Substituted pyrazolo[1,5-a]pyrimidin-7-amine derivatives, compositions thereof and medical uses

Abstract

A substituted pyrazolo[1,5-a]pyrimidine-7-amine derivative having a structure as shown in formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, prodrug, or pharmaceutical composition thereof. The derivative has significant CDK9 selective inhibitory activity. #imgabs0#

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a substituted pyrazolo[1,5-a]pyrimidin-7-amine derivative, a pharmaceutically acceptable salt, a solvate, a stereoisomer, a prodrug, a pharmaceutical composition and medical uses thereof. Background Art

[0002] Cyclin dependent kinase (CDK) protein family is made up of the member (cell cycle CDK) of the key regulator of the cell division cycle that participates in gene transcription regulation (transcription CDK) and the member with other functions.CDK needs to activate the combination with cyclin regulatory subunit.The CDK1 / cyclin B, CDK2 / cyclin A, CDK2 / cyclin E, CDK4 / cyclin D and CDK6 / cyclin D of described cell cycle CDK are activated in succession, to drive cell to enter and pass through described cell division cycle.The CDK9 / cyclin T and CDK7 / cyclin H of described CDK of transcription regulate the activity of RNA polymerase II by the phosphorylation of carboxyl terminal domain (CTD).

[0003] CDK9 is the catalytic subunit of the positive transcription elongation factor b (P-TEFb) complex. It regulates gene transcription elongation by phosphorylating the carbon-terminal region of RNA complex II. It is located on chromosome 9q34.1 and is a RNA transcription elongation regulatory kinase. CDK9 is widely expressed in various eukaryotic cells and human tissues. CDK9 kinase is highly expressed in cardiomyocytes, hepatocytes, hematopoietic tissue, adipocytes, neurons, and muscle cells, and is generally highly expressed in tumor cells. CDK9 is also a key factor in the progression and maintenance of tumor cells. CDK9 inhibitors, by inhibiting gene transcription elongation, downregulate the expression of the oncoprotein MYC and the apoptosis inhibitor protein Mcl-1, thereby promoting apoptosis in cancer cells. CDK9 inhibitors, by regulating the epigenetic factor BRG1, reactivate silenced genes, including ERVs in tumor cells, promote interferon expression, and render tumor cells more sensitive to immunotherapy.

[0004] Currently, several companies are developing CDK9 inhibitors, including the selective CDK9 inhibitor BAY1251152 developed by Bayer, the selective CDK9 inhibitor AZD4573 developed by AstraZeneca, the non-selective CDK9 inhibitor TP-1287 developed by Tolero, and the non-selective CDK9 inhibitor QHRD107 developed by Changzhou Qianhong Pharmaceutical. However, most selective CDK9 inhibitors are still in early clinical development, and their selective inhibitory activity against CDK9 and in vivo pharmacokinetic parameters need further improvement. Therefore, the development of new CDK9 inhibitors with high activity and selectivity, as well as reduced in vivo toxicity, is of great clinical significance. Summary of the Invention

[0005] The purpose of the present invention is to provide a substituted pyrazolo[1,5-a]pyrimidin-7-amine derivative with high CDK9 inhibitory activity, good selectivity and better pharmacokinetic parameters.

[0006] The first aspect of the present invention provides a compound represented by formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a solvate thereof, or a prodrug thereof:

[0007]

[0008] in,

[0009] R1 is C 3-8 Cycloalkyl (preferably C 3-6 Cycloalkyl), C 2-8 Alkenyl (preferably C 2-6 Alkenyl, more preferably C 2-4 alkenyl) or C 2-8 Alkynyl (preferably C 2-6 Alkynyl, more preferably C 2-4 Alkynyl); the C 3-8 Cycloalkyl, C 2-8 Alkenyl and C 2-8 Alkynyl is unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxyl, carboxyl, C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkyl, halogenated C 1-3 Alkoxy, NR a1 R b1 、-SO2C 1-3 Alkyl, -C(O)NR a1 R b1 、-C(O)C 1-3 Alkyl, -C(O)OC 1-3 Alkyl, -OC(O)C1-3 Alkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl; wherein the phenyl is optionally substituted by 1, 2 or 3 substituents each independently selected from Substituent Group S;

[0010] R2 and R3 are each independently hydrogen, C 1-8 Alkyl (preferably C 1-6 Alkyl, more preferably C 1-3 Alkyl), C 3-8 Cycloalkyl (preferably C 3-6 Cycloalkyl), C 1-8 Alkoxy (preferably C 1-6 Alkoxy, more preferably C 1-3 alkoxy), cyano, hydroxy, carboxyl, halogen (preferably fluorine or chlorine), -C(O)NR a0 R b0 、-C(O)C 1-8 Alkyl (preferably -C(O)C 1-6 Alkyl, more preferably -C(O)C 1-3 alkyl), -C(O)OC 1-8 Alkyl (preferably -C(O)OC 1-6 Alkyl, more preferably -C(O)OC 1-3 alkyl) or -OC(O)C 1-8 Alkyl (preferably -OC(O)C 1-6 Alkyl, more preferably -OC(O)C 1-3 Alkyl); wherein said C 1-8 Alkyl, C 1-8 Alkoxy, C 3-8 The cycloalkyl group is unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxyl, carboxyl, C 1-3 Alkyl, C 1-3 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, halo C 1-3 Alkyl, halogenated C 1-3 Alkoxy, NR a1 R b1 、-SO2C 1-3 Alkyl, -S(O)C 1-3 Alkyl, -C(O)NR a1 R b1 、-C(O)OC 1-3 Alkyl, -OC(O)C 1-3 Alkyl, C 3-6 Cycloalkyl, C 3-6Cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl; wherein the phenyl, 5- to 6-membered heteroaryl is optionally substituted by 1, 2 or 3 substituents each independently selected from Substituent Group S;

[0011] R4 and R5 are each independently hydrogen, C 1-8 Alkyl (preferably C 1-6 Alkyl, more preferably C 1-3 alkyl), -C(O)OC 1-8 Alkyl (preferably -C(O)OC 1-6 Alkyl, more preferably -C(O)OC 1-3 alkyl), -C(O)C 1-8 Alkyl (preferably -C(O)C 1-6 Alkyl, more preferably -C(O)C 1-3 alkyl), -(C=N)-C 1-8 Alkyl (preferably -(C=N)-C 1-6 Alkyl, more preferably -(C=N)-C 1-3 alkyl) or -(C=N)-NR a0 R b0 ; or R4, R5 and the nitrogen atom to which they are connected together form a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle; wherein the 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle is unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxyl, carboxyl, C 1-3 Alkyl, C 1-3 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, halo C 1-3 Alkyl, halogenated C 1-3 Alkoxy, NR a1 R b1 、-SO2C 1-3 Alkyl, -S(O)C 1-3 Alkyl, -C(O)NR a1 R b1 、-C(O)OC 1-3 Alkyl, -OC(O)C 1-3 Alkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkyloxy, 3- to 6-membered heterocycloalkyl;

[0012] Z is N or CR Z ; where R Z is hydrogen, cyano, hydroxy, carboxyl, halogen (preferably fluorine or chlorine), C 1-8 Alkyl (preferably C 1-6 Alkyl, more preferably C 1-3 Alkyl), C 1-8Alkoxy (preferably C 1-6 Alkoxy, more preferably C 1-3 Alkoxy), -C(O)C 1-8 Alkyl (preferably -C(O)C 1-6 Alkyl, more preferably -C(O)C 1-3 alkyl), -C(O)OC 1-8 Alkyl (preferably -C(O)OC 1-6 Alkyl, more preferably -C(O)OC 1-3 Alkyl), -C(O)NR a0 R b0 , 5- to 6-membered heteroaryl or 8- to 10-membered heteroaryl; wherein said C 1-8 Alkyl, C 1-8 Alkoxy is unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxy, carboxyl, C 1-3 Alkyl, C 1-3 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, halo C 1-3 Alkyl, halogenated C 1-3 Alkoxy, NR a1 R b1 、-SO2C 1-3 Alkyl, -S(O)C 1-3 Alkyl, -C(O)NR a1 R b1 、-C(O)OC 1-3 Alkyl, -OC(O)C 1-3 Alkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl; wherein the phenyl, 5- to 6-membered heteroaryl is optionally substituted by 1, 2 or 3 substituents each independently selected from Substituent Group S;

[0013] R a 、R b are independently hydrogen, C 1-8 Alkyl (preferably C 1-6 Alkyl, more preferably C 1-3 Alkyl), C 3-8 Cycloalkyl (preferably C 3-6 Cycloalkyl), C 1-8 Alkoxy (preferably C 1-6 Alkoxy, more preferably C 1-3 alkoxy), cyano, hydroxy, carboxyl, halogen (preferably fluorine or chlorine), -C(O)NR a0 R b0 、-C(O)C 1-8Alkyl (preferably -C(O)C 1-6 Alkyl, more preferably -C(O)C 1-3 alkyl), -C(O)OC 1-8 Alkyl (preferably -C(O)OC 1-6 Alkyl, more preferably -C(O)OC 1-3 alkyl), -OC(O)C 1-8 Alkyl (preferably -OC(O)C 1-6 Alkyl, more preferably -OC(O)C 1-3 Alkyl), -SO2C 1-8 Alkyl (preferably -SO2C 1-6 Alkyl, more preferably -SO2C 1-3 Alkyl), -SO2NR a0 R b0 ; or R a 、R b are connected to form a fused 3 to 7-membered saturated or partially unsaturated monocyclic ring or a fused 3 to 7-membered saturated or partially unsaturated monocyclic ring; wherein said C 1-8 Alkyl, C 1-8 The alkoxy, 3- to 7-membered saturated or partially unsaturated monocyclic ring and 3- to 7-membered saturated or partially unsaturated monocyclic ring are unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxyl, carboxyl, C 1-3 Alkyl, C 1-3 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, halo C 1-3 Alkyl, halogenated C 1-3 Alkoxy, NR a1 R b1 、-SO2C 1-3 Alkyl, -S(O)C 1-3 Alkyl, -C(O)NR a1 R b1 、-C(O)OC 1-3 Alkyl, -OC(O)C 1-3 Alkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl; wherein the phenyl, 5- to 6-membered heteroaryl is optionally substituted by 1, 2 or 3 substituents each independently selected from Substituent Group S;

[0014] Ring A is a 3- to 7-membered saturated or partially unsaturated monocyclic heterocyclic ring or a 3- to 7-membered saturated or partially unsaturated monocyclic ring;

[0015] (R0) mIndicates that the hydrogen on ring A is replaced by m R0, m is 0, 1, 2 or 3; each R0 is the same or different and is independently cyano, hydroxyl, carboxyl, halogen (preferably fluorine or chlorine), C 1-8 Alkyl (preferably C 1-6 Alkyl, more preferably C 1-3 Alkyl), C 1-8 Alkoxy (preferably C 1-6 Alkoxy, more preferably C 1-3 Alkoxy), -C(O)C 1-8 Alkyl (preferably -C(O)C 1-6 Alkyl, more preferably -C(O)C 1-3 alkyl), -C(O)OC 1-8 Alkyl (preferably -C(O)OC 1-6 Alkyl, more preferably -C(O)OC 1-3 alkyl), -OC(O)C 1-8 Alkyl (preferably -OC(O)C 1-6 Alkyl, more preferably -OC(O)C 1-3 alkyl) or -C(O)NR a0 R b0 ; Or any two R0 connected to the same ring atom or different ring atoms are connected to form a 3-7 membered saturated or partially unsaturated monocyclic heterocycle or a 3-7 membered saturated or partially unsaturated monocyclic ring; wherein said C 1-8 Alkyl, C 1-8 The alkoxy, 3- to 7-membered saturated or partially unsaturated monocyclic ring and 3- to 7-membered saturated or partially unsaturated monocyclic ring are unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxyl, carboxyl, C 1-3 Alkyl, C 1-3 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, halo C 1-3 Alkyl, halogenated C 1-3 Alkoxy, NR a1 R b1 、-SO2C 1-3 Alkyl, -S(O)C 1-3 Alkyl, -C(O)NR a1 R b1 、-C(O)OC 1-3 Alkyl, -OC(O)C 1-3 Alkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl; wherein the phenyl, 5- to 6-membered heteroaryl is optionally substituted by 1, 2 or 3 substituents each independently selected from Substituent Group S;

[0016] The substituent group S consists of the following: halogen, cyano, hydroxyl, carboxyl, C 1-3 Alkyl, C 1-3 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, halo C 1-3 Alkyl, halogenated C 1-3 Alkoxy, NR a1 R b1 、-SO2C 1-3 Alkyl, -S(O)C 1-3 Alkyl, -C(O)NR a1 R b1 、-C(O)OC 1-3 Alkyl, -OC(O)C 1-3 Alkyl, C 3-6 Cycloalkyl, C 3-6 cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl;

[0017] R a0 、R b0 are independently hydrogen, C 1-3 Alkyl or acetyl; or R a0 、R b0 Together with the nitrogen atom to which it is connected, it forms a 4- to 6-membered saturated monocyclic heterocycle; the 4- to 6-membered saturated monocyclic heterocycle is optionally substituted by 1, 2 or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxyl, carboxyl, C 1-3 Alkyl, C 1-3 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, halo C 1-3 Alkyl, halogenated C 1-3 Alkoxy, -SO2C 1-3 Alkyl, -S(O)C 1-3 Alkyl, -C(O)NH2, -C(O)NH(C 1-3 alkyl), -C(O)N(C 1-3 Alkyl)2, -C(O)OC 1-3 Alkyl, -OC(O)C 1-3 Alkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkyloxy, 3- to 6-membered heterocycloalkyl;

[0018] R a1 、R b1 are independently hydrogen, C 1-3 Alkyl or acetyl; or R a1 、R b1Together with the nitrogen atom to which it is connected, it forms a 4- to 6-membered saturated monocyclic heterocycle; the 4- to 6-membered saturated monocyclic heterocycle is optionally substituted by 1, 2 or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxyl, carboxyl, C 1-3 Alkyl, C 1-3 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, halo C 1-3 Alkyl, halogenated C 1-3 Alkoxy, -SO2C 1-3 Alkyl, -S(O)C 1-3 Alkyl, -C(O)NH2, -C(O)NH(C 1-3 alkyl), -C(O)N(C 1-3 Alkyl)2, -C(O)OC 1-3 Alkyl, -OC(O)C 1-3 Alkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkyloxy, 3- to 6-membered heterocycloalkyl.

[0019] In some embodiments, the compound represented by formula (I) is represented by formula (II):

[0020]

[0021] wherein each group in the formula is as defined above.

[0022] In some embodiments, R1 is C 3-8 Cycloalkyl (preferably C 3-6 Cycloalkyl) or C 2-8 Alkenyl (preferably C 2-6 Alkenyl, more preferably C 2-4 R2, R3 are each independently hydrogen, C 1-8 Alkyl (preferably C 1-6 Alkyl, more preferably C 1-3 Alkyl), C 3-8 Cycloalkyl (preferably C 3-6 Cycloalkyl), halogenated C 1-8 Alkyl (preferably halogenated C 1-6 Alkyl, more preferably halogenated C 1-3 Alkyl) or deuterated C 1-8 Alkyl (preferably deuterated C 1-6 Alkyl, more preferably deuterated C 1-3 Alkyl); the C 3-8 Cycloalkyl, C 2-8 Alkenyl is unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxyl, carboxyl, C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C1-3 Alkyl, halogenated C 1-3 Alkoxy, NR a1 R b1 、-SO2C 1-3 Alkyl, -C(O)NR a1 R b1 、-C(O)C 1-3 Alkyl, -C(O)OC 1-3 alkyl.

[0023] In some embodiments, R1 is C 3-8 Cycloalkyl (preferably C 3-6 Cycloalkyl) or C 2-8 Alkenyl (preferably C 2-6 Alkenyl, more preferably C 2-4 R2 is hydrogen or C 1-8 Alkyl (preferably C 1-6 Alkyl, more preferably C 1-3 alkyl); R3 is hydrogen, C 1-8 Alkyl (preferably C 1-6 Alkyl, more preferably C 1-3 Alkyl), C 3-8 Cycloalkyl (preferably C 3-6 Cycloalkyl), halogenated C 1-8 Alkyl (preferably halogenated C 1-6 Alkyl, more preferably halogenated C 1-3 Alkyl) or deuterated C 1-8 Alkyl (preferably deuterated C 1-6 Alkyl, more preferably deuterated C 1-3 Alkyl); the C 3-8 Cycloalkyl, C 2-8 Alkenyl is unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxyl, carboxyl, C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkyl, halogenated C 1-3 Alkoxy, NR a1 R b1 .

[0024] In some embodiments, R1 is cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 1-methylcyclopropyl, 1-methylcyclobutyl, 1-methylcyclopentyl, 1-methylcyclohexyl, vinyl, propenyl, isopropenyl, n-butenyl, isobutenyl, pentenyl, hexenyl; R2 and R3 are each independently hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, cyclopropyl, cyclobutyl, Cyclopentyl, cyclohexyl, 1-methylcyclopropyl, 1-methylcyclobutyl, 1-methylcyclopentyl, 1-methylcyclohexyl, monochloromethyl, dichloromethyl, trichloromethyl, monochloroethyl, 1,2-dichloroethyl, trichloroethyl, monobromoethyl, monofluoromethyl, difluoromethyl, trifluoromethyl, monofluoroethyl, difluoroethyl, trifluoroethyl, monodeuteromethyl, monodeuteroethyl, dideuteromethyl, dideuteroethyl, trideuteromethyl, trideuteroethyl.

[0025] In some embodiments, Z is CH.

[0026] In some embodiments, the 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle in Ring A is selected from the group consisting of: azetidine ring, oxetane ring, tetrahydrofuran ring, tetrahydrothiophene ring, tetrahydropyrrole ring, piperidine ring, pyrroline ring, oxazolidine ring, piperazine ring, dioxolane ring, dioxane, morpholine ring, thiomorpholine ring, thiomorpholine-1,1-dioxide, tetrahydropyran ring, azetidine-2-one ring, oxetane-2-one ring, pyrrolidine-2-one ring, pyrrolidine-2,5-dione ring, piperidine-2-one ring, dihydrofuran-2(3H)-one ring, dihydrofuran-2,5-dione ring, tetrahydro-2H-pyran-2-one ring, piperazine-2-one ring, ... 1,2-dihydroazetidine ring, 1,2-dihydrooxetadiene ring, 2,5-dihydro-1H-pyrrole ring, 2,5-dihydrofuran ring, 2,3-dihydrofuran ring, 2,3-dihydro-1H-pyrrole ring, 3,4-dihydro-2H-pyran ring, 1,2,3,4-tetrahydropyridine ring, 3,6-dihydro-2H-pyran ring, 1,2,3,6-tetrahydropyridine ring, 4,5-dihydro-1H-imidazole ring, 1,4,5,6-tetrahydropyrimidine ring, 3,4,7,8-tetrahydro-2H-1,4,6-oxadiazolidine ring, 1,6-dihydropyrimidine ring, 4,5,6,7-tetrahydro-1H-1,3-diazepine ring Cyclic, 2,5,6,7-tetrahydro-1,3,5-oxadiazepine ring.

[0027] In some embodiments, the 3- to 7-membered saturated or partially unsaturated monocyclic ring in Ring A is selected from the group consisting of a cyclopropyl ring, a cyclobutyl ring, a cyclopentyl ring, a cyclopentenyl ring, a cyclohexyl ring, a cyclohexenyl ring, a cyclohexadienyl ring, a cycloheptyl ring, a cycloheptatrienyl ring, a cyclopentanone ring, and a cyclopentane-1,3-dione ring.

[0028] In some embodiments, the compound represented by formula (I) is represented by formula (III-a) or formula (III-b):

[0029]

[0030] Among them, R 1a For hydrogen, C 1-6 Alkyl (preferably C 1-3 Alkyl) or deuterated C 1-6 Alkyl (more preferably deuterated C 1-3 alkyl); n is 1, 2, 3, 4, 5 or 6; t is 0, 1, 2, 3 or 4; R 11 、R 12 are independently hydrogen, C 1-3 Alkyl or halogen, and the remaining groups are as defined above.

[0031] In some embodiments, R2 and R3 are each independently hydrogen, C 1-8 Alkyl (preferably C 1-6 Alkyl, more preferably C 1-3 Alkyl), C 3-8 Cycloalkyl (preferably C 3-6 Cycloalkyl), halogenated C 1-8 Alkyl (preferably halogenated C 1-6 Alkyl, more preferably halogenated C 1-3 Alkyl) or deuterated C 1-8 Alkyl (preferably deuterated C 1-6 Alkyl, more preferably deuterated C 1-3 Alkyl); the C 3-8 The cycloalkyl group is unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxyl, carboxyl, C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkyl, halogenated C 1-3 Alkoxy, NR a1 R b1 、-SO2C 1-3 Alkyl, -C(O)NR a1 R b1 、-C(O)C 1-3 Alkyl, -C(O)OC 1-3 alkyl.

[0032] In some embodiments, R2 is hydrogen or C 1-8 Alkyl (preferably C 1-6 Alkyl, more preferably C 1-3 alkyl); R3 is hydrogen, C 1-8 Alkyl (preferably C 1-6 Alkyl, more preferably C1-3 Alkyl), C 3-8 Cycloalkyl (preferably C 3-6 Cycloalkyl), halogenated C 1-8 Alkyl (preferably halogenated C 1-6 Alkyl, more preferably halogenated C 1-3 Alkyl) or deuterated C 1-8 Alkyl (preferably deuterated C 1-6 Alkyl, more preferably deuterated C 1-3 Alkyl); the C 3-8 The cycloalkyl group is unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxyl, carboxyl, C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkyl, halogenated C 1-3 Alkoxy, NR a1 R b1 .

[0033] In some embodiments, R2 and R3 are each independently hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 1-methylcyclopropyl, 1-methylcyclobutyl, 1-methylcyclopentyl, 1-methylcyclohexyl, monochloromethyl, dichloromethyl, trichloromethyl, monochloroethyl, 1,2-dichloroethyl, trichloroethyl, monobromoethyl, monofluoromethyl, difluoromethyl, trifluoromethyl, monofluoroethyl, difluoroethyl, trifluoroethyl, monodeuteromethyl, monodeuteroethyl, dideuteromethyl, dideuteroethyl, trideuteromethyl, trideuteroethyl.

[0034] In some embodiments, the compound represented by formula (III-a) is represented by formula (III-a-1) or formula (III-a-2):

[0035]

[0036] Wherein, R3' is C 1-8 Alkyl (preferably C 1-6 Alkyl, more preferably C 1-3 Alkyl), C 1-8 Alkoxy (preferably C 1-6 Alkoxy, more preferably C 1-3 alkoxy), cyano, hydroxy, carboxyl, halogen (preferably fluorine or chlorine), -C(O)NR a0 R b0 、-C(O)C 1-8 Alkyl (preferably -C(O)C 1-6 Alkyl, more preferably -C(O)C 1-3 alkyl), -C(O)OC 1-8Alkyl (preferably -C(O)OC 1-6 Alkyl, more preferably -C(O)OC 1-3 alkyl) or -OC(O)C 1-8 Alkyl (preferably -OC(O)C 1-6 Alkyl, more preferably -OC(O)C 1-3 Alkyl); wherein said C 1-8 Alkyl, C 1-8 Alkoxy is unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxy, carboxyl, C 1-3 Alkyl, C 1-3 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, halo C 1-3 Alkyl, halogenated C 1-3 Alkoxy, NR a1 R b1 、-SO2C 1-3 Alkyl, -S(O)C 1-3 Alkyl, -C(O)NR a1 R b1 、-C(O)OC 1-3 Alkyl, -OC(O)C 1-3 Alkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl; wherein the phenyl, 5- to 6-membered heteroaryl is optionally substituted by 1, 2 or 3 substituents each independently selected from Substituent Group S; the remaining groups are as defined above.

[0037] In some embodiments, the compound represented by formula (III-a) has a structure represented by formula (III-a-3):

[0038]

[0039] wherein the groups are as defined above.

[0040] In some embodiments, R3' is C 1-8 Alkyl (preferably C 1-6 Alkyl, more preferably C 1-3 Alkyl), halogenated C 1-8 Alkyl (preferably halogenated C 1-6 Alkyl, more preferably halogenated C 1-3 Alkyl) or deuterated C 1-8 Alkyl (preferably deuterated C 1-6 Alkyl, more preferably deuterated C 1-3 alkyl).

[0041] In some embodiments, R3' is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, monochloromethyl, dichloromethyl, trichloromethyl, monochloroethyl, 1,2-dichloroethyl, trichloroethyl, monobromoethyl, monofluoromethyl, difluoromethyl, trifluoromethyl, monofluoroethyl, difluoroethyl, trifluoroethyl, monodeuterated methyl, monodeuterated ethyl, dideuterated methyl, dideuterated ethyl, trideuterated methyl, trideuterated ethyl.

[0042] In some embodiments, in the structure of formula (III-a), formula (III-a-1) or formula (III-a-2), t is 1 or 2, m is 0, R4 is H, R5 is H, -C(O)C 1-3 Alkyl, -C(O)OC 1-3 Alkyl, -(C=N)-C 1-3 Alkyl or -(C=N)-NH2.

[0043] In some embodiments, R a 、R b are independently hydrogen, C 1-6 Alkyl (preferably C 1-3 Alkyl), C 3-6 Cycloalkyl, C 1-6 Alkoxy (preferably C 1-3 Alkoxy), halogenated C 1-6 Alkyl (preferably halogenated C 1-3 alkyl), cyano, hydroxy, carboxyl, halogen (preferably fluorine or chlorine), -C(O)NR a0 R b0 、-C(O)C 1-6 Alkyl (preferably -C(O)C 1-3 alkyl), -C(O)OC 1-6 Alkyl (preferably -C(O)OC 1-3 alkyl), -OC(O)C 1-6 Alkyl (preferably -OC(O)C 1-3 Alkyl), -SO2C 1-6 Alkyl (preferably -SO2C 1-3 Alkyl), -SO2NR a0 R b0 ; wherein said C 1-6 Alkyl, C 1-6 Alkoxy is unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxy, carboxyl, C 1-3 Alkyl, C 1-3 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, halo C 1-3 Alkyl, halogenated C 1-3Alkoxy, NR a1 R b1 、-SO2C 1-3 Alkyl, -S(O)C 1-3 Alkyl, -C(O)NR a1 R b1 、-C(O)OC 1-3 Alkyl, -OC(O)C 1-3 Alkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl; wherein the phenyl, 5- to 6-membered heteroaryl is optionally substituted by 1, 2 or 3 substituents each independently selected from Substituent Group S.

[0044] In some embodiments, R a 、R b are each independently hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, methoxy, ethoxy, n-propoxy, isopropoxy, monochloromethyl, dichloromethyl, trichloromethyl, monochloroethyl, 1,2-dichloroethyl, trichloroethyl, monobromoethyl, monofluoromethyl, difluoromethyl, trifluoromethyl, monofluoroethyl, difluoroethyl, trifluoroethyl, monodeuteromethyl, monodeuteroethyl, dideuteromethyl, dideuteroethyl, trideuteromethyl, trideuteroethyl, cyano, hydroxyl, carboxyl, halogen (preferably fluorine or chlorine), -C(O)NH2, -C(O)NH(C 1-3 alkyl), -C(O)N(C 1-3 Alkyl)2, -C(O)C 1-6 Alkyl (preferably -C(O)C 1-3 alkyl), -C(O)OC 1-6 Alkyl (preferably -C(O)OC 1-3 alkyl), -OC(O)C 1-6 Alkyl (preferably -OC(O)C 1-3 Alkyl), -SO2C 1-6 Alkyl (preferably -SO2C 1-3 Alkyl), -SO2NH2, -SO2NH(C 1-3 Alkyl), -SO2N(C 1-3 Alkyl)2.

[0045] In some embodiments, R4 and R5 are each independently hydrogen, C 1-8 Alkyl (preferably C 1-6 Alkyl, more preferably C 1-3 alkyl), -C(O)OC 1-8 Alkyl (preferably -C(O)OC 1-6Alkyl, more preferably -C(O)OC 1-3 alkyl); or R4, R5 and the nitrogen atom to which they are connected together form a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle; wherein the 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle is unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxyl, carboxyl, C 1-3 Alkyl, C 1-3 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, halo C 1-3 Alkyl, halogenated C 1-3 Alkoxy, NR a1 R b1 、-SO2C 1-3 Alkyl, -S(O)C 1-3 Alkyl, -C(O)NR a1 R b1 、-C(O)OC 1-3 Alkyl, -OC(O)C 1-3 Alkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkyloxy, 3- to 6-membered heterocycloalkyl;

[0046] In some embodiments, the 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle formed by R4, R5 and the nitrogen atom to which they are attached is a 4- to 6-membered saturated monocyclic heterocycle selected from the following: azetidine, tetrahydropyrrole ring, piperidine ring, piperazine ring, morpholine ring, thiomorpholine ring, thiomorpholine-1,1-dioxide, 1,2-dihydroazetadiene ring, 2,5-dihydro-1H-pyrrole ring, 2,3-dihydro-1H-pyrrole ring, 1,2,3,4-tetrahydropyridine ring or 1,2,3,6-tetrahydropyridine ring.

[0047] In some embodiments, R a 、R bThe fused 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle formed by connecting is a fused 4- to 6-membered saturated monocyclic heterocycle selected from the following: fused azetidine, fused oxetane, fused tetrahydrofuran ring, fused tetrahydrothiophene ring, fused tetrahydropyrrole ring, fused piperidine ring, fused piperazine ring, fused morpholine ring, fused thiomorpholine ring, fused thiomorpholine-1,1-dioxide, fused tetrahydropyran ring, fused 1,2-dihydro an azetadiene ring, a fused 1,2-dihydrooxetadiene ring, a fused 2,5-dihydro-1H-pyrrole ring, a fused 2,5-dihydrofuran ring, a fused 2,3-dihydrofuran ring, a fused 2,3-dihydro-1H-pyrrole ring, a fused 3,4-dihydro-2H-pyran ring, a fused 1,2,3,4-tetrahydropyridine ring, a fused 3,6-dihydro-2H-pyran ring or a fused 1,2,3,6-tetrahydropyridine ring.

[0048] In some embodiments, R a 、R b The fused 3- to 7-membered saturated or partially unsaturated monocyclic ring formed by connecting together is a fused 3- to 6-membered saturated monocyclic ring selected from the following: a fused cyclopropyl ring, a fused cyclobutyl ring, a fused cyclopentyl ring, a fused cyclopentenyl ring, a fused cyclohexyl ring, a fused cyclohexenyl ring, and a fused cyclohexadienyl ring.

[0049] In some embodiments, the 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle formed by any two R0s attached to the same ring atom or different ring atoms is a 4- to 6-membered saturated monocyclic heterocycle selected from the following: azetidine, oxetane, tetrahydrofuran ring, tetrahydrothiophene ring, tetrahydropyrrole ring, piperidine ring, piperazine ring, morpholine ring, thiomorpholine ring, thiomorpholine-1,1-dioxide, and tetrahydropyran ring.

[0050] In some embodiments, the 3- to 7-membered saturated or partially unsaturated monocyclic ring formed by any two R0 attached to the same ring atom or different ring atoms is a 3- to 6-membered saturated monocyclic ring selected from the following: cyclopropyl ring, cyclobutyl ring, cyclopentyl ring, and cyclohexyl ring.

[0051] In some embodiments, R a0 、R b0 The 4- to 6-membered saturated monocyclic heterocyclic ring formed together with the connected nitrogen atom is selected from: azetidine, oxetane, tetrahydrofuran ring, tetrahydrothiophene ring, tetrahydropyrrole ring, piperidine ring, piperazine ring, morpholine ring, thiomorpholine ring, thiomorpholine-1,1-dioxide, and tetrahydropyran ring.

[0052] In some embodiments, R a1 、R b1The 4- to 6-membered saturated monocyclic heterocyclic ring formed together with the connected nitrogen atom is selected from: azetidine, oxetane, tetrahydrofuran ring, tetrahydrothiophene ring, tetrahydropyrrole ring, piperidine ring, piperazine ring, morpholine ring, thiomorpholine ring, thiomorpholine-1,1-dioxide, and tetrahydropyran ring.

[0053] In some embodiments, in the above structural formulas, the 5- to 6-membered heteroaryl groups in each group are independently selected from the group consisting of: thienyl, furyl, thiazolyl, isothiazolyl, imidazolyl, oxazolyl, pyrrolyl, pyrazolyl, triazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, tetrazolyl, isoxazolyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl and tetrazinyl.

[0054] In some embodiments, the 5- to 6-membered heteroaryl group is selected from:

[0055]

[0056] The above 5- to 6-membered heteroaryl groups are unsubstituted or substituted by 1, 2 or 3 substituents independently selected from the group consisting of halogen, cyano, hydroxyl, carboxyl, C 1-3 Alkyl, C 1-3 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, halo C 1-3 Alkyl, halogenated C 1-3 Alkoxy, NR a1 R b1 、-SO2C 1-3 Alkyl, -S(O)C 1-3 Alkyl, -C(O)NR a1 R b1 、-C(O)OC 1-3 Alkyl, -OC(O)C 1-3 Alkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl; R a1 、R b1 As defined above.

[0057] In some embodiments, in the above structural formulas, the 8- to 10-membered heteroaryl group in each group is selected from the group consisting of indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzisothiazolyl, benzothiadiazolyl, indanyl, purinyl, pyrido[3,2-d]pyrimidinyl, pyrido[2,3-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrido[4,3-d]pyrimidinyl, 1,8-naphthyridinyl, 1,7-naphthyridinyl, 1,6-naphthyridinyl, 1,5-naphthyridinyl, pteridinyl, quinolyl, isoquinolyl, cinnolinyl, quinoxalinyl, phthalazinyl and quinazolinyl.

[0058] In some embodiments, in the above structural formulas, the 3- to 6-membered heterocycloalkyl groups in each group are independently selected from the following 4- to 6-membered heterocycloalkyl groups: azetidinyl, oxetanyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyrrolyl, oxazolidinyl, dioxolanyl, piperidinyl, piperazinyl, morpholinyl, dioxane, thiomorpholinyl, thiomorpholine-1,1-dioxide, tetrahydropyranyl, pyrrolidin-2-one, dihydrofuran-2(3H)-one, morpholin-3-one, piperazin-2-one, and piperidin-2-one.

[0059] In some embodiments, the compound of formula (I) is any one of the following compounds:

[0060]

[0061]

[0062]

[0063]

[0064]

[0065]

[0066] In some embodiments, the compound of formula (I) is any one of the compounds of the Examples.

[0067] The second aspect of the present invention provides a pharmaceutical composition comprising the compound according to the first aspect of the present invention, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a solvate thereof, or a prodrug thereof, and a pharmaceutically acceptable carrier.

[0068] A third aspect of the present invention provides use of the compound of the first aspect of the present invention, or a pharmaceutically acceptable salt, stereoisomer, solvate, or prodrug thereof, and the pharmaceutical composition of the second aspect of the present invention, in the preparation of a medicament for treating a disease associated with or mediated by CDK9 activity. The disease is preferably a hyperproliferative disease, a virus-induced infectious disease, and / or a cardiovascular disease, more preferably a hyperproliferative disease.

[0069] A fourth aspect of the present invention provides a method for treating a disease associated with or mediated by CDK9 activity, the method comprising administering to a subject an effective amount of the compound of the first aspect of the present invention, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a solvate thereof, or a prodrug thereof, or the pharmaceutical composition of the second aspect of the present invention. In certain embodiments, the disease associated with CDK9 activity is a hyperproliferative disease, particularly cancer, such as solid tumors and hematological tumors.

[0070] In the present invention, diseases associated with CDK9 activity or mediated by CDK9 activity include diseases associated with CDK9 activity (such as excessive activity of CDK9) or related to CDK9 activity, and the conditions associated with these diseases. The excessive activity of CDK9 refers to the increased CDK9 enzymatic activity compared to normal non-disease cells, or refers to the increased CDK9 activity that causes unwanted cell proliferation, or reduced or insufficient programmed cell death (apoptosis), or refers to a mutation that causes constitutive activation of CDK9. Hyperproliferative diseases include diseases involving the undesirable or uncontrolled proliferation of cells, and include diseases involving reduced or insufficient programmed cell death (apoptosis). The compounds of the present invention can be used to prevent, inhibit, block, reduce, reduce, control, etc., and / or produce apoptosis to cell proliferation and / or cell division. The method includes administering to a subject in need (including mammals, including humans) a certain amount of the compound of the present invention or its pharmaceutically acceptable salt, hydrate or solvate that effectively treats or prevents the disease.

[0071] Hyperproliferative diseases in the context of the present invention include, but are not limited to, for example, angiogenesis or angiogenic disorders, mesangial cell proliferative disorders, and solid tumors, such as cancers of the breast, respiratory tract, brain, reproductive organs, digestive tract, urinary tract, eye, liver, skin, head and neck, thyroid, parathyroid glands, and their distant metastases. Those diseases also include lymphomas, sarcomas, and leukemias. In some embodiments, the cancer is selected from pancreatic cancer, breast cancer, ovarian cancer, cervical cancer, and leukemia.

[0072] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. BRIEF DESCRIPTION OF THE DRAWINGS

[0073] Figure 1 The graph shows the changes in tumor volume after oral administration of compound D and compound Z-11-P1 once a day.

[0074] Figure 2 Figure 2 is a graph showing changes in mouse body weight after oral administration of compound D and compound Z-11-P1 once daily.

[0075] Figure 3 The graph shows the changes in tumor volume after oral administration of compound D and compound Z-5-2 once a day.

[0076] Figure 4 The graph shows the changes in body weight of mice after oral administration of compound D and compound Z-5-2 once a day. DETAILED DESCRIPTION

[0077] After extensive and in-depth research, the inventors unexpectedly discovered a class of substituted pyrazolo[1,5-a]pyrimidin-7-amine derivatives that exhibit significant selective CDK9 inhibitory activity and excellent in vivo pharmacokinetic activity. Therefore, this series of compounds is expected to be developed as drugs for the treatment and / or prevention of diseases associated with or mediated by CDK9 activity. Based on this, the inventors completed the present invention.

[0078] Definition of terms

[0079] In order to more clearly understand the technical content of the present invention, the terms of the present invention are further explained below.

[0080] "Alkyl" refers to straight-chain and branched saturated aliphatic hydrocarbon groups. 1-8 "Alkyl" refers to an alkyl group having 1 to 8 carbon atoms, preferably C 1-6 Alkyl, more preferably C 1-3Alkyl; Non-limiting examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl pentyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched-chain isomers thereof.

[0081] "Alkenyl" refers to a straight or branched unsaturated aliphatic hydrocarbon group having one or more carbon-carbon double bonds (C=C). 2-8 "Alkenyl" refers to an alkenyl group having 2 to 8 carbon atoms, preferably C 2-6 Alkenyl, more preferably C 2-4 Alkenyl is similarly defined; non-limiting examples of alkenyl include ethenyl, propenyl, isopropenyl, n-butenyl, isobutenyl, pentenyl, hexenyl, and the like.

[0082] "Alkynyl" refers to a straight-chain or branched unsaturated aliphatic hydrocarbon group having one or more carbon-carbon triple bonds. 2-8 "Alkynyl" refers to an alkynyl group having 2 to 8 carbon atoms, preferably C 2-6 Alkynyl, more preferably C 2-4 Alkynyl is similarly defined; non-limiting examples of alkynyl include ethynyl, propynyl, n-butynyl, isobutynyl, pentynyl, hexynyl, and the like.

[0083] "Cycloalkyl" and "cycloalkyl ring" are used interchangeably and refer to a saturated monocyclic, bicyclic or polycyclic hydrocarbon group, or a group fused with an aryl or heteroaryl group, or an optionally substituted form thereof. In certain embodiments, the cycloalkyl ring contains one or more carbonyl groups, such as an oxo group. "C 3-8"Cycloalkyl" refers to a monocyclic cycloalkyl group having 3 to 8 carbon atoms. Non-limiting examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclobutanone, cyclopentanone, cyclopentane-1,3-dione, etc. Preferably, C 3-6 Cycloalkyl, including cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl. 8-10 "Cycloalkyl" refers to a fused bicyclic hydrocarbon radical having 8 to 10 ring atoms, C 8-10 Non-limiting examples of cycloalkyl groups include

[0084] "Spirocyclyl" and "spirocycle" are used interchangeably and refer to polycyclic hydrocarbon groups in which the monocyclic rings share a carbon atom (called a spiro atom). "7- to 11-membered spirocyclyl" refers to a spirocycle with 7 to 11 ring atoms. Spirocycles are classified as bispirocycles or polyspirocycles based on the number of rings, preferably bispirocycles. More preferably, they are 4-membered / 5-membered, 5-membered / 5-membered, or 5-membered / 6-membered bispirocycles. For example:

[0085]

[0086] "Cycloalkenyl" and "cycloalkenyl ring" are used interchangeably and refer to a monocyclic, bicyclic, or polycyclic hydrocarbon group containing one or more carbon-carbon double bonds within the ring, which may be fused to an aryl or heteroaryl group. The cycloalkenyl ring may be optionally substituted. In certain embodiments, the cycloalkenyl ring contains one or more carbonyl groups, such as an oxo group. "C 3-8 "Cycloalkenyl" refers to a monocyclic cycloalkenyl group having 3 to 8 carbon atoms. Preferably, C 3-6 Cycloalkenyl. Non-limiting examples of cycloalkenyl groups include cyclobutenyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl, cycloheptatrienyl, cyclopentyl-2-en-1-one, cyclohexyl-2,5-dien-1-one, cyclohexyl-2-en-1-one, cyclohex-2-ene-1,4-dione, and the like.

[0087] "Heterocycloalkyl" and "heterocycloalkyl ring" are used interchangeably and refer to a cycloalkyl group containing at least one heteroatom, such as nitrogen, oxygen and sulfur, on the ring atoms, which group may be fused to an aryl or heteroaryl group. The heterocycloalkyl ring may be optionally substituted. In certain embodiments, the heterocycloalkyl ring contains one or more carbonyl or thiocarbonyl groups, such as groups containing oxo and thio. "3 to 8 membered heterocycloalkyl" refers to a monocyclic cyclic hydrocarbon group having 3 to 8 ring atoms, wherein 1, 2 or 3 ring atoms are heteroatoms selected from nitrogen, oxygen and sulfur, preferably 4 to 8 membered heterocycloalkyl. More preferably, it is a 3 to 6 membered heterocycloalkyl group having 3 to 6 ring atoms, wherein 1 or 2 ring atoms are heteroatoms selected from nitrogen, oxygen and sulfur. More preferably, it is a 4 to 6 membered heterocycloalkyl group having 4 to 6 ring atoms, wherein 1 or 2 ring atoms are heteroatoms selected from nitrogen, oxygen and sulfur. Non-limiting examples include aziridine, oxiranyl, azetidinyl, oxetanyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyrrolyl, oxazolidinyl, dioxolanyl, piperidinyl, piperazinyl, morpholinyl, dioxane, thiomorpholinyl, thiomorpholine-1,1-dioxide, tetrahydropyranyl, azetidin-2-onyl, oxetan-2-onyl, dihydrofuran-2(3H)-onyl, pyrrolidin-2-onyl, pyrrolidin-2,5-dionyl, dihydrofuran-2,5-dionyl, piperidin-2-onyl, tetrahydro-2H-pyran-2-onyl, piperazin-2-onyl, morpholin-3-onyl, and the like. "6- to 12-membered heterocycloalkyl" and "6- to 12-membered fused heterocycloalkyl" are used interchangeably and refer to fused bicyclic cyclic hydrocarbon groups having 6 to 12 ring atoms, of which 1, 2, or 3 ring atoms are heteroatoms selected from nitrogen, oxygen, and sulfur. "8- to 10-membered heterocycloalkyl" and "8- to 10-membered fused heterocycloalkyl" are used interchangeably and refer to fused bicyclic cyclic hydrocarbon groups having 8 to 10 ring atoms, of which 1, 2, or 3 ring atoms are heteroatoms selected from nitrogen, oxygen, and sulfur. Non-limiting examples include hexahydro-1H-furo[3,4-c]pyrrole, octahydro-1H-cyclopenta[c]pyridine, hexahydro-1H-pyrrolo[2,1-c][1,4]oxazine, octahydropyrrolo[1,2-a]pyrazine, hexahydropyrrolo[1,2-a]pyrazin-4(1H)-one, octahydrocyclopenta[c]pyrrole, and the like. In fused bicyclic heterocycloalkyl groups containing one or more nitrogen atoms, the point of attachment may be a carbon or nitrogen atom as valence permits.Bicyclic heterocycloalkyl systems may include one or more heteroatoms in one or both rings.

[0088] "Heterospirocyclyl" and "heterospirocycle" are used interchangeably and refer to a monovalent non-aromatic ring system having two monocyclic rings sharing one carbon atom, which consists of carbon atoms and heteroatoms selected from nitrogen, oxygen, sulfur and phosphorus, contains no unsaturation, and is connected to the parent nucleus by a single bond. The heterospirocycle can be optionally substituted. In certain embodiments, the heterospirocycle contains one or more carbonyl or thiocarbonyl groups, such as groups comprising oxo and thio. "7 to 11 membered heterospirocyclyl" refers to a heterospirocyclyl having 7 to 11 ring atoms, of which 1, 2 or 3 ring atoms are heteroatoms. Non-limiting examples of heterospirocyclyl include 2,6-diazaspiro[3.4]octan-5-onyl, 2-oxo-6-azaspiro[3.3]heptan-2-yl, 6-oxaspiro[3.3]heptan-2-yl, 7-methyl-7-azaspiro[3.5]nonan-2-yl, 7-methyl-2,7-diazaspiro[3.5]nonan-2-yl, 9-methyl-9-phosphaspiro[5.5]undec-3-yl, and the like.

[0089] "Heterocycloalkenyl" and "heterocycloalkenyl ring" are used interchangeably and refer to a heterocycloalkyl group containing one or more carbon-carbon double bonds or carbon-nitrogen double bonds in the ring backbone, but are not intended to include heteroaryl moieties as defined herein. This group may be fused to an aryl or heteroaryl group. The heterocycloalkenyl ring may be optionally substituted. In certain embodiments, the heterocycloalkenyl ring contains one or more carbonyl or thiocarbonyl groups, such as groups containing oxo and thio. "5- to 8-membered heterocycloalkenyl ring" refers to a heterocycloalkenyl ring having 5 to 8 ring atoms, wherein 1, 2 or 3 of the ring atoms are heteroatoms selected from nitrogen, oxygen and sulfur. Preferably, the heterocycloalkenyl ring is a 5- to 6-membered heterocycloalkenyl ring. Non-limiting examples of heterocycloalkenyl rings include 4,5-dihydro-1H-imidazole ring, 1,4,5,6-tetrahydropyrimidine ring, 3,4,7,8-tetrahydro-2H-1,4,6-oxadiazolidine ring, 1,6-dihydropyrimidine ring, 4,5,6,7-tetrahydro-1H-1,3-diazepine ring, Cyclic, 2,5,6,7-tetrahydro-1,3,5-oxadiazepine ring.

[0090] "Aryl" and "aromatic ring" are used interchangeably to refer to an all-carbon monocyclic or fused polycyclic (i.e., rings that share adjacent pairs of carbon atoms) group having a conjugated π electron system, which may be fused to a cycloalkyl ring, a heterocycloalkyl ring, a cycloalkenyl ring, a heterocycloalkenyl ring, or a heteroaryl ring. 6-10 The term "aryl" refers to a monocyclic or bicyclic aromatic group having 6 to 10 carbon atoms. Non-limiting examples of aryl include phenyl, naphthyl, and the like.

[0091] "Heteroaryl" and "heteroaryl ring" are used interchangeably and refer to a group of a monocyclic, bicyclic or polycyclic 4n+2 aromatic ring system (e.g., having 6 or 10 π electrons shared in a cyclic arrangement) having ring carbon atoms and ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen and sulfur. In the present invention, heteroaryl also includes a ring system in which the above-mentioned heteroaryl ring is fused to one or more cycloalkyl rings, heterocycloalkyl rings, cycloalkenyl rings, heterocycloalkenyl rings or aromatic rings. The heteroaryl ring may be optionally substituted. "5 to 10 membered heteroaryl" refers to a monocyclic or bicyclic heteroaryl group having 5 to 10 ring atoms, wherein 1, 2, 3 or 4 ring atoms are heteroatoms. "5- to 6-membered heteroaryl" refers to a monocyclic heteroaryl group having 5 to 6 ring atoms, wherein 1, 2, 3 or 4 of the ring atoms are heteroatoms, non-limiting examples of which include thienyl, furanyl, thiazolyl, isothiazolyl, imidazolyl, oxazolyl, pyrrolyl, pyrazolyl, triazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, tetrazolyl, isoxazolyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, and tetrazinyl. "8- to 10-membered heteroaryl" refers to a bicyclic heteroaryl group having 8 to 10 ring atoms, wherein 1, 2, 3 or 4 of the ring atoms are heteroatoms, non-limiting examples of which include indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzisothiazolyl, The term "heteroatom" refers to nitrogen, oxygen, or sulfur. In heteroaryl groups containing one or more nitrogen atoms, the point of attachment may be a carbon or nitrogen atom, as valence permits. Heteroaryl bicyclic ring systems may include one or more heteroatoms in one or both rings.

[0092] "Fused" refers to structures in which two or more rings share one or more bonds.

[0093] "Benzoheterocycloalkyl" refers to a group in which a benzene ring is fused to a heterocycloalkyl ring to form a bicyclic, tricyclic, or polycyclic ring system, wherein the heterocycloalkyl ring is as defined above. "7- to 11-membered phenylheterocycloalkyl" refers to a bicyclic cyclic group having 7 to 11 ring atoms, of which 1, 2, 3, or 4 are heteroatoms selected from nitrogen, oxygen, and sulfur. Preferably, it is an 8- to 10-membered phenylheterocycloalkyl group having 8 to 10 ring atoms, of which 1, 2, or 3 are heteroatoms selected from nitrogen, oxygen, and sulfur. Non-limiting examples include indoline, benzo[d][1,3]dioxazole, 1,2,3,4-tetrahydroisoquinoline, 3,4-dihydro-2H-benzo[b][1,4]oxazine, and the like.

[0094] "Heteroarylheterocycloalkyl" refers to a group in which a heteroaryl ring is fused to a heterocycloalkyl ring to form a bicyclic, tricyclic, or polycyclic ring system, wherein the heterocycloalkyl ring is as defined above. "7- to 11-membered heteroarylheterocycloalkyl" refers to a bicyclic cyclic group having 7 to 11 ring atoms, of which 1, 2, 3, or 4 are heteroatoms selected from nitrogen, oxygen, and sulfur. Preferably, it is an 8- to 10-membered heteroarylheterocycloalkyl group having 8 to 10 ring atoms, of which 1, 2, 3, or 4 are heteroatoms selected from nitrogen, oxygen, and sulfur. Non-limiting examples include 2,3-dihydro-1H-pyrrolo[2,3-b]pyridine, [1,3]dioxolane[4,5-b]pyridine, 2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazine, 2,3,4,6-tetrahydropyrrolo[3,4-b][1,4]oxazine, 2,4,5,6-tetrahydropyrano[2,3-c]pyrazole, 5,6,7,8-tetrahydropyrido[3,4-d]pyrimidine, and the like.

[0095] "Alkoxy" refers to an -O-alkyl group, wherein alkyl is as defined above. Preferably C 1-8 Alkoxy, more preferably C 1-6 Alkoxy, most preferably C 1-3 Alkoxy. Non-limiting examples of alkoxy include methoxy, ethoxy, n-propoxy, isopropoxy, butoxy, tert-butoxy, isobutoxy, pentoxy, and the like.

[0096] "Cycloalkyloxy" refers to an -O-cycloalkyl group, wherein the cycloalkyl group is as defined above. Preferably C 3-8 Cycloalkyloxy, more preferably C 3-6 Cycloalkyloxy. Non-limiting examples of cycloalkyloxy include cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like.

[0097] "A bond" means that the two groups connected thereto are connected by one covalent bond.

[0098] "Halogen" refers to fluorine, chlorine, bromine or iodine.

[0099] "Halo" refers to a group in which one or more (eg, 1, 2, 3, 4, or 5) hydrogen atoms are replaced by a halogen.

[0100] For example, "haloalkyl" refers to an alkyl group substituted with one or more (e.g., 1, 2, 3, 4, or 5) halogens, wherein alkyl is as defined above. 1-8 Alkyl, more preferably halogenated C 1-6 Alkyl, more preferably halogenated C 1-3 Examples of haloalkyl groups include, but are not limited to, monochloromethyl, dichloromethyl, trichloromethyl, monochloroethyl, 1,2-dichloroethyl, trichloroethyl, monobromoethyl, monofluoromethyl, difluoromethyl, trifluoromethyl, monofluoroethyl, difluoroethyl, trifluoroethyl, and the like.

[0101] For another example, "haloalkoxy" refers to an alkoxy group substituted by one or more (such as 1, 2, 3, 4 or 5) halogens, wherein the definition of alkoxy is as described above. 1-8 Alkoxy, more preferably halogenated C 1-6 Alkoxy, more preferably halogenated C 1-3 Alkoxy. Haloalkoxy includes, but is not limited to, trifluoromethoxy, trifluoroethoxy, monofluoromethoxy, monofluoroethoxy, difluoromethoxy, difluoroethoxy, and the like.

[0102] For another example, "halocycloalkyl" refers to a cycloalkyl group substituted by one or more (such as 1, 2, 3, 4 or 5) halogens, wherein the definition of cycloalkyl is as described above. 3-8 Cycloalkyl, more preferably halogenated C 3-6 Cycloalkyl. Halocycloalkyl includes, but is not limited to, trifluorocyclopropyl, monofluorocyclopropyl, monofluorocyclohexyl, difluorocyclopropyl, difluorocyclohexyl, and the like.

[0103] "Deuterated alkyl" refers to an alkyl group substituted with one or more (e.g., 1, 2, 3, 4, or 5) deuterium atoms, wherein the definition of alkyl is as described above. 1-8 Alkyl, more preferably deuterated C 1-6 Alkyl, more preferably deuterated C 1-3 Examples of deuterated alkyl groups include, but are not limited to, monodeuterated methyl, monodeuterated ethyl, dideuterated methyl, dideuterated ethyl, trideuterated methyl, trideuterated ethyl, and the like.

[0104] "Amino" refers to NH2, "cyano" refers to CN, "nitro" refers to NO2, "benzyl" refers to -CH2-phenyl, "oxo" refers to =O, "carboxyl" refers to -C(O)OH, "acetyl" refers to -C(O)CH3, "hydroxymethyl" refers to -CH2OH, "hydroxyethyl" refers to -CH2CH2OH or -CHOHCH3, "hydroxy" refers to -OH, "thiol" refers to SH, and the structure of "cyclopropylene" is:

[0105] "Saturated or partially unsaturated monocyclic ring" refers to a saturated or partially unsaturated all-carbon monocyclic ring system, wherein "partially unsaturated" refers to a ring portion including at least one double bond or triple bond, and "partially unsaturated" is intended to cover rings with multiple unsaturated sites, but is not intended to include aryl or heteroaryl moieties as defined herein. In certain embodiments, the saturated or partially unsaturated monocyclic ring contains one or more carbonyl groups, such as an oxo group. "3 to 7 membered saturated or partially unsaturated monocyclic ring" has 3 to 7 ring carbon atoms, preferably a saturated or partially unsaturated monocyclic ring with 3 to 6 ring carbon atoms, more preferably a saturated monocyclic ring with 3 to 6 ring carbon atoms. Non-limiting examples of saturated or partially unsaturated monocyclic rings include cyclopropyl ring, cyclobutyl ring, cyclopentyl ring, cyclopentenyl ring, cyclohexyl ring, cyclohexenyl ring, cyclohexadienyl ring, cycloheptyl ring, cycloheptatrienyl ring, cyclopentanone ring, cyclopentane-1,3-dione ring, etc.

[0106] "Saturated or partially unsaturated monocyclic heterocycle" means a saturated or partially unsaturated monocyclic ring in which one, two or three carbon atoms are selected from nitrogen, oxygen or S(O) t(wherein t is an integer from 0 to 2) is substituted with a heteroatom, excluding the ring portion of -OO-, -OS- or -SS-, and the remaining ring atoms are carbon. A "3- to 7-membered saturated or partially unsaturated monocyclic heterocycle" has 3 to 7 ring atoms, of which 1, 2 or 3 ring atoms are the above-mentioned heteroatoms. Preferably, the 3- to 6-membered saturated or partially unsaturated monocyclic heterocycle has 3 to 6 ring atoms, of which 1 or 2 ring atoms are the above-mentioned heteroatoms. More preferably, the 5- to 6-membered saturated or partially unsaturated monocyclic heterocycle has 5 to 6 ring atoms, of which 1 or 2 ring atoms are the above-mentioned heteroatoms. Most preferably, the 5- or 6-membered saturated monocyclic heterocycle has 5 to 6 ring atoms, of which 1 or 2 ring atoms are the above-mentioned heteroatoms. Non-limiting examples of saturated monocyclic heterocycles include an oxetane ring, an azetidine ring, an oxetane ring, a tetrahydrofuran ring, a tetrahydrothiophene ring, a tetrahydropyrrole ring, a piperidine ring, a pyrroline ring, an oxazolidine ring, a piperazine ring, a dioxolane ring, a dioxane ring, a morpholine ring, a thiomorpholine ring, a thiomorpholine-1,1-dioxide, a tetrahydropyran ring, an azetidine-2-one ring, an oxetane-2-one ring, a pyrrolidine-2-one ring, a pyrrolidine-2,5-dione ring, a piperidin-2-one ring, a dihydrofuran-2(3H)-one ring, a dihydrofuran-2,5-dione ring, a tetrahydro-2H-pyran-2-one ring, a piperazin-2-one ring, and a morpholine-3-one ring. Non-limiting examples of partially unsaturated monocyclic heterocycles include 1,2-dihydroazetidine ring, 1,2-dihydrooxetadiene ring, 2,5-dihydro-1H-pyrrole ring, 2,5-dihydrofuran ring, 2,3-dihydrofuran ring, 2,3-dihydro-1H-pyrrole ring, 3,4-dihydro-2H-pyran ring, 1,2,3,4-tetrahydropyridine ring, 3,6-dihydro-2H-pyran ring, 1,2,3,6-tetrahydropyridine ring, 4,5-dihydro-1H-imidazole ring, 1,4,5,6-tetrahydropyrimidine ring, 3,4,7,8-tetrahydro-2H-1,4,6-oxadiazolidine ring, 1,6-dihydropyrimidine ring, 4,5,6,7-tetrahydro-1H-1,3-diazepine ring. Cyclic, 2,5,6,7-tetrahydro-1,3,5-oxadiazepine Ring, etc.

[0107] "Substituted" means that one or more hydrogen atoms, preferably 1 to 5 hydrogen atoms, and more preferably 1 to 3 hydrogen atoms, in a group are independently replaced by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) which substitutions are possible or impossible without undue effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom with an unsaturated (e.g., olefinic) bond.

[0108] Unless otherwise defined, the "substituents independently selected from..." described in the present invention means that when more than one hydrogen on a group is replaced by a substituent, the substituents may be the same or different, and the substituents selected are independently of each other.

[0109] Unless otherwise defined, the phrase "...same or different, and each independently is..." in the present invention means that when there are more than one identical substituent group in the general formula, the substituent group may be the same or different and are each independent species. For example, L is (CR 01 R 02 ) s , when s is 2, that is, L is (CR 01 R 02 )-(CR 01 R 02 ), where two R 01 or R 02 They can be the same or different and are independent species. For example, L can be C(CH3)(CN)-C(CH2CH3)(OH), C(CH3)(CN)-C(CH3)(OH) or C(CN)(CH2CH3)-C(OH)(CH2CH3).

[0110] Unless otherwise defined, any group herein may be substituted or unsubstituted. When the above groups are substituted, the substituents are preferably 1 to 5 groups independently selected from the following groups: cyano, halogen (preferably fluorine or chlorine), C 1-8 Alkyl (preferably C 1-6 Alkyl, more preferably C 1-3 Alkyl), C 1-8 Alkoxy (preferably C 1-6 Alkoxy, more preferably C 1-3 Alkoxy), halogenated C 1-8 Alkyl (preferably halogenated C 1-6 Alkyl, more preferably halogenated C 1-3 Alkyl), C 3-8 Cycloalkyl (preferably C 3-6 Cycloalkyl), halogenated C 1-8 Alkoxy (preferably halogenated C 1-6 Alkoxy, more preferably halo C 1-3 Alkoxy), C 1-8 Alkyl-substituted amino, halogenated C 1-8 Alkyl-substituted amino, acetyl, hydroxy, hydroxymethyl, hydroxyethyl, carboxyl, nitro, C 6-10 Aryl (preferably phenyl), C 3-8 Cycloalkyloxy (preferably C 3-6 Cycloalkyloxy), C 2-8 Alkenyl (preferably C 2-6 Alkenyl, more preferably C 2-4alkenyl), C 2-8 Alkynyl (preferably C 2-6 Alkynyl, more preferably C 2-4 Alkynyl), -CONR a0 R b0 、-C(O)OC 1-10 Alkyl (preferably -C(O)OC 1-6 Alkyl, more preferably -C(O)OC 1-3 alkyl), -CHO, -OC(O)C 1-10 Alkyl (preferably -OC(O)C 1-6 Alkyl, more preferably -OC(O)C 1-3 Alkyl), -SO2C 1-10 Alkyl (preferably -SO2C 1-6 Alkyl, more preferably -SO2C 1-3 Alkyl), -SO2C 6-10 Aryl (preferably -SO2C6 aryl, such as -SO2-phenyl), -COC 6-10 Aryl (preferably -COC6 aryl, such as -CO-phenyl), 4 to 6-membered saturated or unsaturated monocyclic heterocyclic ring, 4 to 6-membered saturated or unsaturated monocyclic ring, 5 to 6-membered monocyclic heteroaryl ring, 8 to 10-membered bicyclic heteroaryl ring, spiro ring, spiroheterocycle, bridged ring or bridged heterocycle, wherein R a0 、R b0 are each independently hydrogen or C 1-3 alkyl.

[0111] Each type of substituent group described herein above can itself be substituted with the groups described herein.

[0112] When the 4- to 6-membered saturated monocyclic heterocycle described herein is substituted, the substituents may be positioned at their possible chemical positions. Representative substitutions of exemplary monocyclic heterocycles are shown below:

[0113] Wherein "Sub" represents various substituents described herein; Indicates connections to other atoms.

[0114] Pharmaceutical composition

[0115] Generally, the compounds of the present invention or their pharmaceutically acceptable salts, or solvates, or stereoisomers, or prodrugs can be administered in a suitable dosage form with one or more pharmaceutical carriers. These dosage forms are suitable for oral, rectal, topical, oral, and other parenteral administrations (e.g., subcutaneous, intramuscular, intravenous, etc.). For example, dosage forms suitable for oral administration include capsules, tablets, granules, and syrups. The compounds of the present invention contained in these preparations can be solid powders or granules; solutions or suspensions in aqueous or non-aqueous liquids; water-in-oil or oil-in-water emulsions, etc. The above dosage forms can be prepared by common pharmaceutical methods from active compounds and one or more carriers or excipients. The above carriers need to be compatible with the active compounds or other excipients. For solid preparations, commonly used non-toxic carriers include, but are not limited to, mannitol, lactose, starch, magnesium stearate, cellulose, glucose, sucrose, etc. Carriers for liquid preparations include water, physiological saline, aqueous glucose solution, ethylene glycol, and polyethylene glycol, etc. The active compound can form a solution or suspension with the above carriers.

[0116] "Pharmaceutically acceptable carrier" refers to a non-toxic, inert, solid, semi-solid substance or liquid filler, diluent, encapsulating material or auxiliary formulation or any type of excipient that is compatible with the subject to which it is administered (preferably a mammal, more preferably a human) and is suitable for transporting the active agent to the target site without terminating the activity of the agent.

[0117] "Active substance of the present invention" or "active compound of the present invention" refers to the compound of formula (I) of the present invention, or a pharmaceutically acceptable salt, or a solvate, or a stereoisomer, or a prodrug thereof, which has higher CDK9 selective inhibitory activity.

[0118] The compositions of the present invention are formulated, dosed, and administered in a manner consistent with standard medical practice. The "therapeutically effective amount" of the compound administered is determined by factors such as the specific condition to be treated, the individual being treated, the cause of the condition, the target of the drug, and the mode of administration.

[0119] "Therapeutically effective amount" refers to an amount of a compound of the present invention or a pharmaceutically acceptable salt thereof, or a solvate thereof, or a stereoisomer thereof, or a prodrug thereof that will elicit a biological or medical response in a subject, such as reducing or inhibiting enzyme or protein activity or improving symptoms, alleviating symptoms, slowing or delaying disease progression, or preventing disease.

[0120] The therapeutically effective amount of the compound of the present invention or its pharmaceutically acceptable salt, solvate, stereoisomer, or prodrug contained in the pharmaceutical composition or pharmaceutical use composition of the present invention is preferably 0.1 mg / kg-5 g / kg (body weight).

[0121] "Subject" refers to an animal, preferably a mammal, more preferably a human. The term "mammal" refers to warm-blooded vertebrate mammals, including cats, dogs, rabbits, bears, foxes, wolves, monkeys, deer, mice, pigs and humans.

[0122] "Treatment" refers to alleviating, slowing the progression of, attenuating, preventing, or maintaining an existing disease or condition (eg, cancer). Treatment also includes curing, preventing the development of, or alleviating to some extent, one or more symptoms of a disease or condition.

[0123] The term "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts. Pharmaceutically acceptable acid addition salts are salts formed with inorganic or organic acids that retain the biological effectiveness of the free base without any other side effects. These salts can be prepared by methods known in the art.

[0124] "Pharmaceutically acceptable base addition salts" include, but are not limited to, salts with inorganic bases, such as sodium salts, potassium salts, calcium salts, and magnesium salts. They also include, but are not limited to, salts with organic bases, such as ammonium salts, triethylamine salts, lysine salts, and arginine salts. These salts can be prepared by methods known in the art.

[0125] As used herein, "solvates" refer to complexes formed between a compound of the present invention and a solvent. These complexes react in, precipitate, or crystallize from a solvent. For example, a complex formed with water is referred to as a "hydrate." Solvates of compounds of formula (I) are within the scope of this invention.

[0126] When the compound represented by formula (I) of the present invention contains one or more chiral centers, it may exist in different optically active forms. When the compound of formula (I) contains one chiral center, the compound comprises a pair of enantiomers. The two enantiomers of the compound and the mixture of the pair of enantiomers, such as a racemic mixture, are also within the scope of protection of the present invention. Enantiomers can be resolved by methods known in the art, such as crystallization and chiral chromatography. When the compound of formula (I) contains more than one chiral center, the compound comprises enantiomers and diastereomers. All enantiomers and diastereomers of the compound, as well as mixtures of enantiomers, mixtures of diastereomers, and mixtures of enantiomers and diastereomers are also within the scope of protection of the present invention. Enantiomers and diastereomers can be resolved by methods known in the art, such as crystallization and preparative chromatography.

[0127] The present invention includes prodrugs of the above-mentioned compounds. Prodrugs include known amino protecting groups and carboxyl protecting groups, which are hydrolyzed under physiological conditions or released via enzymatic reactions to yield the parent compound. Specific methods for preparing prodrugs can be found in (Saulnier, MG; Frennesson, DB; Deshpande, MS; Hansel, SB and Vysa, DM Bioorg. Med. Chem Lett. 1994, 4, 1985-1990; and Greenwald, RB; Choe, YH; Conover, CD; Shum, K.; Wu, D.; Royzen, MJ Med. Chem. 2000, 43, 475.).

[0128] Preparation method

[0129] The present invention provides methods for preparing compounds of formula (I), which can be synthesized using standard synthetic techniques known to those skilled in the art or using methods known in the art in combination with the methods described herein. The solvents, temperatures, and other reaction conditions provided herein can be varied according to the skill in the art. The reactions can be used sequentially to provide compounds of the present invention, or they can be used to synthesize fragments that are subsequently added by the methods described herein and / or methods known in the art.

[0130] The compound described in the present invention can use the illustrative method described in the following similar method or embodiment, or the relevant open literature used by those skilled in the art, by using appropriate selectable starting material synthetic compound.The starting material for synthesizing the compound described in the present invention can be synthesized or can be obtained from commercial sources.The compound described in the present invention and other related compounds with different substituents can be synthesized using technology and raw materials known to those skilled in the art.The general method for preparing the compound disclosed in the present invention can be from reaction known in the art, and the reaction can be modified by those skilled in the art to consider appropriate reagents and conditions to introduce the various parts in the molecule provided by the present invention.

[0131] Compared with traditional technologies, the main advantages of the present invention are:

[0132] Provides a series of novel substituted pyrazolo[1,5-a]pyrimidin-7-amine derivatives with high selective inhibitory activity against CDK9 and excellent in vivo pharmacokinetic activity, IC 50 The value is less than 100 nM, preferably less than 50 nM, more preferably less than 10 nM, and thus can be used as a drug for treating and / or preventing diseases associated with or mediated by CDK9 activity.

[0133] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the invention. The experimental methods for which specific conditions are not specified in the following examples are generally based on conventional conditions such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight. Unless otherwise defined, the terms used herein have the same meaning as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein may be applied to the present invention.

[0134] Known starting materials can be synthesized by methods known in the art, or can be purchased from companies such as ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, and Darui Chemicals.

[0135] Unless otherwise specified, the reactions in the examples were carried out under nitrogen or argon atmosphere.

[0136] DMF: dimethylformamide, DMSO: dimethyl sulfoxide, THF: tetrahydrofuran, DIEA: N,N-diisopropylethylamine, EA: ethyl acetate, PE: petroleum ether, BINAP: (2R,3S)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl, NBS: N-bromosuccinimide, NCS: N-chlorosuccinimide, CDI: N,N'-carbonyldiimidazole, Pd2(dba)3: tris(dibenzylideneacetone)dipalladium, Pd(dppf)Cl2: [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium, DPPA: diphenylphosphoryl azide, DBU: 1,8-diazabicycloundec-7-ene, TBAF: tetrabutylammonium fluoride, Na Ascorbate: sodium ascorbate, t-BuXPhosPd-G3: (2-di-tert-butylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate.

[0137] As used herein, room temperature refers to about 20°C-30°C.

[0138] intermediates

[0139] Preparation of intermediate V1

[0140]

[0141] Step 1: Dissolve 1,3-disulfan-2-yl(trimethyl)silane (4.23 g, 21.97 mmol) in tetrahydrofuran (20 mL), cool to -78°C, and add n-butyllithium (2.5 M, 8.79 mL) dropwise. Allow to react for 0.5 hours. Then, add a solution of dicyclopropyl ketone (2.2 g, 19.97 mmol) in tetrahydrofuran (10 mL) dropwise. Stir at -78°C for 1.5 hours, then warm to room temperature and react for 16 hours. Add saturated sodium chloride solution (100 mL), extract with ethyl acetate (100 mL), dry the organic phase over anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue was purified by silica gel column chromatography (40 g, 100% petroleum ether) to afford compound V1-1 (3.0 g, pale yellow liquid) in a 70.7% yield. MS m / z (ESI): 213.0 [M+1].

[0142] Step 2: Compound V1-1 (3.0 g, 14.13 mmol) was dissolved in acetonitrile (50 mL) and water (12.5 mL). Trifluoroacetic acid (4.83 g, 42.38 mmol) and 30% hydrogen peroxide (20.82 g, 183.64 mmol) were then added. Stir at 80°C for 1 hour, cool to 40°C, and add 2N NaOH solution (40 mL). Stirring continued for 0.5 hour, the solution was neutralized with 2N hydrochloric acid (40 mL), and extracted with ethyl acetate (60 mL x 3). The organic phase was dried over anhydrous sodium sulfate and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (40 g, 0-60% ethyl acetate / petroleum ether) to afford compound V1-2 (1.5 g, colorless oil) in a yield of 75.8%. MS m / z (ESI): 139.0 [M-1].

[0143] Step 3: Dissolve 2,2-dicyclopropylacetic acid (1.50 g, 10.70 mmol) in tetrahydrofuran (30 mL) in a reaction flask, add CDI (2.31 g, 16.05 mmol), and react at room temperature for 16 hours (Solution A). In a separate reaction flask, add potassium (3-ethoxy-3-oxopropanoyl)oxy (2.73 g, 16.05 mmol) and magnesium chloride (1.02 g, 10.70 mmol). Heat to 50°C and react for 16 hours (Solution B). Add Solution A dropwise to Solution B (approximately 5 minutes), and stir the mixture at room temperature for 16 hours. Add ethyl acetate (100 mL), wash with saturated sodium chloride solution (80 mL x 3), dry the organic phase over anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue was separated by silica gel column chromatography (40 g, 0-30% ethyl acetate / petroleum ether) to give compound V1 (800 mg, colorless oil) in a yield of 33.6%. MS m / z (ESI): 211.1 [M+1]

[0144] Preparation of intermediate V2

[0145]

[0146] Refer to the method of step 3 in the preparation of intermediate V1, except that 2-cyclopropylacetic acid (CAS.NO: 5239-82-7) is used instead of 2,2-dicyclopropylacetic acid to prepare intermediate V2 (pale yellow oil, yield: 88.2%), MS m / z (ESI): 171.1 [M+1]

[0147] Preparation of intermediate V3

[0148]

[0149] Referring to the preparation method of intermediate V1, except that 1-cyclopropylethanone was used instead of dicyclopropyl ketone, intermediate V3 (light yellow liquid, yield 47.50%) was prepared, MS m / z (ESI): 185.1 [M+1].

[0150] Preparation of intermediate V4

[0151]

[0152] Under nitrogen, add thionyl chloride (919.31 mg, 7.73 mmol) to 2,2-dimethylbut-3-enoic acid (588 mg, 5.15 mmol) and react at 80°C for 2 hours. Concentrate the reaction solution to obtain a yellow oil. Under nitrogen, add n-butyllithium (2.5 M, 6.42 mL) to a solution of 3-ethoxy-3-oxopropanoic acid (1.14 g, 8.60 mmol) and tetrahydrofuran (15 mL) at -65°C and react at -10°C for 1 hour. Add a solution of the yellow oil in tetrahydrofuran (15 mL) and react at -65°C for 1 hour and then at room temperature for 1 hour. Pour the reaction solution into a solution of tetrahydrofuran / 1N hydrochloric acid (20 / 20 mL) and stir at room temperature for 2 hours. The organic layer was separated, washed with saturated sodium carbonate and saturated sodium chloride, then concentrated and purified by column chromatography (petroleum ether:ethyl acetate = 97.5%:2.5%) to give intermediate V4 (380 mg, 2.06 mmol, yield 71.97%) as a colorless oil. MS m / z (ESI): 185.2 [M+1].

[0153] Preparation of intermediate V5

[0154]

[0155] Intermediate V5 can be prepared by referring to the method of step 3 in the preparation of intermediate V1, except that 2-cyclobutylpropionic acid (CAS.NO: 1082453-55-1) is used instead of 2,2-dicyclopropylacetic acid.

[0156] Preparation of intermediate V6

[0157]

[0158] Referring to the preparation method of intermediate V1, the difference is that 1-cyclopentyl ethyl ketone is used instead of dicyclopropyl ketone to prepare intermediate V6.

[0159] Preparation of intermediate V7

[0160]

[0161] Referring to the preparation method of intermediate V4, the difference is that 2-cyclopropyl-2-methylpropionic acid is used instead of 2,2-dimethylbut-3-enoic acid to prepare intermediate V7.

[0162] Preparation of intermediate V8

[0163]

[0164] Step 1: Dissolve 2,2-dicyclopropylacetic acid (2.5 g, 17.83 mmol) in methanol (50 mL), then add thionyl chloride (2.6 mL, 35.67 mmol). The reaction is stirred at 70°C for 6 hours. Evaporate the solvent under reduced pressure, add ethyl acetate (60 mL), and wash with saturated sodium chloride solution (50 mL x 3). The organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue is purified by silica gel column chromatography to afford methyl 2,2-dicyclopropyl acetate (1.6 g, colorless oil) in a yield of 51.2%. 1 H NMR(400MHz,DMSO-d6)δ3.60(s,3H),1.17–1.08(m,1H),1.02–0.88(m,2H), 0.53–0.43(m,2H),0.41–0.31(m,2H),0.26-0.20(m,2H),0.14-0.06(m,2H).

[0165] Step 2: Dissolve methyl 2,2-dicyclopropyl acetate (1.5 g, 9.73 mmol) in tetrahydrofuran (30 mL), cool to -78°C, and add lithium diisopropylamide (2 M, 14.59 mL). The reaction is stirred at -78°C for 0.5 hour, then slowly warmed to -20°C and stirred for 0.5 hour. Methyl iodide (4.14 g, 29.18 mmol) is added dropwise to the mixture, and the mixture is slowly warmed to room temperature for 5 hours. The mixture is then cooled to -78°C and lithium diisopropylamide (2 M, 14.59 mL) is added. The reaction is stirred at -78°C for 0.5 hour, then slowly warmed to -20°C and stirred for 0.5 hour. Methyl iodide (4.14 g, 29.18 mmol) is added dropwise to the mixture, and the mixture is slowly warmed to room temperature for 16 hours. Ethyl acetate (80 mL) was added, and the mixture was washed with saturated sodium chloride solution (80 mL x 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product, methyl 2,2-dicyclopropylpropionate (1.5 g, pale yellow oil), was obtained in a yield of 91.7%. The product was used directly in the next step without further purification.

[0166] Step 3: Dissolve methyl 2,2-dicyclopropylpropanoate (1.5 g, 8.92 mmol) in methanol (10 mL), then add sodium hydroxide (1.43 g, 35.66 mmol) dissolved in water (4 mL). Stir the reaction at room temperature for 16 hours. Stop the reaction, adjust the pH to approximately 2 with 2N hydrochloric acid, and extract with ethyl acetate (50 mL x 2). The organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue is purified by silica gel column chromatography to yield 2,2-dicyclopropylpropionic acid (900 mg, light yellow oil) in a yield of 65.5%. MS m / z (ESI): 155.1 [M+1]. 1 H NMR (400MHz, DMSO-d6) δ1.01–0.89(m,2H),0.66(s,3H),0.33–0.20(m,8H).

[0167] Step 4: In a reaction flask, dissolve 2,2-dicyclopropylpropionic acid (900 mg, 5.84 mmol) in tetrahydrofuran (15 mL), then add N,N'-carbonyldiimidazole (1.26 g, 8.75 mmol) and react at room temperature for 16 hours (Solution A). In a separate reaction flask, dissolve potassium (3-ethoxy-3-oxopropanoyl)oxide (2.48 g, 14.59 mmol) in tetrahydrofuran (25 mL), add magnesium chloride (1.11 g, 11.67 mmol), and heat at 50°C for 16 hours (Solution B). Solution A is then added dropwise to Solution B (approximately 5 minutes), and the mixed solution is stirred at 30°C for 16 hours. Stop the reaction, add ethyl acetate (50 mL), and wash with saturated sodium chloride solution (50 mL x 3). The organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give ethyl 4,4-dicyclopropyl-3-oxo-pentanoate (600 mg, light yellow oil) in a yield of 45.8%. MS m / z (ESI): 225.1 [M+1].

[0168] Preparation of intermediate V9

[0169]

[0170] Step 1: Dissolve 3-methylbut-3-en-1-ol (8.6 g, 99.85 mmol) in dichloromethane (200 mL) at -20°C. Add diethylzinc (1 M, 299.54 mL) and diiodomethane (79.98 g, 299.54 mmol). React at -20°C for 4 h. Saturated aqueous ammonium chloride solution is slowly added to the reaction solution. Extract with 200 mL of dichloromethane, dry over anhydrous sodium sulfate, and spin dry. The crude product is purified by column chromatography (petroleum ether:ethyl acetate = 80%:20%) to obtain 2-(1-methylcyclopropyl)ethanol (5.6 g, yield: 56.0%) as a pale yellow liquid. 1 H NMR (400MHz, DMSO-d6) δ4.27(s,1H),3.46(dd,J=6.8,4.8Hz,2H),1.40–1.32(m,2H),0.97(s,3H),0.23(t,J=4.8Hz,2H),0.15(dd,J=5.2,4.0Hz,2H).

[0171] Step 2: Dissolve 2-(1-methylcyclopropyl)ethanol (4.6 g, 45.93 mmol) in acetone under ice, add Jones reagent (2.2 M, 62.63 mL), and allow to react at room temperature for 16 h. Add 70 mL of water, extract with 2 x 50 mL of dichloromethane, wash twice with saturated brine, dry over anhydrous sodium sulfate, and spin-dry at low temperature. The residue is purified by column chromatography (petroleum ether:ethyl acetate = 70%:30%) to afford 2-(1-methylcyclopropyl)acetic acid (1.7 g, 32.43% yield) as a pale yellow liquid. MS m / z (ESI): 113.1 [M-1].

[0172] Step 3: Under nitrogen at 0°C, add N,N'-carbonyldiimidazole (1.92 g, 11.83 mmol) to a solution of 2-(1-methylcyclopropyl)acetic acid (900 mg, 1.67 mmol) and tetrahydrofuran (30 mL). After complete addition, react at room temperature for 16 hours (Solution 1). Under nitrogen, heat (3-ethoxy-3-oxopropanoyl)oxypotassium (2.01 g, 11.83 mmol), magnesium chloride (749.07 mg, 7.88 mmol), and tetrahydrofuran (30 mL) at 50°C for 16 hours. Cool to 0°C and add one drop of the solution to the reaction mixture. After complete addition, react at room temperature for another 16 hours. Pour the reaction mixture into ice water and extract with ethyl acetate (50 mL x 3). Concentrate the organic layer to dryness to obtain the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 90%:10%) to give ethyl 4-(1-methylcyclopropyl)-3-oxobutanoate (1.2 g, yield 82.61%) as a light yellow liquid. MS m / z (ESI): 185.1 [M+1].

[0173] Preparation of intermediate V10

[0174]

[0175] At 0°C under nitrogen, add N,N'-carbonyldiimidazole (404.67 mg, 2.50 mmol) to a solution of 2-(3,3-difluorocyclobutyl)acetic acid (250 mg, 1.67 mmol) and tetrahydrofuran (15 mL). After complete addition, allow to react at room temperature for 16 hours (Solution 1). Under nitrogen, heat (3-ethoxy-3-oxopropanoyl)oxypotassium (425.16 mg, 2.50 mmol), magnesium chloride (158.20 mg, 1.67 mmol), and tetrahydrofuran (15 mL) at 50°C for 16 hours. Cool to 0°C, then add one drop of the solution to the reaction mixture. After complete addition, allow to react at room temperature for another 16 hours. Pour the reaction mixture into ice water and extract with ethyl acetate (15 mL x 3). Concentrate the organic layer to dryness to obtain the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 90%:10%) to give ethyl 4-(3,3-difluorocyclobutyl)-3-oxobutanoate (290 mg, 79.08% yield) as a light yellow liquid. MS m / z (ESI): 221.1 [M+1].

[0176] Preparation of intermediate V11

[0177]

[0178] Step 1: Dissolve 2-(1-methylcyclopropyl)acetic acid (1.7 g, 14.89 mmol) in N,N-dimethylformamide (15 mL), add potassium carbonate (6.18 g, 44.68 mmol) and benzyl bromide (3.06 g, 17.87 mmol). React at 25°C overnight. Add 50 mL of water to the reaction solution, extract with 2 x 30 mL of ethyl acetate, wash twice with saturated brine, dry over anhydrous sodium sulfate, and spin dry. The crude product is purified by column chromatography (petroleum ether:ethyl acetate = 95%:5%) to obtain benzyl 2-(1-methylcyclopropyl)acetate (2.1 g, yield: 69.03%) as a light yellow liquid. 1 H NMR (400MHz, DMSO-d6) δ7.45–7.24(m,5H),5.10(s,2H),2.29(s,2H),1.06(s,3H),0.41(t,J=4.8Hz,2H),0.30(dd,J=5.6,4.0Hz,2H).

[0179] Step 2: Dissolve benzyl 2-(1-methylcyclopropyl)acetate (1.6 g, 7.83 mmol) in THF (20 mL) at -78°C and add iodomethane (5.52 g, 39.17 mmol). Stir for 30 min, then add lithium bis(trimethylsilyl)amide (1 M, 15.67 mL). Slowly return the mixture to room temperature and stir for 2 h. Add saturated ammonium chloride to the reaction mixture, extract with 2 x 50 mL of ethyl acetate, combine the organic phases, wash twice with saturated brine, dry over anhydrous sodium sulfate, and spin-dry. The residue is purified by column chromatography (petroleum ether:ethyl acetate = 95%:5%) to afford benzyl 2-(1-methylcyclopropyl)propanoate (1.6 g, yield: 93.57%) as a light yellow liquid.

[0180] Step 3: Dissolve benzyl 2-(1-methylcyclopropyl)propanoate (2.1 g, 9.62 mmol) in methanol (20 mL) and add sodium hydroxide (3.85 g, 96.20 mmol) in water (10 mL). React at 30°C for 16 h. Drain the reaction mixture to remove most of the methanol, extract with 2 x 30 mL of methyl tert-butyl ether, adjust the pH to approximately 4 with hydrochloric acid, and then extract with 2 x 40 mL of methyl tert-butyl ether. Wash twice with saturated brine, dry over anhydrous sodium sulfate, and spin dry to obtain 2-(1-methylcyclopropyl)propionic acid (1.1 g, yield: 89.21%) as a pale yellow liquid. MS m / z (ESI): 127.1 [M-1].

[0181] Step 4: Prepared according to Step 4 of Intermediate V8. MS m / z (ESI): 199.1 [M+1].

[0182] Preparation of intermediate V12

[0183]

[0184] Step 1: Dissolve 2-cyclopropylacetic acid (50 g, 499.42 mmol) in N,N-dimethylformamide (500 mL), add potassium carbonate (207.07 g, 1.50 mol) and benzyl bromide (111.04 g, 649.25 mmol), and react at 30°C for 16 h. Add 2 L of water to the reaction solution, extract with 3 x 500 mL of ethyl acetate, combine the organic phases, wash three times with saturated brine, dry over anhydrous sodium sulfate, filter, and spin-dry. The residue is combined with petroleum ether:ethyl acetate in a ratio of 10:1 to yield compound V12-1 (89 g, yield: 93.68%) as a pale yellow liquid. 1 H NMR (400MHz, DMSO-d6) δ7.42–7.25(m,5H),5.08(s,2H),2.27(d,J=4Hz,2H),1.01–0.90(m,1H),0.51–0.38(m,2H),0.19–0.05(m,2H).

[0185] Step 2: Lithium bis(trimethylsilyl)amide (1 M, 78.85 mL) was added to a three-necked flask. Under argon, compound V12-1 (5 g, 26.28 mmol) and iodoethane (41.00 g, 262.83 mmol) in tetrahydrofuran (70 mL) were added dropwise at -78°C. The reaction mixture was allowed to react at -40°C for 2 h. Saturated ammonium chloride solution was added to the reaction solution, and the mixture was extracted with 200 mL of ethyl acetate. The combined organic phases were washed twice with saturated brine. The mixture was dried over anhydrous sodium sulfate, filtered, and dried. The residue was then flash-flashed with petroleum ether:ethyl acetate in a ratio of 10:1 to afford compound V12-2 (2.85 g, yield: 49.67%) as a pale yellow liquid. 1 H NMR(400MHz,DMSO-d6)δ7.40–7.26(m,5H),5.10(s,2H),1.57-1.65(m,3H), 0.90–0.79(m,4H),0.52–0.43(m,1H),0.43–0.35(m,1H),0.21–0.12(m,2H).

[0186] Step 3: Dissolve compound V12-2 (22 g, 100.78 mmol) in methanol (200 mL). Add a solution of sodium hydroxide (32.25 g, 806.26 mmol) in water (100 mL) dropwise under an ice bath. After complete addition, react at 30°C for 16 hours. Remove the methanol from the reaction mixture by vortexing. Add 200 mL of water to the residue and extract with 3 x 150 mL of methyl tert-butyl ether. Adjust the pH of the aqueous phase to 4 and extract with 3 x 150 mL of methyl tert-butyl ether. Combine the organic phases, wash twice with saturated brine, dry over anhydrous sodium sulfate, filter, and dry at low temperature. This yields compound V12-3 (12 g, yield: 90.2%) as a pale yellow liquid. MS m / z (ESI): 127.2 [M-1].

[0187] Step 4: Dissolve carbonyldiimidazole (18.96 g, 117.03 mmol) in tetrahydrofuran (200 mL) at 0°C and add compound V12-3 (10 g, 78.02 mmol). Stir at room temperature for 16 hours. Dissolve potassium (3-ethoxy-3-oxopropanoyl)oxide (39.84 g, 234.07 mmol) in tetrahydrofuran (200 mL) and add magnesium chloride (22.24 g, 234.07 mmol). Stir at 50°C for 16 hours. Then, slowly add the liquid from the first vial dropwise to the second vial. Stir at room temperature for 16 hours. Add water to the reaction mixture and extract with ethyl acetate (3 x 150 mL). Combine the organic phases, wash twice with saturated brine, dry over anhydrous sodium sulfate, and spin dry. The residue is combi-flashed with petroleum ether:ethyl acetate in a ratio of 10:1 to yield compound V12 (14.8 g, yield: 95.68%) as a yellow liquid. MS m / z (ESI): 199 [M+1].

[0188] Preparation of intermediate V13

[0189]

[0190] Step 1: Under nitrogen at 0°C, add oxalyl chloride (1.78 g, 14.02 mmol, 1.20 mL) dropwise to a solution of 2-cyclopropyl-2-oxoacetic acid (1 g, 8.76 mmol) and dichloromethane (15 mL). Add N,N-dimethylformamide (6.41 mg, 87.64 μmol) and allow to react at room temperature for 1 hour. Concentrate the reaction mixture to dryness, then add dichloromethane (15 mL). Add benzyl alcohol (1.33 g, 12.27 mmol, 1.27 mL), triethylamine (2.22 g, 21.91 mmol, 3.06 mL) and dichloromethane (15 mL) dropwise at 0°C, and allow to react at room temperature for 1 hour. Pour the reaction mixture into ice water and extract with dichloromethane (15 mL x 2). Combine the organic layers, wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate to obtain the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate=90%:10%) to give compound V13-1 (1.26 g, 67.56% yield) as a colorless liquid (no ion current).

[0191] Step 2: Under nitrogen at room temperature, bis(2-methoxyethyl)aminosulfur trifluoride (3.90 g, 17.63 mmol) was added to compound V13-1 (900 mg, 4.41 mmol) and dichloromethane (30 mL). The mixture was allowed to react at 50°C for 16 hours. The reaction mixture was poured into ice and extracted with dichloromethane (50 mL x 2). The combined organic layers were washed with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 70%:30%) to obtain compound V13-2 (555 mg, 55.67% yield) as a colorless liquid (no ion flux).

[0192] Step 3: Compound V13-2 (555 mg, 2.45 mmol), sodium hydroxide (490.64 mg, 12.27 mmol), methanol (5 mL), and water (5 mL) were mixed and reacted at room temperature for 16 hours. The methanol was removed under reduced pressure, and the aqueous phase was washed with methyl tert-butyl ether (15 mL x 2). The aqueous phase was collected and cooled to 0°C. Concentrated hydrochloric acid was added to adjust the pH to 2, and the mixture was extracted with dichloromethane (15 mL x 2). The combined organic layers were dried over anhydrous sodium sulfate and concentrated to yield compound V13-3 (320 mg, 2.35 mmol, 95.84% yield) as a yellow liquid. MS m / z (ESI): 135.1 [M-1].

[0193] Step 4: Under nitrogen at 0°C, add N,N'-carbonyldiimidazole (507.70 mg, 3.53 mmol) to a solution of compound V13-3 (320 mg, 2.35 mmol) and tetrahydrofuran (20 mL). After complete addition, react at room temperature for 16 hours. Under nitrogen, heat (3-ethoxy-3-oxopropanoyl)oxypotassium (1.00 g, 5.88 mmol), magnesium chloride (447.73 mg, 4.70 mmol), and tetrahydrofuran (20 mL) at 50°C for 16 hours. Cool to 0°C and add one drop of the solution to the reaction mixture. After complete addition, react at room temperature for another 16 hours. Add ethyl acetate (50 mL) and water (50 mL). Separate the organic layer, wash with saturated sodium bicarbonate solution and saturated sodium chloride solution, dry over anhydrous sodium sulfate, and concentrate to dryness to obtain the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate=80%:20%) to give compound V13 (100 mg, yield 20.63%) as a yellow oil.

[0194] Preparation of intermediate V14

[0195]

[0196] Compound V14 was prepared by referring to the method of intermediate V12. MS m / z (ESI): 213.1 [M+1].

[0197] Preparation of intermediate V15

[0198]

[0199] Step 1: Dissolve triethyl 2-fluoro-2-phosphorylacetate (1.90 g, 7.85 mmol) in 10 mL of THF, replace the atmosphere with nitrogen three times, then cool to -70°C and slowly add n-butyllithium (2.5 M, 3.42 mL) dropwise. Stir at -70°C for 0.5 h. Warm to 0°C and stir for 0.5 h. Then, slowly add cyclobutanone (0.5 g, 7.13 mmol) dropwise at -70°C and stir at room temperature for 3 h. Quench with water (20 mL) and extract with ethyl acetate (20 mL x 2). The organic phase is washed with saturated sodium chloride solution (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue is purified by silica gel column chromatography using 0-20% ethyl acetate in petroleum ether as eluent to afford compound V15-1 (0.8 g, pale yellow liquid) in a 70.9% yield.

[0200] Step 2: Dissolve compound V15-1 (0.8 g, 5.06 mmol) in methanol (15 mL) and add 10% wet Pd / C (538.26 mg, 505.79 μmol). Replace the mixture with hydrogen three times. Stir under hydrogen (15 psi) at 25°C for 1 h. Filter the catalyst and concentrate under reduced pressure to obtain compound V15-2 (0.7 g, colorless liquid) in an 86.40% yield. Use the product directly in the next step without further purification.

[0201] Step 3: Dissolve compound V15-2 (0.7 g, 4.37 mmol) and sodium hydroxide (524.39 mg, 13.11 mmol) in methanol (5 mL) and water (2 mL). Stir at 30°C for 2 h. Adjust the pH to 5 with 2M dilute hydrochloric acid. Extract with dichloromethane (10 mL x 3). Combine the organic phases and dry them in a spin dryer. Purify the resulting residue by silica gel column chromatography with 0-20% ethyl acetate in petroleum ether as eluent to afford compound V15-3 (400 mg, colorless liquid) in a 69.27% ​​yield. MS m / z (ESI): 131.1 [M-1].

[0202] Step 4: Dissolve compound V15-3 (0.4 g, 3.03 mmol) in tetrahydrofuran (6 mL) in a reaction flask, then add DCI (736.30 mg, 4.54 mmol) and react at room temperature for 16 hours (Solution A). In a separate reaction flask, add potassium (3-ethoxy-3-oxopropanoyl)oxy (2.06 g, 12.11 mmol) to anhydrous magnesium chloride (864.68 mg, 9.08 mmol) and tetrahydrofuran (20 mL). Heat to 50°C under argon for 16 hours (Solution B). Solution A is then slowly added dropwise to Solution B, and the mixture is stirred at 30°C for 16 hours. Add 30 mL of water to the reaction mixture, and extract with ethyl acetate (30 mL x 3). The combined organic phases are washed with brine, dried over anhydrous sodium sulfate, and then evaporated to dryness under reduced pressure. The resulting residue was purified by silica gel column chromatography using 0-10% ethyl acetate in petroleum ether as eluent to afford compound V15 (230 mg, light yellow oil) in a yield of 37.57%. MS m / z (ESI): 203.2 [M+1].

[0203] Preparation of intermediate V16

[0204]

[0205] Prepared by following the method of Intermediate V15, except that cyclobutanone was replaced by cyclopentanone. MS m / z (ESI): 217.1 [M+1].

[0206] Preparation of intermediate V17

[0207]

[0208] The preparation was carried out according to the method of intermediate V15, except that triethyl 2-fluoro-2-phosphorylacetate was replaced with ethyl 2-(diethoxyphosphoryl)butyrate, MS m / z (ESI): 213.1 [M+1].

[0209] Preparation of intermediate V18

[0210]

[0211] Step 1: Under nitrogen protection, triethylamine (6.06 g, 59.93 mmol, 8.36 mL) was added dropwise to a solution of 2-cyclopropylacetic acid (5 g, 49.94 mmol) and tetrahydrofuran (100 mL) at -78 °C. After the addition was complete, pivaloyl chloride (6.62 g, 54.94 mmol, 6.76 mL) was added dropwise at -78 °C. The reaction solution changed from a yellow clear solution to a white suspension. After the addition was complete, the reaction solution was raised to 0 °C and reacted for 1.5 hours (Solution 1). Under nitrogen, n-butyllithium (1.6 M, 37.46 mL) was added dropwise to a solution of (R)-4-benzyl-2-oxazolidinone (8.85 g, 49.94 mmol) and tetrahydrofuran (50 mL) at -78°C. The reaction solution turned from a colorless, clear solution to a white suspension. After the addition was complete, the temperature was raised to 0°C, and the solution turned yellow again. The reaction solution was allowed to react at 0°C for 0.5 hours. This solution was then slowly added dropwise to Solution 1 at 0°C, followed by stirring at room temperature for 2 hours. The reaction was quenched with water (200 mL), extracted with ethyl acetate (300 mL x 3), and the combined organic phases were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to yield the crude product. The residue was purified by silica gel column chromatography using 0-20% ethyl acetate in petroleum ether as eluent to afford compound V18-1 (8.5 g, colorless oil) in a yield of 65.64%. MS m / z(ESI):260.1[M+1].

[0212] Step 2: Dissolve compound V18-1 (7.5 g, 28.92 mmol) in dichloromethane (200 mL) and cool to 0°C under argon. Then, slowly add anhydrous titanium tetrachloride (6.03 g, 31.82 mmol, 3.49 mL) dropwise. After the addition, the solution turns yellow. Stir at 0°C for 5 minutes. Then, slowly add N,N-diisopropylethylamine (4.30 g, 33.26 mmol, 5.79 mL) dropwise. After the solution turns black, maintain the temperature and stir for 1 hour. Finally, add benzyl chloromethyl ether (9.06 g, 57.85 mmol, 8.05 mL) dropwise and stir at 0°C for 6 hours. Quench the reaction with saturated aqueous ammonium chloride (150 mL) and extract with dichloromethane (100 mL x 3). The organic phases are combined, washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The resulting residue was purified by silica gel column chromatography using 0-15% ethyl acetate in petroleum ether as eluent to afford compound V18-2 (9 g, yield: 82%) as a white solid. MS m / z (ESI): 380.2 [M+1].

[0213] Step 3: Compound V18-2 (9 g, 23.72 mmol) was dissolved in methanol (120 mL), and 10% wet Pd / C (2.37 g, 4.74 mmol) was added. The mixture was then replaced with hydrogen three times. The mixture was stirred at room temperature under a hydrogen pressure of 15 psi for 2 h. The catalyst was filtered off, and the organic phase was concentrated under reduced pressure to afford compound V18-3 (6 g, white solid) in a yield of 87.43%. MS m / z (ESI): 290.1 ​​[M+H] + .

[0214] Step 4: Dissolve compound V18-3 (3 g, 10.37 mmol) in dichloromethane (50 mL). Cool to 0°C under argon, then slowly add 1,8-bis(dimethylaminonaphthalene) (5.56 g, 25.92 mmol) and trimethyloxonium tetrafluoroborate (3.07 g, 20.74 mmol). Slowly return the mixture to room temperature and stir for 16 h. Quench the reaction with saturated aqueous ammonium chloride (50 mL), extract with dichloromethane (50 mL x 3). The combined organic phases are washed with dilute hydrochloric acid, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to yield the crude product. The resulting residue is purified by silica gel column chromatography using 0-15% ethyl acetate in petroleum ether as eluent to afford compound V18-4 (2.6 g, white solid) in a 66% yield. MS m / z (ESI): 304.2 [M+1].

[0215] Step 5: Dissolve compound V18-4 (2.6 g, 8.57 mmol) in tetrahydrofuran (25 mL) and water (15 mL). Cool to 0°C, then add 33% hydrogen peroxide (3.89 g, 34.28 mmol, 3.44 mL) and lithium hydroxide monohydrate (720.47 mg, 17.14 mmol) dropwise. Return to room temperature and stir for 1 h. Slowly add aqueous sodium sulfite solution under ice to quench the mixture. Methyl tert-butyl ether (20 mL x 2) is then extracted to remove impurities. The aqueous phase is adjusted to pH 5 with dilute hydrochloric acid, and methyl tert-butyl ether (20 mL x 3) is extracted. The combined organic phases are dried over anhydrous sodium sulfate, concentrated, and spun down to afford crude compound V18-5 (1.2 g, colorless oil) in a 97.12% yield. MS m / z (ESI): 143.1 [M-1].

[0216] Step 6: In a reaction flask, compound V18-5 (0.7 g, 4.86 mmol) was dissolved in tetrahydrofuran (10 mL), and CDI (1.18 g, 7.28 mmol) was added. The mixture was allowed to react at room temperature for 16 hours (Solution A). In a separate reaction flask, potassium (3-ethoxy-3-oxopropanoyl)oxide (3.31 g, 19.42 mmol) was added to anhydrous magnesium chloride (1.39 g, 14.57 mmol) and tetrahydrofuran (20 mL). The mixture was heated at 50°C under argon for 16 hours (Solution B). Solution A was then added dropwise to Solution B (approximately 10 minutes), and the mixture was stirred at 70°C for 16 hours. 30 mL of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (30 mL x 3). The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, and then evaporated to dryness under reduced pressure. The resulting residue was purified by silica gel column chromatography using 0-15% ethyl acetate in petroleum ether as eluent to afford compound V18 (0.9 g, pale yellow oil) in a yield of 86.51%. MS m / z (ESI): 215.2 [M+1].

[0217] Preparation of intermediate V19

[0218]

[0219] Step 1: Under nitrogen at 0°C, add oxalyl chloride (3.56 g, 28.05 mmol, 2.39 mL) dropwise to a solution of 2-cyclopropyl-2-oxoacetic acid (2 g, 17.53 mmol) and dichloromethane (50 mL). Add N,N-dimethylformamide (12.81 mg, 175.29 μmol, 13.57 μL) and warm to room temperature for 1 hour. Concentrate the reaction mixture to dryness and dissolve it in dichloromethane (50 mL). Add benzyl alcohol (2.65 g, 24.54 mmol, 2.54 mL), triethylamine (4.43 g, 43.82 mmol, 6.11 mL) and dichloromethane (50 mL) dropwise at 0°C. After addition, react at room temperature for 1 hour. The reaction mixture was poured into ice water and extracted with dichloromethane (30 mL x 2). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to yield the crude product. The residue was purified by silica gel column chromatography using 0-10% ethyl acetate in petroleum ether as eluent to afford compound V19-1 (2.1 g, colorless liquid) in a yield of 58.66%. The desired product was not detected by MS and TLC, and the obtained product was directly carried forward to the next step.

[0220] Step 2: Dissolve compound V19-1 (1.9 g, 9.30 mmol) in methanol (30 mL). Cool to -10°C, then slowly add sodium borohydride (422.38 mg, 11.16 mmol). Stir at -10°C for 30 minutes. Quench the reaction with water, extract with ethyl acetate (40 mL x 3), wash with brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. Purify the resulting residue by silica gel column chromatography with 0-20% ethyl acetate in petroleum ether as eluent to afford compound V19-2 (1.5 g, colorless oil) in a yield of 78.18%. MS m / z (ESI): 207.1 [M+1].

[0221] Step 3: Dissolve compound V19-2 (1.5 g, 7.27 mmol), iodomethane (5.16 g, 36.37 mmol, 2.35 mL), and silver oxide (3.37 g, 14.55 mmol, 472.12 μL) in acetonitrile (15 mL) in a sealed tube. Purge with argon briefly, seal tightly, and heat to 70°C with stirring overnight. Filter the solids and evaporate to dryness under reduced pressure to obtain the crude product. Purify the residue by silica gel column chromatography with 0-10% ethyl acetate in petroleum ether as eluent to afford compound V19-3 (1.1 g, colorless oil) in a yield of 68.66%. MS m / z (ESI): 221.1 [M+1].

[0222] Step 4: Dissolve compound V19-3 (1.1 g, 4.99 mmol) and sodium hydroxide (599.24 mg, 14.98 mmol) in methanol (15 mL) and water (5 mL). Warm to 30°C and stir for 4 h. Remove the solvent under reduced pressure, adjust the pH to 3 with 2M dilute hydrochloric acid, and extract with ethyl acetate (15 mL x 3). Combine the organic phases, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. Purify the resulting residue by silica gel column chromatography with 0-20% ethyl acetate in petroleum ether as eluent to afford compound V19-4 (0.6 g, colorless oil) in a yield of 92.32%. MS m / z (ESI): 129.1 [M-1].

[0223] Step 5: Prepared according to Step 6 of Intermediate V18 to give Compound V19. MS m / z (ESI): 201.1 [M+1].

[0224] Preparation of intermediate V20

[0225]

[0226] Compound V20 was prepared by referring to the method of intermediate V2. MS m / z (ESI): 185.1 [M+1].

[0227] Preparation of intermediate V21

[0228]

[0229] Step 1: Under nitrogen, dissolve ethyl glyoxylate (5 g, 24.49 mmol) in tetrahydrofuran (30 mL). Cool to -78°C and slowly add cyclopropylmagnesium bromide (1.78 g, 12.24 mmol). Incubate the reaction mixture at -20°C for 3 hours. Quench the reaction with ice water and extract with ethyl acetate (30 mL x 2). Combine the organic phases, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. Column chromatography (PE:EA = 4:1) afforded compound V21-1 (620 mg, colorless liquid) in a 17.56% yield. MS m / z (ESI): 145.0 [M+1].

[0230] Step 2: Dissolve compound V21-1 (620 mg, 4.30 mmol) in 8 mL of dichloromethane. Under nitrogen, slowly add DAST (1.39 g, 8.60 mmol) at -70°C. Stir the reaction mixture at room temperature for 12 hours. Add 20 mL of water, extract with dichloromethane (20 mL x 2), combine the organic phases, dry over anhydrous sodium sulfate, and concentrate under reduced pressure to obtain compound V21-2 (448 mg, brown liquid) in a 78% yield. MS m / z (ESI): 147.2 [M+1].

[0231] Step 3: Compound V21-2 (448 mg, 3.07 mmol) was dissolved in water (3 mL) and methanol (1 mL). Lithium hydroxide (293.62 mg, 12.26 mmol) was added and nitrogen atmosphere was applied. The reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction, the mixture was concentrated under reduced pressure, the pH was adjusted to 1 with hydrochloric acid, and the mixture was extracted with dichloromethane (20 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to yield compound V21-3 (380 mg, brown liquid) in a 78% yield. MS m / z (ESI): 117.1 [M-1].

[0232] Step 4: Prepared according to Step 6 of Intermediate V18 to obtain Compound V21. MS m / z (ESI): 189.1 [M+1].

[0233] Preparation of intermediate V22

[0234]

[0235] Compound V22 was prepared by referring to the method of intermediate V4. MS m / z (ESI): 199.1 [M+H + ].

[0236] Preparation of intermediate 3-2

[0237]

[0238] Step 1: Compound V2 (1.2 g, 7.05 mmol) and 1H-pyrazol-5-amine (585.8 mg, 7.05 mmol) were dissolved in acetic acid (8 mL). The reaction was stirred at 70°C for 4 hours. The solvent was evaporated under reduced pressure, and ethyl acetate (15 mL) was added. A solid precipitated, which was filtered and dried to afford compound 3-1 (800 mg, brown solid) in a 60.0% yield. MS m / z (ESI): 190.1 [M+1].

[0239] Step 2: Dissolve compound 3-1 (500 mg, 2.64 mmol) in phosphorus oxychloride (6 mL) and heat to 120°C with stirring for 3 hours. Stop the reaction, cool to room temperature, pour into ice water (60 g), and extract with dichloromethane (80 mL). The organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue is purified by silica gel column chromatography (20 g, 0%-40% ethyl acetate / petroleum ether) to afford compound 3-2 (300 mg, light yellow oil) in a 54.7% yield. MS m / z (ESI): 208.1 [M+1].

[0240] Intermediate 4-2 to Intermediate 23-2

[0241] Intermediates 4-2 to 23-2 were prepared using the corresponding ethyl oxobutyrate as raw materials according to the method of Intermediate 3-2.

[0242]

[0243]

[0244] Preparation of intermediate 24-2

[0245]

[0246] Prepared according to the method of compound 2-1. MS m / z (ESI): 256.0 [M+1].

[0247] Preparation of intermediate 27-3

[0248]

[0249] Step 1: Hydrogen peroxide (818.72 mg, 7.22 mmol, 30% purity) and iodine (1.53 g, 6.02 mmol) were added to a solution of 1H-pyrazol-5-amine (1 g, 12.03 mmol) and water (10 mL) at room temperature. The mixture was allowed to react for 1 hour. 20 mL of saturated sodium sulfite solution was added to the reaction mixture, stirred for 10 minutes, and filtered. The filter cake was washed with water and dried to yield 4-iodo-1H-pyrazol-5-amine (1.3 g, 51.69% yield). The crude product was used directly in the next step. MS m / z (ESI): 210.0 [M+1].

[0250] Step 2: Compound V1 (1 g, 4.76 mmol), 4-iodo-1H-pyrazol-5-amine (993.91 mg, 4.76 mmol), and acetic acid (35 mL) were mixed and reacted at 70°C under nitrogen for 1 hour. The reaction solution was concentrated to dryness to obtain a crude product. The crude product was purified by column chromatography (dichloromethane:dichloromethane / tetrahydrofuran / ethyl acetate (volume ratio 2 / 1 / 1) = 70%:30%) to obtain compound 27-1 (300 mg, 17.76% yield) as a yellow solid. MS m / z (ESI): 356.0 [M+1].

[0251] Step 3: Cuprous iodide (257.38 mg, 1.35 mmol) and potassium fluoride (78.51 mg, 1.35 mmol) were added to a reaction flask. The atmosphere was purged with nitrogen, followed by the addition of compound 27-1 (240 mg, 675.73 μmol), N,N-dimethylformamide (12 mL), and trimethyl(trifluoromethyl)silane (288.25 mg, 2.03 mmol). Nitrogen was then added and the mixture was reacted at 90°C for 16 hours (condenser tube + nitrogen balloon). After cooling, additional cuprous iodide (257.38 mg, 1.35 mmol), potassium fluoride (78.51 mg, 1.35 mmol), and trimethyl(trifluoromethyl)silane (288.25 mg, 2.03 mmol) were added, and the mixture was heated at 90°C for another 16 hours. The reaction vessel was then replaced with a large sealed tube and the reaction was continued at 95°C for 6 hours. The reaction mixture was poured into ice water, and the insoluble material was filtered off. Ethyl acetate (20 mL x 2) was then added for extraction. The combined organic layers were washed with water (10 mL x 3) and saturated brine (15 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 70%:30%) to afford compound 27-2 (137 mg, 293.70 μmol, 43.46% yield) as a tan solid. MS m / z (ESI): 298.2 [M+1].

[0252] Step 4: Compound 27-2 (127 mg, 427.21 μmol) and phosphorus oxychloride (655.06 mg, 4.27 mmol) were heated to 110°C for 3 hours. Excess phosphorus oxychloride was evaporated to dryness. The residue was added to ice water and extracted with dichloromethane (20 mL x 3). The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 70%:30%) to obtain compound 27-3 (86 mg, 39.30% yield) as a light yellow solid. MS m / z (ESI): 316.1 [M+1].

[0253] Intermediate 25-2 to Intermediate 50-2

[0254] Intermediates 25-2 to 50-2 were prepared using the corresponding ethyl oxobutyrate as raw materials according to the above method.

[0255]

[0256]

[0257] Example 1: Preparation of Compound Z-1

[0258]

[0259] Step 1: Dissolve compound V1 (0.7 g, 3.33 mmol) and 1H-pyrazol-5-amine (276.6 mg, 3.33 mmol) in acetic acid (6 mL) and stir at 70°C for 4 hours. Evaporate the solvent under reduced pressure, add ethyl acetate (15 mL), and a solid precipitates. Filter and dry to obtain compound 1-1 (500 mg, brown solid) in a yield of 65.5%. MS m / z (ESI): 230.1 [M+1].

[0260] Step 2: Dissolve compound 1-1 (500.0 mg, 2.18 mmol) in phosphorus oxychloride (6 mL) and heat to 120°C with stirring for 3 hours. Cool to room temperature, pour into ice water (60 g), and extract with dichloromethane (80 mL). The organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue is purified by silica gel column chromatography (20 g, 0%-40% EA / DCM) to afford compound 1-2 (350 mg, light yellow oil) in a yield of 64.8%. MS m / z (ESI): 248.0 [M+1].

[0261] Step 3: Compound 1-2 (150.0 mg, 605.5 μmol) and tert-butyl ((1S,3S)-3-aminocyclopentyl)carbamate (121.27 mg, 605.52 μmol) were dissolved in acetonitrile (20 mL), and potassium carbonate (251.0 mg, 1.81 mmol) was added. The reaction was stirred at 90°C for 16 hours. Ethyl acetate (80 mL) was added to dilute the mixture, and the mixture was washed with saturated sodium chloride solution (80 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford compound 1-3 (180 mg, light yellow oil) in a yield of 72.2%. MS m / z (ESI): 412.2 [M+1].

[0262] Step 4: Compound 1-3 (180 mg, 437.38 μmol) was dissolved in 1,4-dioxane (3 mL), 4 M hydrochloric acid solution (3.0 mL) was added, and the mixture was stirred at room temperature for 3 hours. The solvent was evaporated under reduced pressure, water (60 mL) was added, and the mixture was extracted with ethyl acetate (50 mL). The aqueous phase was adjusted to pH 9-10 with saturated sodium carbonate solution and extracted with ethyl acetate (60 mL x 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Compound Z-1 (43.5 mg) was isolated by preparative HPLC chromatography in a yield of 31.3%. MS m / z (ESI): 312.2 [M+1]; 1 H NMR (400MHz, DMSO-d6) δ7.96(d,J=2.4Hz,1H),7.39(d,J=7.2Hz,1H),6.27(d,J=2.4Hz,1H),6.00(s,1H),4.25–4.10(m,1H),3.42 -3.35(m,1H),2.26–2.10(m,1H),2.04–1.53(m,6H),1.34–1.09(m,4H),0.59-0.47(m,2H),0.36–0.16(m,4H),0.13–0.02(m,2H).

[0263] Example 2: Preparation of Compound Z-2

[0264]

[0265] Step 1: Dissolve compound 1-2 (100 mg, 403.68 μmol) in acetonitrile (6 mL), add NCS (53.9 mg, 403.7 μmol), and stir at 20°C for 2 hours. Add ethyl acetate (100 mL), wash with saturated sodium chloride solution (80 mL x 3), dry the organic phase over anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue was purified by silica gel column chromatography (20 g, 0-30% ethyl acetate / petroleum ether) to afford compound 2-1 (100 mg, light yellow oil) in a yield of 87.8%. MS m / z (ESI): 282.0 [M+1]

[0266] Step 2: Follow the procedure of Step 3 of Example 1, except that Compound 2-1 is used instead of Compound 1-2 to obtain Compound 2-2 (120 mg, light yellow oil) in a yield of 75.9%. MS m / z (ESI): 446.1 [M+1]

[0267] Step 3: Follow the procedure of Step 4 in Example 1, except that compound 2-2 is used instead of compound 1-3 to obtain compound Z-2 (22.3 mg) in a yield of 23.4%. MS m / z (ESI): 346.1 [M+1]; 1 H NMR(400MHz,DMSO-d6)δ8.13(s,1H),7.68(bs,1H),6.12(s,1H),4.28–4.15(m,1H),3.45-3.30(m,1H),2.24–2.13(m,1H),2.13– 1.78(m,4H),1.77-1.70(m,1H),1.68-1.58(m,1H),1.37–1.13(m,4H),0.58-0.49(m,2H),0.40–0.18(m,4H),0.12-0.02(m,2H).

[0268] Example 3: Preparation of Compound Z-3

[0269]

[0270] Referring to the preparation method of compound Z-1, except that compound V2 was used instead of compound V1, compound Z-3 (33.6 mg) was prepared in a yield of 37.7%. MS m / z (ESI): 272.1 [M+1]; 1H NMR (400MHz, DMSO-d6) δ7.96(d,J=2.0Hz,1H),6.80(s,1H),6.26(d,J=2.0Hz,1H),6.08(s,1H),4.19–4.10(m,1H),3.98-3.91(m,1H),2.50(d,J=7 .2Hz,2H),2.17-2.08(m,1H),2.05–1.80(m,3H),1.72-1.62(m,1H),1.50 -1.38(m,1H),1.13–1.02(m,1H),0.49–0.40(m,2H),0.26–0.16(m,2H)..

[0271] Example 4: Preparation of Compound Z-4

[0272]

[0273] Referring to the preparation method of compound Z-1, except that 3-amino-1H-pyrazole-4-carbonitrile was used instead of 1H-pyrazole-5-amine, compound Z-4 (9.4 mg) was prepared in a yield of 15.2%. MS m / z (ESI): 337.1 [M+1]; 1 H NMR (400MHz, DMSO-d6) δ8.55(s,1H),8.20(bs,1H),6.34(s,1H),4.30-4.20(m,1H),3.45-3.35(m,1H),2.26–2.07(m,1 H),2.00–1.82(m,2H),1.79–1.58(m,2H),1.49–1.13(m,4H),0.67–0.44(m,2H),0.43–0.18(m,4H),0.11-0.02(m,2H).

[0274] Example 5: Preparation of Compound Z-5 and Its Stereoconfigurations Compound Z-5-1 and Compound Z-5-2

[0275]

[0276] Referring to the preparation method of compound Z-1, except that compound V3 was used instead of compound V1, compound Z-5 was prepared (yield 41.29%). MS m / z (ESI): 286.1 [M+1]; 1H NMR(400MHz,DMSO-d6)δ7.96(d,J=2.2Hz,1H),7.41(s,1H),6.27(d,J=2.2Hz ,1H),6.03(s,1H),4.24–4.16(m,1H),3.43–3.37(m,1H),2.22–2.13(m,1H), 2.01–1.82(m,3H),1.78–1.71(m,1H),1.68–1.58(m,1H),1.32–1.22(m,4H), 1.06–0.97(m,1H),0.51–0.45(m,1H),0.35–0.30(m,1H),0.22–0.14(m,2H).

[0277] Compound Z-5 (50 mg) was subjected to chiral separation (instrument: SFC-150 (Waters); column: IC 20*250 mm, 10 um (Daicel); co-solvent: CO2 / IPA [0.5% NH3 (7 M in MeOH)] = 60 / 40, wavelength: 214 nm) to obtain two single-configuration products Z-5-1 (retention time: 2.704 min) and Z-5-2 (retention time: 3.175 min): compound Z-5-1 (yield 21.93%), MS m / z (ESI): 286.1 [M+1]; 1 H NMR (400MHz, DMSO-d6) δ8.00(d,J=2.2Hz,1H),7.49(d,J=7.6Hz,1H),6.31(d,J =2.2Hz,1H),6.07(s,1H),4.28–4.19(m,1H),3.49–3.40(m,1H),2.26–2.18(m, 1H),2.05–1.87(m,3H),1.82–1.75(m,1H),1.72–1.63(m,1H),1.37–1.23(m,4H ),1.11–1.00(m,1H),0.56–0.48(m,1H),0.40–0.31(m,1H),0.27–0.15(m,2H);

[0278] Compound Z-5-2 (yield 22.62%), MS m / z (ESI): 286.1 [M+1]; 1H NMR (400MHz, DMSO-d6) δ8.00(d,J=2.2Hz,1H),7.48(d,J=7.4Hz,1H),6.31(d,J =2.2Hz,1H),6.07(s,1H),4.29–4.18(m,1H),3.48–3.40(m,1H),2.25–2.17(m, 1H),2.05–1.87(m,3H),1.81–1.75(m,1H),1.70–1.61(m,1H),1.35–1.23(m,4H ),1.11–1.01(m,1H),0.55–0.48(m,1H),0.40–0.32(m,1H),0.26–0.17(m,2H).

[0279] Example 6: Preparation of Compound Z-6

[0280]

[0281] Compound Z-1 (60.0 mg, 192.66 μmol) was dissolved in dichloromethane (10 mL), and triethylamine (195.0 mg, 1.93 mmol, 0.27 mL) and methyl chloroformate (91.0 mg, 963.32 μmol) were added. The mixture was stirred at room temperature for 1 hour. Saturated sodium chloride solution (50 mL) was added, and the mixture was extracted with ethyl acetate (60 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to afford compound Z-6 (40.3 mg) in a yield of 56.4%. MS m / z (ESI): 370.2 [M+1]; 1 H NMR (400MHz, DMSO-d6) δ7.97(d,J=2.0Hz,1H),7.57(d,J=8.0Hz,1H),7.20(d,J=6. 8Hz,1H),6.28(d,J=2.0Hz,1H),6.01(s,1H),4.18-4.12(m,1H),4.02-3.96(m,1H), 3.48(s,3H),2.19–2.08(m,1H),2.06–1.83(m,3H),1.69-1.64(m,1H),1.52–1.36(m ,1H),1.29–1.10(m,3H),0.58–0.45(m,2H),0.41–0.14(m,4H),0.13–0.02(m,2H).

[0282] Example 7: Preparation of Compound Z-7

[0283]

[0284] Step 1-2: Refer to the method of step 1-2 of Example 1, except that 3-amino-1H-pyrazole-4-carbonitrile is used instead of 1H-pyrazole-5-amine to prepare compound 7-1 (pale yellow oil, yield: 46.6%), MS m / z (ESI): 273.1 [M+1].

[0285] Step 3: Dissolve compound 7-1 (31 mg, 0.11 mmol), tert-butyl ((1S,3S)-3-aminocyclopentyl)carbamate (22.76 mg, 0.11 mmol), and potassium carbonate (22.76 mg, 0.11 mmol) in 3 mL of acetonitrile. The reaction mixture was stirred at 60°C for 3 hours. The reaction mixture was concentrated under reduced pressure, 20 mL of water was added, and the mixture was extracted with dichloromethane (20 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound 7-2 (30 mg, yellow oil) in a yield of 60.46%. MS m / z (ESI): 437.2 [M+1].

[0286] Step 4: Dissolve compound 7-2 (30 mg, 0.068 mmol) and 5 M sodium hydroxide solution (0.5 mL) in ethanol (1 mL) and dimethyl sulfoxide (1 mL). Stir the reaction mixture at 60°C for 3 hours. After completion of the reaction, extract with dichloromethane (20 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, and concentrated under reduced pressure to afford compound 7-3 (30 mg, yellow oil) in a 91.23% yield. MS m / z (ESI): 455.2 [M+1].

[0287] Step 5: Dissolve compound 7-3 (30 mg, 0.07 mmol) in 3 mL of hydrochloric acid / 1,4-dioxane solution and stir at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure and purified by preparative HPLC to afford compound Z-7 (4.57 mg) in a 19.46% yield. MS m / z (ESI): 355.1 [M+1]. 1 H NMR (400MHz, DMSO-d6) δ8.34(s,1H),8.10(s,1H),7.77(s,1H),7.27(s,1H),6.28(s,1H),4.35-4.22(m,1H),3.55-3.43(m,1H),2. 30-2.18(m,1H),2.10-1.90(m,3H),1.89-1.65(m,3H),1.46–1.24(m,4H),0.62-0.53(m,2H),0.41-0.30(,4H),0.20–0.08(m,2H).

[0288] Example 8: Preparation of Compound Z-8

[0289]

[0290] Compound Z-8 was prepared by referring to the preparation method of compound Z-1, except that 5-methyl-1H-pyrazol-3-amine was used instead of 1H-pyrazol-5-amine. (Yield: 33.4%). MS m / z (ESI): 326.2 [M+1]; 1 H NMR(400MHz,DMSO-d6)δ7.23(s,1H),6.07(s,1H),5.95(s,1H),4.24-4.08(m,1H),3.46–3.36(m,1H),2.35(s,3H),2.25-2.1 0(m,1H),1.99–1.80(m,3H),1.80–1.55(m,3H),1.35–1.12(m,4H),0.56-0.48(m,2H),0.39–0.19(m,4H),0.15–0.02(m,2H).

[0291] Example 9: Preparation of Compound Z-9

[0292]

[0293] Compound Z-9 was prepared by referring to the preparation method of compound Z-1, except that 4-methyl-1H-pyrazol-3-amine was used instead of 1H-pyrazol-5-amine. MS m / z (ESI): 326.2 [M+1]; 1 H NMR(400MHz, DMSO-d6)δ7.83(s,1H),7.30(d,J=7.6Hz,1H),5.98(s,1H),4.25-4.16(m,1H),3.43-3.35(m,1H),2.24–2.14 (m,4H),1.96–1.83(m,2H),1.79–1.57(m,4H),1.33–1.16(m,4H),0.57–0.48(m,2H),0.38–0.22(m,4H),0.13-0.03(m,2H).

[0294] Example 10: Preparation of Compound Z-10

[0295]

[0296] Step 1: Dissolve compound 1-3 (200 mg, 0.485 mmol) and potassium thiocyanate (141.68 mg, 1.46 mmol) in methanol (5 mL) and stir at room temperature for 3 hours. Extract with dichloromethane (15 mL x 2). Combine the organic phases, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. Column chromatography (PE:EA = 4:1) afforded compound 10-1 (169 mg, yellow solid) in a 74.21% yield. MS m / z (ESI): 469.2 [M+1].

[0297] Step 2: Dissolve compound 10-1 (169 mg, 0.36 mmol) in 3 mL of tetrahydrofuran. Add 3 M methylmagnesium chloride solution (0.2 mL) at 0°C. Stir the reaction mixture at 0°C for 2 hours, then add acetic acid (43.31 mg, 0.72 mmol). Concentrate the reaction mixture under reduced pressure, add 20 mL of water, and extract with ethyl acetate (15 mL x 3). The combined organic phases are dried over anhydrous sodium sulfate and concentrated under reduced pressure to afford compound 10-2 (105 mg, yellow solid) in a 63.62% yield. MS m / z (ESI): 458.2 [M+1].

[0298] Step 3: Dissolve compound 10-2 (105 mg, 0.229 mmol) in dichloromethane (3 mL). Add m-chloroperbenzoic acid (73.31 mg, 0.482 mmol) with stirring. Stir the reaction mixture at room temperature for 2 hours. After completion of the reaction, extract with ethyl acetate (15 mL x 3). The combined organic phases are dried over anhydrous sodium sulfate and concentrated under reduced pressure. Column chromatography (PE:EA = 4:1) afforded compound 10-3 (83 mg, pale yellow solid) in a yield of 79.44%. MS m / z (ESI): 490.2 [M+1].

[0299] Step 4: Dissolve compound 10-3 (83 mg, 0.17 mmol) in 3 mL of hydrochloric acid / 1,4-dioxane solution and stir at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure and purified by preparative HPLC to afford compound Z-10 (24.27 mg) in a 33.45% yield. MS m / z (ESI): 390.1 [M+1]. 1H NMR (400MHz, DMSO-d6) δ8.37(s,1H),6.85(s,1H),6.43(s,1H),6.05(s,2H),4.31–4.21(m,1H),4.00–4.92(m,1H),3.28(s,3H),2.25–2.09( m,1H),2.06–1.87(m,3H),1.78–1.66(m,1H),1.51–1.34(m,2H),1.29– 1.17(m,2H),0.62-0.50(m,2H),0.45–0.25(m,4H),0.16-0.06(m,2H).

[0300] Example 11: Preparation of Compound Z-11

[0301]

[0302] Compound Z-11 was prepared by referring to the preparation method of compound Z-1, except that intermediate V5 was used instead of intermediate V1. MS m / z (ESI): 300.2 [M+1]; 1 H NMR(400MHz,DMSO-d6)δ7.97(s,1H),7.54(s,1H),6.28(s,1H),5.99(s,1H),4.13-4.22(m,1H),3.90-3.99(m,1H),2.75-2.6.3 (m,1H),2.51-2.57(m,1H),2.20-1.96(m,3H),1.85-1.94(m,2H),1.60-1.78(m,6H),1.40-1.49(m,1H),1.11(d,J=6.8Hz,3H).

[0303] Example 11a: Preparation of Compound Z-11-P1

[0304]

[0305] Compound Z-11 (94.46 mg, 315.48 μmol) was subjected to chiral separation (column type: IC-3 4.6*100 mm 3 μm; cosolvent: IPA [1% NH3 (7 M in MeOH)]; injection volume: 5.00 μl; wavelength: 220.0 nm; run time: 6.0 minutes; flow rate: 3.0 mL / min; pressure: 2000 psi; column temperature: 40°C) to obtain compound Z-11-P1 (9.70 mg, retention time 2.471 min) in a yield of 9.89% and a purity of 96.34%. MS m / z (ESI): 300.2 [M+H + ].1 H NMR(400MHz,DMSO-d6)δ7.96(d,J=2.0Hz,1H),7.41(d,J=7.6Hz,1H), 6.27(d,J=2.0Hz,1H),5.96(s,1H),4.21(q,J=7.2Hz,1H),3.42(q,J=6.0Hz,1H),2.65-2.74(m,1H),2.58–2.50(m,1H) ,2.16-2.24(m,1H),2.03-2.09(m,1H),1.83-1.95(m,2H),1.80–1.61(m,7H),1.35–1.25(m,1H),1.11(d,J=6.8Hz,3H).

[0306] Example 11b: Preparation of Compound Z-11-P2

[0307]

[0308] Compound Z-11 (94.46 mg, 315.48 μmol) was subjected to chiral separation (column type: IC-3 4.6*100 mm 3 μm; cosolvent: IPA [1% NH3 (7 M in MeOH)]; injection volume: 5.00 μl; wavelength: 220.0 nm; run time: 6.0 minutes; flow rate: 3.0 mL / min; pressure: 2000 psi; column temperature: 40°C) to obtain compound Z-11-P2 (9.37 mg, retention time 2.012 min) in a yield of 9.55% and a purity of 96.24%. MS m / z (ESI): 300.2 [M+H + ]. 1 H NMR(400MHz,DMSO-d6)δ7.96(d,J=2.0Hz,1H),7.42(d,J=7.6Hz,1H),6.27( d,J=2.0Hz,1H),5.96(s,1H),4.21(q,J=7.2Hz,1H),3.44-3.37(m,1H),2.65 -2.74(m,1H),2.58–2.50(m,1H),2.15-2.25(m,1H),2.02-2.08(m,1H),1.8 2-1.95(m,2H),1.79–1.60(m,7H),1.35–1.24(m,1H),1.10(d,J=6.8Hz,3H).

[0309] Example 12: Preparation of Compound Z-12

[0310]

[0311] Compound Z-12 was prepared by referring to the preparation method of compound Z-1, except that intermediate V7 was used instead of intermediate V1. MS m / z (ESI): 300.2 [M+1]; 1 H NMR (400MHz, DMSO-d6) δ8.01(d,J=2.0Hz,1H),7.43(d,J=6.0Hz,1H),6.34(d,J=2.0Hz,1H),6.23(s,1H),4.30–4.18(m,1H),3.46–3.40(m,1H),2 .26–2.18(m,1H),1.99–1.86(m,2H),1.83–1.74(m,1H),1.73–1.64(m,1H ),1.37–1.27(m,1H),1.17(s,6H),1.15–1.11(m,1H),0.39–0.34(m,4H).

[0312] Example 13: Preparation of Compound Z-13

[0313]

[0314] Compound Z-13 was prepared by referring to the preparation method of compound Z-1, except that intermediate V6 was used instead of intermediate V1. MS m / z (ESI): 314.2 [M+1]; 1 H NMR (400MHz, DMSO-d6) δ7.99(d,J=2.4Hz,1H),7.50(d,J=38.8Hz,1H),6.30(d,J=2.4Hz,1H),6.02(d,J=8.0Hz,1H),4.23(s,1H),3. 50-3.38(m,1H),2.52-2.55(m,1H),2.07-2.25(m,2H),1.99–1.77(m,4H),1.71–1.29(m,7H),1.26-1.16(m,4H),1.12–1.01(m,1H).

[0315] Example 14: Preparation of Compound Z-14

[0316]

[0317] Compound Z-14 was prepared by referring to the preparation method of compound Z-1, except that 4-cyclopropyl-1H-pyrazol-3-amine was used instead of 1H-pyrazol-5-amine. MS m / z (ESI): 352.2 [M+1]; 1H NMR(400MHz, DMSO-d6)δ7.74(s,1H),7.29(d,J=7.2Hz,1H),5.98(s,1H),4.24-4.12(m,1H),3.45–3.36(m,1H),2.27–1.79(m,6H),1 .76-1.70(m,1H),1.67-1.56(m,1H),1.35–1.14(m,4H),0.89–0.71(m,4H),0.57–0.48(m,2H),0.39–0.20(m,4H),0.15–0.06(m,2H).

[0318] Example 15: Preparation of Compound Z-15

[0319]

[0320] Step 1: Dissolve compound 1-2 (145 mg, 0.585 mmol) and acetic acid (70.24 mg, 1.17 mmol) in acetonitrile (5 mL). Add Selectfluor reagent (207.36 mg, 0.584 mmol) with stirring, and react at 70°C for 4 hours. Extract with ethyl acetate (15 mL x 2). Combine the organic phases, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. Column chromatography (PE:EA = 4:1) afforded compound 15-1 (45 mg, yellow liquid) in a 28.93% yield. MS m / z (ESI): 266.0 [M+1].

[0321] Step 2-3: Refer to the preparation method of step 3 and step 4 of compound Z-1, except that compound 15-1 is used instead of compound 1-2 to prepare compound Z-15. MS m / z (ESI): 330.3 [M+1]; 1 H NMR(400MHz, DMSO-d6)δ8.16(d,J=3.6Hz,1H),7.65(s,1H),6.08(s,1H),4.30-4.15(m,1H),3.48–3.42(m,1H),2.26-2.15(m,1H),2 .00-1.85(m,2H),1.82-1.76(m,1H),1.72-1.60(m,1H),1.36–1.16(m,4H),0.61–0.50(m,2H),0.39–0.25(m,4H),0.15-0.05(m,2H).

[0322] Example 16: Preparation of Compound Z-16

[0323]

[0324] Compound Z-16 was prepared by referring to the preparation method of compound Z-1, except that 4-ethyl-1H-pyrazol-3-amine was used instead of 1H-pyrazol-5-amine. MS m / z (ESI): 340.2 [M+1]; 1 H NMR(400MHz, DMSO-d6)δ7.86(s,1H),7.30(d,J=7.6Hz,1H),5.99(s,1H),4.25-4.15(m,1H),3.37-3.45(m,1H),2.63(q,J=7.6Hz,2H),2.14-2.2 3(m,1H),1.98–1.81(m,2H),1.71-1.79(m,2H),1.69–1.56(m,1H),1.35 –1.17(m,6H),0.47-0.56(m,2H),0.23-0.34(m,4H),0.15–0.03(m,2H).

[0325] Example 17: Preparation of Compound Z-17

[0326]

[0327] Compound Z-17 was prepared by referring to the preparation method of compound Z-1, except that compound V9 was used instead of compound V1. MS m / z (ESI): 286.2 [M+1]; 1 H NMR (400MHz, DMSO-d6) δ8.01(d,J=2.4Hz,1H),7.64(s,1H),6.90(d,J=6.4Hz,1H),6.32(d,J=2.4Hz,1H),6.07(s,2H),4.15-4.12(m,1H),4.03 -3.91(m,1H),2.57(s,2H),2.25-2.10(s,1H),2.06-1.85(m,3H),1.76- 1.63(m,1H),1.53-1.40(m,1H),1.02(s,3H),0.56(s,2H),0.31(s,2H).

[0328] Example 18: Preparation of Compound Z-18

[0329]

[0330] Referring to the preparation method of compound Z-1, the difference is that compound V4 is used instead of compound V1 to prepare compound Z-18 (yield 25.40%). MS m / z (ESI): 286.1 [M+1]; 1H NMR(400MHz, DMSO-d6)δ8.03(d,J=2.2Hz,1H),7.61&6.89(s,1H),6.37(d,J=2.2 Hz,1H),6.17(dd,J=17.2,10.6Hz,1H),6.04(s,1H),5.11(dd,J=17.6,1.2Hz,1H ),5.07(dd,J=10.6,1.2Hz,1H),4.22-4.14(m,1H),4.00–3.93(m,1H),2.20–2.0 7(m,1H),2.03-1.86(m,3H),1.74–1.62(m,1H),1.53–1.41(m,1H),1.41(s,6H).

[0331] Example 19: Preparation of Compound Z-19

[0332]

[0333] Compound Z-19 was prepared by referring to the preparation method of compound Z-1, except that compound V4 was used instead of compound V1, and 4-methyl-1H-pyrazol-3-amine was used instead of 1H-pyrazol-5-amine. MS m / z (ESI): 300.2 [M+1]; 1 H NMR(400MHz, DMSO-d6)δ7.86(s,1H),7.46-7.34(m,1H),6.17(dd,J=17.6,10.4Hz,1H),5.94(d,J=6.0Hz,1H),5.07(dd,J=21.0,14.0Hz,2H),4.19–4.1 2(m,1H),3.43–3.37(m,1H),2.92(bs,2H),2.18(s,3H),2.21-1.96(m,1H), 1.94–1.82(m,2H),1.77–1.66(m,1H),1.67–1.59(m,1H),1.49–1.24(m,7H).

[0334] Example 20 to Example 25

[0335] Compounds Z-20 to Z-25 can be prepared by referring to the method of Example 1.

[0336]

[0337]

[0338] Example 26: Preparation of Compound Z-26

[0339]

[0340] Compound Z-26 was prepared using compound 22-2 as a starting material according to the methods of steps 3 and 4 in Example 1. MS m / z (ESI): 314.1 [M+1]. 1 H NMR(400MHz, DMSO-d6)δ7.85(s,1H),7.28(d,J=6.0Hz,1H),5.97(s,1H),4.26–4 .12(m,1H),3.50–3.38(m,1H),2.35-2.27(m,1H),2.25-2.10(m,5H),1.97–1.86( m,2H),1.83–1.72(m,1H),1.72–1.56(m,2H),1.34(d,J=7.2Hz,1H),1.29(d,J=7 .2Hz,3H),0.96(s,3H),0.75–0.66(m,1H),0.48–0.38(m,1H),0.23-0.18(m,2H).

[0341] Example 26a: Preparation of Compound Z-26-P1

[0342]

[0343] Step 1: Compound Z-26-1 (650 mg) was separated by supercritical fluid chromatography (SFC) (column type: IC-3 4.6*100mm 3um, cosolvent: IPA / ACN=1 / 1 [0.1% DEA] flow rate: 3.0 mL / min, column temperature: 40°C) to obtain 212 mg of compound Z-26-a (retention time 2.468 minutes).

[0344] Step 2: Compound Z-26-a (212 mg, 512.63 μmol) was dissolved in 10 mL of hydrochloric acid / dioxane (4 M). The reaction was allowed to react at room temperature for 2 h. The reaction mixture was evaporated to dryness, and the residue was dissolved in 20 mL of ethyl acetate, washed twice with saturated sodium bicarbonate, dried over anhydrous sodium sulfate, filtered, and evaporated. The residue was purified by preparative liquid chromatography (preparative column: 21.2 × 250 mm C18 column; system: 10 mM NH4HCO3H2O; wavelength: 254 / 214 nm; gradient: 5%-95% acetonitrile) to obtain 60.12 mg of compound Z-26-P1, yield: 36.79%. MS m / z (ESI): 314.1 [M+1]. 1H NMR(400MHz,DMSO-d6)δ7.83(s,1H),7.36(s,1H).5.96(s,1H),4.25–4.05(m,1H),3.50–3.38(m,1H),2.35-2.25(m,1H),2.25-2.07(m,4H),2.05 -1.80(m,3H),1.75-1.60(m,1H),1.53-1.40(m,1H),1.27(d,J=7.2Hz,3H ),0.94(s,3H),0.74–0.64(m,1H),0.44–0.37(m,1H),0.31–0.19(m,2H).

[0345] Example 26b: Preparation of Compound Z-26-P2

[0346]

[0347] Step 1: Compound Z-26-1 (650 mg) was separated by supercritical fluid chromatography (SFC) (method: column type: IC-34.6*100mm 3um, cosolvent: IPA / ACN=1 / 1 [0.1% DEA] flow rate: 3.0 mL / min, column temperature: 40°C) to obtain 120 mg of compound Z-26-b (retention time 2.976 minutes).

[0348] Step 2: Dissolve compound Z-26-b (120 mg, 290.17 μmol) in 10 mL of hydrochloric acid / dioxane (4 M). Reaction was allowed to proceed at room temperature for 2 h. The reaction mixture was evaporated to dryness, and the residue was dissolved in 20 mL of ethyl acetate, washed twice with saturated sodium bicarbonate, dried over anhydrous sodium sulfate, filtered, and evaporated. The residue was purified by preparative liquid chromatography (preparative column: 21.2 × 250 mm C18 column; system: 10 mM NH4HCO3H2O; wavelength: 254 / 214 nm; gradient: 5%-95% acetonitrile) to obtain 34.2 mg of compound Z-26-P2, yield: 36.96%. MS m / z (ESI): 314.1 [M+1]. 1H NMR (400MHz, DMSO-d6) δ7.83 (s, 1H), 7.24 (d, J = 7.2Hz, 1H), 5.96 (s, 1H), 4 .23-4.13(m,1H),3.47–3.37(m,1H),2.35-2.25(m,1H),2.25–2.10(m,4H) ,2.00–1.82(m,2H),1.81-1.72(m,1H),1.70–1.57(m,1H),1.37–1.23(m,4 H),0.94(s,3H),0.73–0.64(m,1H),0.45-0.37(m,1H),0.34–0.19(m,2H).

[0349] Example 27 to Example 30

[0350] Compounds Z-27 to Z-30 can be prepared by referring to the method of Example 1.

[0351]

[0352] Example 27a: Preparation of Compound Z-27-P1

[0353]

[0354] Step 1: Compound Z-27-1 was prepared using compound 51-2 as a starting material according to the method of step 3 in Example 1. MS m / z (ESI): 400.2 [M+1].

[0355] Step 2: 410 mg of compound Z-27-1 was separated by supercritical fluid chromatography (SFC) (column type: IG 4.6*100mm5um, cosolvent: MeOH [0.2% NH3 (7M in MeOH)], injection volume: 5.00ul, run time: 4.0 minutes, flow rate: 3.0mL / min, pressure: 2000psi, column temperature: 40°C) to obtain 160 mg of compound Z-27-a (retention time 1.954 minutes).

[0356] Step 3: Compound Z-27-P1 was prepared by referring to Step 2 of Example 26a. MS m / z (ESI): 300.2 [M+1]. 1H NMR (400MHz, DMSO-d6) δ8.00(d,J=2.4Hz,1H),7.44(s,1H),6.35(d,J=2.0Hz,1H),6.05(s,1H),4.28-4.15(m,1H),3.49–3.38(m,1H),2.38– 2.10(m,2H),2.05–1.88(m,2H),1.87–1.61(m,2H),1.55–1.22(m,4H), 0.94(s,3H),0.75–0.64(m,1H),0.47-0.38(m,1H),0.34–0.13(m,2H).

[0357] Example 27b: Preparation of Compound Z-27-P2

[0358]

[0359] Step 1: 410 mg of compound Z-27-1 was separated by supercritical fluid chromatography (SFC) (column type: IG 4.6*100mm5um, cosolvent: MeOH [0.2% NH3 (7M in MeOH)], injection volume: 1.00ul, run time: 4.0 minutes, flow rate: 3.0mL / min, pressure: 2000psi, column temperature: 40°C) to obtain 120 mg of compound Z-27-b (retention time 2.336 minutes).

[0360] Step 2: Compound Z-27-P2 was prepared by referring to Step 2 of Example 26a. MS m / z (ESI): 300.2 [M+1]. 1 H NMR(400MHz, DMSO-d6)δ8.00(d,J=2.4Hz,1H),7.45(s,1H),6.35(d,J=2.0Hz,1H),6.04(s,1H),4.27-4.15(m,1H),3.50-3.40(m,1H),2.37– 2.11(m,2H),2.05–1.87(m,2H),1.84–1.55(m,2H),1.54–1.19(m,4H), 0.94(s,3H),0.77–0.59(m,1H),0.47-0.39(m,1H),0.35-0.22(m,2H).

[0361] Example 28a: Preparation of Compound Z-28-P1

[0362]

[0363] Step 1: Dissolve compound 23-2 (5 g, 21.21 mmol) in acetonitrile (30 mL), add tert-butyl ((1S,3S)-3-aminocyclopentyl)carbamate (5.10 g, 25.45 mmol) and potassium carbonate (8.80 g, 63.64 mmol). React at 90°C for 16 h. Filter the reaction mixture and spin dry. The residue is combined with petroleum ether:ethyl acetate in a ratio of 3:1 to afford compound Z-28-1 (7 g, yield: 82.6%) as a yellow liquid. MS m / z (ESI): 400.2 [M+1].

[0364] Step 2: Compound Z-28-1 (6.0 g) was separated by supercritical fluid chromatography (SFC) (method: column type: IG4.6*100mm 5um, cosolvent: IPA / ACN=1 / 1 [0.1% DEA] flow rate: 3.0 mL / min column temperature: 40°C) to obtain 2.2 g of compound Z-28-a (retention time 1.931 minutes).

[0365] Step 3: Dissolve compound Z-28-a (2.2 g, 5.51 mmol) in hydrochloric acid / dioxane (10 mL) and react at 30°C for 2 h. The reaction mixture was dried by vortexing. The residue was dissolved in 50 mL of water and extracted twice with ethyl acetate. The aqueous phase was adjusted to pH 8 with aqueous ammonia, extracted twice with ethyl acetate, and dried over anhydrous sodium sulfate. Filter and vortex to obtain compound Z-28-P1 (1.3 g, yield: 79.1%). MS m / z (ESI): 300.2 [M+1]. 1 H NMR(400MHz,DMSO-d6)δ7.98(d,J=2.0Hz,1H),7.41(d,J=7.6Hz,1H),6.29 (d,J=2.0Hz,1H),6.00(s,1H),4.28-4.16(m,1H),3.50-3.40(m,1H),2.27- 2.17(m,1H),2.00–1.57(m,7H),1.35-1.24(m,1H),1.11–0.97(m,1H),0.80 (t,J=7.6Hz,3H),0.60-0.51(m,1H),0.35-0.20(m,2H),0.18-0.08(m,1H).

[0366] Example 28b: Preparation of Compound Z-28-P2

[0367]

[0368] Step 1: Compound Z-28-1 (6.0 g) was separated by supercritical fluid chromatography (SFC) (method: column type: IG4.6*100mm 5um, cosolvent: IPA / ACN=1 / 1 [0.1% DEA] flow rate: 3.0 mL / min column temperature: 40°C) to obtain 2.3 g of compound Z-28-b (retention time 2.566 minutes).

[0369] Step 2: Compound Z-28-b (220 mg, 550.6 μmol) was dissolved in hydrochloric acid / dioxane (10 mL) and reacted at 30°C for 2 h. The reaction mixture was dried by rotary evaporation. The residue was dissolved in ethyl acetate and washed twice with saturated sodium bicarbonate. Drying was performed over anhydrous sodium sulfate. Filtering was performed and the residue was dried by rotary evaporation. The residue was purified by preparative liquid chromatography (preparative column: 21.2 × 250 mm C18 column; system: 10 mM NH4HCO3 H2O; wavelength: 254 / 214 nm; gradient: 5%-95% acetonitrile) to obtain compound Z-28-P2 (99.58 mg, yield: 60.10%). MS m / z (ESI): 300.2 [M+1]. 1 H NMR(400MHz,DMSO-d6)δ7.98(d,J=2.0Hz,1H),7.42(d,J=6.0Hz,1H),6.29 (d,J=2.4Hz,1H),6.01(s,1H),4.26-4.16(m,1H),3.50-3.40(m,1H),2.25- 2.17(m,1H),2.00–1.57(m,7H),1.34-1.24(m,1H),1.10–0.97(m,1H),0.80 (t,J=7.2Hz,3H),0.58-0.48(m,1H),0.32-0.17(m,2H),0.16-0.06(m,1H).

[0370] Example 30a: Preparation of Compound Z-30-P1

[0371]

[0372] Step 1: Compound Z-30-1 was prepared using compound 25-2 as a raw material using the method of step 3 in Example 1. MS m / z (ESI): 414.2 [M+1].

[0373] Step 2: 1.1 g of compound Z-30-1 was separated by supercritical fluid chromatography (SFC) (column type: IC-3 4.6*100 mm3um, co-solvent: IPA [1% NH3 (7M in MeOH)], injection volume: 5.00 ul, run time: 6.0 minutes, flow rate: 3.0 mL / min, pressure: 2000 psi, column temperature: 40°C) to obtain 370 mg of compound Z-30-a (retention time 2.135 minutes).

[0374] Step 3: Compound Z-30-P1 was prepared by referring to Step 2 of Example 26a. MS m / z (ESI): 314.2 [M+1]. 1 H NMR(400MHz, DMSO-d6)δ7.83(s,1H),7.25(d,J=7.6Hz,1H),5.95(s,1H),4.26-4.16(m,1H),3.45–3.36(m,1H),2.26–2.13(m,4H),2 .02–1.54(m,7H),1.33-1.22(m,1H),1.10–0.99(m,1H),0.85-0.78(m,3H),0.59–0.49(m,1H),0.32–0.17(m,2H),0.14-0.08(m,1H).

[0375] Example 30b: Preparation of Compound Z-30-P2

[0376]

[0377] Step 1: 1.1 g of compound Z-30-1 was separated by supercritical fluid chromatography (SFC) (column type: IC-3 4.6*100 mm3um, co-solvent: IPA [1% NH3 (7M in MeOH)], injection volume: 5.00 ul, run time: 6.0 minutes, flow rate: 3.0 mL / min, pressure: 2000 psi, column temperature: 40°C) to obtain 380 mg of compound Z-30-b (retention time 2.416 minutes).

[0378] Step 2: Compound Z-30-P2 was prepared by referring to Step 2 of Example 26a. MS m / z (ESI): 314.2 [M+1]. 1H NMR(400MHz, DMSO-d6)δ7.83(s,1H),7.25(d,J=7.6Hz,1H),5.96(s,1H),4.25-4.15(m,1H),3.48–3.35(m,1H),2.24–2.09(m,4H),2.01–1.70 (m,6H),1.68-1.56(m,1H),1.35-1.25(m,1H),1.10-1.00(m,1H),0.86 -0.79(m,3H),0.57–0.47(m,1H),0.35-0.19(m,2H),0.15-0.08(m,1H).

[0379] Example 31 Preparation of Compound Z-31

[0380]

[0381] Step 1: Dissolve compound 5-2 (400 mg, 1.80 mmol) and acetic acid (216.70 mg, 3.61 mmol) in acetonitrile (20 mL). Add 1-chloromethyl-4-fluoro-1,4-diazoniabicyclo[2.2.2]octane bis(tetrafluoroborate) (639.2 mg, 1.80 mmol) and stir at 70°C for 4 hours. Stop the reaction, evaporate the solvent under reduced pressure, and dissolve the mixture in ethyl acetate (50 mL). The organic phase is washed with saturated sodium bicarbonate solution (50 mL) and saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification by silica gel column chromatography affords compound 31-1 (200 mg, yield: 46.3%) as a light yellow oil. MS m / z (ESI): 240.1 [M+1].

[0382] Step 2: Dissolve compound 31-1 (100 mg, 417.23 μmol) and tert-butyl 3-aminopyrrolidine-1-carboxylate (116.6 mg, 625.84 μmol) in acetonitrile (20 mL), then add potassium carbonate (173.00 mg, 1.25 mmol). Stir the reaction at 90°C for 16 hours. Stop the reaction, dissolve the mixture in ethyl acetate (50 mL), wash with saturated sodium chloride solution (50 mL x 3), dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue is purified by silica gel column chromatography to afford compound 31-2 (120 mg, yield: 73.85%) as a light yellow oil. MS m / z (ESI): 389.6 [M+1].

[0383] Step 3: Compound 31-2 (120 mg, 308.11 μmol) was dissolved in a 4 M hydrogen chloride / 1,4-dioxane solution (4 M, 7.70 mL). The reaction was stirred at room temperature for 3 hours. The solvent was evaporated under reduced pressure, purified water (30 mL) was added, and impurities were extracted with ethyl acetate (30 mL). The aqueous phase was adjusted to pH 9-10 with saturated sodium carbonate solution and extracted with ethyl acetate (50 mL x 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to yield compound 31-3 (89 mg, yield: 99.83%) as a light yellow oil. MS m / z (ESI): 290 [M+1].

[0384] Step 4: Dissolve compound 31-3 (90 mg, 311.04 μmol) in water (2 mL) and acetic acid (2 mL). Slowly add a solution of sodium nitrite (31.73 mg, 466.56 μmol) in water (2 mL) dropwise under ice. Return to room temperature and stir for 2 h. Adjust the pH to 8 with saturated sodium bicarbonate. Extract with 2 x 30 mL of ethyl acetate, dry over anhydrous sodium sulfate, and spin dry. The residue was combi-flashed with petroleum ether:ethyl acetate in a 1:1 ratio to afford compound 31-4 (55 mg, yield: 55.54%) as a pale yellow solid. MS m / z (ESI): 319 [M+1].

[0385] Step 5: Compound 31-4 (55 mg, 172.77 μmol) was dissolved in methanol (3 mL) and acetic acid (0.3 mL). Zinc (225.94 mg, 3.46 mmol) was added and allowed to react at room temperature for 1 h. Aqueous ammonia was added to adjust the reaction mixture to pH 9. Extraction was performed with 2 x 30 mL of ethyl acetate. The organic phases were combined, washed twice with saturated brine, dried over anhydrous sodium sulfate, filtered, and the residue was dried by spin drying. The crude product was purified by preparative liquid chromatography (preparative column: 21.2 x 250 mm C18 column; system: 10 mM NH₄HCO₃H₂O; wavelength: 254 / 214 nm; gradient: 5%-95% acetonitrile) to afford compound Z-31 (4.98 mg, yield: 9.07%). MS m / z (ESI): 304.6 [M+1]. 1H NMR (400MHz, DMSO-d6) δ8.16 (dd, J=6.0, 3.6Hz, 1H), 7.58 (d, J=6.8Hz, 1H), 6.12 (s,1H),4.28-4.16(m,1H),3.01–2.91(m,1H),2.91–2.68(m,2H),2.67–2.52(m, 1H),2.33–2.18(m,1H),2.07-1.95(m,1H),1.93-1.81(m,1H),1.30(d,J=6.8Hz, 3H),1.10-0.98(m,1H),0.55-0.47(m,1H),0.39-0.30(m,1H),0.27-0.15(m,2H).

[0386] Example 32: Preparation of Compound Z-32

[0387]

[0388] Compound Z-32 was prepared using compound 26-2 as a starting material according to the methods of steps 3 and 4 in Example 1. MS m / z (ESI): 300.2 [M+1]. 1 H NMR(400MHz,DMSO-d6)δ7.83(s,1H),7.43(s,1H),6.01(s,1H),4.25–4.13(m,1H),3.99–3.91(m,1H),2.24–1.81(m,4H),2.16(s,3H),1.79–1.72 (m,1H),1.67–1.58(m,1H),1.50–1.40(m,1H),1.30(d,J=6.8Hz,3H),1.1 0–1.01(m,1H),0.54–0.46(m,1H),0.38–0.30(m,1H),0.23–0.16(m,2H).

[0389] Example 32a: Preparation of Compound Z-32-P1

[0390]

[0391] Step 1: 590 mg of compound Z-32-1 was separated by supercritical fluid chromatography (SFC) (AD-3 4.6*100 mm 3 μm column, cosolvent: EtOH [1% NH3 (7 M in MeOH)], injection volume: 2.00 μl, run time: 4.0 minutes, flow rate: 3.0 mL / min, pressure: 2000 psi, column temperature: 40°C) to afford compound Z-32-a (218 mg, 545.64 μmol, yield 36.95%, retention time 2.592 minutes) as a yellow oil. MS m / z (ESI): 400.3 [M+1]. %ee = 100%.

[0392] Step 2: Compound Z-32-P1 was prepared by referring to Step 2 of Example 26a. MS m / z (ESI): 300.2 [M+1]. 1 H NMR(400MHz,DMSO-d6)δ7.83(s,1H),7.26(d,J=4.8Hz,1H),6.00(s,1H),4.24– 4.15(m,1H),3.44–3.36(m,1H),2.24–2.17(m,1H),2.17(s,3H),2.05–1.82(m,3 H),1.80–1.73(m,1H),1.67–1.58(m,1H),1.31(d,J=7.0Hz,3H),1.29–1.23(m,1 H),1.11–1.01(m,1H),0.53–0.48(m,1H),0.38–0.31(m,1H),0.24–0.16(m,2H).

[0393] Example 32b: Preparation of Compound Z-32-P2

[0394]

[0395] Step 1: 590 mg of compound Z-32-1 was separated by supercritical fluid chromatography (SFC) (AD-3 4.6*100 mm 3 μm column, cosolvent: EtOH [1% NH3 (7 M in MeOH)], injection volume: 2.00 μl, run time: 4.0 minutes, flow rate: 3.0 mL / min, pressure: 2000 psi, column temperature: 40°C) to afford compound Z-32-b (241 mg, 603.21 μmol, yield 40.85%, retention time 3.167 minutes) as a yellow oil. MS m / z (ESI): 400.3 [M+1]. ee% = 99.6%.

[0396] Step 2: Compound Z-32-P2 was prepared by referring to Step 2 of Example 26a. MS m / z (ESI): 300.2 [M+1]. 1 H NMR(400MHz,DMSO-d6)δ7.83(s,1H),7.27(s,1H),6.00(s,1H),4.26–4.15(m,1 H),3.44–3.38(m,1H),2.25–2.18(m,1H),2.17(s,3H),2.04–1.82(m,4H),1.79 –1.71(m,1H),1.68–1.59(m,1H),1.31–1.30(d,J=6.8Hz,4H),1.28–1.24(m,1H ),1.10–1.01(m,1H),0.53–0.47(m,1H),0.37–0.32(m,1H),0.24–0.16(m,2H).

[0397] Example 33 to Example 44

[0398] Compounds Z-33 to Z-44 can be prepared by referring to the method of Example 1.

[0399]

[0400]

[0401]

[0402] Example 38a: Preparation of Compound Z-38-P1

[0403]

[0404] Step 1: Compound Z-38-1 was prepared using compound 35-2 as a raw material using the method of step 3 in Example 1. MS m / z (ESI): 414.2 [M+1].

[0405] Step 2: 850 mg of compound Z-38-1 was separated by supercritical fluid chromatography (SFC) (column type: IC-3 4.6*100mm 3um, co-solvent: IPA [1% NH3 (7M in MeOH)], injection volume: 2.00ul, run time: 6.0 minutes, flow rate: 3.0mL / min, pressure: 2000psi, column temperature: 40°C) to obtain 230 mg of compound Z-38-a (retention time 3.138 minutes).

[0406] Step 3: Compound Z-38-P1 was prepared by referring to Step 2 of Example 26a. MS m / z (ESI): 314.2 [M+1]. 1 H NMR(400MHz, DMSO-d6)δ7.84(s,1H),7.29(d,J=7.6Hz,1H),5.90(s,1H),4.25–4.17(m,1H),3.45-3.38(m,1H),2.74–2.67(m,1H),2.61–2.52( m,1H),2.20(d,J=8.0Hz,1H),2.18(s,3H),2.12-2.05(m,1H),1.97–1. 82(m,2H),1.81–1.56(m,7H),1.35–1.25(m,1H),1.13(d,J=6.8Hz,3H).

[0407] Example 38b: Preparation of Compound Z-38-P2

[0408]

[0409] Step 1: 850 mg of compound Z-38-1 was separated by supercritical fluid chromatography (SFC) (column type: IC-3 4.6*100mm 3um, co-solvent: IPA [1% NH3 (7M in MeOH)], injection volume: 2.00ul, run time: 6.0 minutes, flow rate: 3.0mL / min, pressure: 2000psi, column temperature: 40°C) to obtain 230 mg of compound Z-38-b (retention time 3.507 minutes).

[0410] Step 2: Compound Z-38-P2 was prepared by referring to Step 2 of Example 26a. MS m / z (ESI): 314.2 [M+1]. 1 H NMR(400MHz, DMSO-d6)δ7.84(s,1H),7.30(d,J=7.6Hz,1H),5.91(s,1H),4.25–4.17(m,1H),3.46-3.39(m,1H),2.76-2.66(m,1H),2.61–2.5 2(m,1H),2.25-2.19(m,1H),2.18(s,3H),2.12-2.05(m,1H),1.98–1.8 2(m,2H),1.81–1.58(m,7H),1.36–1.26(m,1H),1.13(d,J=6.8Hz,3H).

[0411] Example 41a: Preparation of Compound Z-41-P1

[0412]

[0413] Compound Z-41 (76 mg, 250.51 μmol) was subjected to chiral separation (IC-3 4.6*100 mm 3 μm column; cosolvent: IPA [1% NH3 (7 M in MeOH)]; injection volume: 5.00 μl; run time: 6.0 minutes; flow rate: 3.0 mL / min; pressure: 2000 psi; column temperature: 40°C) to afford compound Z-41-P1 (9.76 mg, retention time 2.153 minutes) in a yield of 12.84% and a purity of 100%. MS m / z (ESI): 304.2 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δ8.08(d,J=2.4Hz,1H),7.82(s,1H),6.39(d,J=2.4Hz,1H),6.15(s,1H),5.34(dd,J=48.8,6.0Hz, 1H),4.22-4.32(m,1H),3.40-3.47(m,1H),2.86-2.99(m,1H),2.24–2.13(m,1H),2.12–1.60(m,12H),1.27-1.36(m,1H).

[0414] Example 41b: Preparation of Compound Z-41-P2

[0415]

[0416] Compound Z-41 (76 mg, 250.51 μmol) was subjected to chiral separation (IC-3 4.6*100 mm 3 μm column; cosolvent: IPA [1% NH3 (7 M in MeOH)]; injection volume: 5.00 μl; run time: 6.0 minutes; flow rate: 3.0 mL / min; pressure: 2000 psi; column temperature: 40°C) to afford compound Z-41-P2 (11.42 mg, retention time: 1.448 minutes) in a yield of 14.98% and a purity of 99.72%. MS m / z (ESI): 304.2 [M+H + ]. 1H NMR (400MHz, DMSO-d6) δ8.08(d,J=2.4Hz,1H),7.83(s,1H),6.39(d,J=2.4Hz,1H),6.15(s,1H),5.34(dd,J=48.8,6.0Hz, 1H),4.21-4.31(m,1H),3.48-3.41(m,1H),3.05-2.86(m,1H),2.15-2.25(m,1H),2.13-1.59(m,12H),1.27-1.37(m,1H).

[0417] Example 45: Preparation of Compound Z-45

[0418]

[0419] Step 1: Dissolve compound 5-2 (200 mg, 902.18 μmol), tert-butyl 3-aminopyrrolidine-1-carboxylate (201.64 mg, 1.08 mmol), and potassium carbonate (374.06 mg, 2.71 mmol) in acetonitrile (5 mL), warm to 90°C, and stir overnight. Add 5 mL of water to the reaction mixture, and extract with ethyl acetate (10 mL x 3). The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, and then dried under reduced pressure. The resulting residue was purified by silica gel column chromatography using 0-40% ethyl acetate in petroleum ether as eluent to afford compound 45-1 (0.28 g, light yellow oil) in a yield of 83.55%. MS m / z (ESI): 372.2 [M+1].

[0420] Step 2: Dissolve compound 45-1 (0.28 g, 753.75 μmol) in 4M hydrochloric acid-dioxane (5 mL) and stir at room temperature for 2 h. Remove the solvent under reduced pressure to afford crude compound 45-2 (230 mg, pale yellow solid hydrochloride salt) in a 99.13% yield. MS m / z (ESI): 308.2 [M+1].

[0421] Step 3: Dissolve compound 45-2 (150 mg, 487.30 μmol, HCl) in water (1 mL) and acetic acid (2 mL) and cool to 0°C under argon. Then, slowly add sodium nitrite (67.25 mg, 974.59 μmol) dissolved in water (1 mL) to the solution dropwise. Return to room temperature and stir for 1.5 h. Add water (10 mL) and extract with ethyl acetate (10 mL x 3). The combined organic phases are washed with aqueous sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, and spin-dried to afford crude compound 45-3 (140 mg, brown oil) in a yield of 95.65%. MS m / z (ESI): 301.1 [M+1].

[0422] Step 4: Dissolve compound 45-3 (140 mg, 466.11 μmol) in methanol (3 mL) and acetic acid (0.3 mL), then add zinc powder (609.58 mg, 9.32 mmol). Stir at room temperature for 1 h. Filter off the zinc powder, spin dry the solvent, dissolve in ethyl acetate (5 mL), wash with aqueous sodium bicarbonate and brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure to obtain the crude product. Preparative HPLC chromatography yielded compound Z-45 (36.59 mg) in a yield of 23.78% and a purity of 86.74%. MS m / z (ESI): 287.1 [M+1]. 1 H NMR(400MHz,DMSO-d6)δ7.99(d,J=2.4Hz,1H),7.40(s,1H),6.31(d,J=2.4Hz,1H),6.0 9(s,1H),4.24-4.15(m,1H),2.98-2.92(m,1H),2.84-2.77(m,1H),2.76-2.77(m,1H),2 .60-2.53(m,1H),2.31-2.19(m,1H),2.03-1.94(m,1H),1.93–1.80(m,1H),1.29(d,J=6 .8Hz,3H),1.10–0.99(m,1H),0.54–0.44(m,1H),0.37-0.30(m,1H),0.25–0.14(m,2H).

[0423] Examples 46 to 59, 64

[0424] Compounds Z-46 to Z-59 and Z-64 can be prepared by referring to the method of Example 1.

[0425]

[0426]

[0427]

[0428] Example 60: Preparation of Compound Z-60

[0429]

[0430] Step 1: Under nitrogen, N-bromosuccinimide (562.01 mg, 3.16 mmol) was dissolved in acetonitrile (5 mL) and added dropwise to a solution of compound 5-2 (700 mg, 3.16 mmol) in acetonitrile (10 mL) at 0°C. The mixture was allowed to react at room temperature for 0.5 hours. The reaction mixture was poured into water and extracted with ethyl acetate (5 mL x 2). The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 90%:10%) to obtain compound 49-2 (946 mg, 96.57% yield) as a yellow oil. MS m / z (ESI): 300.0 [M+1].

[0431] Step 2: Under nitrogen, compound 49-2 (300 mg, 998.06 μmol), potassium carbonate (344.84 mg, 2.50 mmol), tert-butyl ((1S,3S)-3-aminocyclopentyl)carbamate (239.87 mg, 1.20 mmol), and acetonitrile (10 mL) were mixed and heated to 90°C for 16 hours. The reaction mixture was filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 70%:30% to 65%:35%) to obtain compound Z-60-1 (433 mg, 93.42% yield) as a colorless oil. MS m / z (ESI): 464.2 [M+1].

[0432] Step 3: Under nitrogen, compound Z-60-1 (383 mg, 824.72 μmol), potassium vinyl trifluoroborate (331.41 mg, 2.47 mmol), 1,1'-bis(diphenylphosphinoferrocenepalladium) dichloride (120.69 mg, 164.94 μmol), potassium carbonate (455.93 mg, 3.30 mmol), 1,4-dioxane (10 mL), and water (2.5 mL) were mixed and microwaved at 120°C for 2 hours. The reaction mixture was filtered, and the filtrate was added with water and extracted with ethyl acetate (10 mL x 2). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 90%:10% to 70%:30%) to obtain compound Z-60-2 (320 mg, 94.28% yield) as a yellow oil. MS m / z(ESI):412.2[M+1].

[0433] Step 4: Under nitrogen, zinc bromide (1.05 g, 4.67 mmol) was added to a solution of compound Z-60-2 (320 mg, 777.57 μmol) and dichloromethane (18 mL). The mixture was allowed to react at room temperature for 16 hours. An additional zinc bromide (1.05 g, 4.67 mmol) was added, and the reaction was continued at room temperature for another 16 hours. Water and a small amount of methanol were added to the reaction solution, which was then extracted with dichloromethane (20 mL x 2). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by preparative liquid chromatography (preparative column: 21.2 × 250 mm C18 column; system: 10 mM NH4HCO3H2O; wavelength: 254 / 214 nm) to obtain compound Z-60 (24.73 mg, 9.82% yield). MS m / z (ESI): 312.2 [M+1]. 1 H NMR (400MHz, DMSO-d6) δ8.18(s,1H),6.86(s,1H),6.76(dd,J=18.0,11.6Hz,1H),6.15(s,1H ),5.93(dd,J=17.6,2.0Hz,1H),5.07(dd,J=11.2,2.4Hz,1H),4.25–4.18(m,1H),4.03–3.92 (m,1H),2.22–2.12(m,1H),2.09–1.90(m,4H),1.75–1.64(m,1H),1.51–1.45(m,1H),1.34(d ,J=7.2Hz,3H),1.13–1.04(m,1H),0.56–0.50(m,1H),0.42–0.35(m,1H),0.28–0.19(m,2H).

[0434] Example 61: Preparation of Compound Z-61

[0435]

[0436] Compound Z-9 (150 mg, 352.47 μmol) and ethylacetimidate hydrochloride (262.0 mg, 2.12 mmol) were dissolved in acetonitrile (10 mL), followed by the addition of N,N-diisopropylethylamine (0.68 mL, 3.91 mmol). The reaction was stirred at 80°C for 2 hours, cooled to room temperature, and 5 mL of 7N methanolic ammonia solution was added, followed by stirring at room temperature for 1 hour. The mixture was diluted with ethyl acetate (50 mL) and washed with saturated sodium chloride solution (50 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Preparative HPLC chromatography yielded compound Z-61 (33.58 mg) in a yield of 25.11%. MS m / z (ESI): 367.2 [M+1]. 1H NMR(400MHz, DMSO-d6)δ7.86(s,1H),7.42(s,1H),5.99(s,1H),4.24–4.10(m,1H),4.00–3.90(m,1H),2.29–2.12(m,4H),2.11–1.93(m,2 H),1.90–1.72(m,4H),1.68-1.59(m,1H),1.48-1.40(m,1H),1.37–1.12(m,3H),0.58-0.51(m,2H),0.38-0.24(m,4H),0.15-0.05(m,2H).

[0437] Example 62: Preparation of Compound Z-62

[0438]

[0439] Step 1: Dissolve compound 5-2 (100 mg, 451.1 μmol) and tert-butyl ((1R,3R)-3-aminocyclobutyl)carbamate (84.0 mg, 451.1 μmol) in acetonitrile (20 mL), then add potassium carbonate (187.0 mg, 1.35 mmol). The reaction is stirred at 90°C for 16 hours. Ethyl acetate (80 mL) is added, and the mixture is washed with saturated sodium chloride solution (80 mL x 3). The organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford compound Z-62-1 (120 mg, light yellow oil) in a yield of 71.6%. MS m / z (ESI): 372.2 [M+1].

[0440] Step 2: Compound Z-62-1 (120 mg, 323.04 μmol) was dissolved in 1,4-dioxane (2 mL), and a 4 M solution of hydrochloric acid in 1,4-dioxane (5 mL) was added. The reaction was stirred at room temperature for 5 hours. The solvent was evaporated under reduced pressure, and water (50 mL) was added. The mixture was extracted with ethyl acetate (50 mL x 2). The aqueous phase was adjusted to pH 9-10 with saturated sodium carbonate solution and extracted with ethyl acetate (60 mL x 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Preparative HPLC chromatography gave compound Z-62 (47.9 mg) in a yield of 54.3%. MS m / z (ESI): 272.2 [M+1]. 1H NMR (400MHz, DMSO-d6) δ8.00(s,1H),7.85(s,1H),6.30(d,J=2.0Hz,1H),5.85(s,1H),4.26-4.17(m,1H),3.56-3.47(s,1H),2.45- 2.26(m,3H),2.15–1.96(m,2H),1.28(d,J=7.2Hz,3H),1.05-0.96(m,1H),0.55-0.46(m,1H),0.37–0.27(m,1H),0.24-0.15(m,2H).

[0441] Example 62a: Preparation of Compound Z-62-P1

[0442]

[0443] Compound Z-62 (45 mg, 165.83 μmol) was subjected to chiral separation (Separation conditions: column type: IC-3 4.6*100 mm 3 μm; cosolvent: IPA (0.5% DEA); wavelength: 220.0 nm; flow rate: 3.0 mL / min; column temperature: 40°C) to afford compound Z-62-P1 (7.47 mg, retention time: 3.394 minutes) in a yield of 15.4%. MS m / z (ESI): 272.2 [M+1]. 1 HNMR (400MHz, DMSO-d6) δ8.00(d,J=2.4Hz,1H),7.85(s,1H),6.30(d,J=2.4Hz,1H),5.85(s,1H),4.24-4.16(m,1H),3.56-3.50(m,1H) ,2.49-2.5(m,2H),2.15–1.92(m,3H),1.35–1.18(m,3H),1.07–0.95(m,1H),0.54-0.45(m,1H),0.37-0.28(m,1H),0.22–0.15(m,2H).

[0444] Example 62b: Preparation of Compound Z-62-P2

[0445]

[0446] Compound Z-62 (45 mg, 165.83 μmol) was subjected to chiral separation (Separation conditions: column type: IC-3 4.6*100 mm 3 μm; cosolvent: IPA (0.5% DEA); wavelength: 220.0 nm; flow rate: 3.0 mL / min; column temperature: 40°C) to afford compound Z-62-P2 (7.74 mg, retention time: 2.850 minutes) in a yield of 16.3%. MS m / z (ESI): 272.2 [M+1]. 1 HNMR(400MHz, DMSO-d6)δ8.00(d,J=2.4Hz,1H),7.84(s,1H),6.30(d,J=2.4Hz,1H),5.85(s,1H),4.26-4.18(m,1H),3.56-3.47(m,1H) ,2.45–2.29(m,2H),2.15–1.89(m,3H),1.32-1.20(m,3H),1.06–0.96(m,1H),0.55-0.45(m,1H),0.37-0.29(m,1H),0.22-0.14(m,2H).

[0447] Example 63: Preparation of Compound Z-63

[0448]

[0449] Step 1: Dissolve compound 49-2 (550 mg, 2.15 mmol) and tert-butyl ((1S,3S)-3-aminocyclopentyl)carbamate (645.10 mg, 3.22 mmol) in acetonitrile (20 mL), then add potassium carbonate (890.32 mg, 6.44 mmol). Stir the reaction at 90°C for 16 hours. Stop the reaction and dilute with ethyl acetate (60 mL). The organic phase is washed with saturated sodium chloride solution (60 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford crude product Z-63-1 (850 mg, light yellow oil) in a yield of 94.3%. MS m / z (ESI): 420.2 [M+1].

[0450] Step 2: Compound Z-63-1 (550 mg, 1.31 mmol) was dissolved in 1,4-dioxane (2 mL), and a 4 M hydrogen chloride / 1,4-dioxane solution (4 M, 9.82 mL) was added. The reaction was stirred at room temperature for 3 hours. The solvent was evaporated under reduced pressure, and purified water (50 mL) was added. Impurities were extracted with ethyl acetate (50 mL). The aqueous phase was adjusted to pH 9-10 with aqueous ammonia solution and extracted with ethyl acetate (60 mL x 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford compound Z-63-2 (410 mg, light yellow oil) in a yield of 97.9%. MS m / z (ESI): 320.2 [M+1].

[0451] Step 3: Compound Z-63-2 (60 mg, 187.60 μmol) was dissolved in dichloromethane (10 mL), and acetyl chloride (73.63 mg, 937.99 μmol) was added, followed by N,N-diisopropylethylamine (0.33 mL, 1.88 mmol). The reaction was stirred at room temperature for 0.5 hours. The reaction was stopped and diluted with ethyl acetate (50 mL). The organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The residue was purified by preparative HPLC to afford compound Z-63 (47.0 mg) in a yield of 67.9%. MS m / z (ESI): 362.2 [M+1]. 1 H NMR(400MHz,DMSO-d6)δ8.15(s,1H),7.90-7.83(m,2H),6.14(s,1H), 4.25-4.14(m,2H),2.21–2.12(m,1H),2.08–1.95(m,3H),1.90–1.81(m,1H),1.72(s,3H),1.74–1.65(m,1H),1.47 -1.36(m,1H),1.31(d,J=6.8Hz,3H),1.11–1.02(m,1H),0.55-0.47(m,1H),0.39-0.31(m,1H),0.25–0.17(m,2H).

[0452] Example 64a: Preparation of Compound Z-64-P1

[0453]

[0454] Step 1: 1.2 g of compound Z-63-1 was separated by supercritical fluid chromatography (SFC) (column type: AD-3 4.6*100 mm3um, cosolvent: EtOH [1% NH3 (7M in MeOH)], injection volume: 1.00 ul, run time: 4.0 minutes, flow rate: 3.0 mL / min, pressure: 2000 psi, column temperature: 40°C) to obtain 440 mg of compound Z-63-a (retention time 1.329 minutes).

[0455] Step 2: Dissolve compound Z-63-a (170.00 mg, 404.81 μmol) in 1,4-dioxane (2 mL), then add a 4M hydrogen chloride / 1,4-dioxane solution (6.07 mL). Stir the reaction at room temperature for 3 hours. Evaporate the solvent under reduced pressure to yield the crude product, compound Z-63-P1 (120 mg, pale yellow solid hydrochloride salt), in a yield of 92.68%. MS m / z (ESI): 320.2 [M+1].

[0456] Step 3: Dissolve compound Z-63-P1 (120 mg, 375.20 μmol) and ethyl acetimidate (163.44 mg, 1.88 mmol) in acetonitrile (15 mL), then add N,N-diisopropylethylamine (0.65 mL, 3.75 mmol). Stir the reaction at 80°C for 2 hours, cool to room temperature, and add 2 mL of 7N methanolic ammonia. Stir at room temperature for 1 hour. Stop the reaction, dilute with ethyl acetate (50 mL), and wash the organic phase with saturated sodium chloride solution (50 mL), dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue was purified by preparative liquid chromatography (preparative column: 21.2 x 250 mm C18 column; system: 10 mM NH4HCO3H2O; wavelength: 254 / 214 nm; gradient: 5% to 95% acetonitrile) to obtain compound Z-64-P1 (63.5 mg) in a yield of 46.2%. MS m / z (ESI): 361.2 [M+1]. 1 H NMR(400MHz, DMSO-d6)δ8.17(s,1H),6.17(s,1H),4.30–4.15(m,1H),3.99–3.89(m,1H),2.26–2.17(m,1H),2.12–1.93(m,3H),1.89–1.70(m, 4H),1.70-1.60(m,1H),1.50-1.38(m,1H),1.33(d,J=6.8Hz,3H),1.10 -1.02(m,1H),0.57-0.48(m,1H),0.42-0.31(m,1H),0.28-0.17(m,2H).

[0457] Example 64b: Preparation of Compound Z-64-P2

[0458]

[0459] Step 1: 1.2 g of compound Z-63-1 was separated by supercritical fluid chromatography (SFC) (column type: AD-3 4.6*100 mm3um, cosolvent: EtOH [1% NH3 (7M in MeOH)], injection volume: 5.00 ul, run time: 6.0 minutes, flow rate: 3.0 mL / min, pressure: 2000 psi, column temperature: 40°C) to obtain 440 mg of compound Z-63-b (retention time 1.512 minutes).

[0460] Step 2-3: Compound Z-64-P2 was prepared by referring to the methods of Steps 2 and 3 of Example 64a. MS m / z (ESI): 361.2 [M+1]. 1 H NMR(400MHz, DMSO-d6)δ8.17(s,1H),6.16(s,1H),4.24–4.12(m,1H),3.98–3.88(m,1H),2.27–2.16(m,1H),2.12–1.92(m,3H),1.86–1.71(m, 4H),1.70-1.60(m,1H),1.50-1.38(m,1H),1.33(d,J=6.8Hz,3H),1.12 -1.02(m,1H),0.56-0.48(m,1H),0.41-0.30(m,1H),0.28-0.18(m,2H).

[0461] Example 65 Preparation of Compound Z-65

[0462]

[0463] Compound Z-63-2 (60 mg, 187.60 μmol) and 1H-pyrazole-1-carboximidamide hydrochloride (620 mg, 4.23 mmol) were dissolved in acetonitrile (12 mL), and N,N-diisopropylethylamine (1.31 mL, 7.50 mmol) was added. The reaction was stirred at room temperature for 16 hours. The reaction was terminated, and saturated sodium chloride solution (30 mL) was added. The mixture was extracted with ethyl acetate (30 mL x 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Preparative HPLC chromatography gave the desired product, compound Z-65 (26.21 mg), in a yield of 36.68%. MS m / z (ESI): 362.1 [M+1]. 1H NMR (400MHz, DMSO-d6) δ7.96 (s, 1H), 7.60 (s, 1H), 5.99 (d, J = 2.8Hz, 1H), 4.10-4.00 (m, 1H), 3.85-3.70 (m, 1H), 2.09–1.77(m,4H),1.75-1.60(m,1H),1.55-1.46(m,1H),1.36-1.25(m,1H),1.07(d,J= 6.8Hz,3H),0.92–0.75(m,1H),0.33-0.25(m,1H),0.17-0.08(m,1H),0.05-0.01(m,2H).

[0464] Example 66 to Example 74

[0465] Compounds Z-66, Z-68, Z-69, Z-72, and Z-73 can be prepared by following the method of Example 64a. Compounds Z-67 and Z-71 can be prepared by following the method of Example 65. Compounds Z-70 and Z-74 can be prepared by following the method of Example 1.

[0466]

[0467]

[0468] Preparation of comparative compounds D and E

[0469]

[0470] Compounds D and E can be prepared by referring to existing patent documents.

[0471] Test Example 1: Activity inhibition test of CDK family kinases

[0472] In the following LANCE Ultra test method, kinase reagents were purchased from Carna Bioscience, reaction substrates and detection reagents were purchased from PerkinElmer, and the remaining reagents were purchased from Thermo Scientific.

[0473] The LANCE Ultra method was used to determine the inhibitory effect of the test substances on the kinase activities of CDK1 / CycB (Carna bioscience, #04-102), CDK2 / CycA (Carna bioscience, #04-103), and CDK9 / CycT (Carna bioscience, #04-110).

[0474] The kinase activity assay uses a 10 μL system containing the following components: CDK kinase diluent, a substrate diluent containing a mixture of Ulight-Myelicbasic protein (Perkin Elmer, #TRF-0109, hereinafter referred to as U-MBP) and ATP (Thermo Scientific, #PV3227), and the compound prepared in the above examples of the present invention (i.e., the test substance). Each kinase in the assay consists of three test groups: a background group (Blank), a non-inhibitory group (PC), and a compound test group (Test). The components of each test group are as follows:

[0475] Kinase substrate Compound Blank 1.33x reaction buffer Substrate dilution 2% DMSO only PC Kinase diluent Substrate dilution 2% DMSO only Test Kinase diluent Substrate dilution Compound dilution

[0476] The working concentrations of the components of the Test group in different kinase reactions are as follows:

[0477] Compounds: Dissolve the test compound at 10 mM at room temperature and serially dilute it in DMSO. Then dilute it to a 4x working solution in deionized water with 2% DMSO. The highest compound concentration used in the CDK1 and CDK2 assays is 10 μM, and CDK9 is 1 μM.

[0478] 1.33x reaction buffer: Components: 26.7 mM MOPS, 6.67 mM MgCl2, and 0.0133% Tween-20. Store in a 4°C refrigerator away from light. Add freshly prepared DTT to a final concentration of 5.33 mM before use.

[0479]

[0480] The working concentration of DMSO in the reaction was 0.5%.

[0481] After mixing the above components, place them on a shaker and incubate them in the dark at room temperature for 1 hour. Then, 10 μL of detection solution was added to all test groups (including Blank, PC and Test groups).

[0482] 10 μL of detection solution contains the following components: 16 mM EDTA (Thermo scientific, #15575), 1 nM phosphorylated U-MBP protein antibody (Perkin Elmer, #TRF-0201) and 1x detection buffer (Perkin Elmer, #CR97-100).

[0483] After adding the test solution, place the tube on a shaker and continue incubating at room temperature in the dark for 1 hour. After incubation, read the signal using a PerkinElmer Victor X5 fluorescence microplate reader with an excitation wavelength of 320 nm and emission wavelengths of 615 nm and 665 nm. The inhibition rate is calculated as follows:

[0484] 1. The 665nm / 615nm value (hereinafter referred to as the Ratio value) was calculated for all groups, and the inhibition rate was calculated based on the Ratio value of each group;

[0485] 2. Inhibition rate = (PC Ratio –Test Ratio ) / (PC Ratio –Blank Ratio )*100%;

[0486] 3. XLFIT 5.0 software (IDBS, UK) was used for fitting, with the logarithm of compound concentration as the X-axis and the inhibition rate as the Y-axis, and the half-maximal inhibitory concentration IC of the compound was calculated using a four-parameter model. 50 .

[0487] Table 1 Inhibitory activity of compounds against CDK9

[0488]

[0489]

[0490] Table 2 Inhibitory activity of compounds against CDK1 and CDK2

[0491]

[0492]

[0493] As can be seen from Tables 1 and 2, the compounds of the present invention have high inhibitory activity against CDK9 and high CDK9 inhibition selectivity.

[0494] Test Example 2: In vivo pharmacokinetic study in mice

[0495] The LC / MS / MS method was used to determine the drug concentration in the plasma of mice at different times after intravenous injection and oral gavage of the compound of the present invention, to study the pharmacokinetic behavior of the compound of the present invention in mice and to evaluate its pharmacokinetic characteristics.

[0496] Experimental plan:

[0497] Experimental animals: Healthy adult male ICR mice (weight 30-40 g, 12 mice; mice in the intravenous injection group had free access to water and food; mice in the oral administration group were fasted overnight and had free access to water and food 4 h after administration) were provided by Beijing Vital River Laboratory Animal Co., Ltd.

[0498] Administration and dosage: ICR mice were administered via tail vein (2 mg / kg, 5% DMSO, pH 4.5 20% Captisol) and oral administration (10 mg / kg, 5% DMSO, pH 4.5 20% Captisol).

[0499] Blood Sampling: Before dosing, select animals that meet the experimental requirements, weigh them, and label them. Before blood sampling, restrain the mice. Each dosed mouse receives blood at the designated time points (intravenous administration: 0.083, 0.25, 0.5, 1, 2, 4, 6, 7.5, and 24 hours after dosing, for a total of nine time points; oral administration: 0.083, 0.25, 0.5, 1, 2, 4, 6, 7.5, and 24 hours after dosing, for a total of nine time points). Blood is transferred to a 1.5 mL tube pre-spiked with K2EDTA and centrifuged for 4 minutes (8000 rpm, 4°C) to remove plasma. This entire process should be completed within 15 minutes of blood collection. All samples should be stored at -20°C until analysis. The drug concentration was determined by LC / MS / MS. The pharmacokinetic parameters of some of the compounds of the present invention in mice at the same dose and administration method are shown in Table 3:

[0500] Table 3 Pharmacokinetic parameters of compounds in mice

[0501]

[0502] Test Example 3: In vivo efficacy experiment

[0503] In vivo efficacy experiments were performed on BALB / c nude mice subcutaneously implanted with MV4-11 acute myeloid leukemia patient-derived human tumor cell line-based xenografts (CDX).

[0504] Experimental protocol: BALB / c nude mice, female, 6-10 weeks old, weighing approximately 20-23 g, were kept in a special pathogen-free environment in single ventilated cages (5 mice per cage, 10 mice per group, 2 cages). All cages, bedding, and water were disinfected before use. All animals had free access to standard certified commercial laboratory diet. A total of 80 mice purchased from the Laboratory Animal Management Department of Shanghai Institute of Family Planning Science (3577 Jinke Road, Pudong, Shanghai) were used for the study. Each mouse was implanted with tumor cells (1×10 70.1 ml + 0.1 ml Matrigel) was used for tumor growth. When the average tumor volume reached approximately 165 cubic millimeters, the animals were randomly divided into groups based on body weight and tumor volume, and dosing began. The test compound was administered orally by gavage daily. Antitumor efficacy was determined by dividing the average tumor volume increase in animals treated with the compound by the average tumor volume increase in untreated animals.

[0505] Tumor volume was measured twice a week using a two-dimensional caliper and measured in cubic millimeters. Tumor volume TV = 0.5a × b 2 Where a is the long diameter of the tumor and b is the short diameter of the tumor.

[0506] The relative tumor growth rate T / C (%) is the percentage of the relative tumor volume of the treatment group and the control group at a certain time point. The calculation formula is as follows: T / C% = T RTV / C RTV ×100%(T RTV : Average RTV of treatment group; C RTV : Average RTV of vehicle control group; RTV=V t / V0, V0 is the tumor volume of the animal when grouped, and Vt is the tumor volume of the animal after treatment).

[0507] The body weight change (%) of tumor-bearing animals was calculated as follows: (body weight at measurement - body weight at grouping) / body weight at grouping×100.

[0508] Table 4 Tumor inhibition results of the compounds in mice

[0509]

[0510] Table 5 Tumor inhibition results of compounds in mice

[0511]

[0512] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. A compound represented by formula (III-a) or formula (III-b), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof: in, R 1a For hydrogen, C 1-6 Alkyl or deuterated C 1-6 Alkyl; n is 1, 2, 3 or 4; t is 1, 2 or 3; R 11 、R 12 are each independently hydrogen; R2 is hydrogen, C 1-6 Alkyl or halogen; R3 is hydrogen, C 1-6 Alkyl, C 3-6 Cycloalkyl, C 1-6 Alkoxy or halogen; wherein the C 1-6 The alkyl group is unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of: 1-3 alkoxy; R4 is hydrogen, -C(O)C 1-3 Alkyl, -(C=N)-C 1-3 Alkyl or -(C=N)-NR a0 R b0 ; R5 is hydrogen; R a For hydrogen, C 1-6 Alkyl, C 3-6 Cycloalkyl, cyano or halogen; wherein said C 1-6 Alkyl is unsubstituted or substituted with 1, 2 or 3 substituents each independently selected from the group consisting of halogen; R b is hydrogen; (R0) m Indicates that the hydrogen on ring A is replaced by m R0, where m is 0; R a0 、R b0 are each independently hydrogen.

2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: The compound is a structure shown in formula (III-a): Among them, R 1a is hydrogen or C 1-3 alkyl; n is 1, 2, or 3; t is 1, 2, or 3; R2 is hydrogen, C 1-3 Alkyl or halogen; R3 is hydrogen, C 1-3 Alkyl, C 3-6 Cycloalkyl, C 1-3 Alkoxy or halogen; wherein said C 1-3 The alkyl group is unsubstituted or substituted with one substituent selected from the group consisting of: C 1-3 alkoxy; R4 is hydrogen, -(C=N)-C 1-3 Alkyl or -(C=N)-NR a0 R b0 ; R a For hydrogen, C 1-3 Alkyl, C 3-6 Cycloalkyl, cyano or halogen; wherein said C 1-3 Alkyl is unsubstituted or substituted with 1, 2 or 3 substituents each independently selected from the group consisting of halogen.

3. The compound according to claim 2, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: R2 is hydrogen or C 1-3 alkyl; R3 is hydrogen, C 1-3 Alkyl or C 3-6 Cycloalkyl.

4. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a solvate thereof, or a prodrug thereof, wherein: t is 1 or 2.

5. The compound according to claim 2, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: n is 1 or 2.

6. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: R 1a is hydrogen; n is 1 or 2; t is 1 or 2; R2 is hydrogen or C 1-3 alkyl; R3 is C 1-3 Alkyl or C 3-6 Cycloalkyl; R4 is hydrogen; R a For hydrogen, C 1-3 Alkyl or halogen.

7. The compound according to claim 2, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: R 1a is hydrogen; n is 1 or 2; R2 is hydrogen; R3 is C 1-3 Alkyl or C 3-6 Cycloalkyl; R4 is hydrogen; R a is hydrogen or C 1-3 alkyl.

8. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: The compound is a structure shown in formula (III-b): in, t is 2; R2 is hydrogen or C 1-3 alkyl; R3 is C 1-3 Alkyl or C 3-6 Cycloalkyl; R4 is hydrogen; R a For hydrogen, C 1-3 Alkyl or halogen.

9. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: The compound represented by formula (III-a) is a structure represented by formula (III-a-1) or formula (III-a-2): Wherein, R3' is C 1-3 alkyl; R 1a is hydrogen or C 1-3 alkyl; n is 1, 2, or 3; t is 1 or 2; R4 is hydrogen, -(C=N)-C 1-3 Alkyl or -(C=N)-NR a0 R b0 ; R a For hydrogen, C 1-3 Alkyl, C 3-6 Cycloalkyl, cyano or halogen; wherein said C 1-3 Alkyl is unsubstituted or substituted with 1, 2 or 3 substituents each independently selected from the group consisting of halogen.

10. The compound according to claim 9, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: R3' is a methyl group.

11. The compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: R a For hydrogen.

12. The compound according to any one of claims 1 to 5, 9 to 10, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: R4 is hydrogen.

13. The compound according to any one of claims 1 to 7, 9 to 10, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: t is 2.

14. The compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: t is 2, R2 is hydrogen, and R3 is C 1-3 alkyl.

15. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: The compound of formula (III-a) or formula (III-b) is any one of the following compounds:

16. A compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: The compound is any one of the following:

17. A pharmaceutical composition comprising: A compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof; and a pharmaceutically acceptable carrier.

18. Use of the compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or the pharmaceutical composition according to claim 17 in the preparation of a medicament for treating or preventing a disease associated with or mediated by CDK9 activity.

19. The use according to claim 18, wherein: The disease associated with CDK9 activity or mediated by CDK9 activity is cancer.

20. The use according to claim 19, wherein: The cancer is selected from pancreatic cancer, breast cancer, ovarian cancer, cervical cancer and leukemia.

21. Use of the compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or the pharmaceutical composition according to claim 17 in the preparation of a CDK9 inhibitor.

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