Nitrogen-containing heterocyclic cell cycle inhibitor compounds, methods of making and uses

Abstract

The application discloses a kind of nitrogen-containing heterocyclic compounds, preparation method and purposes, specifically, a kind of nitrogen-containing heterocyclic compounds as shown in general formula I, or its pharmaceutically acceptable salt, or its enantiomer, diastereoisomer, tautomer, torsion isomer, solvate, polymorph or prodrug, its preparation method and pharmaceutical application, wherein the definition of each group is described in the specification.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of pharmaceutical chemistry, and discloses a kind of nitrogen-containing heterocyclic cell cycle inhibitor compound, pharmaceutical composition and purposes thereof. BACKGROUND

[0002] Cell cycle abnormality is a hallmark feature of cancer, cyclin-dependent kinase (CDK) is a class of serine / threonine kinases, plays a central role in the cell cycle, dominates the start, progress and end of cell cycle. CDK family is an important signal transduction molecule in cells, and the CDK-cyclin complex formed by CDK and cyclin is involved in cell growth, proliferation, dormancy and apoptosis.

[0003] In the past 20 years, drug development targeting CDK kinases for tumor treatment has received extensive attention, such as Flavopiridol (Alvocidib), Seliciclib (CYC202), Dinaciclib (SCH727965) and Milciclib (PHA-848125) have entered different stages of clinical research. However, due to the low inhibitory activity of early discovered CDK inhibitors on each CDK family subtype, or lack of certain selectivity, or poor in vivo absorption, etc., the clinical application is limited. In recent years, due to the improved selectivity of CDK inhibitors for each CDK family subtype or the improved inhibitory activity of CDK kinases, especially the discovery of selective inhibitors targeting CDK4 / 6, drug research in this field has become a hot spot again.

[0004] Recent studies have found that CDK7 kinase of CDK family has dual functions of regulating kinase and transcription: 1) in cytoplasm, CDK7 exists in the form of heterotrimeric complex and acts as the activating kinase (CAK) of CDK1 / 2, whereby the phosphorylation of conserved residues in CDK1 / 2 by CDK7 is essential for complete catalytic CDK activity and cell cycle progression; 2) in nucleus, CDK7 forms the kinase core of RNA polymerase (RNAP) II general transcription factor complex and is responsible for phosphorylating the C-terminal domain (CTD) of RNAP II, which is an essential step in gene transcription initiation. The two functions of CDK7, CAK and CTD phosphorylation, support critical aspects of cell proliferation, cell cycle, and transcription. Studies have shown that CDK7 kinase plays a very important role in the regulation of triple-negative breast cancer, and inhibition of CDK7 kinase has a significant killing effect on the growth of triple-negative breast cancer cells.

[0005] Although the research and development of CDK inhibitors has made very significant progress, there are still some unsolved problems, such as drug resistance of existing CDK inhibitors and subtype selectivity of CDK kinase family targets, and therefore, there is an urgent need in the art to research and develop new CDK inhibitors with high efficiency, low toxicity, anti-drug resistance and clinical application value, such as specific CDK7 kinase inhibitors. SUMMARY

[0006] One of the technical problems to be solved by the present application is to provide a new CDK7 inhibitor for preparing a tumor treatment drug.

[0007] The present application solves the above technical problems by the following technical solutions:

[0008] The present application provides a nitrogen-containing heterocyclic compound as shown in general formula I, or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereoisomer, tautomer, rotamer, solvate, polymorph or prodrug thereof,

[0009]

[0010] In the formula:

[0011] W, X and M are each independently selected from CR w or N; R w is independently selected from H, deuterium, halogen, cyano, C1-C3 alkyl or C1-C3 haloalkyl;

[0012] Y and Z are independently selected from C or N;

[0013] R 1 is independently selected from C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkyl-(C=O)-, C1-C3 alkyl, C1-C3 haloalkyl, 3-6 membered cycloalkyl, 3-6 membered halocycloalkyl, 3-6 membered heterocycloalkyl, C1-C 10 alkoxy, C1-C 10 haloalkoxy, 3-6 membered cycloalkyl-O-, 3-6 membered heterocycloalkyl-O-, C1-C 10 alkyl-NH-, C1-C 10 haloalkyl-NH-, 3-6 membered cycloalkyl-NH- or 3-6 membered heterocycloalkyl-NH-; the above-mentioned alkenyl, alkynyl, alkyl, cycloalkyl, heterocycloalkyl can be substituted by one or several R 1-1 , R 1-1independently selected from: halogen, deuterium, hydroxyl, amino, substituted amino, C1-C6alkyl, hydroxyl substituted C1-C6alkyl, amino substituted C1-C6alkyl, C1-C6alkoxy, 3-10 membered cycloalkyl, or 3-10 membered heterocycloalkyl substituted C1-C6alkyl; the substituents in the substituted amino are 1-3 independently selected from: C1-C6alkyl, C1-C6alkoxy, 3-6 membered cycloalkyl, or 3-6 membered heterocycloalkyl;

[0014] R 2 independently selected from C1-C3alkyl, C1-C3haloalkyl, 3-6 membered cycloalkyl, 3-6 membered halocycloalkyl, or 3-6 membered heterocycloalkyl; R 2 may be further substituted by one or more R 2-1 independently selected from: deuterium, halogen, hydroxyl, C1-C3alkyl, C1-C3haloalkyl; 2-1 independently selected from: deuterium, halogen, hydroxyl, C1-C3alkyl, C1-C3haloalkyl;

[0015] R 3 independently selected from acryloyl, substituted acryloyl, propynoyl, substituted propynoyl, ethenesulfonyl, substituted ethenesulfonyl, or cyano; R 3 independently selected from: deuterium, halogen, hydroxyl, C1-C3alkyl, C1-C3haloalkyl; 3-1 independently selected from: deuterium, halogen, hydroxyl, C1-C3alkyl, C1-C3haloalkyl; 3-1 independently selected from: halogen, deuterium, hydroxyl, cyano, amino, C1-C6alkyl, C1-C6alkoxy, 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, 3-6 membered heterocycloalkyl substituted C1-C3alkyl, amino substituted C1-C3alkyl, mono C1-C3alkyl substituted amino-C1-C3alkyl, or di C1-C3alkyl substituted amino-C1-C3alkyl; or, two R 3-1 together with the carbon atom to which they are attached form a 3-8 membered carbocyclic ring or a 3-8 membered heterocyclic ring, preferably

[0016] R4, R5are independently selected from hydrogen, deuterium, halogen, C1-C3alkyl, C1-C3haloalkyl, hydroxyl, amino, substituted amino, or C1-C3alkoxy; the substituents in the substituted amino are 1-3 independently selected from: C1-C6alkyl, C1-C6alkoxy, 3-6 membered cycloalkyl, or 3-6 membered heterocycloalkyl;

[0017] R a , R b are independently selected from hydrogen, deuterium, halogen, C1-C3alkyl; or R a , R b form a 3-6 membered saturated carbocyclic ring, preferably cyclopropane or cyclobutane, by carbon chain;

[0018] Ring A and Ring B are each independently selected from a 4-10 membered nitrogen-containing heterocycle, preferably a 5-6 membered nitrogen-containing heterocycle;

[0019] n is independently selected from an integer from 0-3;

[0020] The above alkyl, substituted alkyl or alkenyl can be substituted with substituents independently selected from the group consisting of, but not limited to, deuterium, halogen, hydroxyl, monoalkylamino, dialkylamino, C1-C6 alkyl or haloalkyl, 3-10 membered cycloalkyl or heterocycloalkyl, cyano, C1-C6 alkoxy or haloalkoxy;

[0021] wherein the heterocycle contains 1-3 heteroatoms selected from the group consisting of N, O, P, S or Se, the heterocycloalkyl contains 1-3 heteroatoms selected from the group consisting of N, O, P or S, and the ring system contains saturated or partially unsaturated ring system such as spiro, bridged, fused, annelated, etc.

[0022] In certain preferred embodiments of the present application, the nitrogen-containing heterocyclic compound as shown in Formula I, or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, rotamer, solvate, polymorph or prodrug thereof,

[0023]

[0024] wherein:

[0025] W, X and M are each independently selected from CR w or N; R w is independently selected from H, halogen, cyano, C1-C3 alkyl or C1-C3 haloalkyl;

[0026] Y, Z are each independently selected from C or N;

[0027] R 1 is independently selected from C1-C3 alkyl, C1-C3 haloalkyl, 3-6 membered cycloalkyl, 3-6 membered halocycloalkyl, 3-6 membered heterocycloalkyl, C1-C 10 alkoxy, C1-C 10 haloalkoxy, 3-6 membered cycloalkyl-O-, 3-6 membered heterocycloalkyl-O-, C1-C 10 alkyl-NH-, C1-C 10 haloalkyl-NH-, 3-6 membered cycloalkyl-NH- or 3-6 membered heterocycloalkyl-NH-; the above alkyl, cycloalkyl, heterocycloalkyl can be substituted with one or several R 1-1 substituents, R 1-1The substituents are independently selected from: halogens, deuterium, hydroxyl groups, amino groups, substituted amino groups, C1-C6 alkyl groups, hydroxyl-substituted C1-C6 alkyl groups, amino-substituted C1-C6 alkyl groups, C1-C6 alkoxy groups, 3-10 membered cycloalkyl groups, or 3-10 membered heterocycloalkyl groups; the substituted amino group has 1-3 substituents, independently selected from: C1-C6 alkyl groups, C1-C6 alkoxy groups, 3-6 membered cycloalkyl groups, or 3-6 membered heterocycloalkyl groups;

[0028] R 2 It is independently selected from C1-C3 alkyl, C1-C3 haloalkyl, 3-6 membered cycloalkyl or 3-6 membered heterocycloalkyl;

[0029] R 3 Independently selected from acryloyl or substituted acryloyl, propynyl, vinylsulfonyl or substituted vinylsulfonyl, cyano, etc.; R 3 It can be further divided into one or more R 3-1 The R that was replaced 3-1 Independently selected from: halogen, deuterium, hydroxyl, cyano, amino, C1-C6 alkyl, C1-C6 alkoxy, 3-6 membered cycloalkyl, 3-6 membered heterocyclic alkyl-substituted C1-C3 alkyl, amino-substituted C1-C3 alkyl, mono-C1-C3 alkyl-substituted amino-C1-C3 alkyl, or bis-C1-C3 alkyl-substituted amino-C1-C3 alkyl; or, two R 3-1 Together with the attached carbon atom, it forms a 3-8 membered carbon ring or a 3-8 membered heterocycle, preferably.

[0030] R4 and R5 are independently selected from hydrogen, halogen, C1-C3 alkyl, C1-C3 haloalkyl, hydroxyl, amino, substituted amino, and C1-C3 alkoxy; the substituted amino group has 1 to 3 substituents, independently selected from: C1-C6 alkyl, C1-C6 alkoxy, 3-6 membered cycloalkyl or 3-6 membered heterocycloalkyl.

[0031] R a R b Independently selected from hydrogen, halogen, C1-C3 alkyl; or R a R b The carbon chain forms a 3-6 member saturated carbon ring, preferably cyclopropane or cyclobutane;

[0032] Ring A and ring B are selected from 4-10 member nitrogen-containing heterocycles, preferably 5-6 member nitrogen-containing heterocycles;

[0033] n is an integer independently selected from 0 to 3;

[0034] The alkyl or substituted alkyl groups mentioned above may be substituted with substituents, wherein each substituent is independently selected from the group consisting of, but not limited to, deuterium, halogen, hydroxyl, monoalkylamino, dialkylamino, C1-C6 alkyl or haloalkyl, 3-10 membered cycloalkyl or heterocycloalkyl, cyano, C1-C6 alkoxy or haloalkoxy; wherein the heteroaryl group comprises 1-3 heteroatoms selected from the group consisting of N, O, P or S, the heterocycloalkyl group comprises 1-3 heteroatoms selected from the group consisting of N, O, P or S, and the ring system comprises saturated or partially unsaturated ring systems such as spirocyclic, bridged ring, fused ring, and fused ring.

[0035] In certain preferred embodiments of the present invention, certain groups in the nitrogen-containing heterocyclic compound represented by general formula I, or its pharmaceutically acceptable salt, or its enantiomers, diastereomers, tautomers, torsional isomers, solvates, polymorphs, or prodrugs, are defined as follows, and groups not mentioned are as described in any embodiment of the present invention (hereinafter referred to as "in some preferred embodiments").

[0036]

[0037] In the formula:

[0038] W, X, and M are each independently selected from CR w Or N; R w Independently selected from H, halogen, cyano, C1-C3 alkyl or C1-C3 alkyl haloalkyl;

[0039] Y and Z are selected from C or N respectively;

[0040] R 1 Independently selected from C1-C3 alkyl, C1-C3 haloalkyl, 3-6 membered cycloalkyl, 3-6 membered halocycloalkyl, 3-6 membered heterocycloalkyl, C1-C 10 Alkoxy, C1-C 10 Haloalkoxy, 3-6 membered cycloalkyl-O-, 3-6 membered heterocycloalkyl-O-, C1-C 10 Alkyl-NH-, C1-C 10 Halogenated alkyl group -NH-, 3-6 membered cycloalkyl group -NH-, or 3-6 membered heterocycloalkyl group -NH-; the above alkyl, cycloalkyl, and heterocycloalkyl groups can be represented by one or more R groups. 1-1 Replaced by, R 1-1 The substituents are independently selected from: halogens, deuterium, hydroxyl groups, amino groups, substituted amino groups, C1-C6 alkyl groups, hydroxyl-substituted C1-C6 alkyl groups, amino-substituted C1-C6 alkyl groups, C1-C6 alkoxy groups, 3-10 membered cycloalkyl groups, or 3-10 membered heterocycloalkyl groups; the substituted amino group has 1-3 substituents, independently selected from: C1-C6 alkyl groups, C1-C6 alkoxy groups, 3-6 membered cycloalkyl groups, or 3-6 membered heterocycloalkyl groups;

[0041] R 2 independently selected from C1-C3 alkyl, C1-C3 haloalkyl, 3-6 membered cycloalkyl, 3-6 membered halocycloalkyl, or 3-6 membered heterocycloalkyl; R 2 may be further substituted by one or more R 2-1 substituents, said R 2-1 are independently selected from halogen, hydroxyl, C1-C3 alkyl, C1-C3 haloalkyl;

[0042] R 3 are independently selected from acryloyl or substituted acryloyl, propioloyl, ethenesulfonyl or substituted ethenesulfonyl, cyano, and the like; R 3 may be further substituted by one or more R 3-1 substituents, said R 3-1 are independently selected from halogen, deuterium, hydroxyl, cyano, amino, C1-C6 alkyl, C1-C6 alkoxy, 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl substituted C1-C3 alkyl, amino substituted C1-C3 alkyl, mono C1-C3 alkyl substituted amino-C1-C3 alkyl, or di C1-C3 alkyl substituted amino-C1-C3 alkyl; or, two R 3-1 together with the carbon atom to which they are attached form a 3-8 membered carbocyclic ring or a 3-8 membered heterocyclic ring, preferably

[0043] R4, R5are independently selected from hydrogen, halogen, C1-C3 alkyl, C1-C3 haloalkyl, hydroxyl, amino, substituted amino, C1-C3 alkoxy; the substituents in the substituted amino are 1-3, independently selected from C1-C6 alkyl, C1-C6 alkoxy, 3-6 membered cycloalkyl, or 3-6 membered heterocycloalkyl;

[0044] R a , R b are independently selected from hydrogen, halogen, C1-C3 alkyl; or R a , R b form a 3-6 membered saturated carbocyclic ring, preferably cyclopropane, cyclobutane, by a carbon chain;

[0045] Ring A and Ring B are each selected from 4-10 membered nitrogen-containing heterocyclic ring, preferably 5-6 membered nitrogen-containing heterocyclic ring;

[0046] n is independently selected from an integer from 0-3;

[0047] The alkyl or substituted alkyl, alkenyl groups described above can be substituted with substituents independently selected from the group consisting of, but not limited to, deuterium, halogen, hydroxyl, monoalkylamino, dialkylamino, C1-C6 alkyl or haloalkyl, 3-10 membered cycloalkyl or heterocycloalkyl, cyano, C1-C6 alkoxy or haloalkoxy; wherein the heteroaryl comprises 1-3 heteroatoms selected from the group consisting of N, O, P or S, the heterocycloalkyl comprises 1-3 heteroatoms selected from the group consisting of N, O, P or S, and the ring system comprises a saturated or partially unsaturated ring system of spiro, bridged, fused, annelated, etc.

[0048] In some preferred embodiments, is not

[0049] In some preferred embodiments, when Y is N, at least one of W, X and M is N.

[0050] In some preferred embodiments, the compound having the general formula (I), or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, rotamer, solvate, polymorph or prodrug thereof, wherein R 1 is CH3, CF3, CH3O-, NH2or CH3NH-, CH3CH2,

[0051] or R 2 is methyl, ethyl, isopropyl, cyclopropyl;

[0052] or R 3 is

[0053] or R a , R b is preferably hydrogen, methyl;

[0054] or R4is hydrogen, halogen, hydroxyl, methyl;

[0055] or R5is hydrogen, hydroxyl, halogen, methyl;

[0056] or ring A is preferably wherein the * end is attached to -NH-; and the N end is attached to -(CH2) n- end;

[0057] or ring B is preferably wherein the N end is attached to R 3 ; and the * end is attached to -(CH2)n- end;

[0058] or the configuration of the carbon atom attached to the carbon chain in ring B is preferably in the R configuration.

[0059] In some preferred embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, rotamer, solvate, polymorph, or prodrug thereof, is preferably a compound of general formula (II), or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, rotamer, solvate, polymorph, or prodrug thereof:

[0060]

[0061] wherein W, X, Y, Z, M, R 1 , R 2 , R 3 , R4, R a , R b are in the range as defined above.

[0062] In some preferred embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, rotamer, solvate, polymorph, or prodrug thereof, is preferably a compound of general formula (III-1)-(III-9),

[0063]

[0064] wherein the other groups are in the range as defined above.

[0065] In some preferred embodiments, the nitrogen-containing heterocyclic compound of general formula I is a compound of general formula (VI-1)-(VI-9),

[0066]

[0067]

[0068] wherein the other groups (R 1 , R 2 , R 3 , R a , R b ) are in the range as defined above.

[0069] In some preferred embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, rotamer, solvate, polymorph, or prodrug thereof, is preferably a compound of general formula (IV-1)-(IV-2),

[0070] wherein R 1preferably selected from halogen, C1-C3 alkyl or haloalkyl, C3-C6 cycloalkyl or halocycloalkyl; R 3 is preferably selected from halogen, C1-C3 alkyl or haloalkyl, C3-C6 cycloalkyl or halocycloalkyl; R is preferably selected from halogen, C1-C3 alkyl or haloalkyl, C3-C6 cycloalkyl or halocycloalkyl; R c is preferably selected from halogen, C1-C3 alkyl or haloalkyl, C3-C6 cycloalkyl or halocycloalkyl; R d is preferably selected from halogen, C1-C3 alkyl or haloalkyl, C3-C6 cycloalkyl or halocycloalkyl; R e is preferably selected from halogen, C1-C3 alkyl or haloalkyl, C3-C6 cycloalkyl or halocycloalkyl; R f is preferably selected from halogen, C1-C3 alkyl or haloalkyl, C3-C6 cycloalkyl or halocycloalkyl; R g is preferably selected from halogen, C1-C3 alkyl or haloalkyl, C3-C6 cycloalkyl or halocycloalkyl; R h is preferably selected from halogen, C1-C3 alkyl or haloalkyl, C3-C6 cycloalkyl or halocycloalkyl; R k is preferably selected from halogen, C1-C3 alkyl or haloalkyl, C3-C6 cycloalkyl or halocycloalkyl; R m is independently selected from hydrogen, deuterium, halogen, C1-C6 alkyl, or cyano; or any two adjacent groups of the above can form a 3-8 membered carbocyclic ring or a 3-8 membered heterocyclic ring through a carbon chain or a heteroatom.

[0071] In some preferred embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, rotamer, solvate, polymorph or prodrug thereof, is preferably a compound of general formula (IV-1)-(IV-2):

[0072] wherein R 1 is preferably selected from halogen, C1-C3 alkyl or haloalkyl, C3-C6 cycloalkyl or halocycloalkyl; R

[0073] R 3 is preferably selected from halogen, C1-C3 alkyl or haloalkyl, C3-C6 cycloalkyl or halocycloalkyl; R Rc, R d is preferably selected from halogen, C1-C3 alkyl or haloalkyl, C3-C6 cycloalkyl or halocycloalkyl; R e is preferably selected from halogen, C1-C3 alkyl or haloalkyl, C3-C6 cycloalkyl or halocycloalkyl; R f is preferably selected from halogen, C1-C3 alkyl or haloalkyl, C3-C6 cycloalkyl or halocycloalkyl; R g is preferably selected from halogen, C1-C3 alkyl or haloalkyl, C3-C6 cycloalkyl or halocycloalkyl; R h is preferably selected from halogen, C1-C3 alkyl or haloalkyl, C3-C6 cycloalkyl or halocycloalkyl; R k is preferably selected from halogen, C1-C3 alkyl or haloalkyl, C3-C6 cycloalkyl or halocycloalkyl; R m is independently selected from hydrogen, deuterium, halogen, C1-C6 alkyl, or cyano; or any two adjacent groups of the above can form a 3-8 membered carbocyclic ring or a 3-8 membered heterocyclic ring through a carbon chain or a heteroatom.

[0074] In some preferred embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, rotamer, solvate, polymorph or prodrug thereof, is a compound of general formula (V-1)-(V-2):

[0075] wherein R 1 is preferably selected from halogen, C1-C3 alkyl or haloalkyl, C3-C6 cycloalkyl or halocycloalkyl; R3 is selected from the group consisting of: R c , R d , R e , R f , R g , R h , R k , R m independently selected from hydrogen, deuterium, halogen, C1-C6alkyl, cyano; or any two adjacent groups of the above can form a 3-8 membered carbocyclic ring or a 3-8 membered heterocyclic ring through a carbon chain or a heteroatom; the range of other groups is defined as above.

[0076] In some preferred embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, rotamer, solvate, polymorph or prodrug thereof, is a compound represented by general formula (V-1)-(V-2):

[0077] wherein R 1 is preferably selected from C2-C6alkenyl, C2-C6alkynyl, C1-C6alkyl-(C=O)-, halogen, C1-C3alkyl, haloalkyl, C3-C6cycloalkyl or halocycloalkyl; more preferably from ethenyl, ethynyl, C1-C3alkyl, haloalkyl, C3-C6cycloalkyl or halocycloalkyl;

[0078] R 3 is selected from the group consisting of: R c , R d , R e , R f , R g , R h , R k , R m independently selected from hydrogen, deuterium, halogen, C1-C6alkyl, cyano; or any two adjacent groups of the above can form a 3-8 membered carbocyclic ring or a 3-8 membered heterocyclic ring through a carbon chain or a heteroatom.

[0079] In some preferred embodiments, C2-C6alkenyl can be C2-C4alkenyl; preferably ethenyl or propenyl.

[0080] In some preferred embodiments, C2-C6alkynyl can be C2-C4alkynyl, preferably

[0081] In some preferred embodiments, C1-C3alkyl can be methyl, ethyl, n-propyl or isopropyl.

[0082] In some preferred embodiments, the C1-C6 alkyl group may be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, primary butyl, secondary butyl, or tert-butyl; preferably isopropyl or tert-butyl.

[0083] In some preferred embodiments, the halogen in the C1-C3 haloalkyl group is preferably fluorine, chlorine, bromine or iodine; the C1-C3 haloalkyl group is preferably a C1-C3 fluoroalkyl group.

[0084] In some preferred embodiments, the 3-6 membered cycloalkyl group may be cyclopropyl, cyclobutyl, cyclopentyl, cyclohexenyl, or cyclohexyl; preferably cyclopropyl.

[0085] In some preferred embodiments, the halogen may be fluorine, chlorine, bromine or iodine; preferably fluorine.

[0086] In some preferred embodiments, the 3-6 membered heterocyclic alkyl group may be

[0087] In some preferred embodiments, the 4-10 member nitrogen-containing heterocycle can be

[0088] In some preferred embodiments, R w Selected independently from H.

[0089] In some preferred embodiments, R 1 It can be C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkyl-(C=O)-, C1-C3 alkyl, C1-C3 haloalkyl or 3-6 membered cycloalkyl.

[0090] In some preferred embodiments, R 1-1 The number of R is 1, 2, or 3; 1-1 It is independently selected from halogens or hydroxyl groups; preferably hydroxyl groups.

[0091] In some preferred embodiments, R 1 It can be C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkyl-(C=O)-, C1-C3 alkyl, C1-C3 haloalkyl or 3-6 membered cycloalkyl; the above alkyl groups can be one or more R groups. 1-1 Replaced by, R 1-1 It is independently selected from hydroxyl groups.

[0092] In some preferred embodiments, R 2 It can be a C1-C3 alkyl group; preferably isopropyl.

[0093] In some preferred embodiments, R 3 It can be acryloyl, substituted acryloyl, propynyl, substituted propynyl or ethylene sulfonyl.

[0094] In some preferred embodiments, R 3-1 It is independently selected from: halogen, cyano, C1-C6 alkyl, 3-6 membered heterocyclic alkyl-substituted C1-C3 alkyl, mono-C1-C3 alkyl-substituted amino-C1-C3 alkyl or bis-C1-C3 alkyl-substituted amino-C1-C3 alkyl.

[0095] In some preferred embodiments, R 3 It can be acryloyl, substituted acryloyl, propynyl, substituted propynyl, or vinylsulfonyl; the substituted acryloyl group is formed by one or more R groups. 3-1 The R that was replaced 3-1 It is independently selected from: halogen, cyano, C1-C6 alkyl, 3-6 membered heterocyclic alkyl-substituted C1-C3 alkyl, mono-C1-C3 alkyl-substituted amino-C1-C3 alkyl or bis-C1-C3 alkyl-substituted amino-C1-C3 alkyl.

[0096] In some preferred embodiments, R4 may be a halogen, a hydroxyl group, or a C1-C3 alkyl group.

[0097] In some preferred embodiments, R5 may be a halogen, a hydroxyl group, or a C1-C3 alkyl group.

[0098] In some preferred embodiments, R a R b It is independently selected from hydrogen or C1-C3 alkyl.

[0099] In some preferred embodiments, ring A may be The * terminal is connected to -NH-; the N terminal is connected to... The ends are connected.

[0100] In some preferred embodiments, ring B may be The N-terminus and R 3 Connected, *end and The ends are connected.

[0101] In some preferred embodiments, the carbon atoms in ring B that are connected to the carbon chain are preferably in the R configuration.

[0102] In some preferred embodiments, R 1 It can be vinyl, ethynyl, CH3(C=O)-, -CH3, -CHF2, CF3, CH3O-, -NH2, CH3NH-, CH3CH2-,

[0103] In some implementations, R 2 It can be methyl, ethyl, isopropyl or cyclopropyl; preferably isopropyl.

[0104] In some preferred embodiments, R 3 may be

[0105] In some preferred embodiments, R may be

[0106] In some preferred embodiments, R may be

[0107] In some preferred embodiments, R may be

[0108] In some preferred embodiments, the nitrogen-containing heterocyclic compound of formula I is any one of the following structures:

[0109]

[0110]

[0111]

[0112] A method for preparing a compound of formula I, characterized in that a compound of formula (A) is subjected to steps a-c to form a compound of formula (I):

[0113] a) substituting or reductive amination of a compound of formula (A) with a suitable alkylating agent to form an intermediate compound (B);

[0114] b) removing the protecting group on the amino group of a compound of formula (B) under suitable reaction conditions to form an intermediate compound of formula (C);

[0115] c) reacting a compound of formula (C) with a suitable reagent under catalysis of a base or a suitable reagent to form a compound of formula (I).

[0116]

[0117] Pg is a protecting group for the amino group, such as t-butyl carbonate, benzyl carbonate, benzyl, etc., and the definitions of the various groups are as described above;

[0118] Preferably, the reaction is carried out in a solvent, and the solvent is selected from the group consisting of water, methanol, ethanol, isopropanol, butanol, ethylene glycol, ethylene glycol methyl ether, N-methylpyrrolidone, dimethyl sulfoxide, tetrahydrofuran, toluene, dichloromethane, 1,2-dichloroethane, acetonitrile, N,N-dimethylformamide, N,N-dimethylacetamide, dioxane, or a combination thereof.

[0119] Preferably, the inorganic base is selected from the group consisting of sodium hydride, potassium hydroxide, sodium acetate, potassium acetate, potassium tert-butoxide, sodium tert-butoxide, potassium fluoride, cesium fluoride, potassium phosphate, potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, or a combination thereof; and the organic base is selected from the group consisting of pyridine, triethylamine, N,N-diisopropylethylamine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), lithium hexamethyldisilazide, sodium hexamethyldisilazide, dimethylpyridine, or a combination thereof.

[0120] Preferably, the acid is selected from the group consisting of hydrochloric acid, sulfuric acid, phosphoric acid, methanesulfonic acid, toluenesulfonic acid, trifluoroacetic acid, formic acid, acetic acid, trifluoromethanesulfonic acid, or a combination thereof.

[0121] Another object of the present application is to provide a medicament for treating or preventing a tumor or an autoimmune disease and a composition thereof. The technical solutions of achieving the above object are as follows:

[0122] In one aspect, the present application provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula I, a stereoisomer, a geometric isomer, a tautomer, a pharmaceutically acceptable salt, or a prodrug thereof, and a pharmaceutically acceptable carrier. In certain embodiments of the pharmaceutical composition, the pharmaceutical composition is formulated for intravenous administration, intramuscular administration, oral administration, rectal administration, inhalation administration, nasal administration, topical administration, ocular administration, or aural administration. In other embodiments of the pharmaceutical composition, the pharmaceutical composition is a tablet, a pill, a capsule, a liquid, an inhaler, a nasal spray solution, a suppository, a solution, an emulsion, an ointment, an eye drop, or an ear drop. In other embodiments of the pharmaceutical composition, the pharmaceutical composition further comprises one or more additional therapeutic agents.

[0123] In another aspect, the present application provides use of a compound of Formula I, a stereoisomer, a geometric isomer, a tautomer, a pharmaceutically acceptable salt, or a prodrug thereof, in the manufacture of a medicament for preventing, treating, or alleviating a disorder or a disease mediated by abnormal activity of a CDK kinase, particularly a CDK7 kinase.

[0124] In another aspect, the present application provides use of a substance Z in the manufacture of a medicament for preventing, treating, or alleviating a disorder or a disease mediated by abnormal activity of a CDK kinase;

[0125] The CDK kinase is preferably a CDK7 kinase.

[0126] said substance Z is a compound of Formula I, a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, or prodrug thereof, or a pharmaceutical composition as described above.

[0127] In another aspect, the present application provides a use of a substance Z in the manufacture of a medicament for treating or preventing a proliferative disease (e.g., a cancer (e.g., leukemia, melanoma, multiple myeloma), a benign neoplasm, angiogenesis, an inflammatory disease, an autoinflammatory disease, and an autoimmune disease) in a subject, said tumor is independently selected from the group consisting of non-small cell lung cancer, small cell lung cancer, lung adenocarcinoma, lung squamous carcinoma, breast cancer, prostate cancer, liver cancer, skin cancer, stomach cancer, intestinal cancer, cholangiocarcinoma, brain cancer, leukemia, lymphoma, fibroma, sarcoma, basal cell carcinoma, glioma, kidney cancer, melanoma, bone cancer, thyroid cancer, nasopharyngeal cancer, pancreatic cancer, etc.; said autoimmune disease is independently selected from the group consisting of rheumatoid arthritis, systemic lupus erythematosus, idiopathic thrombocytopenic purpura, hemolytic anemia, or psoriasis; said inflammatory disease is independently selected from the group consisting of osteoarthritis, gouty arthritis, ulcerative colitis, and / or inflammatory bowel disease, etc.; said infectious disease is independently selected from the group consisting of sepsis, septic shock, endotoxic shock, gram-negative sepsis, and / or toxic shock syndrome.

[0128] In another aspect, the present application provides a use of a substance Z in the manufacture of a medicament for treating or preventing a proliferative disease (e.g., a cancer (e.g., leukemia, melanoma, multiple myeloma), a benign neoplasm, angiogenesis, an inflammatory disease, an autoinflammatory disease, and an autoimmune disease) in a subject, said tumor is independently selected from the group consisting of non-small cell lung cancer, small cell lung cancer, lung adenocarcinoma, lung squamous carcinoma, breast cancer, prostate cancer, liver cancer, skin cancer, stomach cancer, intestinal cancer, cholangiocarcinoma, brain cancer, leukemia, lymphoma, fibroma, sarcoma, basal cell carcinoma, glioma, kidney cancer, melanoma, bone cancer, thyroid cancer, nasopharyngeal cancer, pancreatic cancer, etc.; said autoimmune disease is independently selected from the group consisting of rheumatoid arthritis, systemic lupus erythematosus, idiopathic thrombocytopenic purpura, hemolytic anemia, or psoriasis; said inflammatory disease is independently selected from the group consisting of osteoarthritis, gouty arthritis, ulcerative colitis, and / or inflammatory bowel disease, etc.; said infectious disease is independently selected from the group consisting of sepsis, septic shock, endotoxic shock, gram-negative sepsis, and / or toxic shock syndrome;

[0129] Said substance Z is a nitrogen-containing heterocyclic compound of general formula (I), or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, rotamer, solvate, polymorph or prodrug thereof or a pharmaceutical composition as described above.

[0130] In another aspect, the present application provides a method of treating or preventing a disorder or disease mediated by abnormal activity of a CDK kinase, comprising administering to a patient in need thereof a therapeutically effective amount of a substance Z as described above;

[0131] Said CDK kinase is preferably a CDK7 kinase;

[0132] Said substance Z is a nitrogen-containing heterocyclic compound of general formula (I), or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, rotamer, solvate, polymorph or prodrug thereof or a pharmaceutical composition as described above.

[0133] The term

[0134] 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 the subject matter of this application belongs. All patents, patent applications, published materials referred to throughout the entire disclosure herein, unless otherwise indicated, are incorporated by reference herein in their entirety.

[0135] It should be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the subject matter claimed. In this application, the use of the singular includes the plural unless specifically stated otherwise. It must be noted that as used herein and in the appended claims, the singular forms "a", "an" and "the" include plural

[0136] Definitions of standard chemical terminology can be found in reference works, including Carey and Sundberg "ADVANCED ORGANIC CHEMISTRY 4TH ED." Vols. A (2000) and B (2001), Plenum Press, New York. Conventional methods within the skill in the art, such as mass spectrometry, NMR, IR and UV / VIS spectroscopy and pharmacological methods are employed, unless otherwise indicated. Unless specific definitions are provided, the nomenclature employed in connection with the description herein, in the analytical chemistry, synthetic organic or medicinal chemistry arts and the like, is intended to have the meanings commonly understood by those of ordinary skill in the art. Standard techniques can be used for chemical synthesis, chemical analysis, pharmaceutical formulation, and delivery, and treatment of patients. For example, reactions and purification can be performed according to manufacturers' instructions or according to published procedures in the literature. Standard techniques can be used for chemical synthesis, chemical analysis, pharmaceutical formulation, and delivery, and treatment of patients. For example, reactions and purification can be performed according to manufacturers' instructions or according to published procedures in the literature. In general, the techniques and procedures can be performed according to conventional methods well known in the art in the light of the descriptions in the numerous exemplary and more specific references cited herein. In the present description, groups and substituents thereof can be selected by one of ordinary skill in the art to provide stable moieties and compounds.

[0137] When a substituent is described by a conventional chemical formula written from left to right, the substituent is also meant to include chemically equivalent substituents resulting from writing the formula from right to left. For example, -CH2O- is equivalent to -OCH2-.

[0138] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. All documents, or portions of documents, cited in this application, including but not limited to patents, patent applications, articles, books, manuals, and treatises, are hereby expressly incorporated by reference in their entirety.

[0139] Certain chemical groups defined herein are preceded by a shorthand notation indicating the total number of carbon atoms in the group. For example, C1-6alkyl refers to an alkyl radical as defined below having from one to six carbon atoms. The total number of carbon atoms in the shorthand notation does not include carbon atoms that can be present in substituents of the group.

[0140] In addition to the foregoing, the following terms, as used herein in the specification and claims, have the meanings indicated below.

[0141] In the present application, the term "halogen" means fluorine, chlorine, bromine or iodine; "hydroxy" means the -OH group; "hydroxyalkyl" means an alkyl group as defined below, substituted with a hydroxy (-OH) group; "carbonyl" means the -C(=O)- group; "nitro" means -NO2; "cyano" means -CN; "amino" means -NH2; "substituted amino" means an amino group substituted with one or two alkyl, alkylcarbonyl, aralkyl, heteroaralkyl groups as defined below, for example, monoalkylamino, dialkylamino, alkylamido, aralkylamino, heteroaralkylamino; "carboxy" means -COOH.

[0142] In the present application, the term "alkyl" as a group or part of a group (for example in the groups halo-substituted alkyl and the like) means a straight or branched chain hydrocarbon group consisting only of carbon and hydrogen atoms, containing no unsaturation, having, for example, 1 to 12 (preferably 1 to 8, more preferably 1 to 6) carbon atoms and being attached to the rest of the molecule by a single bond. Examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, n-pentyl, 2-methylbutyl, 2,2-dimethylpropyl, n-hexyl, heptyl, 2-methylhexyl, 3-methylhexyl, octyl, nonyl, decyl and the like.

[0143] In the present application, the term "alkenyl" as a group or part of a group means a straight or branched chain hydrocarbon group consisting only of carbon and hydrogen atoms, containing at least one double bond, having, for example, 2 to 14 (preferably 2 to 10, more preferably 2 to 6) carbon atoms and being attached to the rest of the molecule by a single bond, for example, but not limited to, ethenyl, propenyl, allyl, but-1-enyl, but-2-enyl, pent-1-enyl, pent-1,4-dienyl and the like.

[0144] In the present application, the term "alkynyl" as a group or part of a group means a straight or branched chain hydrocarbon group consisting only of carbon and hydrogen atoms, containing at least one triple bond and optionally one or more double bonds, having, for example, 2 to 14 (preferably 2 to 10, more preferably 2 to 6) carbon atoms and being attached to the rest of the molecule by a single bond, for example, but not limited to, ethynyl, prop-1-ynyl, but-1-ynyl, pent-1- en-4-ynyl and the like.

[0145] In the present application, the term "cycloalkyl" as a group or part of a group refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon group consisting solely of carbon and hydrogen atoms, which can include fused ring systems, bridged ring systems or spirocyclic ring systems, having, for example, from 3 to 15 carbon atoms, preferably having from 3 to 10 carbon atoms, more preferably having from 3 to 8 carbon atoms, and which is saturated or unsaturated and can be attached to the remainder of the molecule through a single bond via any suitable carbon atom. Unless otherwise specifically noted in the specification, the carbon atoms in a cycloalkyl group can optionally be oxidized. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclooctyl, 1H-indenyl, 2,3-dihydroindenyl, 1,2,3,4-tetrahydro-naphthyl, 5,6,7,8-tetrahydro-naphthyl, 8,9-dihydro-7H-benzocyclohepten-6-yl, 6,7,8,9-tetrahydro-5H-benzocycloheptenyl, 5,6,7,8,9,10-hexahydro-benzocyclooctenyl, fluorenyl, bicyclo[2.2.1]heptyl, 7,7-dimethyl-bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, bicyclo[2.2.2]octyl, bicyclo[3.1.1]heptyl, bicyclo[3.2.1]octyl, bicyclo[2.2.2]octenyl, bicyclo[3.2.1]octenyl, adamantyl, octahydro-4,7-methano-1H-indenyl, and octahydro-2,5-methano-indenyl, and the like.

[0146] In the present application, the term "heterocyclyl" as a group or as part of a group means a stable 3- to 20-membered non-aromatic ring radical consisting of 2 to 14 carbon atoms and 1 to 6 heteroatoms selected from the group consisting of nitrogen, phosphorus, oxygen, sulfur and selenium. Unless otherwise particularly specified herein, the heterocyclyl radical can be a monocyclic, bicyclic, tricyclic or more ring ring system, which can include fused ring systems, bridged ring systems or spirocyclic ring systems; the nitrogen, carbon or sulfur atoms in the heterocyclyl radical thereof can optionally be oxidized; the nitrogen atom can optionally be quaternized; and the heterocyclyl radical can be partially or fully saturated. The heterocyclyl radical can be attached to the remainder of the molecule via a carbon atom or a heteroatom and by a single bond. In a heterocyclyl radical comprising fused rings, one or more of the rings can be an aryl or heteroaryl group as defined below, provided that the point of attachment to the remainder of the molecule is a non-aromatic ring atom. For the purposes of the present application, the heterocyclyl radical is preferably a stable 4- to 11-membered non-aromatic monocyclic, bicyclic, bridged or spirocyclic radical comprising 1 to 3 heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur, more preferably a stable 4- to 8-membered non-aromatic monocyclic, bicyclic, bridged or spirocyclic radical comprising 1 to 3 heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur. Examples of heterocyclyl radicals include, but are not limited to: pyrrolidinyl, morpholinyl, piperazinyl, homopiperazinyl, piperidinyl, thiomorpholinyl, 2,7-diaza-spiro[3.5]nonan-7-yl, 2-oxa-6-aza-spiro[3.3]heptan-6-yl, 2,5-diaza-bicyclo[2.2.1]heptan-2-yl, azetidinyl, pyranyl, tetrahydropyranyl, thiopyranyl, tetrahydrofuranyl, oxazinyl, dioxolanyl, tetrahydroisoquinolinyl, decahydroisoquinolinyl, imidazolinyl, imidazolidinyl, quinolizinyl, thiazolidinyl, isothiazolidinyl, isoxazolidinyl, indolinyl, octahydroindolinyl, octahydroisoindolinyl, pyrrolidinyl, pyrazolidinyl, phthalimido and the like.

[0147] In the present application, the term "aryl" as a group or as part of a group means a conjugated hydrocarbon ring system radical having 6 to 18 carbon atoms, preferably having 6 to 10 carbon atoms. For the purposes of the present application, the aryl radical can be a monocyclic, bicyclic, tricyclic or more ring ring system, which can also be fused with a cycloalkyl or heterocyclyl radical as defined above, provided that the aryl radical is attached to the remainder of the molecule via an atom on the aromatic ring by a single bond. Examples of aryl radicals include, but are not limited to: phenyl, naphthyl, anthryl, phenanthryl, fluorenyl, 2,3-dihydro-1 H-isoindolyl, 2-benzoxazolinonyl, 2H-1,4-benzoxazin-3(4H)-on-7-yl and the like.

[0148] In the present application, the term "arylalkyl" means an alkyl radical as defined above which is substituted by an aryl radical as defined above.

[0149] In the present application, the term "heteroaryl" as a group or part of a group means a 5- to 16-membered, conjugated ring system having 1 to 15 carbon atoms (preferably having 1 to 10 carbon atoms) and 1 to 6 heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur. Unless specifically indicated otherwise in the present specification, the heteroaryl group can be a monocyclic, bicyclic, tricyclic or more ring ring system, and can also be fused with a cycloalkyl or heterocyclyl group as defined above, provided that the heteroaryl group is attached to the rest of the molecule by a single bond via an atom of the aromatic ring. The nitrogen, carbon or sulfur atoms in the heteroaryl group can optionally be oxidized; the nitrogen atoms can optionally be quaternized. For the purposes of the present application, the heteroaryl group is preferably a stable 5- to 12-membered aromatic radical containing 1 to 5 heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur, more preferably a stable 5- to 10-membered aromatic radical containing 1 to 4 heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur or a 5- to 6-membered aromatic radical containing 1 to 3 heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur. Examples of heteroaryl groups include, but are not limited to, thienyl, imidazolyl, pyrazolyl, thiazolyl, oxazolyl, oxadiazolyl, isoxazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, benzimidazolyl, benzopyrazolyl, indolyl, furanyl, pyrrolyl, triazolyl, tetrazolyl, triazinyl, indolizinyl, isoindolyl, indazolyl, isoindazolyl, purinyl, quinolyl, isoquinolyl, naphthyridinyl, cinnolinyl, quinazolinyl, benzothiophenyl, indolizinyl, phenoxazinyl, phenothiazinyl, 4,5,6,7-tetrahydrobenzo[b]thiophenyl, naphthpyridinyl, [l,2,4]triazolo[4,3-b]pyridazine, [l,2,4]triazolo[4,3-a]pyrazine, [l,2,4]triazolo[4,3-c]pyrimidine, [l,2,4]triazolo[4,3-a]pyridine, imidazo[l,2-a]pyridine, imidazo[l,2-b]pyridazine, imidazo[l,2-a]pyrazine, and the like.

[0150] In the present application, the term "heteroarylalkyl" means an alkyl group as defined above which is substituted by a heteroaryl group as defined above.

[0151] In the present application, "optionally" or "may" means that the subsequently described event or circumstance can or can not occur, and the description includes both the occurrence and non-occurrence of the event or circumstance. For example, "optionally substituted aryl" means that the aryl group is substituted or unsubstituted, and the description includes both substituted aryl groups and unsubstituted aryl groups.

[0152] The terms "moiety," "structural moiety," "chemical moiety," "group," "chemical group" as used herein refer to a specific fragment or functional group within a molecule. Chemical moieties are generally recognized chemical entities that are embedded or appended to a molecule.

[0153] "Stereoisomers" refer to compounds which have the same atomic constituents, bonded by the same sequence of bonds, but have different three-dimensional structures. The present invention will encompass all such stereo isomers and mixtures thereof.

[0154] When the compounds of the present invention contain an olefinic double bond, the compounds of the present invention are intended to include both E- and Z- geometric isomers, unless otherwise noted.

[0155] "tautomers" refer to isomers that differ in the protonation site of a functional group. All tautomeric forms of the compounds of the present invention are also intended to be included within the scope of the present invention.

[0156] The compounds of the present invention, or pharmaceutically acceptable salts thereof, can contain one or more chiral carbon atoms and, therefore, can give rise to enantiomers, diastereomers, and other stereoisomeric forms. Each chiral carbon atom can be defined, based on its stereochemistry, as either an (R)- or (S)-. The present invention is intended to include all possible isomers, as well as their racemic and optically pure forms. The preparation of the compounds of the present invention can select a racemic, diastereomeric, or enantiomeric form as starting material or intermediate. The optically active isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, such as crystallization and chromatography on chiral supports.

[0157] Conventional techniques for preparing / isolating individual isomers include chiral synthesis from a suitable optically pure precursor, or resolution of a racemate (or racemate of a salt or derivative) using, for example, chiral high-performance liquid chromatography, see, for example, Gerald Gϋbitz and Martin G. Schmid (Eds.), Chiral Separations, Methods and Protocols, Methods in Molecular Biology, Vol. 243, 2004; A. M. Stalcup, Chiral Separations, Annu. Rev. Anal. Chem. 3:341-63, 2010; Fumiss et al. (eds.), VOGEL'S ENCYCLOPEDIA OF PRACTICAL ORGANIC CHEMISTRY 5.sup.TH ED., Longman Scientific and Technical Ltd., Essex, 1991, 809-816; Heller, Acc. Chem. Res. 1990, 23, 128.

[0158] In the present application, the term "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

[0159] A "pharmaceutically acceptable acid addition salt" means a salt with an inorganic or organic acid that retains the biological effectiveness of the free base and that does not impart undesired toxicological effects to the subject in need of treatment. Inorganic acid salts include, but are not limited to, hydrochlorides, hydrobromides, sulfates, nitrates, phosphates, and the like; organic acid salts include, but are not limited to, formates, acetates, 2,2-dichloroacetates, trifluoroacetates, propionates, hexanoates, octanoates, decanoates, undecylenates, glycolates, gluconates, lactates, sebacates, adipates, glutarates, malonates, oxalates, maleates, succinates, fumarates, tartrates, citrates, palmitates, stearates, oleates, cinnamates, laurates, malates, glutamates, pyroglutamates, aspartates, benzoates, mesylates, besylates, tosylates, alginates, ascorbates, salicylates, 4-aminosalicylates, naphthalene-2- disulfonates, and the like. These salts can be prepared by methods known in the art.

[0160] "Pharmaceutically acceptable base addition salt" refers to those salts which retain the biological effectiveness and non-toxicity of the free acids and are formed with inorganic or organic bases. Salts derived from inorganic bases include but are not limited to sodium salts, potassium salts, lithium salts, ammonium salts, calcium salts, magnesium salts, iron salts, zinc salts, copper salts, manganese salts, aluminum salts and the like. Preferred inorganic salts are ammonium, sodium, potassium, calcium and magnesium salts. Salts derived from organic bases include but are not limited to salts of primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, triethanolamine, dimethyl ethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, theobromine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins and the like. Preferred organic bases include isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline and caffeine. These salts can be prepared by methods known to those skilled in the art.

[0161] "Polymorph" refers to different solid crystalline phases of certain compounds of the present application that arise due to the presence of two or more different molecular arrangements in the solid state. Certain compounds of the present application can exist in more than one crystal form, and the present application is intended to include all such polymorphs and mixtures thereof.

[0162] In general, crystallization will produce solvates of the compounds of the present application. The term "solvate", as used herein, refers to an aggregate that comprises one or more molecules of a compound of the present application with one or more molecules of solvent. The solvent can be water, in which case the solvate is a hydrate. Alternatively, the solvent can be an organic solvent. Thus, the compounds of the present application can exist as a hydrate, including a monohydrate, dihydrate, hemihydrate, sesquihydrate, trihydrate, tetrahydrate and the like, as well as the corresponding solvated forms. The compounds of the present application can form true solvates, but in some cases can only retain a mixture of water or solvent that is not stoichiometrically determined. The compounds of the present application can be reacted or precipitated out of a solvent or crystallized from a solvent. Solvates of the compounds of the present application are also within the scope of the present application.

[0163] The present application also includes prodrugs of the above compounds. In the present application, the term "prodrug" means a compound that is convertible in vivo into a biologically active compound of the present application. Thus, the term "prodrug" refers to a pharmacologically acceptable metabolic precursor of a compound of the present application. When administered to a subject in need thereof, a prodrug can not be active, but is converted in vivo to the active compound of the present application. Prodrugs are typically rapidly transformed in vivo to yield the parent compound, for example, by hydrolysis in blood. The prodrug compound often offers advantages of solubility, tissue compatibility or sustained release compared to the parent compound. Prodrugs include known amino-protecting groups and carboxy-protecting groups. Specific methods for preparing prodrugs are found in Saulnier, M.G., et al., Bioorg. Med. Chem. Lett. 1994, 4, 1985-1990; Greenwald, R.B., et al., J. Med. Chem. 2000, 43, 475.

[0164] In the present application, "pharmaceutical composition" means a formulation of a compound of the present application and a medium generally accepted in the art for the delivery of biologically active compounds to mammals, e.g., humans. The medium includes a pharmaceutically acceptable carrier. The object of a pharmaceutical composition is to facilitate administration of the active ingredient to the organism and to facilitate its absorption into the organism to thereby exert its biological activity.

[0165] The term "pharmaceutically acceptable" is used in the present application to mean a substance (such as a carrier or diluent) that does not affect the biological activity or properties of a compound of the present application, and is relatively nontoxic, i.e., the material is administered to an individual without causing undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained.

[0166] In the present application, "pharmaceutically acceptable carrier" includes, but is not limited to, any adjuvant, carrier, excipient, flow regulating agent, sweetening agent, diluent, preservative, dye / colorant, flavor agent, surface-active agent, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier approved by a regulatory agency of the relevant government for use in humans or animals.

[0167] The "tumor", "cell proliferation abnormality-related disease", and the like described in the present application include, but are not limited to, leukemia, gastrointestinal stromal tumor, histiocytic lymphoma, non-small cell lung cancer, small cell lung cancer, pancreatic cancer, lung squamous cell carcinoma, lung adenocarcinoma, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell cancer, cervical cancer, ovarian cancer, intestinal cancer, nasopharyngeal cancer, brain cancer, bone cancer, esophageal cancer, melanoma, renal cancer, oral cancer, and the like.

[0168] The terms "preventative", "prevention", and "preventing" as used herein include reducing the likelihood of the occurrence or worsening of a disease or condition in a subject.

[0169] The terms "treat" and other similar synonymous terms as used herein include the following meanings:

[0170] (i) preventing the disease or condition from occurring in a mammal, in particular, when such mammal is predisposed or has a predisposition to the disease or condition but has not yet been diagnosed as having it;

[0171] (ii) inhibiting the disease or condition, i.e., arresting its development;

[0172] (iii) relieving the disease or condition, i.e., causing the condition to regress; or

[0173] (iv) alleviating the symptoms of the disease or condition.

[0174] The terms "effective amount", "therapeutically effective amount", or "pharmaceutically effective amount" as used herein refer to the amount of at least one pharmaceutical agent or compound that, upon administration to a subject, is effective to alleviate to some extent one or more of the symptoms of the disease or condition being treated. The result can be reduction and / or alleviation of signs, symptoms, or causes of a disease or condition, or any other desired alteration of a biological system. For example, an "effective amount" for therapy is the amount of a composition comprising a compound disclosed herein that is required to provide clinically significant relief of symptoms of a condition. Techniques for

[0175] The terms "administration", "administering" and the like, as used herein, refer to methods allowing delivery of a compound or composition to the desired site of biological action. These methods include, but are not limited to, oral routes, transduodenal routes, parenteral injections (including intravenous, subcutaneous, intraperitoneal, intramuscular, intraarterial injection or infusion), topical administration and transrectal administration. Administration techniques useful for the compounds and methods described herein are well known to those skilled in the art, for example, as discussed in Goodman and Gilman, The Pharmacological Basis of Therapeutics, current ed.; Pergamon; and Remington's, Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton, Pa. In preferred embodiments, the compounds and compositions discussed herein are administered orally.

[0176] The terms "pharmaceutical combination," "pharmaceutical combinations," "combination therapy," "administration of additional therapies," "administration of additional therapeutic agents," and the like, as used herein refer to the association or combination of more than one active ingredient in single entity or separate entities in the form of a single dosage. The term "fixed combination" means that at least one compound described herein and at least one co-agent are both administered to a patient simultaneously in the form of a single entity or dosage. The term "non-fixed combination" means that at least one compound described herein and at least one co-agent are administered to a patient as separate entities either simultaneously, concurrently or sequentially with variable intervening time periods. These also apply to cocktail therapies, e.g. the administration of three or more active ingredients.

[0177] Those skilled in the art will further appreciate that in the methods described below, the functional groups of intermediate compounds can need to be protected as appropriate. Such functional groups include hydroxyl, amino, mercapto, and carboxylic acid. Suitable protecting groups for hydroxyl include trialkylsilyl or diarylalkylsilyl groups (e.g., tert-butyldimethylsilyl, tert-butyldiphenylsilyl, or trimethylsilyl), tetrahydropyranyl, benzyl, and the like. Suitable protecting groups for amino, amidino, and guanidino include tert-butoxycarbonyl, benzyloxycarbonyl, and the like. Suitable protecting groups for mercapto include -C(O)-R" (where R" is alkyl, aryl, or aralkyl), p-methoxybenzyl, trityl, and the like. Suitable protecting groups for carboxylic acid include alkyl, aryl, or aralkyl esters.

[0178] Protecting groups can be introduced and removed in accordance with standard techniques known to those skilled in the art and as described herein. The use of protecting groups is thoroughly described in Greene, T.W. and P.G.M. Wuts, Protective Groups in Organic Synthesis, (1999), 4th Ed., Wiley. The protecting group can also be a polymeric resin.

[0179] On the basis of common general knowledge in the art, the above-mentioned preferred conditions can be combined arbitrarily, i.e. to obtain each preferred embodiment of the present application.

[0180] The reagents and starting materials used in the present application are commercially available.

[0181] The positive progress effect of the present application is that the present application relates to a novel compound having the structural characteristics of general formula (I), selectively inhibits the enzyme activity of CDK7, significantly inhibits the growth of various tumor cells, and is a therapeutic drug with a completely new mechanism of action. DETAILED DESCRIPTION

[0182] The application will be further described in the following examples without limiting the application to the examples. The experimental methods in the following examples, unless otherwise specified, are selected according to the conventional methods and conditions, or according to the commercial instruction. Unless otherwise specified, the percentages and parts are weight percentages and weight parts.

[0183] In each of the examples, the experimental instrumentations are described (e.g. 1 H NMR was recorded on a Varian Mercury-300 or Varian Mercury-400 NMR spectrometer,1 13 C NMR was recorded on a Varian Mercury-400 or Varian Mercury-500 or Varian Mercury-600 NMR spectrometer, chemical shifts are expressed in δ (ppm); mass spectra were recorded on a Finnigan / MAT-95 (EI) and Finnigan LCQ / DECA and Micromass Ultra Q-TOF (ESI) mass spectrometers; preparative HPLC was performed using silica gel (200-300 mesh).

[0184] In which, the reagents represented by chemical formula or English abbreviation are listed in the following table:

[0185] iPrOH: isopropyl alcohol; EtOH: ethanol; DCM: dichloromethane; TFA: trifluoroacetic acid; MeOH: methanol; NaOH: sodium hydroxide; HCl: hydrogen chloride; TEA: triethylamine; Raney Ni: Raney nickel; 1,4-dioxane: 1,4-dioxane; NaH: sodium hydride; H2O: water; Pd / C: palladium on carbon; H2: hydrogen; HATU: 2-(7-oxabenzo-triazol-l-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; DMF: N,N-dimethylformamide; THF: tetrahydrofuran; Boc2O: di-tert-butyl dicarbonate; NBS: N-bromosuccinimide; NCS: N-chlorosuccinimide; NIS: N-iodosuccinimide; MeCN: acetonitrile; DIPEA / DIEA: N,N-diisopropylethylamine; NaBH4: sodium borohydride; AcOH: acetic acid; ethyl acetate: ethyl acetate; NaBH3CN: sodium cyanoborohydride; K2CO3: potassium carbonate; Cs2CO3: cesium carbonate; nBuLi: n-butyllithium; LiAlH4: lithium aluminum hydride; Pd(dppf)Cl2: [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium; KOAc: potassium acetate. Fumaronitrile: fumaric acid nitrile; P(nBu)3: tri-n-butylphosphine; LDA: lithium diisopropylamide; LiOH: lithium hydroxide; MeI: methyl iodide; EtI: ethyl iodide; (CH2O)n : paraformaldehyde; HCO2H: formic acid; CH3COCl: acetyl chloride; LCMS: liquid chromatography mass spectrometry; Xantphos: 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene; TLC: thin layer chromatography; eq.: equivalent; DCE: 1,2-dichloroethane; HEPES: 4-(2-hydroxyethyl)piperazineethanesulfonic acid; EGTA: ethylene glycol bis(2-aminoethylether)tetraacetic acid; DTT: dithiothreitol

[0186] Preparation of key intermediates:

[0187]

[0188]

[0189] Example Preparation

[0190] Example 1: (R)-1-(2-((4-((8-isopropyl-2-methylpyrimido[1,5,-a][1,3,5]triazin-4-yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1-ketone

[0191]

[0192] First step: Intermediate A (490 mg, 2.02 mmol), 4-aminopiperidine-1-carboxylic tert-butyl ester (421 mg, 2.12 mmol), DIEA (800 mg, 6.21 mmol) were dissolved in acetonitrile (50 mL) under nitrogen protection, and the reaction was carried out at 70 °C for 4 h. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE:EA = 2:1) to obtain the intermediate product (710 mg) as a white solid. LC-MS [M+H] + : m / z 407.6.

[0193] Second step: To the above intermediate product (700 mg, 1.72 mmol) in dichloromethane (50 mL), m-chloroperoxybenzoic acid (596 mg, 3.48 mmol) was added under ice-water bath cooling. The reaction mixture was stirred at room temperature for 2 h. LC-MS detection showed that the starting material was almost consumed. The reaction mixture was washed with saturated aqueous sodium bicarbonate solution (30 mL) twice, and then with aqueous sodium thiosulfate solution (30 mL) and saturated brine (30 mL) in turn. The separated organic phase was concentrated under reduced pressure, and the obtained crude product was purified by silica gel column chromatography (PE:EA = 1:1) to obtain the intermediate product (320 mg) as a yellow solid. LC-MS [M+H] + : m / z 439.6.

[0194] Step 3: To the above intermediate product (180 mg, 0.41 mmol) in anhydrous tetrahydrofuran (20 mL) was added methylmagnesium bromide in tetrahydrofuran (1 M, 1.6 mL, 1.6 mmol) at room temperature. The reaction was stirred at room temperature for 3 hours. The reaction was quenched with saturated aqueous sodium bicarbonate (30 mL) at 0 °C. Ethyl acetate (30 mL) was added to the reaction mixture. The organic phase was separated and the aqueous phase was extracted with ethyl acetate (30 mL) twice. The combined organic phase was dried over MgSO4, filtered and concentrated. The crude product was purified by HPLC to give the product as a yellow solid (84 mg). LC-MS [M+H] + m / z 375.5.

[0195] Step 4: To the above intermediate product (80 mg, 0.21 mmol) in anhydrous dichloromethane (10 mL) was added trifluoroacetic acid (1 mL) at room temperature. The reaction mixture was stirred at room temperature for 1 hour. The reaction was checked by LC-MS and was essentially complete. The reaction was directly concentrated under reduced pressure to give the crude intermediate product as a white solid (58 mg). LC-MS [M+H] + m / z 275.4.

[0196] Step 5: To the above intermediate product (58 mg, 0.21 mmol) in DMF (10 mL) was added (S)-2-((p-toluenesulfonyloxy)methyl)morpholine-4-carboxylic acid tert-butyl ester (78 mg, 0.21 mmol) at room temperature. The reaction mixture was stirred at 80 °C for 72 hours. The reaction was checked by LC-MS and was essentially complete. Saturated aqueous sodium bicarbonate (30 mL) was added to the reaction and extracted with ethyl acetate (30 mL) twice. The combined organic phase was concentrated under reduced pressure to give the crude intermediate product as a white solid (25 mg). LC-MS [M+H] + m / z 474.6.

[0197] Step 6: To the above intermediate product (25 mg, 0.05 mmol) in anhydrous dichloromethane (5 mL) was added trifluoroacetic acid (1 mL) at room temperature. The reaction mixture was stirred at room temperature for 1 hour. The reaction was checked by LC-MS and was essentially complete. The reaction was directly concentrated under reduced pressure to give the crude intermediate product as a white solid (15 mg). LC-MS [M+H] + m / z 374.5.

[0198] Seventh step: To a solution of above intermediate product (8 mg, 0.04 mmol) and triethylamine (20 mg, 0.20 mmol) in anhydrous dichloromethane (5 mL) was added acryloyl chloride (10 mg, 0.11 mmol) at room temperature. The reaction mixture was stirred at room temperature for 1 hour. LC-MS indicated the reaction was essentially complete. The reaction was directly concentrated under reduced pressure, and the crude product was prepared by HPLC to give the target product (10 mg) as a white solid. LC-MS [M+H] + : m / z 428.2. 1 H NMR (400 MHz, DMSO-d6): δ 9.51 (s, 1H), 8.76 (d, J = 8.0 Hz, 1H), 8.02 (d, J = 6.8 Hz, 1H), 6.85-6.77 (m, 1H), 6.16 (d, J = 16.8 Hz, 1H), 5.75 (d, J = 12.4 Hz, 1H), 4.37-4.32 (m, 2H), 4.28-4.04 (m, 5H), 3.97-3.88 (m, 3H), 3.55-3.12 (m, 5H), 2.40 (s, 3H), 2.07-1.97 (m, 4H), 1.27 (d, J = 7.2 Hz, 6H).

[0199] Example 2: (R)-1-(2-((4-((-8-isopropyl-2-(trifluoromethyl)pyrimido[1,5- a][1,3,5]triazin-4-yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1- one

[0200]

[0201] First step: To a solution of intermediate B (500 mg, 2.25 mmol) in DMF (10 mL) was added N-iodosuccinimide (506 mg, 2.25 mmol) under nitrogen protection. The reaction mixture was stirred at room temperature for 20 hours. LC-MS indicated the reaction was essentially complete. To the reaction mixture was added saturated aqueous sodium bicarbonate solution (30 mL), and the organic phase was separated. The organic phase was washed once with saturated brine, and then dried over anhydrous sodium sulfate. The organic phase was filtered, and concentrated under reduced pressure. The resulting crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 4:1) to give the intermediate product (528 mg) as a yellow solid. LC-MS [M+H] + : m / z 331.0.

[0202] Second Step: To a solution of the above intermediate compound (525 mg, 1.59 mmol) in acetonitrile (10 mL) was added phosphorous oxychloride (5 mL) under nitrogen. The reaction mixture was heated to 80 °C for 2 h. The reaction mixture was concentrated under reduced pressure to give a yellow solid (480 mg). LC-MS [M+H] + : m / z 349.5.

[0203] Third Step: The above intermediate compound (480 mg, 1.38 mmol), 4-aminopiperidine-1- carboxylic acid tert-butyl ester (410 mg, 2.05 mmol), and DIEA (800 mg, 6.21 mmol) were dissolved in acetonitrile (50 mL) under nitrogen and stirred at 70 °C for 4 h. The reaction mixture was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 2:1) to give the intermediate product as a white solid (560 mg). LC-MS [M+H] + : m / z 513.3.

[0204] Fourth Step: To a solution of the above intermediate (550 mg, 1.09 mmol) in 1,4-dioxane / water (12 mL / 4 mL) was added isopropenylboronic acid (96 mg, 1.11 mmol), tetrakis(triphenylphosphine)palladium (240 mg, 0.2 mmol), and sodium carbonate powder (560 mg, 5.3 mmol) at room temperature. The reaction mixture was stirred at 100 °C under argon overnight. After the reaction was completed, the reaction mixture was extracted with ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The product was purified by silica gel flash column chromatography to give the product as a white solid (310 mg). LC-MS m / z: 427.2 [M+H] + .

[0205] Fifth Step: To a solution of the above intermediate compound (310 mg, 0.73 mmol) in methanol (20 mL) was added 5% palladium on carbon (50 mg) at room temperature. The reaction mixture was stirred at room temperature under 1 atm of hydrogen for 6 h. The reaction was checked by LC-MS. The reaction mixture was filtered through celite and washed with methanol twice. The combined organic phase was concentrated under reduced pressure to give the product as a white solid (210 mg). LC-MS m / z: 429.4 [M+H] + .

[0206] Sixth Step: To a solution of the above intermediate product (201 mg, 0.47 mmol) in anhydrous dichloromethane (10 mL) was added trifluoroacetic acid (3 mL) at room temperature. The reaction mixture was stirred at room temperature for 2 h. The reaction was checked by LC-MS. The reaction mixture was directly concentrated under reduced pressure to give the product as a white solid (140 mg). LC-MS [M+H]+ m / z 329.3.

[0207] Step 7: To a solution of the above intermediate product (135 mg, 0.41 mmol) in DMF (10 mL) was added (S)-2-((p-toluenesulfonyloxy)methyl)morpholine-4-carboxylic acid tert-butyl ester (156 mg, 0.42 mmol) at room temperature. The reaction mixture was stirred at 80 °C for 72 h. LC-MS indicated the reaction was essentially complete. To the reaction was added saturated aqueous sodium bicarbonate solution (30 mL) and extracted with ethyl acetate (30 mL) twice. The combined organic phase was concentrated under reduced pressure and the resulting crude product was purified by HPLC prep to give the intermediate product as a white solid (65 mg). LC-MS [M+H] + m / z 528.6.

[0208] Step 8: To a solution of the above intermediate product (65 mg, 0.12 mmol) in anhydrous dichloromethane (10 mL) was added trifluoroacetic acid (3 mL) at room temperature. The reaction mixture was stirred at room temperature for 1 h. LC-MS indicated the reaction was essentially complete. The reaction was directly concentrated under reduced pressure to give the crude intermediate product as a white solid (35 mg). LC-MS [M+H] + m / z 528.5.

[0209] Step 9: To a solution of the above intermediate compound (35 mg, 0.08 mmol) and triethylamine (20 mg, 0.20 mmol) in anhydrous dichloromethane (5 mL) was added acryloyl chloride (10 mg, 0.11 mmol) at room temperature. The reaction mixture was stirred at room temperature for 1 h. LC-MS indicated the reaction was essentially complete. The reaction was concentrated under reduced pressure and the crude product was purified by HPLC prep to give the target product as a white solid (20 mg). LC-MS [M+H] + m / z 482.5. 1 H NMR (400 MHz, MeOD) δ 7.56 (s, 1H), 6.75 (dd, J = 16.7, 9.5 Hz, 1H), 6.27 (dd, J = 16.8, 1.9 Hz, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74 - 3.49 (m, 3H), 3.26 (m, 1H), 2.98 (m, 3H), 2.66 - 2.51 (m, 2H), 2.35 (m, 2H), 2.15 - 2.03 (m, 2H), 1.88 - 1.49 (m, 2H), 1.31 (d, J = 6.9 Hz, 6H).

[0210] Example 3: (R)-1-(2-((4-((2-cyclopropyl-8-isopropyl-pyrimido[1,5-a][1,3,5]triazin-4- yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1-one

[0211]

[0212] Reference Example 1 was prepared using cyclopropylbromomagnesium instead of methylbromomagnesium according to the synthetic procedure of Reference Example 1. LC-MS [M+H] + : m / z 454.2. 1 H NMR (400 MHz, DMSO-d6): δ 9.51 (s, 1H), 8.76 (d, J = 8.0 Hz, 1H), 7.98 (d, J = 6.8 Hz, 1H), 6.95-6.65 (m, 1H), 6.17 (m, 1H), 5.75 (m, 1H), 4.37-4.30 (m, 2H), 4.28-4.04 (m, 5H), 3.97-3.88 (m, 3H), 3.20-2.98 (m, 5H), 2.49-1.97 (m, 5H), 1.27 (d, J = 7.2 Hz, 6H), 1.18-0.96 (m, 2H), 0.95-0.85 (m, 2H).

[0213] Example 4: (R)-1-(2-((4-((3-isopropyl-5-methylpyrazolo[1,5-a]pyrimidin-7-yl)amino)piperidin-1- yl)methyl)morpholine)prop-2-enyl-1-one

[0214]

[0215] First Step: To a solution of compound C (500 mg, 2.18 mmol) in DMF (15 mL) was added 4-aminopiperidine-1-carboxylic tert-butyl ester (480 mg, 2.40 mmol) and DIEA (844 mg, 6.55 mmol) at room temperature. The reaction mixture was stirred at room temperature overnight. LCMS showed the reaction was complete. The reaction was extracted with ethyl acetate (30 mL) twice after saturated aqueous sodium bicarbonate (30 mL) was added. The combined organic phase was concentrated under reduced pressure, the resulting crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1:2) to give the product yellow solid intermediate (790 mg). LC-MS [M+H] + : m / z 394.2 / 396.2.

[0216] Second Step: To the above intermediate compound (700 mg, 1.78 mmol) in dichloromethane (20 mL), trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 2 hours. LCMS indicated the reaction was complete. The reaction was concentrated under reduced pressure to give the crude intermediate product as a yellow solid (520 mg). LC-MS [M+H] + : m / z 294.1 / 296.1.

[0217] Third Step: To the above intermediate compound (520 mg, 1.77 mmol) in DMF (15 mL), (S)-2-((p-toluenesulfonyloxy)methyl)morpholine-4-carboxylic acid tert-butyl ester (659 mg, 1.77 mmol) and DIEA (1.52 g, 11.78 mmol) were added. The reaction mixture was stirred at 80 °C for 70 hours. LCMS indicated the reaction was substantially complete. After the reaction mixture was added to saturated aqueous sodium bicarbonate solution (30 mL), it was extracted twice with ethyl acetate (30 mL). The combined organic phase was concentrated under reduced pressure. The resulting crude product was subjected to HPLC preparation to give the product as a yellow solid intermediate (210 mg). LC-MS [M+H] + : m / z 493.3.

[0218] Fourth Step: To the above intermediate compound (190 mg, 0.38 mmol) in dichloromethane (20 mL), dimethylzinc (222 mg, 2.31 mmol) and Pd(dppf)Cl2(29 mg, 0.04 mmol) were added under nitrogen protection. The reaction mixture was heated to 80 °C and stirred overnight. LCMS indicated the reaction was substantially complete. To the reaction mixture was added saturated aqueous sodium bicarbonate solution (10 mL). The separated organic phase was concentrated under reduced pressure. The resulting crude product was subjected to HPLC preparation to give the product as a yellow solid intermediate (52 mg). LC-MS [M+H] + : m / z 473.3.

[0219] Fifth Step: To the above intermediate compound (52 mg, 0.11 mmol) in dichloromethane (5 mL), trifluoroacetic acid (1 mL) was added at room temperature. The reaction mixture was stirred at room temperature for 2 hours. LCMS indicated the reaction was substantially complete. The reaction was directly concentrated under reduced pressure to give the product as a yellow solid intermediate (40 mg). LC-MS [M+H] + : m / z 373.3.

[0220] Sixth Step: To a solution of the above intermediate compound (40 mg, 0.11 mmol) in tetrahydrofuran / water mixture (1:1, 5 mL) was added acryloyl chloride (20 mg, 0.21 mmol) and sodium bicarbonate powder (47 mg, 0.56 mmol) at room temperature. The reaction mixture was stirred at room temperature overnight. LC-MS indicated the reaction was essentially complete. Ethyl acetate (20 mL) and water (20 mL) were added to the reaction mixture. The separated organic phase was concentrated under reduced pressure, and the resulting crude product was purified by HPLC to give the compound of Example 4 (11 mg) as a white solid. LC-MS [M+H] + : m / z 427.1. 1 H NMR (400 MHz, MeOD) δ 7.87 (s, 1H), 6.75 (dd, J = 16.5, 9.5 Hz, 1H), 6.23 (dd, J = 16.8, 1.9 Hz, 1H), 6.05 (s, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.41 (m, 1H), 3.99 (m, 2H), 3.72 - 3.44 (m, 3H), 3.24 (d, J = 6.8 Hz, 1H), 2.98 (m, 3H), 2.66 - 2.53 (m, 2H), 2.48 (s, 3H), 2.35 (m, 2H), 2.12 - 2.01 (m, 2H), 1.88 - 1.49 (m, 2H), 1.31 (d, J = 6.9 Hz, 6H).

[0221] Example 5: (R)-1-(2-((4-((3-isopropyl-5-(trifluoromethyl)pyrazolo[1,5- a]pyrimidin-7-yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-en-1-one

[0222]

[0223] First Step: 4-isopropyl-1H-pyrazole-5-amine (1.25 g, 10.0 mmol) and ethyl 4,4,4-trifluoro-3-oxobutanoate (2.31 g, 12.5 mmol) were dissolved in glacial acetic acid solution (100 mL). The reaction mixture was heated to 110 degrees for 6 hours. LCMS indicated the reaction was essentially complete. The reaction was concentrated under reduced pressure, and the resulting crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1:4) to give the product yellow solid intermediate (890 mg). LC-MS [M+H] + : m / z 246.2.

[0224] Second Step: To a solution of the above intermediate compound (890 mg, 3.63 mmol) in DMF (20 mL) was added phosphorous oxychloride (5 mL) under nitrogen. The reaction mixture was heated to 110 °C for 2 hours. The reaction was concentrated under reduced pressure. The resulting crude product was purified by HPLC to give the product as a yellow oil (580 mg). LC-MS [M+H] + : m / z 264.6.

[0225] Third Step: To a solution of the above intermediate compound (550 mg, 2.09 mmol) in DMF (20 mL) was added 4-aminopiperidine-1-carboxylic tert-butyl ester (480 mg, 2.40 mmol) and DIEA (844 mg, 6.55 mmol) at room temperature. The reaction mixture was stirred at room temperature overnight. After the reaction was completed, the reaction mixture was diluted with saturated aqueous sodium bicarbonate solution (30 mL) and extracted with ethyl acetate (40 mL) twice. The combined organic phase was concentrated under reduced pressure. The resulting crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1:2) to give the product as a yellow solid (670 mg). LC-MS [M+H] + : m / z 428.5.

[0226] Fourth Step: To a solution of the above intermediate compound (670 mg, 1.57 mmol) in dichloromethane (20 mL) was added trifluoroacetic acid (3 mL). The reaction mixture was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The reaction mixture was concentrated under reduced pressure to give the product as a yellow solid (510 mg). LC-MS [M+H] + : m / z 328.3.

[0227] Fifth Step: To a solution of the above intermediate compound (510 mg, 1.56 mmol) in DMF (15 mL) was added (S)-2-((p-toluenesulfonyl) methyl)morpholine-4-carboxylic tert-butyl ester (579 mg, 1.56 mmol) and DIEA (1.52 g, 11.78 mmol). The reaction mixture was stirred at 80 °C for 70 hours. The reaction was monitored by LCMS. After the reaction was completed, the reaction mixture was diluted with saturated aqueous sodium bicarbonate solution (30 mL) and extracted with ethyl acetate (30 mL) twice. The combined organic phase was concentrated under reduced pressure. The resulting crude product was purified by HPLC to give the product as a yellow solid (130 mg). LC-MS [M+H] + : m / z 527.6.

[0228] Sixth Step: To a solution of the above intermediate compound (65 mg, 0.12 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (1 mL) at room temperature. The reaction mixture was stirred at room temperature for 2 hours. LCMS indicated the reaction was essentially complete. The reaction was concentrated directly under reduced pressure to give the intermediate product as a yellow solid (51 mg). LC-MS [M+H] + : m / z 426.5.

[0229] Seventh Step: To a solution of the above intermediate compound (51 mg, 0.12 mmol) in tetrahydrofuran / water mixture (1:1, 5 mL) was added acryloyl chloride (20 mg, 0.21 mmol) and sodium bicarbonate powder (47 mg, 0.56 mmol) at room temperature. The reaction mixture was stirred at room temperature overnight. LC-MS indicated the reaction was essentially complete. To the reaction mixture was added ethyl acetate (20 mL) and water (20 mL). The separated organic phase was concentrated under reduced pressure. The resulting crude product was purified by HPLC prep to give the compound of Example 5 as a white solid (15 mg). LC-MS [M+H] + : m / z 481.3. 1 H NMR (400 MHz, MeOD) d 7.97 (s, 1H), 6.75 (dd, J = 16.5, 9.5 Hz, 1H), 6.27 (dd, J = 16.8, 1.9 Hz, 1H), 6.15 (s, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.41 (m, 1H), 3.99 (m, 2H), 3.72 - 3.44 (m, 3H), 3.24 (d, J = 6.8 Hz, 1H), 2.98 (m, 3H), 2.66 - 2.53 (m, 2H), 2.35 (m, 2H), 2.12 - 2.01 (m, 2H), 1.88 - 1.45 (m, 2H), 1.29 (d, J = 7.2 Hz, 6H).

[0230] Example 6: (R)-1-(2-((4-((5-cyclopropyl-3-isopropylpyrazolo[1,5-a]pyrimidin-7- yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1-ketone

[0231]

[0232] First Step: To a solution of (R)-tert-butyl 2-((4-((5-chloro-3- isopropylpyrazolo[l,5-a]pyrimidin-7-yl)amino)piperidin-l-yl)methyl)morpholine-4- carboxylate (95 mg, 0.19 mmol) in 1,4-dioxane / water (10 mL / 2 mL) was added cyclopropylboronic acid (23 mg, 0.26 mmol), tetrakis(triphenylphosphine)palladium (240 mg, 0.2 mmol) and sodium carbonate powder (560 mg, 5.3 mmol) at room temperature. The reaction mixture was stirred at 80 °C under nitrogen overnight. The reaction was complete, extracted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous Na2S04, filtered, concentrated under reduced pressure, and purified by silica gel flash column chromatography to give the product as a white solid (45 mg). LC-MS m / z: 499.6 [M+H] + .

[0233] Second Step: To a solution of the above intermediate compound (45 mg, 0.09 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (1 mL) at room temperature. The reaction mixture was stirred at room temperature for 2 hours. LCMS showed the reaction was almost complete. The reaction mixture was directly concentrated under reduced pressure to give the intermediate product as a yellow solid (35 mg). LC-MS [M+H] + : m / z 398.5.

[0234] Third Step: To a solution of the above intermediate compound (36 mg, 0.09 mmol) in tetrahydrofuran / water mixture (1:1, 5 mL) was added acryloyl chloride (15 mg, 0.15 mmol) and sodium bicarbonate powder (47 mg, 0.56 mmol) at room temperature. The reaction mixture was stirred at room temperature overnight. LC-MS showed the reaction was almost complete. To the reaction mixture was added ethyl acetate (20 mL) and water (20 mL). The separated organic phase was concentrated under reduced pressure, and the crude product was purified by HPLC preparative purification to give the compound of Example 6 as a white solid (5 mg). LC-MS [M+H] + : m / z 453.6. 1H NMR (400 MHz, MeOD): δ 7.83 (s, 1H), 6.75 (dd, J = 16.5, 9.5 Hz, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 6.05 (s, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.41 (m, 1H), 3.99 (m, 2H), 3.72-3.44 (m, 3H), 3.24 (d, J = 6.8 Hz, 1H), 2.98 (m, 3H), 2.66-2.53 (m, 2H), 2.48 (m, 1H), 2.35 (m, 2H), 2.12-2.01 (m, 2H), 1.88-1.49 (m, 2H), 1.31 (d, J = 6.9 Hz, 6H), 0.68 (m, 2H), 0.54 (m, 2H).

[0235] Example 7: (R)-1-(2-((4-((3-isopropyl-6-methylimidazo[1,2-b]pyrimidin-8- yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1-one

[0236]

[0237] First Step: To a solution of compound D (500 mg, 1.82 mmol) in 1,4-dioxane (15 mL) was added 4-aminopiperidine-1-carboxylic acid tert-butyl ester (385 mg, 1.92 mmol) and DIEA (844 mg, 6.55 mmol) at room temperature. The reaction mixture was heated to 90 degree and stirred for 8 hours. LCMS indicated the reaction was complete. The reaction was diluted with saturated aqueous sodium bicarbonate solution (30 mL) and extracted with ethyl acetate (30 mL) twice. The combined organic phase was concentrated under reduced pressure and the resulting crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1:2) to give the product yellow solid intermediate (620 mg). LC-MS [M+H] + : m / z 394.2 / 396.2.

[0238] Second Step: To a solution of the above intermediate compound (300 mg, 0.76 mmol) in 1,4-dioxane / water (3:1, 4 mL) was added trimethyltriazene (190 mg, 1.51 mmol), tetrakis(triphenylphosphine)palladium (240 mg, 0.2 mmol) and sodium carbonate powder (280 mg, 2.64 mmol) under nitrogen atmosphere. The reaction mixture was heated to 90 °C in a microwave for 6 h. After the reaction mixture was added water (10 mL), it was extracted with ethyl acetate (10 mL) twice. The combined organic phase was washed with saturated brine, dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The resulting crude product was purified by silica gel flash column chromatography to give the product as a white solid (145 mg). LC-MS m / z: 374.5 [M+H] + .

[0239] Third Step: To a solution of the above intermediate compound (145 mg, 0.39 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (1 mL) at room temperature. The reaction mixture was stirred at room temperature for 2 h. The reaction was checked by LCMS and was found to be complete. The reaction mixture was directly concentrated under reduced pressure to give the intermediate product as a yellow solid (95 mg). LC-MS [M+H] + : m / z 274.3.

[0240] Fourth Step: To a solution of the above intermediate compound (95 mg, 0.35 mmol) in DMF (15 mL) was added (S)-2-((p-toluenesulfonyloxy)methyl)morpholine-4-carboxylic acid tert-butyl ester (130 mg, 0.35 mmol) and DIEA (200 mg, 1.55 mmol). The reaction mixture was stirred at 80 °C for 64 h. The reaction was checked by LCMS and was found to be complete. After the reaction mixture was added saturated aqueous sodium bicarbonate solution (30 mL), it was extracted with ethyl acetate (30 mL) twice. The combined organic phase was concentrated under reduced pressure and the resulting crude product was purified by HPLC to give the product as a yellow solid (45 mg). LC-MS [M+H] + : m / z 473.6.

[0241] Fifth Step: To a solution of the above intermediate compound (44 mg, 0.09 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (1 mL) at room temperature. The reaction mixture was stirred at room temperature for 2 h. The reaction was checked by LCMS and was found to be complete. The reaction mixture was directly concentrated under reduced pressure to give the intermediate product as a yellow solid (30 mg). LC-MS [M+H] + : m / z 373.5.

[0242] Sixth Step: To a solution of the above intermediate compound (30 mg, 0.08 mmol) in tetrahydrofuran / water mixture (1:1, 5 mL) was added acryloyl chloride (15 mg, 0.15 mmol) and sodium bicarbonate powder (24 mg, 0.29 mmol) at room temperature. The reaction mixture was stirred at room temperature overnight. LC-MS indicated the reaction was essentially complete. To the reaction mixture was added ethyl acetate (20 mL) and water (20 mL). The separated organic phase was concentrated under reduced pressure, and the resulting crude product was purified by HPLC to give the white compound of Example 7 (9 mg). LC-MS [M+H] + : m / z 427.3. 1 H NMR (400 MHz, MeOD) δ 7.57 (s, 1H), 6.75 (dd, J = 16.8, 9.5 Hz, 1H), 6.65 (s, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74 - 3.49 (m, 3H), 3.26 (m, 1H), 2.98 (m, 3H), 2.66 - 2.51 (m, 2H), 2.37 (m, 2H), 2.24 (s, 3H), 2.15 - 2.01 (m, 2H), 1.88 - 1.49 (m, 2H), 1.28 (d, J = 6.9 Hz, 6H).

[0243] Example 8: (R)-1-(2-((4-((3-isopropyl-6-(trifluoromethyl)imidazo[1,2-b]pyrimidin-8- yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-en-1-one

[0244]

[0245] First Step: To a solution of 6-(trifluoromethyl)pyridazin-3-amine (960 mg, 5.89 mmol) and sodium bicarbonate (600 mg, 7.14 mmol) in methanol (10 mL) was added liquid bromine (950 mg, 5.94 mmol) at room temperature. The reaction mixture was stirred at room temperature for 2 hours. After the reaction was quenched with saturated aqueous sodium bicarbonate solution (50 mL), it was extracted with ethyl acetate (50 mL) twice. The combined organic phase was washed with saturated brine, and concentrated under reduced pressure to give the yellow crude intermediate compound (1.3 g). LC-MS [M+H] + : m / z 243.

[0246] Second Step: The above intermediate compound (1.3 g, 5.37 mmol) and 2-bromo-3- methylbutyraldehyde (886 mg, 5.37 mmol) were dissolved in anhydrous ethanol (100 mL), and the reaction mixture was heated to 100 °C and refluxed overnight. The reaction was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 2: 1) to obtain a white solid compound (1.25 g). LC-MS [M+H] + : m / z 308.1 / 310.1.

[0247] Third Step: The above intermediate (600 mg, 1.94 mmol) was reacted with 4- aminopiperidine-1-carboxylic acid tert-butyl ester according to the synthesis method of the first step of Example 7 to obtain a white solid intermediate compound (480 mg). LC-MS [M+H] + : m / z 428.4.

[0248] Fourth Step: The above intermediate (480 mg, 1.12 mmol) was reacted with trifluoroacetic acid according to the synthesis method of the third step of Example 7 to obtain a white solid intermediate compound (360 mg). LC-MS [M+H] + : m / z 328.3.

[0249] Fifth Step: The above intermediate (360 mg, 1.10 mmol) was reacted with (S)-2- ((p-toluenesulfonyl)methyl)morpholine-4-carboxylic acid tert-butyl ester according to the synthesis method of the fourth step of Example 7 to obtain a white solid intermediate compound (120 mg). LC-MS [M+H] + : m / z 527.6.

[0250] Sixth Step: The above intermediate (42 mg, 0.08 mmol) was reacted with trifluoroacetic acid according to the synthesis method of the fifth step of Example 7 to obtain a white solid intermediate compound (30 mg). LC-MS [M+H] + : m / z 427.5.

[0251] Seventh Step: The above intermediate (30 mg, 0.07 mmol) was reacted with acryloyl chloride according to the synthesis method of the sixth step of Example 7 to obtain a white solid compound of Example 8 (10 mg). LC-MS [M+H] + : m / z 481.2. 1H NMR (400 MHz, MeOD) δ 7.67 (s, 1H), 6.73 (dd, J = 16.8, 9.5 Hz, 1H), 6.35 (s, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74-3.49 (m, 3H), 3.26 (m, 1H), 2.98 (m, 3H), 2.66-2.51 (m, 2H), 2.37 (m, 2H), 2.15-2.01 (m, 2H), 1.88-1.49 (m, 2H), 1.28 (d, J = 6.9 Hz, 6H).

[0252] Example 9: (R)-1-(2-((4-((6-cyclopropyl-3-isopropylimidazo[1,2-b]pyrimidin-8- yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1-one

[0253]

[0254] The compound of Example 9 was prepared as a white solid according to the synthetic procedure of Reference Example 7, using cyclopropylmagnesium bromide instead of trimethylsilylborate. LC-MS [M+H] + : m / z 453.2. 1 H NMR (400 MHz, MeOD) δ 7.55 (s, 1H), 6.78 (dd, J = 16.8, 9.5 Hz, 1H), 6.69 (s, 1H), 6.27 (dd, J = 16.8, 1.9 Hz, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74-3.49 (m, 3H), 3.26 (m, 1H), 2.98 (m, 3H), 2.66-2.51 (m, 2H), 2.37 (m, 2H), 2.29 (s, 3H), 2.15-2.01 (m, 2H), 1.88-1.49 (m, 2H), 1.28 (d, J = 6.9 Hz, 6H), 0.65 (m, 2H), 0.52 (m, 2H).

[0255] Example 10: (R)-1-(2-((4-((3-isopropyl-6-methylimidazo[1,2-a]pyrazin-8- yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1-one

[0256]

[0257] The compound of Example 10 was prepared as a white solid according to the procedure of Reference Example 7, using Intermediate E in place of Intermediate D. LC-MS [M+H] + : m / z 427.2. 1 H NMR (400 MHz, MeOD) δ 7.65 (s, 1H), 6.73 (dd, J = 16.8, 9.5 Hz, 1H), 6.73 (s, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74 - 3.49 (m, 3H), 3.26 (m, 1H), 2.98 (m, 3H), 2.66 - 2.51 (m, 2H), 2.48 (s, 3H), 2.37 (m, 2H), 2.15 - 2.01 (m, 2H), 1.88 - 1.45 (m, 2H), 1.32 (d, J = 6.9 Hz, 6H).

[0258] Example 11: (R)-1-(2-((4-((3-isopropyl-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-8- yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-en-1-one

[0259]

[0260] First Step: To a solution of 3-chloro-5-(trifluoromethyl)pyridazin-2-amine (580 mg, 2.93 mmol) in ethanol (20 mL), chloroacetaldehyde aqueous solution (50%, 10 mL) was added slowly. The reaction mixture was heated to 150 degree under microwave for 30 minutes. LC-MS indicated the reaction was essentially complete. The reaction was concentrated under reduced pressure, the residue was dissolved in dichloromethane (20 mL), then saturated aqueous sodium bicarbonate solution (30 mL) was added, the organic phase was separated. The organic phase was washed with saturated brine once, then dried over anhydrous sodium sulfate. The organic phase was filtered, concentrated under reduced pressure, the crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 4:1) to give the intermediate product (428 mg) as a yellow solid. LC-MS [M+H] + : m / z 427.2.

[0261] Second Step: To a solution of the above intermediate compound (428 mg, 1.93 mmol) in DMF (10 mL) was added N-iodosuccinimide (434 mg, 1.93 mmol) under nitrogen atmosphere. The reaction mixture was stirred at room temperature for 20 hours. LC-MS indicated the reaction was essentially complete. The reaction was quenched by the addition of saturated aqueous sodium bicarbonate solution (30 mL). The organic phase was separated and washed with saturated brine once, then dried over anhydrous sodium sulfate. The organic phase was filtered and concentrated under reduced pressure. The resulting crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 4: 1) to give the intermediate compound as a yellow solid (520 mg). LC-MS [M+H] + : m / z 347.8 / 349.8.

[0262] Third Step: Refer to the third step of Example 2 for synthesis, the above intermediate (520 mg, 1.50 mmol) was reacted with 4-aminopiperidine-1-carbonyl tert-butyl ester to give the intermediate compound as a white solid (420 mg). LC-MS [M+H] + : m / z 512.3.

[0263] Fourth Step: Refer to the fourth step of Example 2 for synthesis, the above intermediate (420 mg, 0.82 mmol) was reacted with isopropenyl boronic acid to give the intermediate compound as a white solid (220 mg). LC-MS [M+H] + : m / z 426.4.

[0264] Fifth Step: Refer to the fifth step of Example 2 for synthesis, the above intermediate (220 mg, 0.52 mmol) was hydrogenated in the presence of palladium on carbon to give the intermediate compound as a white solid (130 mg). LC-MS [M+H] + : m / z 428.4.

[0265] Sixth Step: Refer to the sixth step of Example 2 for synthesis, the above intermediate (120 mg, 0.28 mmol) was reacted with trifluoroacetic acid to give the intermediate compound as a white solid (88 mg). LC-MS [M+H] + : m / z 328.2.

[0266] Seventh Step: Refer to the seventh step of Example 2 for synthesis, the above intermediate (85 mg, 0.26 mmol) was reacted with (S)-2-((p-toluenesulfonyl) methyl)morpholine-4-carbonyl tert-butyl ester to give the intermediate compound as a white solid (43 mg). LC-MS [M+H] + : m / z 527.6.

[0267] Eighth Step: Synthesized according to the eighth step of Example 2 using the above intermediate (43 mg, 0.08 mmol) to react with trifluoroacetic acid to produce an intermediate compound as a white solid (23 mg). LC-MS [M+H] + : m / z 427.4.

[0268] Ninth Step: Synthesized according to the ninth step of Example 2 using the above intermediate (23 mg, 0.05 mmol) to react with propenyl chloride to produce an intermediate compound as a white solid (12 mg). LC-MS [M+H] + : m / z 481.4. 1 H NMR (400 MHz, MeOD) δ 7.69 (s, 1H), 6.73 (dd, J = 16.8, 9.5 Hz, 1H), 6.53 (s, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74 - 3.49 (m, 3H), 3.26 (m, 1H), 2.98 (m, 3H), 2.66 - 2.51 (m, 2H), 2.37 (m, 2H), 2.15 - 2.01 (m, 2H), 1.88 - 1.45 (m, 2H), 1.30 (d, J = 7.2 Hz, 6H).

[0269] Example 12: (R)-1-(2-((4-((3-isopropyl-6-(trifluoromethyl)imidazo[1,2-a]pyrazin-8- yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-en-1-one

[0270]

[0271] Prepared according to the synthetic procedure of Reference Example 10 using cyclopropylboronic acid instead of trimethylboroxine to produce Example 12 as a white solid. LC-MS [M+H] + : m / z 427.2. 1H NMR (400 MHz, MeOD) δ 7.65 (s, 1H), 6.73 (dd, J = 16.8, 9.5 Hz, 1H), 6.73 (s, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74-3.49 (m, 3H), 3.26 (m, 1H), 2.98 (m, 3H), 2.66-2.51 (m, 2H), 2.40-2.37 (m, 3H), 2.15-2.01 (m, 2H), 1.88-1.45 (m, 2H), 1.30 (d, J = 6.9 Hz, 6H), 0.68 (m, 2H), 0.53 (m, 2H).

[0272] Example 13: (R)-1-(2-((4-((7-isopropyl-6-methylimidazo[2,1-f][1,2,4]triazin-4- yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1-one

[0273]

[0274] First Step: To a solution of ethyl 1-amino-1H-imidazole-2-carboxylate (1.61 g, 10.4 mmol) in ethanol (20 mL) was added ethylamidine (10 mL). The reaction mixture was heated to 95 °C for 96 hours. LC-MS indicated the reaction was essentially complete. The reaction was allowed to cool to room temperature and a white solid precipitated. The solid was filtered and dried to give the intermediate product as a white solid (528 mg). LC-MS [M+H] + : m / z 151.1.

[0275] Second Step: The above intermediate compound (528 mg, 3.52 mmol) was reacted with N-iodosuccinimide according to the procedure described in the first step of Example 2 to give the intermediate as a white solid (620 mg). LC-MS [M+H] + : m / z 277.0.

[0276] Third Step: The above intermediate compound (652 mg, 2.36 mmol) was reacted with phosphorus oxychloride according to the procedure described in the second step of Example 2 to give the intermediate as a white solid (660 mg). LC-MS [M+H] + : m / z 294.9.

[0277] Fourth Step: Using the synthetic method of the third step of Example 2, react the above intermediate compound (660 mg, 2.24 mmol) with 4-aminopiperidine-1-carboxylic tert-butyl ester to give the intermediate as a white solid (760 mg). LC-MS [M+H] + : m / z 459.1.

[0278] Fifth Step: Using the synthetic method of the fourth step of Example 2, react the above intermediate compound (760 mg, 1.66 mmol) with isopropenyl boronic acid to give the intermediate as a white solid (460 mg). LC-MS [M+H] + : m / z 373.2.

[0279] Sixth Step: Using the synthetic method of the fifth step of Example 2, hydrogenate the above intermediate compound (460 mg, 1.23 mmol) in the presence of palladium on carbon to give the intermediate as a white solid (310 mg). LC-MS [M+H] + : m / z 375.2.

[0280] Seventh Step: Using the synthetic method of the sixth step of Example 2, react the above intermediate compound (310 mg, 0.83 mmol) with trifluoroacetic acid to give the intermediate as a white solid (225 mg). LC-MS [M+H] + : m / z 275.2.

[0281] Eighth Step: Using the synthetic method of the seventh step of Example 2, react the above intermediate compound (310 mg, 0.83 mmol) with (S)-2-((p-toluenesulfonyl) methyl)morpholine-4-carboxylic tert-butyl ester to give the intermediate as a white solid (72 mg). LC-MS [M+H] + : m / z 474.3.

[0282] Ninth Step: Using the synthetic method of the eighth step of Example 2, react the above intermediate compound (72 mg, 0.15 mmol) with trifluoroacetic acid to give the intermediate as a white solid (43 mg). LC-MS [M+H] + : m / z 374.2.

[0283] Tenth Step: Using the synthetic method of the ninth step of Example 2, react the above intermediate compound (38 mg, 0.10 mmol) with trifluoroacetic acid to give the intermediate as a white solid (16 mg). LC-MS [M+H] + : m / z 428.3. 1H NMR (400 MHz, MeOD) δ 7.65 (s, 1H), 6.73 (dd, J = 16.8, 9.5 Hz, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74-3.49 (m, 3H), 3.26 (m, 1H), 2.98 (m, 3H), 2.63-2.51 (m, 2H), 2.37 (m, 2H), 2.15-2.01 (m, 2H), 1.88-1.45 (m, 2H), 1.30 (d, J = 6.9 Hz, 6H).

[0284] Example 14: (R)-1-(2-((4-((7-isopropyl-6-(trifluoromethyl)imidazo[2,1- b] [1,2,4]triazin-4-yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1- one

[0285]

[0286] The compound of Example 14 was prepared as a white solid according to the synthetic procedure of Reference Example 13, using trifluoroethylcarbamidine instead of ethylcarbamidine. LC-MS [M+H] + : m / z 482.2. 1 H NMR (400 MHz, MeOD) δ 7.65 (s, 1H), 6.73 (dd, J = 16.8, 9.5 Hz, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74-3.49 (m, 3H), 3.26 (m, 1H), 2.98 (m, 3H), 2.63-2.51 (m, 2H), 2.37 (m, 2H), 2.15-2.01 (m, 2H), 1.88-1.45 (m, 2H), 1.30 (d, J = 6.9 Hz, 6H).

[0287] Example 15: (R)-1-(2-((4-((7-isopropyl-6-(trifluoromethyl)imidazo[2,1- b] [1,2,4]triazin-4-yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1- one

[0288]

[0289] The compound of Example 15 was prepared as a white solid according to the synthetic procedure of Reference Example 13, using cyclopropylcarbamidine instead of ethylcarbamidine. LC-MS [M+H] +m / z 482.2. 1 H NMR (400 MHz, MeOD) δ 7.69 (s, 1H), 6.75 (dd, J = 16.8, 9.5 Hz, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74 - 3.49 (m, 3H), 3.26 (m, 1H), 2.96 (m, 3H), 2.69 - 2.51 (m, 3H), 2.37 (m, 2H), 2.15 - 2.01 (m, 2H), 1.88 - 1.45 (m, 2H), 1.30 (d, J = 6.9 Hz, 6H), 0.65 (m, 2H), 0.52 (m, 2H).

[0290] Example 16: (R)-1-(2-((4-((3-isopropyl-6-methyl-[1,2,4]triazolo[4,3,-a]pyrazin-8- yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1-ketone

[0291]

[0292] The compound of Example 16 was prepared as a white solid according to the synthetic procedure of Reference Example 7, using Intermediate F in place of Intermediate D. LC-MS [M+H] + m / z 428.2. 1 H NMR (400 MHz, MeOD) δ 7.69 (s, 1H), 6.75 (dd, J = 16.8, 9.5 Hz, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74 - 3.49 (m, 3H), 3.26 (m, 1H), 2,98 (m, 3H), 2.66 - 2.51 (m, 2H), 2.58 (s, 3H), 2.37 (m, 2H), 2.15 - 2.01 (m, 2H), 1.90 - 1.42 (m, 2H), 1.31 (d J = 6.9 Hz, 6H).

[0293] Example 17: (R)-1-(2-((4-((6-cyclopropyl-3-isopropyl-[1,2,4]triazolo[4,3,-a]pyrazin-8- yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1-ketone

[0294]

[0295] The compound of Example 17 was prepared as a white solid according to the procedure of Reference Example 9, using Intermediate F in place of Intermediate D. LC-MS [M+H] + : m / z 428.2. 1 H NMR (400 MHz, MeOD) δ 7.95 (s, 1H), 6.78 (dd, J = 16.8, 9.5 Hz, 1H), 6.27 (dd, J = 16.8, 1.9 Hz, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74 - 3.49 (m, 3H), 3.26 (m, 1H), 2.98 (m, 3H), 2.66 - 2.51 (m, 2H), 2.37 (m, 2H), 2.29 (s, 3H), 2.15 - 2.01 (m, 2H), 1.78 - 1.49 (m, 2H), 1.25 (d, J = 6.9 Hz, 6H), 0.62 (m, 2H), 0.48 (m, 2H).

[0296] Example 18: (R)-1-(2-((4-((3-isopropyl-6-methyl-[1,2,4]triazolo[4,3,-b]pyridazin-8- yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1-ketone

[0297]

[0298] First Step: To a solution of 3,4,6-trichloropyridazine (460 mg, 2.49 mmol) in N- methylpyrrolidone (20 mL) was added (4-aminopiperidine-1-carboxylic acid tert-butyl ester (385 mg, 1.92 mmol) and DIEA (422 mg, 3.27 mmol). The reaction mixture was allowed to react at room temperature for 24 hours. After adding water (50 mL) to the reaction, it was extracted with ethyl acetate (50 mL) twice, and the combined organic phase was concentrated under reduced pressure. The resulting crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3:1) to give the intermediate compound (650 mg) as a white solid. LC-MS [M+H] + : m / z 347.1 / 349.1.

[0299] Second Step: To a solution of the above intermediate compound (650 mg, 1.87 mmol) in ethanol (20 mL) was added hydrazine hydrate (10 mL). The reaction mixture was heated to 60 degrees for 2 hours. LC-MS was used to monitor the reaction, which was found to be substantially complete. The reaction was allowed to cool to room temperature, and a white solid precipitated. The solid was filtered off and dried to give the intermediate product (560 mg) as a white solid. LC-MS [M+H] + : m / z 344.2 / 346.2.

[0300] Step 3: To a solution of the above intermediate compound (550 mg, 1.60 mmol) in isobutyric acid (10 mL) was added isobutyric acid anhydride (1.61 g, 10.2 mmol). The reaction mixture was stirred at room temperature for 3 hours. To the reaction was added saturated aqueous sodium bicarbonate solution (50 mL) and stirred at room temperature for 20 minutes, then extracted with ethyl acetate (100 mL). The separated organic phase was concentrated under reduced pressure, and the resulting crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1:1) to give the intermediate compound (480 mg) as a white solid. LC-MS [M+H] + : m / z 395.1 / 397.1.

[0301] Step 4: Refer to the reaction of Step 2 of Example 4, the above intermediate compound (475 mg, 1.21 mmol) was reacted with trifluoroacetic acid to give the intermediate compound (350 mg) as a white solid. LC-MS [M+H] + : m / z 295.0 / 297.0.

[0302] Step 5: Refer to the reaction of Step 3 of Example 4, the above intermediate compound (350 mg, 1.19 mmol) was reacted with (S)-2-((p-toluenesulfonyloxy)methyl)morpholine-4-carboxylic acid tert-butyl ester to give the intermediate (120 mg) as a white solid. LC-MS [M+H] + : m / z 494.2 / 496.2.

[0303] Step 6: Refer to the reaction of Step 4 of Example 4, the above intermediate compound (120 mg, 0.24 mmol) was reacted with dimethylzinc to give the intermediate compound (30 mg) as a white solid. LC-MS [M+H] + : m / z 474.3.

[0304] Step 7: Refer to the reaction of Step 5 of Example 4, the above intermediate compound (30 mg, 0.06 mmol) was reacted with trifluoroacetic acid to give the intermediate compound (15 mg) as a white solid. LC-MS [M+H] + : m / z 374.2.

[0305] Step 8: Refer to the reaction of Step 6 of Example 4, the above intermediate compound (15 mg, 0.04 mmol) was reacted with acryloyl chloride to give the intermediate compound (5 mg) as a white solid. LC-MS [M+H] + : m / z 428.2. 1H NMR (400 MHz, MeOD) δ 6.89 (s, 1H), 6.75 (dd, J = 16.8, 9.5 Hz, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74-3.49 (m, 3H), 3.26 (m, 1H), 2.98 (m, 3H), 2.69 (s, 3H), 2.66-2.51 (m, 2H), 2.37 (m, 2H), 2.15-2.01 (m, 2H), 1.90-1.42 (m, 2H), 1.26 (d, J = 6.9 Hz, 6H).

[0306] Example 19: (R)-1-(2-((4-((6-cyclopropyl-3-isopropyl-[1,2,4]triazolo[4,3,-b]pyridazin-8- yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1-one

[0307]

[0308] The compound of Example 19 was prepared as a white solid according to the synthetic procedure of Reference Example 6, using cyclopropylboronic acid in place of methylboronic acid. LC-MS [M+H] + : m / z 454.2. 1 H NMR (400 MHz, MeOD) δ 6.95 (s, 1H), 6.75 (dd, J = 16.8, 9.5 Hz, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74-3.49 (m, 3H), 3.26 (m, 1H), 2.98 (m, 3H), 2.69-2.51 (m, 3H), 2.37 (m, 2H), 2.15-2.01 (m, 2H), 1.90-1.42 (m, 2H), 1.26 (d, J = 6.9 Hz, 6H), 0.65 (m, 2H), 0.48 (m, 2H).

[0309] Example 20: (R)-1-(2-((4-((3-isopropyl 6-(trifluoromethyl)-[1,2,4]triazolo[4,3,-a]pyridin-8- yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1-one

[0310]

[0311] First Step: Dissolve 3-chloro-5-(trifluoromethyl)pyridin-2-ylhydrazine (443 mg, 2.1 mmol) and isobutyric acid (185 mg, 2.1 mmol) in phosphorus oxychloride (2 mL). Heat the reaction mixture to 140 °C under microwave for 20 minutes. Add the reaction mixture to aqueous sodium bicarbonate solution (20 mL) and extract with ethyl acetate (50 mL) twice. Concentrate the combined organic phase under reduced pressure. Purify the resulting crude product by silica gel column chromatography (petroleum ether / ethyl acetate = 4: 1) to give the intermediate compound (310 mg) as a white solid. LC-MS [M+H] + : m / z 264.0 / 266.0.

[0312] Second Step: To a solution of the above intermediate compound (310 mg, 1.17 mmol) and (4-aminopiperidine-l-formyl tert-butyl ester (600 mg, 3.01 mmol) in toluene (20 mL) under nitrogen, add 4,5-bis(diphenylphosphino)-9,9-dimethylxantphos (240 mg, 0.42 mmol), tris(dibenzylideneacetone)dipalladium (201 mg, 0.22 mmol) and sodium tert-butoxide (490 mg, 5.0 mmol). Heat the reaction mixture to 120 °C for 4 hours. Add the reaction mixture to water (40 mL) and extract with ethyl acetate (50 mL) twice. Concentrate the combined organic phase under reduced pressure. Purify the resulting crude product by silica gel column chromatography (petroleum ether / ethyl acetate = 1: 1) to give the intermediate compound (110 mg) as a yellow solid. LC-MS [M+H] + : m / z 428.2.

[0313] Third Step: Refer to the synthesis procedure of the fourth step of Example 5 to react the above intermediate compound (110 mg, 0.26 mmol) with trifluoroacetic acid to give the compound (80 mg) as a yellow solid. LC-MS [M+H] + : m / z 328.2.

[0314] Fourth Step: Refer to the synthesis procedure of the fifth step of Example 5 to react the above intermediate compound (80 mg, 0.24 mmol) with (S)-2-((p-toluenesulfonyl)methyl)morpholine-4-carbonyl tert-butyl ester to give the intermediate product (30 mg) as a white solid. LC-MS [M+H] + : m / z 527.3.

[0315] Fifth Step: Refer to the synthesis procedure of the sixth step of Example 5 to react the above intermediate compound (30 mg, 0.06 mmol) with trifluoroacetic acid to give the compound (22 mg) as a yellow solid. LC-MS [M+H] + : m / z 426.2.

[0316] Step 6: Refer to the synthesis method of Step 7 of Example 5, react the above intermediate compound (22 mg, 0.05 mmol) with acryloyl chloride to obtain the compound of Example 20 as a white solid (4 mg). LC-MS [M+H] + m / z 481.3. 1 H NMR (400 MHz, MeOD) δ 6.97 (s, 1H), 6.73 (dd, J = 16.5, 9.5 Hz, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 6.35 (s, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.41 (m, 1H), 3.99 (m, 2H), 3.72 - 3.44 (m, 3H), 3.24 (d, J = 6.8 Hz, 1H), 2.98 (m, 3H), 2.66 - 2.53 (m, 2H), 2.35 (m, 2H), 2.12 - 2.01 (m, 2H), 1.88 - 1.45 (m, 2H), 1.24 (d, J = 6.8 Hz, 6H).

[0317] Example 21: (R)-3-isopropyl-5-methyl-N-(1-((4-(vinylsulfonyl)morpholin-2- yl)methyl)piperidin-4-yl)pyrazolo[1,5-a]pyrimidin-7-amine

[0318]

[0319] Refer to the synthesis method of Step 7 of Reference Example 1, replace acryloyl chloride with vinylsulfonyl chloride to prepare the compound of Example 21 as a white solid. LC-MS [M+H] + m / z 463.2. 1 H NMR (400 MHz, MeOD) δ 7.57 (s, 1H), 6.25 (m, 1H), 5.77 (m, 1H), 4.48 (m, 1H), 4.05 - 3.95 (m, 2H), 3.74 - 3.49 (m, 3H), 3.26 (m, 1H), 2.98 - 2.86 (m, 3H), 2.66 - 2.51 (m, 2H), 2.48 (s, 3H), 2.37 (m, 2H), 2.15 - 2.01 (m, 2H), 1.88 - 1.49 (m, 2H), 1.32 (d, J = 6.9 Hz, 6H).

[0320] Refer to the synthesis method of Step 7 of Reference Example 1, react with substituted acryloyl chloride (or acid) or substituted butenoyl chloride (or acid) to prepare the compounds of Examples 22-25;

[0321]

[0322]

[0323] Example 26: (R)-1-(2-((4-((3-isopropyl-5-ethylpyrazolo[1,5-a]pyrimidin-7- yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1-one

[0324]

[0325] The compound of Example 26 was prepared as a white solid according to the synthetic procedure of Example 4, using diethylzinc reagent instead of dimethylzinc. LC-MS [M+H] + : m / z 441.20. 1 H NMR (400 MHz, MeOD) δ 7.90 (s, 1H), 6.76 (m, 1H), 6.25 (m, 1H), 6.12 (s, 1H), 5.79 (m, 1H), 4.71-4.36 (m, 2H), 4.01 (d, J = 12.4 Hz, 2H), 3.85 (m, 2H), 3.60 (m, 1H), 3.26 (d, J = 6.9 Hz, 1H), 3.03 (m, 2H), 2.94 (m, 2H), 2.76 (m, 2H), 2.66 (m, 1H), 2.20 (m, 2H), 1.99 (m, 2H), 1.32 (d, J = 6.6 Hz, 6H).

[0326] Example 27: 1-((2R)-2-((4-((5-(1-hydroxyethyl)-3-isopropylpyrazolo[1,5-a]pyrimidin-7- yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1-one

[0327]

[0328] First Step: To a solution of intermediate compound (R)-tert-butyl 2-4-((5-chloro-3- isopropylpyrazolo[l,5-a]pyrimidin-7-yl)amino)piperidin-l-yl)methyl)morpholine-4- carboxylate (390 mg, 0.791 mmol) in 1,4-dioxane (20 mL) was added tributyl(l- ethoxyvinyl)tin (2.6 mL, 1.187 mmol), Pd(PPh3)2Cl2(111 mg, 0.158 mmol) and triethylamine (240 mg, 2.373 mmol). The reaction mixture was heated to 80 °C under nitrogen and stirred for 12 h. LCMS indicated the reaction was complete. The reaction mixture was concentrated under reduced pressure. The crude product was dissolved in ethyl acetate (20 mL) and washed with water twice. The separated organic phase was concentrated. The resulting crude product was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 30: 1) to give the crude intermediate product as yellow oil (410 mg). LC-MS [M+H] + : m / z 529.5.

[0329] Second Step: To a solution of the above intermediate compound (80 mg, 0.15 mmol) in acetonitrile (3 mL) was added 1 M aqueous hydrochloric acid (1 mL). The reaction mixture was stirred at room temperature for 2 h. LCMS indicated the reaction was complete. The reaction mixture was concentrated under reduced pressure to give the crude intermediate compound as yellow oil (60 mg). LC-MS [M+H] + : m / z 501.3.

[0330] Third Step: To a solution of the above intermediate compound (50 mg, 0.10 mmol) in methanol (4 mL) was added NaBH4(10 mg, 0.23 mmol) under ice water bath cooling. The reaction mixture was slowly warmed to room temperature and stirred for 30 min. LCMS indicated the reaction was complete. The reaction mixture was concentrated under reduced pressure. The crude product was dissolved in ethyl acetate (20 mL) and washed with saturated brine twice. The separated organic phase was concentrated under reduced pressure to give the crude product as yellow oil (50 mg). LC-MS [M+H] + : m / z 503.2.

[0331] Fourth Step: To a solution of the above intermediate compound (50 mg, 0.10 mmol) in dichloromethane (4 mL) was added trifluoroacetic acid (1 mL). The reaction mixture was stirred at room temperature for 30 min. LCMS indicated the reaction was complete. The reaction mixture was concentrated under reduced pressure to give the crude intermediate compound as yellow solid (30 mg). LC-MS [M+H] + : m / z 403.2.

[0332] Step 5: The above intermediate compound (30 mg, 0.07 mmol) was reacted with acryloyl chloride according to the procedure described in Step 7 of Example 5 to give the compound of Example 27 (5 mg) as a white solid. LC-MS [M+H] + m / z 457.2. 1 H NMR (400 MHz, MeOD) δ 7.89 (s, 1H), 6.76 (m, 1H), 6.28 (s, 1H), 6.27-6.21 (m, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.41 (m, 2H), 3.99 (m, 2H), 3.76-3.60 (m, 2H), 3.54 (m, 1H), 3.02-2.45 (m, 6H), 2.37 (m, 2H), 2.10 (m, 2H), 1.80 (m, 2H), 1.49 (d, J = 6.6 Hz, 3H), 1.33 (d, J = 6.9 Hz, 6H).

[0333] Example 28: l-((2R)-2-((4-((5-(l-fluoroethyl)-3-isopropylpyrazolo[l,5- a]pyrimidin-7-yl)amino)piperidin-l-yl)methyl)morpholine)prop-2-en-l-one

[0334]

[0335] Step 1 : To a solution of intermediate compound tert-butyl (2R)-2-4-((5-(l- hydroxyethyl)-3-isopropylpyrazolo[l,5-a]pyrimidin-7-yl)amino)piperidin-l-yl)methyl)morpholine-4-carboxylate (50 mg, 0.10 mmol) in dichloromethane (4 mL) was added diethylamine trifluoride (0.4 mL, 2.3 mmol) under ice water bath cooling. The reaction mixture was stirred at room temperature for 2 hours. LC-MS indicated the reaction was complete. After diluting the reaction solution with 20 mL of dichloromethane, 10 mL of saturated aqueous sodium bicarbonate solution was added. The separated organic phase was washed with water, then concentrated under reduced pressure to give the crude intermediate as a yellow oil (42 mg). LC-MS [M+H] + m / z 505.2.

[0336] Step 2: To a solution of the above intermediate compound (42 mg, 0.08 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (1 mL) under ice water bath cooling. The reaction mixture was stirred at room temperature for 2 hours. LCMS indicated the reaction was complete, the reaction solution was concentrated under reduced pressure to give the crude intermediate as a yellow oil (30 mg). LC-MS [M+H] + m / z 405.1.

[0337] Step 3: To a solution of the above intermediate compound (30 mg, 0.07 mmol) in dichloromethane (5 mL) was added acryloyl chloride (10 mg, 0.11 mmol) and triethylamine (23 mg, 0.22 mmol). The reaction mixture was stirred at room temperature for 2 hours. LC-MS indicated the reaction was complete. The reaction was concentrated under reduced pressure. The resulting crude product was subjected to HPLC to afford compound of Example 28 (6 mg) as a white solid. LC-MS [M+H] + : m / z 459.15. 1 H NMR (400 MHz, DMSO) δ 7.96 (s, 1H), 7.71-7.60 (m, 1H), 6.79 (dd, J = 16.4, 10.5 Hz, 1H), 6.23 (s, 1H), 6.13 (dd, J = 16.7, 2.2 Hz, 1H), 5.70 (d, J = 10.6 Hz, 1H), 5.58 (m, 1H), 4.27 (m, 1H), 3.94 (m, 1H), 3.84 (m, 1H), 3.63 (m, 1H), 3.55-3.34 (m, 2H), 3.14 (m, 2H), 3.00-2.73 (m, 3H), 2.41 (m, 2H), 2.26-2.09 (m, 2H), 1.79 (m, 4H), 1.62 (m, 3H), 1.30 (d, J = 6.9 Hz, 6H).

[0338] Example 28 - P1 and 28 - P2 1 - ((R) - 2 - ((4 - ((5 - ((S or R) - 1 - fluoroethyl) - 3 - isopropylpyrazolo [1, 5 - a] pyrimidin - 7 - yl) amino) piperidin - 1 - yl) methyl) morpholin) prop - 2 - en - 1 - one and 1 - ((R) - 2 - ((4 - ((5 - ((R or S) - 1 - fluoroethyl) - 3 - isopropylpyrazolo [1, 5 - a] pyrimidin - 7 - yl) amino) piperidin - 1 - yl) methyl) morpholin) prop - 2 - en - 1 - one

[0339]

[0340] Step one: racemic mixture (2R) - 2 - ((4 - ((5 - (1 - fluoroethyl) - 3 - isopropylpyrazolo [1, 5 - a] pyrimidin - 7 - yl) amino) piperidin - 1 - yl) methyl) morpholin) - 4 - formyl tert - butyl ester (28a) (150 mg, 0.3 mmol) was subjected to chiral preparation by supercritical fluid chromatography to afford two components, single configuration compound 28a - P1 (60 mg, shorter retention time) and single configuration compound 28a - P2 (53 mg, longer retention time), LC - MS (ESI) : m / z: 505.3 [M+H] + .

[0341] Chiral SFC preparation conditions of 28a:

[0342] Column type: IC column specification: 250*25mm 10pm

[0343] Mobile phase A: Supercritical CO2 Mobile phase B: Isopropylamine (0.1% 7.0 mol / l ammonia / methanol solution added)

[0344] Mobile phase gradient ratio: A:B = 80:20 Detection wavelength: 214 nM Mobile phase flow rate: 70 mL / Min

[0345] Cycle time: 5 Min Column temperature: room temperature Pressure: 100 bar

[0346] Chiral analysis conditions of 28a-P1 (Rt = 6.684 min) and 28a-P2 (Rt = 7.607 min):

[0347] Column type: IC column specification: 100*3.0mm 3.0pm

[0348] Mobile phase A: Supercritical CO2 Mobile phase B: Isopropylamine (0.1% diethylamine added)

[0349] Mobile phase gradient ratio: A:B = 90:10 Detection wavelength: 214 nM Mobile phase flow rate: 1.5 mL / Min

[0350] Run time: 10 Min Column temperature: 35 degrees Pressure: 1800 psi

[0351] Step two: According to the method of the second step of Reference Example 28, 28a-P1 (60 mg, 0.12 mmol) and 28a-P2 (53 mg, 0.1 mmol) were reacted with TFA respectively to generate white solid compounds 28b-P1 (33 mg) and 28b-P2 (25 mg). LC-MS (ESI): m / z: 405.2 [M+H] + .

[0352] Step three: According to the method of the third step of Reference Example 28, 28b-P1 (33 mg, 0.12 mmol) and 28b-P2 (25 mg, 0.1 mmol) were reacted with acryloyl chloride respectively to generate white solid compounds 28-P1 (17 mg) and 28-P2 (13 mg). LC-MS (ESI): m / z: 459.5 [M+H] + .

[0353] 1H NMR for 28-P2 (400 MHz, DMSO) δ 7.96 (s, 1H), 7.65 (dd, J = 15.4, 9.2 Hz, 1H), 6.88 - 6.71 (m, 1H), 6.23 (s, 1H), 6.13 (dd, J = 16.7, 1.8 Hz, 1H), 5.70 (d, J = 10.7 Hz, 1H), 5.64 (m, 1H), 5.52 (m, 1H), 4.27 (m, 1H), 3.94 (m, 1H), 3.84 (m, 1H), 3.69 - 3.58 (m, 1H), 3.56 - 3.35 (m, 2H), 3.14 (m, 2H), 3.00 - 2.72 (m, 3H), 2.41 (m, 2H), 2.25 - 2.10 (m, 2H), 1.88 - 1.71 (m, 4H), 1.62 (m, 3H), 1.30 (d, J = 6.9 Hz, 6H).

[0354] 1 H NMR for 28-P2 (400 MHz, DMSO) δ 7.96 (s, 1H), 7.65 (dd, J = 15.4, 9.2 Hz, 1H), 6.88 - 6.71 (m, 1H), 6.23 (s, 1H), 6.13 (dd, J = 16.7, 1.8 Hz, 1H), 5.70 (d, J = 10.7 Hz, 1H), 5.64 (m, 1H), 5.52 (m, 1H), 4.27 (m, 1H), 3.94 (m, 1H), 3.84 (m, 1H), 3.69 - 3.58 (m, 1H), 3.56 - 3.35 (m, 2H), 3.14 (m, 2H), 3.00 - 2.72 (m, 3H), 2.41 (m, 2H), 2.25 - 2.10 (m, 2H), 1.88 - 1.71 (m, 4H), 1.62 (m, 3H), 1.30 (d, J = 6.9 Hz, 6H).

[0355] Example 29: (R)-1-(2-((4-((5-(2-hydroxyisoprop-2-yl)-3-isopropylpyrazolo[1,5- a]pyrimidin-7-yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1-ketone

[0356]

[0357] First Step: To a solution of intermediate compound tert-butyl (R)-2-((4-((5-acetyl-3- isopropylpyrazolo[l,5-a]pyrimidin-7-yl)amino)piperidin-l-yl)methyl)morpholine-4- carboxylate (70 mg, 0.140 mmol) in tetrahydrofuran (5 mL) under nitrogen and ice water bath cooling, methyl magnesium bromide (33 mg, 0.28 mmol) was added. The reaction mixture was slowly warmed to room temperature and the reaction continued to stir for 30 minutes. LCMS indicated the reaction was complete. To the reaction was added 10 mL water and the reaction was concentrated under reduced pressure to remove tetrahydrofuran. The crude yellow oil was extracted with ethyl acetate (20 mL) twice and the combined organic phase was concentrated under reduced pressure to give the intermediate compound (50 mg). LC-MS [M+H] + : m / z 517.5.

[0358] Second Step: The above intermediate compound (50 mg, 0.10 mmol) was reacted with trifluoroacetic acid according to the procedure described in the second step of Example 29 to give the compound (40 mg) as a yellow solid. LC-MS [M+H] + : m / z 417.3.

[0359] Third Step: The above intermediate compound (40 mg, 0.10 mmol) was reacted with trifluoroacetic acid according to the procedure described in the third step of Example 29 to give the compound (7 mg) as a yellow solid. LC-MS [M+H] + : m / z 471.20. 1 H NMR (400 MHz, MeOD) δ 7.88 (s, 1H), 6.76 (dd, J = 16.8, 10.6 Hz, 1H), 6.38 (s, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 5.79 (dd, J = 10.6, 1.9 Hz, 1H), 4.44 (m, 1H), 4.16 - 3.95 (m, 2H), 3.84 (m, 2H), 3.61 (m, 1H), 3.36 (m, 2H), 3.24 (m, 1H), 3.02 (m, 2H), 2.90 (m, 2H), 2.87 - 2.62 (m, 2H), 2.23 (m, 2H), 1.93 (s, 2H), 1.57 (s, 6H), 1.36 (d, J = 6.9 Hz, 6H).

[0360] Example 30: (R)-l-(2-((4-((5-(2-fluoroisoprop-2-yl)-3-isopropylpyrazolo[l,5- a]pyrimidin-7-yl)amino)piperidin-l-yl)methyl)morpholine)prop-2-enyl-l-one

[0361]

[0362] First Step: Refer to the preparation method of the first step of Example 28, react the above intermediate (R)-tert-butyl 2-((4-((5-(2-hydroxyisoprop-2-yl)-3-isopropylpyrazolo[l,5- a]pyrimidin-7-yl)amino)piperidin-l-yl)methyl)morpholine-4-carboxylate (80 mg, 0.15 mmol) with DAST to obtain the compound (60 mg) as a yellow solid. LC-MS [M+H] + : m / z 519.3.

[0363] Second Step: Refer to the preparation method of the second step of Example 28, react the above intermediate compound (58 mg, 0.11 mmol) with trifluoroacetic acid to obtain the compound (45 mg) as a yellow solid. LC-MS [M+H] + : m / z 419.3.

[0364] Third Step: Refer to the preparation method of the third step of Example 28, react the above intermediate compound (42 mg, 0.10 mmol) with trifluoroacetic acid to obtain the compound of Example 30 (9 mg) as a yellow solid. LC-MS [M+H] + : m / z 473.30. 1 H NMR (400 MHz, MeOD) δ 7.89 (s, 1H), 6.77 (m, 1H), 6.38 (s, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 5.79 (dd, J = 10.6, 1.9 Hz, 1H), 4.44 (m, 1H), 4.16-3.95 (m, 2H), 3.84 (m, 2H), 3.61 (m, 1H), 3.36-3.02 (m, 5H), 2.90 (m, 2H), 2.87-2.62 (m, 2H), 2.25 (m, 2H), 1.95 (m, 2H), 1.91 (s, 6H), 1.36 (d, J = 6.9 Hz, 6H).

[0365] Example 31: (R)-l-(2-((4-((2-ethyl-8-isopropylpyrimido[l,5,-a][l,3,5]triazin-4- yl)amino)piperidin-l-yl)methyl)morpholine)prop-2-enyl-l-one

[0366]

[0367] Refer to the synthetic method of Reference Example 1, replace methyl magnesium bromide with ethyl magnesium bromide to prepare the compound of Example 31 as a white solid. LC-MS [M+H] + : m / z 442.15. 1H NMR (400 MHz, MeOD) δ 7.90 (s, 1H), 6.75 (d, J = 6.2 Hz, 1H), 6.23 (dd, J = 16.8, 1.9 Hz, 1H), 5.77 (d, J = 10.3 Hz, 1H), 4.57 (s, 1H), 4.41 (m, 1H), 4.18 (m, 1H), 4.06-3.96 (m, 1H), 3.94 (m, 1H), 3.57 (m, 2H), 3.22 (m, 1H), 3.24-3.12 (m, 1H), 2.99 (m, 3H), 2.72 (q, J = 7.6 Hz, 2H), 2.67-2.46 (m, 3H), 2.35 (m 2H), 2.05 (m, 2H), 1.92-1.74 (m, 2H), 1.35-1.30 (m, 9H).

[0368] Example 32: 1-((2R)-2-((4-((2-(1-hydroxyethyl-8-isopropylpyrimido[1,5- a][1,3,5]triazin-4-yl)amino)piperidin-1-yl)methyl)morpholine-4-carbonyl)pyrrolidin- 1-yl)methyl prop-2-en-1-one

[0369]

[0370] First Step: To a solution of intermediate compound G (230 mg, 1.0 mmol) in DMF (15 mL) was added 4-aminopiperidine-1-carboxylic tert-butyl ester (200 mg, 1.0 mmol) and DIEA (420 mg, 3.3 mmol) at room temperature. The reaction mixture was stirred at room temperature overnight. LCMS indicated the reaction was complete. The reaction was diluted with saturated aqueous sodium bicarbonate solution (30 mL) and extracted with ethyl acetate (30 mL) twice. The combined organic phase was concentrated under reduced pressure and the resulting crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1:2) to give the product as yellow solid intermediate (240 mg). LC-MS [M+H] + : m / z 395.2 / 397.2.

[0371] Second Step: To a solution of the above intermediate compound (240 mg, 0.6 mmol) in 1,4-dioxane (20 mL) was added tributyl(1-ethoxyvinyl)tin (1.4 mL, 0.7 mmol), Pd(PPh3)2Cl2(111 mg, 0.158 mmol) and triethylamine (126 mg, 1.2 mmol). The reaction mixture was heated to 80 °C under nitrogen and stirred for 12 hours. LCMS indicated the reaction was complete. After the reaction mixture was cooled to room temperature, 2 M aqueous hydrochloric acid (5 mL) was added to the reaction solution. The reaction solution was stirred for another 3 hours. The reaction mixture was concentrated under reduced pressure. The crude product was dissolved in ethyl acetate (20 mL) and washed with water twice. The separated organic phase was concentrated. The resulting crude product was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 10:1) to give the crude intermediate product as yellow oil (110 mg). LC-MS [M+H] + : m / z 303.2.

[0372] Third Step: To a solution of the above intermediate product (125 mg, 0.41 mmol) in DMF (10 mL) was added (S)-2-((p-toluenesulfonyl)methyl)morpholine-4-carbonyl tert-butyl ester (156 mg, 0.42 mmol) at room temperature. The reaction mixture was stirred at 80 °C for 72 hours. LCMS indicated the reaction was almost complete. Saturated aqueous sodium bicarbonate (30 mL) was added to the reaction mixture and extracted with ethyl acetate (30 mL) twice. The combined organic phase was concentrated under reduced pressure. The resulting crude product was purified by HPLC prep to give the intermediate product as white solid (63 mg). LC-MS [M+H] + : m / z 502.3.

[0373] Fourth Step: To a solution of the above intermediate compound (60 mg, 0.12 mmol) in methanol (4 mL) was added NaBH4(10 mg, 0.23 mmol) under ice water bath cooling. The reaction mixture was slowly warmed to room temperature and stirred for another 30 minutes. LCMS indicated the reaction was complete. The reaction mixture was concentrated under reduced pressure. The crude product was dissolved in ethyl acetate (20 mL) and washed with saturated brine twice. The separated organic phase was concentrated under reduced pressure to give the crude product as yellow oil (48 mg). LC-MS [M+H] + : m / z 504.3.

[0374] Fifth Step: The above intermediate compound (48 mg, 0.07 mmol) was reacted with trifluoroacetic acid according to the procedure described in the fourth step of Example 27 to give the product as white solid (29 mg). LC-MS [M+H] + : m / z 404.2.

[0375] Step 6: Refer to the synthesis method of Step 5 of Example 27, react the above intermediate compound (29 mg, 0.07 mmol) with acryloyl chloride to obtain the compound of Example 32 as a white solid (5 mg). LC-MS [M+H] + m / z 458.3. 1 H NMR (400 MHz, DMSO) δ 8.53 (m, 1H), 8.03 (s, 1H), 6.82-6.76 (m, 1H), 6.15-6.11 (d, J = 16.6 Hz, 1H), 5.72-5.69 (d, J = 12.4 Hz, 1H), 4.92 (m, 1H), 4.54-4.51 (m, 1H), 4.09-3.99 (m, 1H), 3.94-3.83 (m, 2H), 3.49-3.37 (m, 2H), 3.24-2.91 (m, 4H), 2.40 (m, 2H), 2.11-2.08 (m, 2H), 1.79 (m, 4H), 1.39 (d, J = 4.8 Hz, 3H), 1.29-1.27 (d, J = 6.8 Hz, 6H).

[0376] Example 33: 1-((2R)-2-((4-((2-(1-fluoroethyl-8-isopropylpyrimido[1,5-a][1,3,5]triazin-4-yl)amino)piperidin-1-yl)methyl)morpholin-4-ium)prop-2-enyl-1-oxo

[0377]

[0378] Refer to the synthesis method of Reference Example 28, use the intermediate (2R)-2-((4-((2-(1-hydroxyethyl)-8-isopropylpyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)piperidin-1-yl)methyl)morpholin-4-ium tert-butyl carbonate as the raw material, and prepare the compound of Example 33 as a white solid through three-step reaction. LC-MS [M+H] + m / z 460.2. 1H NMR (400 MHz, DMSO) δ 8.68 (m, 1H), 8.08 (s, 1H), 6.92 - 6.64 (m, 1H), 6.13 (d, J = 16.6 Hz, 1H), 5.70 (d, J = 10.4 Hz, 1H), 5.45 (m, 1H), 4.27 (m, 1H), 4.11 - 3.99 (m, 1H), 3.98 - 3.89 (m, 1H), 3.88 - 3.80 (m, 1H), 3.57 - 3.37 (m, 2H), 3.24 - 3.02 (m, 2H), 2.86 (m, 3H), 2.40 (m, 2H), 2.19 - 2.01 (m, 2H), 1.80 (m, 4H), 1.61 (m, 3H), 1.29 (d, J = 6.9 Hz, 6H).

[0379] Example 33 - P1 and 33 - P2 1 - ((R) - 2 - ((4 - ((2 - ((S or R) - 1 - fluoroethyl) - 8 - isopropylpyrazolo [1, 5 - a] triazin - 4 - yl) amino) piperidin - 1 - yl) methyl) morpholin) prop - 2 - en - 1 - one and 1 - ((R) - 2 - ((4 - ((2 - ((R or S) - 1 - fluoroethyl) - 8 - isopropylpyrazolo [1, 5 - a] triazin - 4 - yl) amino) piperidin - 1 - yl) methyl) morpholin) prop - 2 - en - 1 - one

[0380]

[0381] Following the synthetic procedure of Reference Example 28 - P1 and 28 - P2, compound 33a (100 mg) was prepared by chiral SFC separation to isolate two single configuration intermediate compounds 33a - P1 (shorter retention time) and 33a - P2 (longer retention time), which were then separately prepared to synthesize white solid compounds 33 - P1 (15 mg) and 33 - P2 (11 mg) via two step reactions. LC - MS (ESI): m / z: 460.2 [M + H] + .

[0382] 1H NMR for 33-P2 (400 MHz, DMSO) δ 8.69 (m, 1H), 8.05 (s, 1H), 6.95 - 6.64 (m, 1H), 6.15 (d, J = 16.6 Hz, 1H), 5.70 (d, J = 10.4 Hz, 1H), 5.44 (m, 1H), 4.27 (m, 1H), 4.12 - 3.95 (m, 1H), 3.98 - 3.89 (m, 1H), 3.89 - 3.75 (m, 1H), 3.54 - 3.35 (m, 2H), 3.24 - 3.01 (m, 2H), 2.86 (m, 3H), 2.40 (m, 2H), 2.19 - 2.05 (m, 2H), 1.80 (m, 4H), 1.61 (m, 3H), 1.28 (d, J = 6.9 Hz, 6H).

[0383] 1 H NMR for 33-P2 (400 MHz, DMSO) δ 8.69 (m, 1H), 8.05 (s, 1H), 6.95 - 6.64 (m, 1H), 6.15 (d, J = 16.6 Hz, 1H), 5.70 (d, J = 10.4 Hz, 1H), 5.44 (m, 1H), 4.27 (m, 1H), 4.12 - 3.95 (m, 1H), 3.98 - 3.89 (m, 1H), 3.89 - 3.75 (m, 1H), 3.54 - 3.35 (m, 2H), 3.24 - 3.01 (m, 2H), 2.86 (m, 3H), 2.40 (m, 2H), 2.19 - 2.05 (m, 2H), 1.80 (m, 4H), 1.61 (m, 3H), 1.28 (d, J = 6.9 Hz, 6H).

[0384] Example 34: (R)-1-(2-((4-((2-(2-hydroxyisoprop-2-yl)-8-isopropylpyrimido[1,5- a][1,3,5]triazin-4-yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1- one

[0385]

[0386] The compound of Example 34 was prepared as a white solid according to the synthetic method of Reference Example 29, using (R)-tert-butyl 2-((4-((2-acetyl-8- isopropylpyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)piperidin-1-yl)methyl)morpholine-4- carboxylate as the starting material. LC-MS [M+H] + : m / z 472.20. 1H NMR (400 MHz, DMSO) δ 8.58 (m, 1H), 8.04 (s, 1H), 6.77 (m, 1H), 6.13 (m 1H), 5.71 (m, 1H), 4.82 (s, 1H), 4.28 (m, 1H), 4.07 (m, 1H), 4.00 - 3.80 (m, 2H), 3.51 - 3.39 (m, 2H), 3.24 - 3.03 (m, 2H), 2.99 - 2.76 (m, 3H), 2.41 (m, 2H), 2.21 - 2.04 (m, 2H), 1.79 (m, 4H), 1.46 (s, 6H), 1.30 (d, J = 6.9 Hz, 6H).

[0387] Example 35: (R)-1-(2-((4-((2-(2-hydroxyisoprop-2-yl)-8-isopropylpyrimido[1,5- a][1,3,5]triazin-4-yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1- one

[0388]

[0389] The compound of Example 35 was prepared as a white solid according to the synthetic procedure of Reference Example 30, using (R)-tert-butyl 2-((4-((2-(2- hydroxyisoprop-2-yl)-8-isopropylpyrimido[1,5-a][1,3,5]triazin-4-yl)amino)piperidin-1- yl)methyl)morpholine-4-carboxylate as the starting material. LC-MS [M+H] + : m / z 474.2. 1 H NMR (400 MHz, DMSO) δ 8.65 (m, 1H), 8.06 (s, 1H), 6.87 - 6.72 (m, 1H), 6.13 (d, J = 16.7 Hz, 1H), 5.70 (d, J = 10.5 Hz, 1H), 4.27 (m, 1H), 4.06 - 3.80 (m, 3H), 3.45 (m, 3H), 3.12 (m, 1H), 2.90 (m, 3H), 2.40 (m, 2H), 2.19 - 2.01 (m, 2H), 1.81 (m, 4H), 1.72 (s, 3H), 1.66 (s, 3H), 1.29 (d, J = 6.9 Hz, 6H).

[0390] Example 36: (R)-1-(2-((4-((6-(1-hydroxyethyl)-3-isopropylimidazo[1,2-b]pyrimidin-8- yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1-one

[0391]

[0392] The compound of Example 36 was prepared as a white solid according to the procedure of Reference Example 32, using tert-butyl 4-((6-chloro-3-isopropylimidazo[l,2-b]pyridazin-8- yl)amino)piperidine- 1 -carboxylate as the starting material. LC-MS [M+H] + : m / z 457.30. 1 H NMR (400 MHz, MeOD) δ 7.57 (s, 1H), 6.75 (dd, J = 16.8, 9.5 Hz, 1H), 6.65 (s, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74 - 3.42 (m, 4H), 3.26 (m, 1H), 2.98 (m, 3H), 2.66 - 2.51 (m, 2H), 2.37 (m, 2H), 2.15 - 2.01 (m, 2H), 1.94 (d, J = 6.6 Hz, 3H), 1.88 - 1.49 (m, 2H), 1.28 (d, J = 6.9 Hz, 6H).

[0393] Example 37: (R)-1-(2-((4-((6-ethyl-3-isopropylimidazo[l,2-b]pyrimidin-8-yl)amino)piperidin-l- yl)methyl)morpholine-4-carbonyl)pyrrolidine-3-carboxylic acid

[0394]

[0395] First Step: To a solution of intermediate compound tert-butyl (2R)-2-((4-((6-(l- hydroxyethyl)-3-isopropylimidazo[l,2-b]pyridazin-8-yl)amino)piperidin-l-yl)methyl)morpholine- 4-carboxylate (90 mg, 0.16 mmol) in methanol (5 mL) was added 5% Pd / C (10 mg). The reaction mixture was stirred at room temperature under hydrogen atmosphere (1 atm) for 5 hours. LC-MS indicated the reaction was complete, and 20 mL of methanol was added to the reaction mixture and filtered through celite. The filtrate was concentrated under reduced pressure to give a gray solid compound (50 mg). LC-MS [M+H] + : m / z 487.3.

[0396] Second Step: The above intermediate compound (48 mg, 0.07 mmol) was reacted with trifluoroacetic acid according to the procedure of Reference Example 27, Step 4 to give a white solid (29 mg). LC-MS [M+H] + : m / z 387.3.

[0397] Step 3: Refer to the synthesis method of Example 27, Step 5, to react the above intermediate compound (29 mg, 0.07 mmol) with acryloyl chloride to obtain the compound of Example 37 as a white solid (5 mg). LC-MS [M+H] + m / z 441.3. 1 H NMR (400 MHz, MeOD) δ 7.57 (s, 1H), 6.75 (dd, J = 16.8, 9.5 Hz, 1H), 6.65 (s, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74-3.42 (m, 4H), 3.26 (m, 1H), 2.98 (m, 3H), 2.66-2.37 (m, 5H), 2.15-2.01 (m, 2H), 1.94-1.39 (m, 5H), 1.28 (d, J = 6.9 Hz, 6H).

[0398] Example 38: (R)-1-(2-((4-((6-(1-fluoroethyl)-3-isopropylimidazo[1,2-b]pyridazin-8- yl)amino)piperidin-1-yl)methyl)morpholine

[0399]

[0400] Refer to the synthesis method of Reference Example 28 to use the intermediate (2R)-2-((4-((6-(1- hydroxyethyl)-3-isopropylimidazo[1,2-b]pyridazin-8-yl)amino)piperidin-1-yl)methyl)morpholine- 4-carboxylate as the starting material to prepare the compound of Example 38 as a white solid through three steps. LC-MS [M+H] + m / z 459.3. 1 H NMR (400 MHz, MeOD) δ 7.57 (s, 1H), 6.75 (dd, J = 16.8, 9.5 Hz, 1H), 6.65 (s, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.45-3.99 (m, 4H), 3.74-3.42 (m, 3H), 3.26 (m, 1H), 2.98 (m, 3H), 2.66-2.51 (m, 2H), 2.37 (m, 2H), 2.15-2.01 (m, 2H), 1.94 (m, 3H), 1.88-1.49 (m, 2H), 1.28 (d, J = 6.9 Hz, 6H).

[0401] Example 39: (R)-1-(2-((4-((6-(2-hydroxyisoprop-2-yl)-3-isopropylimidazo[1,2- b]pyrimidin-8-yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1-one

[0402]

[0403] The compound of Example 39 was prepared as a white solid according to the synthetic method of Reference Example 29, using (R)-tert-butyl 2-((4-((6-acetyl-3- isopropylimidazo[1,2-b]pyridazin-8-yl)amino)piperidin-1-yl)methyl)morpholine-4- carboxylate as the starting material. LC-MS [M+H] + m / z 471.30. 1 H NMR (400 MHz, MeOD) d 7.56 (s, 1H), 6.72 (m, 1H), 6.63 (s, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74-3.42 (m, 4H), 3.26 (m, 1H), 2.98 (m, 3H), 2.66-2.51 (m, 2H), 2.37 (m, 2H), 2.15-2.01 (m, 2H), 1.89 (s, 6H), 1.85-1.49 (m, 2H), 1.34 (d, J = 6.9 Hz, 6H).

[0404] Example 40: (R)-1-(2-((4-((6-(2-fluoroisoprop-2-yl)-3-isopropylimidazo[1,2- b]pyrimidin-8-yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1-one

[0405]

[0406] The compound of Example 40 was prepared as a white solid according to the synthetic method of Reference Example 30, using (R)-tert-butyl 2-((4-((6-(2-hydroxyisoprop-2-yl)-3- isopropylimidazo[1,2-b]pyridazin-8-yl)amino)piperidin-1-yl)methyl)morpholine-4- carboxylate as the starting material. LC-MS [M+H] + m / z 473.30. 1H NMR (400 MHz, MeOD) δ 7.56 (s, 1H), 6.72 (m, 1H), 6.65 (s, 1H), 6.27 (dd, J = 16.8, 1.9 Hz, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74-3.42 (m, 4H), 3.26 (m, 1H), 2.98 (m, 3H), 2.66-2.51 (m, 2H), 2.37 (m, 2H), 2.15-2.01 (m, 2H), 1.93-1.49 (m, 8H), 1.33 (d, J = 6.8 Hz, 6H).

[0407] Example 41: l-((2R)-2-((4-((6-(l-hydroxyethyl)-3-isopropylimidazo[l,2- a]pyrazin-8-yl)amino)piperidin-l-yl)methyl)morpholine)prop-2-enyl-l-one

[0408]

[0409] The compound of Example 41 was prepared as a white solid according to the procedure of Reference Example 32, using tert-butyl 4-((6-chloro-3-isopropylimidazo[l,2- a]pyrazin-8-yl)amino)piperidine- 1-carboxylate as the starting material. LC-MS [M+H] + : m / z 457.3. 1 H NMR (400 MHz, MeOD) δ 7.65 (s, 1H), 6.73 (m, 1H), 6.75 (s, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 5.76 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74-3.49 (m, 4H), 3.26 (m, 1H), 2.98 (m, 3H), 2.66-2.41 (m, 2H), 2.37 (m, 2H), 2.15-2.01 (m, 2H), 1.89 (d, J = 7.2 Hz, 3H), 1.85-1.45 (m, 2H), 1.32 (d, J = 6.9 Hz, 6H).

[0410] Example 42: l-((2R)-2-((4-((6-ethyl-3-isopropylimidazo[l,2- a]pyrazin-8-yl)amino)piperidin-l-yl)methyl)morpholine)prop-2-enyl-l-one

[0411]

[0412] The compound of Example 42 was prepared as a white solid according to the synthetic procedure of Reference Example 37, using intermediate tert-butyl (2R)-2-((4-((6-(1- hydroxyethyl)-3-isopropylimidazo[1,2-a]pyrazin-8-yl)amino)piperidin-1-yl)methyl)morpholine- 4-carboxylate as the starting material in a three-step reaction. LC-MS [M+H] + m / z 441.3. 1 H NMR (400 MHz, MeOD) δ 7.65 (s, 1H), 6.73 (dd, J = 16.8, 9.5 Hz, 1H), 6.73 (s, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74 - 3.49 (m, 3H), 3.26 (m, 1H), 2.98 (m, 3H), 2.66 - 2.45 (m, 4H), 2.37 (m, 2H), 2.15 - 2.01 (m, 2H), 1.88 - 1.45 (m, 5H), 1.32 (d, J = 6.9 Hz, 6H).

[0413] Example 43: 1-((2R)-2-((4-((6-(1-fluoroethyl)-3-isopropylimidazo[1,2,-a]pyrazin-8- yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1-one

[0414]

[0415] The compound of Example 43 was prepared as a white solid according to the synthetic procedure of Reference Example 38, using intermediate tert-butyl (2R)-2-((4-((6-(1- hydroxyethyl)-3-isopropylimidazo[1,2-a]pyrazin-8-yl)amino)piperidin-1-yl)methyl)morpholine- 4-carboxylate as the starting material in a three-step reaction. LC-MS [M+H] + m / z 459.3. LC-MS [M+H] + m / z 457.3. 1H NMR (400 MHz, MeOD) δ 7.65 (s, 1H), 6.73 (m, 1H), 6.75 (s, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 5.76 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74-3.49 (m, 3H), 3.26 (m, 1H), 2.98 (m, 3H), 2.66-2.41 (m, 2H), 2.37 (m, 2H), 2.15-2.01 (m, 2H), 1.85-1.65 (m, 2H), 1.48 (s, 6H), 1.32 (d, J = 6.9 Hz, 6H).

[0416] Example 44: l-((2R)-2-((4-((6-(2-hydroxyisoprop-2-yl)-3-isopropylimidazo[l,2- a]pyrazin-8-yl)amino)piperidin-l-yl)methyl)morpholine)prop-2-enyl-l-one

[0417]

[0418] The compound of Example 44 was prepared as a white solid according to the procedure described in Reference Example 39, using (2R)-2-((4-((6-acetyl-3- isopropylimidazo[l,2-a]pyrazin-8-yl)amino)piperidin-l-yl)methyl)morpholine-4- carboxylate as the starting material. LC-MS [M+H] + : m / z 471.3. 1 H NMR (400 MHz, MeOD) δ 7.65 (s, 1H), 6.73 (m, 1H), 6.75 (s, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 5.76 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74-3.49 (m, 3H), 3.26 (m, 1H), 2.98 (m, 3H), 2.66-2.41 (m, 2H), 2.37 (m, 2H), 2.15-2.01 (m, 2H), 1.85-1.65 (m, 2H), 1.48 (s, 6H), 1.32 (d, J = 6.9 Hz, 6H).

[0419] Example 45: l-((2R)-2-((4-((6-(2-fluoroisoprop-2-yl)-3-isopropylimidazo[l,2- a]pyrazin-8-yl)amino)piperidin-l-yl)methyl)morpholine)prop-2-enyl-l-one

[0420]

[0421] The compound of Example 45 was prepared as a white solid according to the synthetic procedure of Reference Example 40, using tert-butyl (2R)-2-((4-((6-(2-hydroxyisopropyl-2-yl)-3- isopropylimidazo[l,2-a]pyrazin-8-yl)amino)piperidin-l-yl)methyl)morpholine-4-carboxylate as the starting material. LC-MS [M+H] + m / z 473.3. 1 H NMR (400 MHz, DMSO) δ 8.65 (s, 1H), 8.06 (s, 1H), 6.87 - 6.72 (m, 1H), 6.13 (d, J = 16.7 Hz, 1H), 5.70 (d, J = 10.5 Hz, 1H), 4.27 (m, 1H), 4.06 - 3.80 (m, 3H), 3.45 (m, 3H), 3.12 (m, 1H), 2.90 (m, 3H), 2.40 (m, 2H), 2.19 - 2.01 (m, 2H), 1.81 (m, 4H), 1.72 (s, 3H), 1.66 (s, 3H), 1.29 (d, J = 6.9 Hz, 6H).

[0422] Example 46: (R)-1-(2-((4-((7-isopropyl-6-ethylimidazo[2,1-f][l,2,4]triazin-4- yl)amino)piperidin-l-yl)methyl)morpholine)prop-2-enyl-l-one

[0423]

[0424] The compound of Example 46 was prepared as a white solid according to the synthetic procedure of Reference Example 13, using propylcarbamidine instead of ethylcarbamidine. LC-MS [M+H] + m / z 442.3. 1 H NMR (400 MHz, MeOD) δ 7.65 (s, 1H), 6.73 (m, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 5.77 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74 - 3.49 (m, 3H), 3.26 (m, 1H), 2.98 (m, 3H), 2.63 - 2.51 (m, 4H), 2.37 (m, 2H), 2.15 - 2.01 (m, 2H), 1.88 - 1.45 (m, 5H), 1.30 (d, J = 6.9 Hz, 6H).

[0425] Example 47: l-((2R)-2-((4-((2-(l-hydroxyethyl)-7-isopropylimidazo[2,l-f][l,2,4]triazin-4- yl)amino)piperidin-l-yl)methyl)morpholine)prop-2-enyl-l-one

[0426]

[0427] First Step: Synthesized according to the procedure described in Example 2, Step 3, using intermediate compound H (537 mg, 2.01 mmol) and 4-aminopiperidine-1-carboxylic tert-butyl ester to produce an intermediate compound as a white solid (480 mg). LC-MS [M+H] + m / z 431.0 / 433.0.

[0428] Second Step: Synthesized according to the procedure described in Example 2, Step 4, using the above intermediate compound (480 mg, 1.11 mmol) and isopropenylboronic acid pinacol ester to produce an intermediate compound as a white solid (395 mg). LC-MS [M+H] + m / z 393.0 / 395.0.

[0429] Third Step: Synthesized according to the procedure described in Example 32, Step 2, using the above intermediate compound (398 mg, 1.01 mmol) and tributyl(l-ethoxyvinyl)tin to produce an intermediate compound as a white solid (278 mg). LC-MS [M+H] + m / z 301.1.

[0430] Fourth Step: Synthesized according to the procedure described in Example 32, Step 3, using the above intermediate compound (278 mg, 0.93 mmol) and (S)-2-((p-toluenesulfonyl) methyl)morpholine-4-carboxylic tert-butyl ester to produce an intermediate compound as a white solid (128 mg). LC-MS [M+H] + m / z 500.3.

[0431] Fifth Step: Synthesized according to the procedure described in Example 2, Step 5, using the above intermediate compound (128 mg, 0.26 mmol) and hydrogenation reduction using Pd / C to produce an intermediate compound as a white solid (95 mg). LC-MS [M+H] + m / z 502.3.

[0432] Sixth Step: Synthesized according to the procedure described in Example 32, Step 4, using the above intermediate compound (95 mg, 0.19 mmol) and sodium borohydride to produce an intermediate compound as a white solid (78 mg). LC-MS [M+H] + m / z 504.3.

[0433] Seventh Step: Synthesized according to the procedure described in Example 32, Step 5, using the above intermediate compound (78 mg, 0.15 mmol) and trifluoroacetic acid to produce an intermediate compound as a white solid (45 mg). LC-MS [M+H] +m / z 404.3.

[0434] Eighth Step: Refer to the sixth step of the synthesis of Example 32, the above intermediate compound (45 mg, 0.11 mmol) was reacted with acryloyl chloride to produce the compound of Example 47 as a white solid (15 mg). LC-MS [M+H] + m / z 458.3. 1 H NMR (400 MHz, DMSO): δ 8.53 (m, 1H), 8.03 (s, 1H), 6.82-6.76 (m, 1H), 6.15-6.11 (m, 1H), 5.72-5.69 (d, J = 12.4 Hz, 1H), 4.92-4.91 (d, J = 4.8 Hz, 1H), 4.54-4.51 (m, 1H), 4.28-4.09 (m, 2H), 3.91-3.83 (m, 2H), 3.49-3.41 (m, 2H), 3.14-3.09 (m, 1H), 2.91-2.55 (m, 3H), 2.41 (m, 2H), 2.11-2.08 (m, 2H), 1.79 (m, 4H), 1.38 (d, J = 4.8 Hz, 3H), 1.29-1.27 (d, J = 6.9 Hz, 6H).

[0435] Example 48: 1-((2R)-2-((4-((2-(1-fluoroethyl)-7-isopropylimidazo[2,1- f][1,2,4]triazin-4-yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1- one

[0436]

[0437] Refer to the synthetic method of Reference Example 38, the intermediate (2R)-2-((4-((2-(1-hydroxyethyl)-7-isopropylimidazo[2,1-f][1,2,4]triazin-4-yl)amino)piperidin-1-yl)methyl)morpholine-4-carboxylate as the starting material, a white solid of the compound of Example 48 was prepared by a three-step reaction. LC-MS [M+H] + m / z 460.3. 1H NMR (400 MHz, MeOD) δ 7.65 (s, 1H), 6.73 (m, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 5.76 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74-3.49 (m, 3H), 3.26 (m, 1H), 2.98 (m, 3H), 2.66-2.35 (m, 4H), 2.15-2.01 (m, 2H), 1.85-1.65 (m, 2H), 1.48 (s, 6H), 1.35 (d, J = 6.8 Hz, 6H).

[0438] Example 49: (R)-1-(2-((4-((2-(2-hydroxyisoprop-2-yl)-7-isopropylimidazo[2,1- f][1,2,4]triazin-4-yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1- one

[0439]

[0440] The compound of Example 49 was prepared as a white solid according to the synthetic procedure of Reference Example 39, using (R)-tert-butyl 2-((4-((2-acetyl-7- isopropylimidazo[2,1-f][1,2,4]triazin-4-yl)amino)piperidin-1-yl)methyl)morpholine-4- carboxylate as the starting material. LC-MS [M+H] + : m / z 472.3. 1 H NMR (400 MHz, MeOD) δ 7.65 (s, 1H), 6.73 (m, 1H), 6.25 (dd, J = 16.8, 1.9 Hz, 1H), 5.76 (d, J = 10.8 Hz, 1H), 4.45 (m, 1H), 3.99 (m, 2H), 3.74-3.49 (m, 3H), 3.26 (m, 1H), 2.98 (m, 3H), 2.66-2.35 (m, 4H), 2.15-2.01 (m, 2H), 1.85-1.65 (m, 2H), 1.48 (s, 6H), 1.35 (d, J = 6.8 Hz, 6H).

[0441] Example 50: (R)-1-(2-((4-((2-(2-fluoroisoprop-2-yl)-7-isopropylimidazo[2,1- f][1,2,4]triazin-4-yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1- one

[0442]

[0443] The compound of Example 50 was prepared as a white solid according to the synthetic procedure of Reference Example 40, using (R)-2-((4-((2-(2-hydroxyisoprop-2-yl)-7- isopropylimidazo[2,1-f][1,2,4]triazin-4-yl)amino)piperidin-1-yl)methyl)morpholine-4- carboxylic acid tert-butyl ester as the starting material. LC-MS [M+H] + : m / z 474.2. 1 H NMR (400 MHz, DMSO) δ 8.65 (m, 1H), 8.06 (s, 1H), 6.87 - 6.72 (m, 1H), 6.13 (d, J = 16.7 Hz, 1H), 5.70 (d, J = 10.5 Hz, 1H), 4.27 (m, 1H), 4.06 - 3.80 (m, 3H), 3.45 (m, 3H), 3.12 (m, 1H), 2.90 (m, 3H), 2.40 (m, 2H), 2.19 - 2.01 (m, 2H), 1.81 (m, 4H), 1.72 (s, 3H), 1.66 (s, 3H), 1.29 (d, J = 6.9 Hz, 6H).

[0444] The compounds of Examples 51-65 were prepared according to the synthetic procedures of Reference Examples 4 and 26, using different reagents as starting materials instead of 4-aminopiperidine-1- carboxylic acid tert-butyl ester or (S)-2-((p-toluenesulfonyl) methyl)morpholine-4- carboxylic acid tert-butyl ester.

[0445]

[0446]

[0447]

[0448] The compounds of Examples 66-70 were prepared according to the synthetic procedure of Reference Example 38, using different reagents as starting materials instead of 4-aminopiperidine-1- carboxylic acid tert-butyl ester or (S)-2-((p-toluenesulfonyl) methyl)morpholine-4- carboxylic acid tert-butyl ester.

[0449]

[0450] The compounds of Examples 71-75 were prepared according to the synthetic procedure of Reference Example 28, using different reagents as starting materials instead of 4-aminopiperidine-1- carboxylic acid tert-butyl ester or (S)-2-((p-toluenesulfonyl) methyl)morpholine-4- carboxylic acid tert-butyl ester.

[0451]

[0452]

[0453] The compounds of Examples 76-80 were prepared according to the synthetic procedure of Reference Example 33, using a different reagent in place of 4-aminopiperidine-1-carboxylic tert-butyl ester or (S)-2-((p-toluenesulfonyl)methyl)morpholine-4-carboxylic tert-butyl ester as starting material:

[0454]

[0455]

[0456] Example 81: (R)-1-(2-((4-((8-isopropyl-2-vinylpyrimido[1,5,-a][1,3,5]triazin-4- yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1-one

[0457]

[0458] The compound of Example 81 was prepared according to the synthetic procedure of Reference Example 1, using vinylmagnesium bromide in place of methylmagnesium bromide. LC-MS [M+H] + m / z 440.2. 1 H NMR (400 MHz, DMSO): δ 8.19 (s, 1H), 6.78 (dd, J = 16.8, 12.4 Hz, 1H), 6.63 (m, 1H), 6.15 (d, J = 16.6 Hz, 1H), 5.89 (d, J = 16.4 Hz, 1H), 5.72 (d, J = 12.4 Hz, 1H), 5.44 (d, J = 12.2 Hz, 1H), 4.38-4.14 (m, 2H), 3.94-3.85 (m, 2H), 3.49-3.39 (m, 3H), 3.19-3.16 (m, 1H), 2.94 (m, 3H), 2.41 (m, 2H), 2.13-2.10 (m, 2H), 1.81 (m, 4H), 1.32 (d, J = 6.9 Hz, 6H).

[0459] Example 82: (R)-2-fluoro-1-(2-((4-((8-isopropyl-2-vinylpyrimido[1,5,-a][1,3,5]triazin-4- yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1-one

[0460]

[0461] The compound of Example 82 was prepared according to the synthetic procedure of Example 81, using 2-fluoropropenoyl chloride in place of acryloyl chloride. LC-MS [M+H] + m / z 458.2. 1H NMR (400 MHz, DMSO): δ 8.17 (s, 1H), 6.63 (m, 1H), 6.15 (m, 1H), 6.05 (d, J = 16.4 Hz, 1H), 5.72 (m, 1H), 5.44 (d, J = 12.2 Hz, 1H), 4.38-4.14 (m, 2H), 3.94-3.85 (m, 2H), 3.49-3.39 (m, 3H), 3.19-3.16 (m, 1H), 2.94 (m, 3H), 2.41 (m, 2H), 2.13-2.10 (m, 2H), 1.81 (m, 4H), 1.32 (d, J = 6.9 Hz, 6H).

[0462] Example 83: (R)-1-(2-((4-((2-ethynyl-8-isopropylpyrimido[1,5-a][1,3,5]triazin-4- yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1-one

[0463]

[0464] The compound of Example 83 was prepared according to the synthetic procedure of Example 1, using ethynylmagnesium bromide instead of methylmagnesium bromide. LC-MS [M+H] + : m / z 438.2. 1 H NMR (400 MHz, DMSO): δ 8.17 (s, 1H), 6.78 (dd, J = 16, 12 Hz, 1H), 6.17-6.13 (d, J = 16.6 Hz, 1H), 5.72 (d, J = 12.4 Hz, 1H), 4.38-4.16 (m, 2H), 3.94-3.85 (m, 3H), 3.49-3.39 (m, 3H), 3.19-3.16 (m, 1H), 2.94 (m, 3H), 2.40 (m, 2H), 2.13-2.11 (m, 2H), 1.82-1.79 (m, 4H), 1.32 (d, J = 6.9 Hz, 6H).

[0465] Example 84: (R)-1-(2-((4-((2-(difluoromethyl)-8-isopropylpyrimido[1,5-a][1,3,5]triazin-4- yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1-one

[0466]

[0467] Step one: Refer to the synthetic method of Step three of Reference Example 1, use 4-((8- isopropyl-2-(methylsulfone)pyrimido[l,5-a][l,3,5]triazin-4-yl)amino)piperidine- 1 -carboxylic acid tert-butyl ester (1.5 g, 3.42 mmol) to react with vinyl magnesium bromide (1 M in THF, 3.5 mL, 3.5 mmol) to obtain the compound as a light yellow oil (0.83 g). LC-MS [M+H] + : m / z 387.2.

[0468] Step two: Bubble ozone into a solution of the above intermediate (0.83 g, 2.15 mmol) in dichloromethane (30 mL) for 4 hours. The reaction was substantially complete as determined by LC-MS. Add 30 mL of saturated aqueous sodium bicarbonate solution to the reaction mixture. Separate the organic phase and concentrate under reduced pressure. Purify the crude product by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 3: 1) to obtain the compound as a light yellow oil (560 mg). LC-MS [M+H] + : m / z 389.2.

[0469] Step three: Refer to the synthetic method of Step one of Reference Example 29, use the above intermediate (560 mg, 1.44 mmol) to react with methyl magnesium bromide to prepare the compound as a white solid (455 mg). LC-MS [M+H] + : m / z 405.2.

[0470] Step four: Add Dess-Martin oxidizing agent (708 mg, 1.76 mmol) to a solution of the above intermediate (450 mg, 1.11 mmol) in tetrahydrofuran (30 mL). Stir the reaction mixture at room temperature overnight. Concentrate the reaction under reduced pressure. Dissolve the residue in ethyl acetate (30 mL). Wash successively with saturated aqueous sodium bicarbonate solution and saturated brine. Separate the organic phase and concentrate under reduced pressure. Purify the crude product by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 4: 1) to obtain the compound as a white solid (387 mg). LC-MS [M+H] + : m / z 403.2.

[0471] Step five: Refer to the synthetic method of Step four of Reference Example 1, use the above intermediate (385 mg, 0.96 mmol) to react with trifluoroacetic acid to prepare the compound as a white solid (242 mg). LC-MS [M+H] + : m / z 303.2.

[0472] Step six: Refer to the synthetic method of example 1, step five, react the above intermediate (240 mg, 0.79 mmol) with (S)-2-((p-toluenesulfonyl)methyl)morpholine-4-carbonyl tert-butyl ester to give the compound (128 mg) as a white solid. LC-MS [M+H] + : m / z 502.3.

[0473] Step seven: Refer to the synthetic method of example 1, step six, react the above intermediate (125 mg, 0.25 mmol) with trifluoroacetic acid to give the compound (85 mg) as a white solid. LC-MS [M+H] + : m / z 402.2.

[0474] Step eight: Refer to the synthetic method of example 1, step seven, react the above intermediate (125 mg, 0.25 mmol) with acryloyl chloride to give the compound (23 mg) as a white solid. LC-MS [M+H] + : m / z 456.2. 1 H NMR (400 MHz, DMSO) δ 8.19 (s, 1H), 6.85-6.70 (m, 1H), 6.13 (d, J = 16.7 Hz, 1H), 5.70 (d, J = 10.2 Hz, 1H), 4.41-4.06 (m, 2H), 4.00-3.79 (m, 2H), 3.58-3.32 (m, 3H), 3.20-3.14 (m, 1H), 2.94 (s, 3H), 2.61 (s, 3H), 2.41 (m, 2H), 2.12 (m, 2H), 1.81 (m, 4H), 1.33 (d, J = 6.9 Hz, 6H).

[0475] Example 85: (R)-1-(2-((4-((2-(Difluoromethyl)-8-isopropylpyrimido[1,5- a][1,3,5]triazin-4-yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1- one

[0476]

[0477] Step 1: At 0°C, diethylaminosulfur trifluoride (664 mg, 4.12 mmol) was added to dichloromethane (5 mL) containing 4-((2-aldehyde-8-isopropylpyrimidino[1,5,-a][1,3,5]triazine-4-yl)amino)piperidine-1-formyl tert-butyl ester (320 mg, 0.824 mmol). The reaction mixture was brought to room temperature and stirred for 2 hours. The reaction was quenched with saturated sodium bicarbonate solution. The reaction mixture was washed with saturated sodium chloride solution, and the aqueous phase was extracted with dichloromethane. The combined organic phases were concentrated under reduced pressure, and the crude product was subjected to silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 10:1) to give a white solid intermediate compound (300 mg). LCMS [M+H] + :m / z 411.2

[0478] Step 2: At room temperature, add 5 mL of trifluoroacetic acid to a dichloromethane solution (300 mg, 0.731 mmol) of the above intermediate compound. React the reaction mixture at room temperature for 2 hours. Concentrate the reaction solution under reduced pressure, and purify the crude product by silica gel column chromatography (eluent: dichloromethane / anhydrous methanol = 20:1) to obtain a pale yellow solid compound (200 mg). LCMS [M+H] + :m / z 311.0.

[0479] Step 3: At room temperature, potassium carbonate (270 mg, 1.95 mmol) and (S)-2-((p-toluenesulfonic acid)methyl)morpholino-4-formyl tert-butyl ester (480 mg, 1.29 mmol) were added to a 5 mL solution of the above intermediate compound (200 mg, 0.65 mmol) in N,N-dimethylformamide. The reaction mixture was heated to 90 °C for 16 hours under nitrogen protection. The reaction was confirmed to be complete by LCMS. After adding 20 mL of water to the reaction mixture, it was extracted three times with ethyl acetate (10 mL). The combined organic phases were concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (eluent: dichloromethane / anhydrous methanol = 20:1) to give a pale yellow solid compound (100 mg). LCMS [M+H] + :m / z 510.3.

[0480] Step 4: At room temperature, add 2 mL of trifluoroacetic acid to a 2 mL solution of the above intermediate compound (100 mg, 0.196 mmol) in dichloromethane. React the reaction mixture at room temperature for 2 hours. Concentrate the reaction mixture under reduced pressure, and purify the crude product by silica gel column chromatography (eluent: dichloromethane / anhydrous methanol = 20:1) to obtain a pale yellow solid compound (70 mg). LCMS [M+H] + :m / z 410.0.

[0481] Step five: To a solution of the above intermediate compound (70 mg, 0.171 mmol) in dichloromethane (5 mL) was added triethylamine (35 mg, 0.342 mmol) and acryloyl chloride (20 mg, 0.222 mmol) at zero degree. The reaction mixture was warmed to room temperature and stirred for 2 hours. The reaction was washed with saturated sodium chloride solution and the separated organic phase was concentrated under reduced pressure. The resulting crude product was purified by HPLC preparative to give the compound of Example 85 as a white solid (18.7 mg). LCMS [M+H] + : m / z 464.4. 1 H NMR (400 MHz, DMSO- d6 ): δ 9.03 (m, 1H), 8.18 (s, 1H), 6.84 - 6.54 (m, 2H), 6.13 (d, J = 16.6 Hz, 1H), 5.70 (d, J = 10.7 Hz, 1H), 4.27 (m, 1H), 4.06 (m, 1H), 3.99 - 3.80 (m, 2H), 3.55 - 3.36 (m, 2H), 3.30 - 3.05 (m, 2H), 2.86 (m, 3H), 2.43 (m, 2H), 2.12 (m, 2H), 1.89 - 1.74 (m, 4H), 1.30 (d, J = 6.9 Hz, 6H).

[0482] 19 F NMR (377 MHz, DMSO- d6 ): δ -118.97 (s, 2H).

[0483] Reference to the synthesis method of Example 85, the compounds of Examples 86-93 were prepared by using different reagents as starting material instead of acryloyl chloride:

[0484]

[0485]

[0486] Example 94: 2-Fluoro-1-((2R)-2-((4-((2-(1-fluoroethyl)-8-isopropylpyrimido[1,5- a][1,3,5]triazin-4-yl)amino)piperidin-1-yl)methyl)morpholine)prop-2-enyl-1- one

[0487]

[0488] Step one: A solution of 4-((2-formyl-8-isopropylpyrimido[l,5-a][l,3,5]triazin-4- yl)amino)piperidine-l-carboxylate (1.1 g, 2.83 mmol) in tetrahydrofuran (30 mL) was cooled to -70 °C under nitrogen. Methylmagnesium bromide in tetrahydrofuran (3 M, 2.36 mL, 7.08 mmol) was added slowly at this temperature and the reaction was continued at this temperature for 1 h. The reaction was checked by LCMS and was found to be substantially complete. The reaction was quenched by the addition of saturated ammonium chloride solution (50 mL) and extracted with ethyl acetate (30 mL) twice. The organic phase was separated and concentrated under reduced pressure. The crude product was purified by column chromatography on silica gel (eluent: petroleum ether / ethyl acetate = 10:1 to 4:1) to give the compound as a light yellow oil (1.05 g). LCMS [M+H] + : m / z 405.2.

[0489] Step two: To a solution of the above intermediate compound (1.05 g, 2.60 mmol) in dichloromethane (10 mL) was added trifluoroacetic acid (5 mL) at room temperature. The reaction mixture was stirred at room temperature for 2 h. The reaction was concentrated under reduced pressure. The crude product was purified by column chromatography on silica gel (eluent: dichloromethane / methanol = 7:1) to give the compound as a white oil (900 mg). LCMS [M+H] + : m / z 305.2.

[0490] Step three: To a solution of the above intermediate compound (900 mg, 2.96 mmol) in N,N-dimethylformamide (25 mL) was added potassium carbonate (270 mg, 1.95 mmol) and (S)-2-((p-toluenesulfonyloxy)methyl)morpholine-4-carboxylate (2.75 g, 7.39 mmol) at room temperature. The reaction mixture was heated to 90 °C for 16 h under nitrogen. The reaction was checked by LCMS and was found to be substantially complete. The reaction mixture was quenched by the addition of saturated aqueous sodium chloride solution (50 mL) and extracted with ethyl acetate (20 mL) three times. The organic phase was combined and concentrated under reduced pressure. The crude product was purified by column chromatography on silica gel (eluent: dichloromethane / anhydrous methanol = 10:1) to give the compound as a light yellow solid (914 mg). LCMS [M+H] + : m / z 504.2.

[0491] Step four: To a solution of the above intermediate compound (300 mg, 0.596 mmol) in dichloromethane (5 mL) was added diethylaminosulfur trifluoride (288 mg, 1.787 mmol) at -70 °C. The reaction mixture was stirred at room temperature for 2 h. To the reaction mixture was added saturated aqueous sodium bicarbonate solution (30 mL) and dichloromethane (30 mL). The separated organic phase was washed with saturated aqueous sodium chloride solution and concentrated under reduced pressure. The resulting crude product was purified by column chromatography on silica gel (eluent: petroleum ether / ethyl acetate = 10:1) to give the compound (110 mg) as colorless oil. LCMS [M+H] + : m / z 506.4.

[0492] Step five: To a solution of the above intermediate compound (110 mg, 0.218 mmol) in dichloromethane (4 mL) was added trifluoroacetic acid (2 mL) at room temperature. The reaction mixture was stirred at room temperature for 2 h. The reaction mixture was concentrated under reduced pressure. The resulting crude product was purified by column chromatography on silica gel (eluent: dichloromethane / methanol = 9:1) to give the compound (45 mg) as white solid. LCMS [M+H] + : m / z 406.3.

[0493] Step six: To a solution of the above intermediate compound (45 mg, 0.11 mmol) in dichloromethane (5 mL) was added 2-fluoropropenoyl chloride (12 mg, 0.13 mmol) and triethylamine (22 mg, 0.22 mmol) at 0 °C. The reaction mixture was stirred at room temperature for 2 h. To the reaction mixture was added dichloromethane (10 mL) and the organic phase was washed with saturated aqueous sodium chloride solution. The separated organic phase was concentrated under reduced pressure. The resulting crude product was purified by HPLC to give the compound of Example 94 (9.73 mg) as white solid. LCMS [M+H] + : m / z 478.2. 1 H NMR (400 MHz, DMSO) δ 8.68 (m, 1H), 8.08 (s, 1H), 6.33 (m, 1H), 5.87 (m, 1H), 5.45 (m, 1H), 4.27 (m, 1H), 4.11-3.99 (m, 1H), 3.98-3.89 (m, 1H), 3.88-3.80 (m, 1H), 3.57-3.37 (m, 2H), 3.24-3.05 (m, 2H), 2.86 (m, 3H), 2.40 (m, 2H), 2.19-2.01 (m, 2H), 1.80 (m, 4H), 1.61 (m, 3H), 1.27 (d, J = 6.9 Hz, 6H).

[0494] Reference to the synthetic method of Example 94, the compounds of Examples 95-100 were prepared by using different reagents instead of 2-fluoropropenoyl chloride:

[0495]

[0496]

[0497] Enzyme activity test

[0498] The inhibitory rate of the compounds of the examples on CDK7 / CycH / MAT1 (Carna) and CDK9 / CycT1 (Carna) kinases was detected, with PHA-793887 and Dinaciclib as positive control compounds. The activity test of the compounds of the examples on the two kinases was carried out by using the method of Mobility shift assay.

[0499] The specific operation process is as follows: (1) preparation of 1xKinase buffer; (2) preparation of compound concentration gradient: the test concentration of the tested compound is 10000nM starting, and is diluted into 100% DMSO solution of 100-fold final concentration in 384 source plate, 3-fold dilution of the compound, 10 concentrations. Use liquid dispenser Echo 550 to transfer 250nL 100-fold final concentration of the compound to the target plate 384-well-plate. Prepare 2.5-fold final concentration of the kinase solution with 1xKinase buffer. (3) add 10μL of 2.5-fold final concentration of the kinase solution to the compound well and the positive control well respectively; add 10μL of 1xKinase buffer to the negative control well. (4) centrifuge at 1000rpm for 30 seconds, shake the reaction plate to mix, and then incubate at room temperature for 10 minutes. (5) prepare a mixed solution of 5 / 3-fold final concentration of ATP and Kinase substrate with 1xKinase buffer. (6) add 15μl of 5 / 3-fold final concentration of the mixed solution of ATP and substrate to start the reaction. (7) centrifuge the 384-well plate at 1000rpm for 30 seconds, shake to mix, and then incubate at room temperature for the corresponding time. (8) add 30μl of termination detection solution to stop the kinase reaction, centrifuge at 1000rpm for 30 seconds, and shake to mix. (9) read the conversion rate with Caliper EZ Reader. (10) calculation formula %Inhibition=Conversion% _max -Conversion% _sample / Conversion% _max -Conversion% _min ×100, wherein: Conversion% _sampleConversion % is the conversion reading of the sample; Conversion % _min Conversion % is the conversion reading of the negative control well, representing the conversion reading of the wells without enzyme activity; Conversion % _max Conversion % is the conversion reading of the positive control well, representing the conversion reading of the wells without compound inhibition. The IC50values of each compound on enzyme activity were obtained by fitting the dose-response curve with the log value of concentration as X axis and the percentage inhibition as Y axis using the log(inhibitor) vs. response - Variable slope fitting dose-response curve of the analysis software GraphPad Prism 5. The calculation formula is Y = Bottom + (Top-Bottom) / (1+10^((LogIC 50 -X)*Hill Slope)).

[0500] Results (see Table 1): Most of the compounds of the present application have higher CDK7 kinase inhibitory activity, still showing higher inhibitory activity (inhibition rate greater than 95%) at a concentration as low as 100 nM; and most of the compounds of the present application have weaker inhibitory activity on CDK9, still showing lower inhibitory activity (inhibition rate less than 10%) at a concentration of 1000 nM. Most of the compounds of the present application show higher CDK7 / CDK9 kinase selectivity. (Wherein the inhibition rate represents ++++ ≥ 90%; 80% < +++ ≤ 60%; 60% < ++ ≤ 30%; + < 10%; IC 50 A represents ≤ 50 nM; 50 nM < B ≤ 500 nM; 500 nM < C ≤ 1000 nM; D > 1000 nM.

[0501] Table 1: CDK7 / CDK9 kinase inhibitory activity and kinase inhibition rate of the compounds of the present application

[0502]

[0503]

[0504] NT represents not tested.

[0505] Most of the compounds of the present application have stronger inhibitory activity on CDK7 kinase, and most of the compounds of the present application have CDK7 kinase IC 50 less than 50 nM, and some compounds even less than 20 nM, such as Example 4 (IC50, 7.1 nM), 27 (IC50, 13.5 nM), 28 (IC 50 , 19.8 nM), 28-P2 (IC50, 2.8 nM), etc.

[0506] Test Example 2 Anti-OVCAR-3 cell proliferation activity test

[0507] 1) After digesting and centrifuging healthy OVCAR-3 cells (ATCC), resuspend them, count them, and seed them into 96-well plates (Corning), 1000 cells per well.

[0508] 2) After pre-culturing in an incubator for 24 hours (37°C, 5% CO2), the cells were treated with different concentrations of the compounds from the examples for 72 hours. Both the experimental group and the control group had 6 replicates, and control wells with added DMSO solvent and blank wells (pure culture medium) without added cells were also included.

[0509] 3) After drug treatment, utilize Cell viability was assessed using a luminescence assay. Before measurement, the culture plate was equilibrated to room temperature, and 50 μL of solution was added to each well. The reagent (Promega) was mixed on an orbital oscillator for 2 minutes to induce cell lysis, and then incubated at room temperature for 60 minutes to stabilize the luminescence signal. The luminescence value was then recorded on an Envision (PerkinElmer) instrument.

[0510] 4) Calculate the cell viability inhibition rate (%) = [A (DMSO) -A (加药) ] / [A (DMSO) -A (空白) ]×100%. (A (加药) : Absorbance of pores containing cells and drug solutions; A (空白) : Absorbance of pores containing culture medium but without cells; A (DMSO) (Absorbance of wells seeded with cells and DMSO added). Three replicate experiments were performed, and the results were statistically analyzed using Graghpad software.

[0511] Results (see Table 2): Most of the compounds in the embodiments of this invention exhibited strong cell proliferation inhibitory activity, IC50... 50 Less than 1000 nM, the cell proliferation inhibitory activity IC50 of some of the compounds in the examples is... 50 Less than 50 nM, and in some examples, the IC50 of the cell proliferation inhibitory activity of the compounds was even less than 10 nM. Specific results are shown in the table below: (IC50) 50 This means A ≤ 50nM; 50nM <B≤500nM;500nM<C≤1000nM;D> 1000nM).

[0512] Table 2: Results of the anti-OVCAR-3 cell proliferation activity data of the compounds of this invention

[0513]

[0514]

[0515] The control compound is a compound reported in patent of Eli Lilly (see WO2021242602, Example 8).

[0516]

[0517] Test Example 3: ADME test of the compound of the example

[0518] (1) Metabolic stability test: metabolic stability incubation was performed in a system of 150 μL of liver microsomes (final concentration 0.5 mg / mL) containing NADPH (final concentration 1 mM), 1 μM of the test compound and positive control midazolam or negative control atenolol, and the reaction was terminated at 0 min, 5 min, 10 min, 20 min and 30 min with acetonitrile containing tinidazole, vortexed for 10 min, centrifuged at 15000 rpm for 10 min, and 50 μL of supernatant was taken for injection into a 96-well plate. The metabolic stability of the compound was calculated by determining the relative decrease in the amount of the original drug.

[0519] Results (see Table 3): The compounds of the examples of the present application have high stability in liver microsomes of various species (rat, mouse, dog, monkey, human), and the data of some of the compounds of the examples are as follows:

[0520] Table 3: Test results of liver microsomal metabolic stability of the compounds of the present patent

[0521]

[0522] NT means not tested.

[0523] Test Example 4: Test of pharmacokinetic parameters of the compound of the example in mice

[0524] 6 male SPF Balbc mice (Shanghai Xipu-Bike Experimental Animal) were divided into two groups, and the test compound was prepared into a suitable solution or suspension; one group was administered intravenously (1 mg / kg), and the other group was administered orally (5 mg / kg). Blood was collected by neck venipuncture, about 0.2 mL / sample / time point, heparin sodium anticoagulation, and the blood sampling time points were as follows: before administration and 5, 15 and 30 min, 1, 2, 4, 6, 8 and 24 h after administration; the blood samples were placed on ice after collection, centrifuged to separate plasma (centrifugation conditions: 8000 rpm / min, 6 min, 2-8℃), and the collected plasma was stored at -80℃ before analysis. Plasma samples were analyzed by LC-MS / MS.

[0525] According to the blood drug concentration data of the drug, the pharmacokinetic calculation software WinNonlin 5.2 was used to calculate the pharmacokinetic parameters AUC 0-t , AUC 0-∞ , MRT 0-∞ , Cmax , T max , T 1 / 2 and V d and their mean and standard deviation. In addition, the bioavailability (F) will be calculated by the following formula.

[0526]

[0527] For samples with concentrations below the lower limit of quantification, samples taken before reaching Cmax should be calculated as zero and samples taken after reaching C max max should be calculated as below the limit of quantification (BLQ) when calculating pharmacokinetic parameters.

[0528] Table 4: Oral pharmacokinetic parameters results of the compound of the present application in mice

[0529]

[0530] Conclusion: The compound of the present application exhibits high drug exposure and bioavailability, and has good in vivo pharmacokinetic properties.

[0531] Test Example 5: Effect of the example compound on the growth of xenograft tumors in nude mice

[0532] To evaluate the in vivo efficacy of the test compound, a variety of xenograft tumor models (such as HCT116, HCC1806, OVCAR-3) were used. 1) HCT116, HCC1806, OVCAR-3 cells were cultured in culture medium containing 10% fetal bovine serum. Exponentially growing tumor cells were collected and resuspended in PBS to an appropriate concentration for subcutaneous tumor inoculation in nude mice. 2) Experimental nude mice were inoculated subcutaneously with 2-8x10 7 tumor cells on the right flank. The tumor cells were resuspended in 1:1 PBS and Matrigel, and the tumor growth was observed regularly. When the tumors grew to an average volume of 120mm 3 -180mm 3 , the mice were randomly divided into groups according to tumor size and body weight for drug administration. 3) The day of tumor cell inoculation was defined as day 0. Before the start of drug administration, the body weight of all animals was measured, and the tumor volume was measured with a vernier caliper. 3) The example compound (prepared to the desired concentration with purified water containing 1% hydroxyethyl cellulose, 0.25% polysorbate 80, and 0.05% antifoaming agent) was administered orally at the given dose every day for three consecutive weeks, and the solvent control group was given an equal amount of solvent. During the entire experiment, the diameter of the transplanted tumor was measured twice a week, and the body weight of the mice was measured.

[0533] The formula for calculating the tumor volume (TV) is: TV = 1 / 2 x a x b 2wherein a, b represent length, width respectively. According to the measured results, relative tumor volume (RTV) is calculated, and the calculation formula is: RTV=V t / V0. Wherein V0 is the tumor volume measured at the time of caging (i.e. d0), and V t is the tumor volume at each time of measurement. The evaluation index of anti-tumor activity is 1) relative tumor proliferation rate T / C (%), and the calculation formula is as follows: T / C (%)=(T RTV / C RTV )×100%, T RTV : RTV of the treatment group; C RTV : RTV of the negative control group; 2) tumor volume growth inhibition rate GI%, and the calculation formula is as follows: GI%= [1-(TV t -TV0) / (CV t -CT0)]×100%, TVt is the tumor volume of the treatment group at each time of measurement; TV0 is the tumor volume of the treatment group at the time of caging; CV t is the tumor volume of the control group at each time of measurement; and CV0 is the tumor volume of the control group at the time of caging.

[0534] The results show that some of the compounds of the present application, such as Examples 28 and 33, all show obvious anti-tumor activity in HCT116, HCC1806, and OVCAR-3 tumor models. Compared with the control group, the tumor inhibition rates of the compounds of Examples 28 and 33 at the dosages of 15 mg / kg, 30 mg / kg, and 60 mg / kg are greater than 70%, and the tumor inhibition rates in the medium and high dosage groups are even greater than 90%, and the test animals can be well tolerated.

[0535] All the documents mentioned in the present application are cited as references in the present application, just as each document is cited as a reference individually. In addition, it should be understood that, after reading the above teaching of the present application, those skilled in the art can make various modifications or amendments to the present application, and these equivalent forms also fall within the scope defined by the claims attached to the present application.

Claims

1. A nitrogen-containing heterocyclic compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof, in the formula: To R 1 independently selected from C2-C6alkenyl, C2-C6alkynyl, C1-C6alkyl-(C=O)-, C1-C3haloalkyl or C1-C3alkyl substituted by one or several R 1-1 substituted C1-C3alkyl, R 1-1 independently selected from: hydroxy or hydroxy-substituted C1-C6alkyl; R 2 independently selected from C1-C3alkyl; R 3 independently selected from acryloyl, substituted acryloyl, propynoyl, substituted propynoyl, ethenesulfonyl, or substituted ethenesulfonyl; R 3 substituted” in R 3-1 is substituted with one or more R 3-1 independently selected from halogen, deuterium, hydroxyl, cyano, amino, C1-C6alkyl, C1-C6alkoxy, 3-6 membered heterocycloalkyl substituted C1-C3alkyl, amino substituted C1-C3alkyl, mono C1-C3alkyl substituted amino-C1-C3alkyl, or di C1-C3alkyl substituted amino-C1-C3alkyl; R4and R5are independently selected from the group consisting of hydrogen, deuterium, halogen, C1-C3alkyl, C1-C3haloalkyl, hydroxyl, and amino; R a , R b is independently selected from hydrogen, deuterium, halogen or C1-C3alkyl; Ring A is wherein the * end is attached to -NH-; the N end is attached to the end. ring B is a 5-6 membered nitrogen-containing heterocycle; n is independently selected from an integer of 0-3; wherein the 5-6 membered nitrogen-containing heterocycle contains 1-3 heteroatoms selected from the group consisting of N and O, and the 3-6 membered heterocycloalkyl group contains 1-3 heteroatoms selected from the group consisting of N, O, P, and S.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein which satisfies one or more of the following conditions: (1) the C2-C6alkenyl group is a C2-C4alkenyl group; (2) the C2-C6alkynyl group is a C2-C4alkynyl group; (3) the C1-C3alkyl group is methyl, ethyl, n-propyl, or i-propyl; (4) the C1-C6alkyl group is methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, sec-butyl, or t-butyl; (5) the halogen in the C1-C3haloalkyl group is fluorine, chlorine, bromine, or iodine; (6) the 3-6 membered cycloalkyl group is cyclopropyl, cyclobutyl, cyclopentyl, cyclohexenyl, or cyclohexyl; (7) the halogen is fluorine, chlorine, bromine, or iodine; (8) 3-6 membered heterocycloalkyl is (9) 5-6 membered nitrogen-containing heterocycle is 3. The compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein which satisfies one of the following conditions: (1) the C2-C6alkenyl group is a vinyl group or a propenyl group; (2) C2-C6alkynyl is (3) the C1-C6alkyl group is i-propyl or t-butyl; (4) the C1-C3haloalkyl group is a C1-C3fluoroalkyl group; (5) the halogen is fluorine.

4. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein which satisfies one or more of the following conditions: (1) R 1 C2-C6alkenyl, C2-C6alkynyl, C1-C6alkyl-(C=0)-, or C1-C3haloalkyl; (2) R 1-1 the number of which is 1, 2 or 3; (3) R 3 is acryloyl, substituted acryloyl, propioloyl, substituted propioloyl or ethenesulfonyl; (4) R 3-1 independently selected from deuterium, halogen, cyano, C1-C6alkyl, 3-6 membered heterocycloalkyl substituted with one or more halogens, mono- and di-C1-C3alkyl substituted amino-C1-C3alkyl; (5) R4is deuterium, halogen, hydroxyl, or C1-C3alkyl; (6) R5is deuterium, halogen, hydroxyl, or C1-C3alkyl; (7) R a , R b independently hydrogen, deuterium, or C1-C3alkyl; (8) Ring B is wherein the N terminus is attached to R 3 and the * terminus is attached to the R terminus. (9) the carbon atom of ring B connected to the carbon chain is in the R configuration.

5. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R 2 is isopropyl.

6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: R 1 is vinyl, ethynyl, CH3(C=0)-, CH2F, CHF2, CF3, or R 2 is methyl, ethyl or isopropyl; or R 3 is or R a , R b is hydrogen, deuterium or methyl; or R4is hydrogen, deuterium, halogen, hydroxyl, or methyl; or R5is hydrogen, deuterium, hydroxyl, halogen, or methyl.

7. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein which satisfies one or more of the following conditions: (1) To (2) To 8. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein: the nitrogen-containing heterocyclic compound represented by the general formula (I) is a compound represented by the general formula (II), wherein W, X, Y, Z, M, R 1 , R 2 , R 3 , R4, R a , R b groups are as defined in claim 1.

9. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein: the nitrogen-containing heterocyclic compound represented by the general formula (I) is a compound represented by the general formula (VI-1)-(VI-3) or (VI-6), R 1 , R 2 , R 3 , R a , R b are as defined in claim 1.

10. The compound of claim 9, or a pharmaceutically acceptable salt thereof, wherein: the nitrogen-containing heterocyclic compound represented by the general formula (I) is a compound represented by the general formula (III-1)-(III-3) or (III-6), 11. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein: the nitrogen-containing heterocyclic compound represented by the general formula (I) is a compound represented by the general formula (IV-1)-(IV-2), wherein R 1 is selected from C2-C6alkenyl, C2-C6alkynyl, C1-C6alkyl-(C=O)- or C1-C3haloalkyl; R 3 selected from the group consisting of: R c , R d , R e , R f , R g , R h , R k , R m is independently selected from hydrogen, deuterium, halogen, Ci-C6alkyl, and cyano.

12. The compound of claim 11, or a pharmaceutically acceptable salt thereof, wherein: R 1 is selected from the group consisting of vinyl, ethynyl and C1-C3haloalkyl.

13. The compound of claim 11, or a pharmaceutically acceptable salt thereof, wherein: the nitrogen-containing heterocyclic compound represented by the general formula (I) is a compound represented by the general formula (V-1)-(V-2), wherein R 1 is selected from C2-C6alkenyl, C2-C6alkynyl, C1-C6alkyl-(C=O)- or C1-C3haloalkyl; R 3 selected from the group consisting of: R c , R d , R e , R f , R g , R h , R k , R m is independently selected from hydrogen, deuterium, halogen, Ci-C6alkyl, and cyano.

14. A compound, or a pharmaceutically acceptable salt thereof, characterized in that: the compound is any one of the following structures:

15. A pharmaceutical composition comprising, the pharmaceutical composition comprises: (i) an effective amount of the compound according to any one of claims 1-14 or a pharmaceutically acceptable salt thereof; and (ii) a pharmaceutically acceptable carrier.

16. Use of a substance Z in the manufacture of a medicament for preventing, treating, or alleviating a disorder or disease mediated by abnormal activity of a CDK kinase. the substance Z is the compound according to any one of claims 1-14 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 15.

17. The use according to claim 16, characterized in that, the CDK kinase is a CDK7 kinase.

18. Use of a substance Z in the manufacture of a medicament for the treatment or prophylaxis of a disease; the disease is cancer, benign neoplasm, angiogenesis, inflammatory disease, infectious disease, autoinflammatory disease or autoimmune disease; the cancer is independently selected from non-small cell lung cancer, small cell lung cancer, lung adenocarcinoma, lung squamous carcinoma, breast cancer, prostate cancer, liver cancer, skin cancer, gastric cancer, intestinal cancer, cholangiocarcinoma, brain cancer, leukemia, lymphoma, fibroma, sarcoma, basal cell carcinoma, glioma, kidney cancer, bone cancer, thyroid cancer, nasopharyngeal cancer or pancreatic cancer; the autoimmune disease is independently selected from rheumatoid arthritis, systemic lupus erythematosus, idiopathic thrombocytopenic purpura, hemolytic anemia or psoriasis; the inflammatory disease is independently selected from osteoarthritis, gouty arthritis, ulcerative colitis and / or inflammatory bowel disease; the infectious disease is independently selected from sepsis, septic shock, endotoxic shock, gram-negative sepsis and / or toxic shock syndrome; the substance Z is a compound according to any one of claims 1 to 14 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 15.

19. The application as described in claim 18, characterized in that, the skin cancer is melanoma.

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