and 4-(((5'-chloro-2'-(((1S,4r)-4-(((S)-1-((5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)methoxy)propyl-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carboxylon (compound 112).

[0720] Example 72: CDK9 Biochemical Assay

[0721] Add 5x kinase assay buffer to each well of a 96-well plate. This buffer contains 10 mM DTT (6 μL), 500 μM ATP (1 μL), 5x CDK substrate (10 μL), and water (8 μL). Add 5 μL of the compound to the test and positive control groups, and 5 μL of the solvent to the blank group. Add 100 ng of CDK9 / CyclinT (in 20 μL of water) to the test and positive control groups, and 20 μL of 1x kinase assay buffer to the blank group. Incubate the reaction mixture at 30 °C for 45 minutes, and add 50 μL of the 5x kinase assay buffer to each well. Max. Cover the plate with light and incubate at room temperature for 15 minutes. Measure the luminescence on a microplate reader and calculate the IC50 using Prism 9 software. 50 value.

[0722] IC obtained according to the above procedure 50 The values ​​are summarized in Table 1:

[0723]

[0724]

[0725] A: <2.50nM; 2.50nM≤B<5.00nM; 5.00nM≤C<10.00nM; 10.00

[0726] nM <D

[0727] Example 73: Cancer Cell Viability Assay

[0728] The human hepatoma cell lines HepG2, Hep3B, Huh7, and SK-HEP-1 cells were rinsed and trypsinized with 0.25% trypsin (Corning #25-053-CI) in an incubator at 37 °C until detached. The cells were resuspended and seeded in a 96-well plate at a density of 5000 cells per well. After cell adhesion, the compound was added to a final concentration of 2-fold dilution. After 72 hours of compound treatment, 2.0 was added to the wells at a ratio of 2:1 of medium / 2.0. The plate was covered, shaken for 2 minutes, and incubated for 10 minutes. Luminescence was measured on a microplate reader, and the EC 50 value was calculated using Prism 9 software.

[0729] The EC 50 values obtained according to the above procedure are summarized in Table 2:

[0730]

[0731] A: <100 nM; 100 nM ≤ B < 500 nM; 500 nM ≤ C < 1,000 nM; 1,000 nM < D; ND = no data

[0732] Example 74: Pharmacokinetics and Tissue Distribution Studies.

[0733] For mouse experiments, after a single oral administration of a compound suspension at a dose of 5 mg / kg, the hepatic and blood pharmacokinetics of the compound in CD-1 mice were analyzed. Liver and blood samples for determination of compound concentration were obtained 2 hours after compound administration (n = 3 for each compound). Figure 1 The ratio of compound concentration in the liver to that in the blood of CD-1 mice after a single oral administration of a compound suspension at a dose of 5 mg / kg was summarized. Figure 1 The data in

[0734] For rat studies, the pharmacokinetics of the compound in blood samples collected from the jugular and portal veins of Sprague-Dawley (SD) rats were analyzed after a single oral administration of a 5 mg / kg suspension of the compound. Blood samples (n = 3 per compound) were obtained from the jugular and portal veins at 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h after compound administration for determining compound concentration. The collected liver and blood samples were analyzed using LC-MS / MS to quantify the compound concentration. Figure 2 The ratio of compound concentrations in blood samples collected from the jugular vein and portal vein of Sprague-Dawley (SD) rats after a single oral administration of a 5 mg / kg suspension of the compound was summarized. Figure 2 Data from the study showed that compounds 6 and 18 had lower liver efflux / efflux ratios than NVP-2, demonstrating that these compounds have improved liver selectivity.

[0735] Example 75: Tolerance and Safety Assessment Study

[0736] Based on body weight, BALB / c nude mice were randomly assigned to groups using a computer-generated randomization procedure. The body weight of all animals was measured daily to record changes in body weight (relative to day 1) in BALB / c nude mice treated with the medium and compound. For routine monitoring, the behavior of all study animals was monitored, including activity, food and water consumption, body weight, eye / hair adhesion, and any other abnormalities. Any mortality and / or abnormal clinical signs were recorded. Animals were euthanized when they experienced significant weight loss (emaciation, a significant weight loss >20%). Figure 3 The mean weight changes (relative to day 1) of BALB / c nude mice treated with the medium and compounds were summarized. Figure 3 Data from the study showed that compound 18 exhibited lower toxicity than NVP-2.

[0737] While preferred embodiments of the invention have been shown and described herein, it will be apparent to those skilled in the art that these embodiments are provided by way of example only. Many variations, modifications, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be used in carrying out the invention. The appended claims are intended to define the scope of the invention and thereby cover the methods and structures within the scope of these claims and their equivalents.

Claims

1. A compound of formula (IB): Or its pharmaceutically acceptable salt, wherein: Ring A is C 3-6 cycloalkyl; R 1 It is H; R 2 It is C 1-6 alkyl; R 3 It is H; R 4 It is -O(C 0-4 Alkyl)C(O)OR 18 ; R 4’ and R 4” Each is H; R 5 It is H; R 8 and R 9 Each is H; R 11 R 12 R 13 and R 14 Each is independently selected from H and halogens; Each R 18 Independently selected from H and C 1-4 alkyl; and n is 0.

2. The compound according to claim 1, wherein ring A is selected from:

3. The compound according to claim 1, wherein R 11 It is chlorine, and R 12 R 13 and R 14 Each is H.

4. The compound according to claim 1, wherein R 2 It's me.

5. The compound according to claim 1, wherein the compound of formula (IB) is represented by formula (IC), formula (ID), formula (IE) or formula (IF):

6. Selected from the following compounds: Or its pharmaceutically acceptable salt.

7. The compound according to claim 6 or a pharmaceutically acceptable salt thereof, wherein the compound is: Or its pharmaceutically acceptable salt.

8. The compound according to claim 6 or a pharmaceutically acceptable salt thereof, wherein the compound is: Or its pharmaceutically acceptable salt.

9. The compound according to claim 6, or a pharmaceutically acceptable salt thereof, is: Or its pharmaceutically acceptable salt.

10. The compound according to claim 6 or a pharmaceutically acceptable salt thereof, wherein the compound is: Or its pharmaceutically acceptable salt.

11. The compound according to claim 6 or a pharmaceutically acceptable salt thereof, wherein the compound is: Or its pharmaceutically acceptable salt.

12. The compound according to claim 6 or a pharmaceutically acceptable salt thereof, wherein the compound is: Or its pharmaceutically acceptable salt.

13. A pharmaceutical composition comprising a compound according to any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

14. Use of the compound of any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 13 in the preparation of a medicament for treating CDK-9 kinase-mediated diseases.

15. The use according to claim 14, wherein the CDK-9 kinase-mediated disease is a proliferative disorder.

16. The use according to claim 15, wherein the proliferative condition is cancer.

17. The use according to claim 16, wherein the cancer is selected from leukemia, breast cancer, prostate cancer, ovarian cancer, colon cancer, cervical cancer, lung cancer, lymphoma, and liver cancer.

18. The use according to claim 17, wherein the cancer is liver cancer.

Images