Heterocyclic compound capable of inhibiting PRMT5MTA and application thereof

Abstract

The invention relates to a nitrogen-containing heterocyclic compound as shown in a formula (I-a), or a stereoisomer, a deuterated compound, a solvate, pharmaceutically acceptable salt, a co-crystal and a pharmaceutical composition of the nitrogen-containing heterocyclic compound, and application of the nitrogen-containing heterocyclic compound in preparation of drugs for treating / preventing PRMT5. MTA-mediated diseases, and groups in the formula (I-a) are defined in the specification. # imgabs0 #

Description

A heterocyclic compound capable of inhibiting PRMT5·MTA and its use Technical Field

[0001] The present invention relates to an inhibitor of PRMT5·MTA complex and use thereof in preparing an inhibitor for treating tumors. Background Art

[0002] Protein arginine methyltransferases (PRMTs) catalyze protein arginine methylation, a key post-translational modification. PRMTs, using S-adenosylmethionine (SAM) as a methyl donor, methylate the nitrogen atom of the arginine side chain of proteins. The nine identified human PRMTs (PRMT1–9) are further divided into types I, II, and III based on their arginine methylation status. Type I PRMTs, including PRMT1, PRMT2, PRMT3, PRMT4, PRMT6, and PRMT8, catalyze the formation of monomethylarginine (MMA) and asymmetric dimethylarginine (aDMA); type II PRMTs, including PRMT5 and PRMT9, catalyze the formation of MMA and symmetric dimethylarginine (sDMA); and type III PRMTs, including PRMT7, catalyze the formation of MMA. PRMT5 belongs to type II PRMT, which transfers methyl group from SAM to the guanidine nitrogen of arginine residues to generate MMA and sDMA, thereby regulating the expression of target genes in this epigenetic manner or regulating key signaling molecules through post-translational methylation modification pathways.

[0003] Homozygous deletions of p16 / CDKN2a are common in cancer. These mutations often involve co-deletions of adjacent genes, including the gene encoding methylthioadenosine phosphorylase (MTAP). It is estimated that approximately 15% of human cancers have homozygous deletions of the MTAP gene (e.g., see Firestone & Schramm (2017) J. Am. Chem Soc. 139(39):13754-13760. doi:10.1021 / jacs.7b05803. Epub 2017 Sep 20).

[0004] In MTAP-deficient cells, methylthioadenosine (MTA) overaccumulates and forms a complex with PRMT5, PRMT5·MTA, partially inhibiting PRMT5 enzymatic activity and increasing the sensitivity of cell proliferation to PRMT5 deficiency or inactivity. Consequently, MTAP deficiency reduces PRMT5 methylation activity, rendering cells selectively dependent on PRMT5 activity. In MTAP-deficient cancers, the synergistic inhibition of PRMT5 activity with MTA could provide therapeutic benefits for a variety of cancers.

[0005] Therefore, it is necessary to develop new MTA-synergistic PRMT5 inhibitors that can inhibit PRMT5 activity under elevated MTA concentrations, especially in MTAP-deficient cells.

[0006] Summary of the Invention

[0007] The present invention provides a compound of formula (Ia), formula (Ib), formula (I), formula (II-a) or formula (II), and its stereoisomers, deuterated forms, solvates, or pharmaceutically acceptable salts. The compound has good activity, excellent physicochemical properties, ease of preparation, high bioavailability, and low toxic and side effects.

[0008] The compound of formula (Ia), formula (Ib), formula (I), formula (II-a) or formula (II), its stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt,

[0009] Wherein, D is selected from: In some embodiments, D is selected from: In some embodiments, D is selected from: In some embodiments, D is selected from In some embodiments, D is selected from In some embodiments, D is selected from In some embodiments, D is selected from

[0010] In some embodiments, D is selected from In some embodiments, D is selected from In some embodiments, D is selected from In some embodiments, D is selected from In some embodiments, D is selected from In some embodiments, D is selected from In some embodiments, D is selected from

[0011] D1 is selected from: In some embodiments, D1 is selected from:

[0012] Ld is selected from C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl; in some embodiments, Ld is selected from C 2-4 Alkenyl, C 2-4 Alkynyl; in some embodiments, Ld is selected from C 2-4 alkenyl; in some embodiments, Ld is selected from

[0013] Rd1 is selected from 5-membered heteroaryl, wherein the heteroaryl is optionally substituted by 1-3 selected from halogen, D, CN, OH, C 1-4 Alkyl, C 1-4 In some embodiments, Rd1 is selected from 5-membered heteroaryl, wherein the heteroaryl is optionally substituted with 1 or 2 atoms selected from halogen, D, CN, OH, C 1-2 Alkyl, C 1-2 Alkoxy and NH2 groups are substituted; in some embodiments, Rd1 is selected from 5-membered heteroaryl, and the heteroaryl is optionally substituted with 1 or 2 groups selected from NH2; in some embodiments, Rd1 is selected from

[0014] X is selected from CR1 or N; in some embodiments, X is CR1; in some embodiments, X is N; in some embodiments, X is CH;

[0015] Y, Z and V are independently CR5, C(R5)2, NR6, N, O or S, and the total number of O and S is no greater than 1; in some embodiments, Y, Z and V are independently CR5, N, O or S, and the total number of O and S is no greater than 1; in some embodiments, Y, Z and V are independently CR5, N or S, and the total number of S is no greater than 1;

[0016] Ring A is phenyl, a 5-7 membered heterocycloalkyl group containing 1-3 heteroatoms selected from N, O, and S, or a 5-6 membered heteroaryl group containing 1-3 heteroatoms selected from N, O, and S; in some embodiments, ring A is phenyl, or a 5-6 membered heteroaryl group containing 1-3 heteroatoms selected from N, O, and S; in some embodiments, ring A is phenyl, thienyl, thiazolyl, pyrrolyl, imidazolyl, oxazolyl, pyridinyl, pyrazinyl, pyrimidinyl, or pyridazinyl;

[0017] Ring B is C 3-12 A carbon ring or a 4-12 membered heterocyclic ring containing 1-3 heteroatoms selected from N, O, and S; in some embodiments, Ring B is C 9-12 Cycloaryl, C 6-12 cycloalkyl, 8-12 membered cycloheterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, or 9-12 membered cycloheteroaryl containing 1-3 heteroatoms selected from N, O, S; in some embodiments, ring B is phenyl, C 3-6 cycloalkyl, 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, and S, or 5-6 membered aryl containing 1-3 heteroatoms selected from N, O, and S; in some embodiments, ring B is thienyl, thiazolyl, pyrrolyl, imidazolyl, or pyrazolyl; in some embodiments, ring B is pyrazolyl;

[0018] Ring C is C 3-12 carbon ring or a 4-12 membered heterocyclic ring containing 1-3 heteroatoms selected from N, O, and S; in some embodiments, ring C is phenyl, C 3-6 cycloalkyl, 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, and S, or 5-6 membered aryl containing 1-3 heteroatoms selected from N, O, and S; in some embodiments, ring C is phenyl, thienyl, thiazolyl, pyrrolyl, imidazolyl, oxazolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, or pyridazinyl; in some embodiments, ring C is phenyl;

[0019] R1 is H, D, C 1-4 Alkyl, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, halogen, CN, OH, NH2 or COOH, the alkyl, alkoxy, alkenyl, alkynyl is optionally substituted with 1-3 groups selected from halogen, D, CN, OH and NH2; in some embodiments, R1 is H, D, C 1-4 Alkyl, halogen, CN, wherein the alkyl is optionally substituted with 1-3 groups selected from halogen, D, CN and OH; in some embodiments, R1 is H, D, C 1-4Alkyl, F, Cl, Br, I or CN, wherein the alkyl group is optionally substituted with 1-3 groups selected from F, Cl, Br, I, D, CN and OH;

[0020] R2 is OH, NH2, -CH2NH2, or -NH-OH; in some embodiments, R2 is OH, NH2, or -NH-OH; in some embodiments, R2 is NH2;

[0021] R3 and R4 are independently H, D, halogen, C 1-4 Alkyl, CN, OH, NH2, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl or C 3-6 Cycloalkyl; the alkyl, alkoxy, alkenyl, alkynyl or cycloalkyl is optionally substituted by 1-3 groups selected from halogen, D, CN, OH and NH2; in some embodiments, R3, R4 are independently H, D, halogen, C 1-4 alkyl, CN, OH or NH2; the alkyl is optionally substituted by 1-3 groups selected from halogen, D, CN, OH and NH2; in some embodiments, R3 and R4 are independently H, D, F, Cl, Br, I, C 1-4 Alkyl or CN; the alkyl group is optionally substituted with 1-3 groups selected from F, Cl, Br, I, D, CN, OH and NH2; in some embodiments, R3, R4 are independently H or D;

[0022] Each R5 is independently H, D, =O, OH, C 1-4 Alkyl, CN, halogen, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, 5-6 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S or C 3-6 Cycloalkyl, wherein the alkyl, alkoxy, alkenyl, alkynyl, heteroaryl or cycloalkyl is optionally substituted by 1-3 groups selected from halogen, D, CN, OH, C 1-4 Alkyl, C 1-4 In some embodiments, each R5 is independently H, D, OH, C 1-4 Alkyl, CN, halogen, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, 5-membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, 4-5-membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S or C 3-4Cycloalkyl, the alkyl, alkoxy, alkenyl, alkynyl, heteroaryl, heterocycloalkyl, cycloalkyl is optionally substituted by 1-3 groups selected from halogen, D, CN, OH, C 1-4 Alkyl, C 1-4 In some embodiments, each R5 is independently H, D, OH, C 1-4 Alkyl, CN, F, Cl, Br, I, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, 5-membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, 4-5-membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S or C 3-4 Cycloalkyl, wherein the alkyl, alkoxy, alkenyl, alkynyl, heteroaryl, heterocycloalkyl, cycloalkyl is optionally substituted with 1-3 groups selected from F, Cl, Br, I, D, CN, OH, methoxy, and NH2; in some embodiments, each R5 is independently H, D, =O, OH, C 1-4 Alkyl, CN, halogen, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl or C 3-6 Cycloalkyl, the alkyl, alkoxy, alkenyl, alkynyl or cycloalkyl is optionally substituted by 1-3 groups selected from halogen, D, CN, OH and NH2; in some embodiments, each R5 is independently H, D, OH, C 1-4 Alkyl, CN, halogen, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, 5-membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, 4-5-membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S or C 3-4 Cycloalkyl, wherein the alkyl, alkoxy, alkenyl, alkynyl, heteroaryl, heterocycloalkyl, cycloalkyl is optionally substituted by 1-3 groups selected from halogen, D, CN, OH and NH2; in some embodiments, each R5 is independently H, D, OH, C 1-4 Alkyl, CN, F, Cl, Br, I, C 1-4 Alkoxy, C 2-4 Alkenyl, or C 2-4 Alkynyl, 5-membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, 4-5-membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S or C 3-4 Cycloalkyl, wherein the alkyl, alkoxy, alkenyl, alkynyl, heteroaryl, heterocycloalkyl, cycloalkyl is optionally substituted with 1-3 groups selected from F, Cl, Br, I, D, CN, OH and NH2; in some embodiments, each R5 is independently H, D, CN, F, Cl, C1-2 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, pyrazolyl, azetidinyl, cyclopropyl, cyclobutyl, the alkyl, alkenyl, alkynyl, pyrazolyl, azetidinyl, cyclopropyl, cyclobutyl are optionally substituted by 1, 2, or 3 groups selected from F, Cl, Br, I, D, CN, OH, and NH2; in some embodiments, each R5 is independently H, D, OH, C 1-4 Alkyl, CN, halogen, C 1-4 Alkoxy, C 2-4 Alkenyl, or C 2-4 Alkynyl, said alkyl, alkoxy, alkenyl or alkynyl being optionally substituted by 1-3 groups selected from halogen, D, CN, OH and NH2; In some embodiments, each R5 is independently H, D, OH, C 1-4 Alkyl, CN, F, Cl, Br, I, C 1-4 Alkoxy, C 2-4 Alkenyl, or C 2-4 Alkynyl, wherein the alkyl, alkoxy, alkenyl or alkynyl group is optionally substituted with 1-3 groups selected from F, Cl, Br, I, D, CN, OH and NH2;

[0023] R6 is H, D, C 1-4 Alkyl or C 3-6 Cycloalkyl; said alkyl or cycloalkyl is optionally substituted by 1-3 groups selected from halogen, D, CN, OH and NH2; in some embodiments, R6 is H;

[0024] Each R7 is independently H, =O, D, OH, CN, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, N(R7')2, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-6 Cycloalkyl or 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl or heterocycloalkyl is optionally substituted by 1-3 halogen, D, OH, NH2, CN, C 1-4 Alkyl, -C(O)C 1-4 Alkyl, -C(O)NHC 1-4 Alkyl, -C(O)NH2, -NHC(O)C 1-4 Alkyl and C 1-4 In some embodiments, each R7 is independently H, =O, D, OH, CN, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, N(R7')2, C 2-4 Alkenyl, or C2-4 Alkynyl, the alkyl, alkoxy, alkenyl, alkynyl is optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-4 Alkyl, -C(O)C 1-4 Alkyl, -C(O)NHC 1-4 Alkyl, -C(O)NH2, -NHC(O)C 1-4 Alkyl and C 1-4 In some embodiments, each R7 is independently H, D, OH, CN, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, N(R7')2, C 2-4 Alkenyl, or C 2-4 Alkynyl, the alkyl, alkoxy, alkenyl, alkynyl is optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-4 Alkyl and C 1-4 Alkoxy group substitution;

[0025] Each R8 is independently H, D, nitro, amino, C 1-6 Alkoxy, =O, OH, CN, halogen, C 2-6 Alkenyl, C 2-6 Alkynyl, -SF5, N3, C 1-6 Alkyl, -COC 1-4 Alkyl, -N(R7')2, -C(=O)N(R7')2, -NR7'C(=O)-R7', -C(=O)-R7', -(CH2) r -O-(CH2) r -R8'、-SC 1-6 Alkyl, -S(O)-C 1-6 Alkyl, -S(O)2-C 1-6 Alkyl, -S-(CH2) r -R8'、-NR7'-(CH2) r -R8' or -(CH2) r -R8', the alkyl, alkenyl, alkynyl is optionally substituted by 1-5 selected from R8"; in some embodiments, each R8 is independently H, D, =O, OH, CN, halogen, C 2-4 Alkenyl, C 2-4 Alkynyl, N3, C 1-4 Alkyl, -COC 1-4 Alkyl, -CONR7'C 1-4 Alkyl, -NR7'COC 1-4Alkyl, N(R7')2, -O-(CH2)r-R8', or -(CH2)r-R8', wherein the alkyl, alkenyl, alkynyl is optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-4 Alkyl and C 1-4 In some embodiments, each R8 is independently D, =O, OH, CN, halogen, C 2-4 Alkenyl, C 2-4 Alkynyl, N3, C 1-4 Alkyl, -COC 1-4 Alkyl, -CONR7'C 1-4 Alkyl, -NR7'COC 1-4 Alkyl, N(R7')2, -O-(CH2)r-R8', or -(CH2)r-R8', wherein the alkyl, alkenyl, alkynyl is optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-2 Alkyl and C 1-2 In some embodiments, each R8 is independently D, CN, F, Cl, Br, C 2-4 Alkenyl, C 2-4 Alkynyl, N3, C 1-4 Alkyl, -O-(CH2)r-R8', or -(CH2)r-R8', wherein the alkyl, alkenyl, alkynyl is optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-2 Alkyl and C 1-2 Alkoxy group substitution; or

[0026] Each R8 is independently H, D, nitro, amino, C 1-6 Alkoxy, =O, OH, CN, halogen, C 2-6 Alkenyl, C 2-6 Alkynyl, -SF5, C 1-6 Alkyl, -COC 1-4 Alkyl, -N(R7')2, -C(=O)N(R7')2, -NR7'C(=O)-R7', -C(=O)-R7', -(CH2) r -O-(CH2) r -R8'、-SC 1-6 Alkyl, -S(O)-C 1-6 Alkyl or -S(O)2-C 1-6 Alkyl, -S-(CH2) r -R8'、-NR7'-(CH2) r -R8' or -(CH2) r-R8', the alkyl, alkenyl, alkynyl is optionally substituted by 1-5 selected from R8"; in some embodiments, each R8 is independently halogen, deuterium, nitro, cyano, amino, hydroxyl, =O, C 1-6 Alkyl, C 1-6 Alkoxy, C 2-6 Alkenyl, C 2-6 Alkynyl, -SF5, -(CH2) r -O-(CH2) r -R8'、-(CH2) r -C 3-12 Cycloalkyl, -(CH2) r -(3-12 membered heterocycloalkyl), -O-(CH2) r -C 3-12 Cycloalkyl, -O-(CH2) r -(3-12 membered heterocycloalkyl), -NR7'-C 3-12 Cycloalkyl, -NR7'-(3-12 membered heterocycloalkyl), -SC 3-12 Cycloalkyl, -S-(3-12 membered heterocycloalkyl), -(CH2) r -(5- to 12-membered heteroaryl), -(CH2) r -(6- to 12-membered aryl), -O-(CH2) r -(5-12 membered heteroaryl), -O-(CH2) r -(6-12 membered aryl), -NR7'-(CH2) r -(5-12 membered heteroaryl), -NR7'-(CH2) r -(6-12 membered aryl), -N(R7')2, -C(=O)N(R7')2, -NR7'C(=O)-R7', -C(=O)-R7', -SC 1-6 Alkyl, -S(O)-C 1-6 Alkyl or -S(O)2-C 1-6 Alkyl, wherein the alkyl, alkoxy, CH2, heterocycloalkyl, cycloalkyl, heteroaryl or aryl is optionally further substituted with 1-5 R8"; or

[0027] Each R8 is independently H, D, =O, OH, CN, halogen, C 2-4 Alkenyl, C 2-4 Alkynyl, -SF5, C 1-4 Alkyl, -COC 1-4 Alkyl, -CONR7'C 1-4 Alkyl, -NR7'COC 1-4 Alkyl, N(R7')2, -O-(CH2) r -R8'、-S-(CH2) r-R8'、-NR7'-(CH2) r -R8' or -(CH2) r -R8', the alkyl, alkenyl, alkynyl are optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-4 Alkyl and C 1-4 In some embodiments, each R8 is independently H, D, =O, OH, CN, halogen, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkyl, -COC 1-4 Alkyl, -CONR7'C 1-4 Alkyl, -NR7'COC 1-4 Alkyl, N(R7')2, -O-(CH2) r -R8', or -(CH2) r -R8', the alkyl, alkenyl, alkynyl are optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-4 Alkyl and C 1-4 In some embodiments, each R8 is independently H, D, OH, CN, F, Cl, Br, I, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkyl, -COC 1-4 Alkyl, -CONR7'C 1-4 Alkyl, -NR7'COC 1-4 Alkyl, N(R7')2, -O-(CH2) r -R8', or -(CH2) r -R8', the alkyl, alkenyl, alkynyl are optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-4 Alkyl and C 1-4 Alkoxy group substitution; or

[0028] Each R8 is independently CN, F, Cl, N3, vinyl, fluorovinyl, -O-CH2CH2-OCH3, -N(CH3)2,

[0029] Each R7' is independently H, D, amino, hydroxyl, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, C 3-6 Cycloalkyl, C 3-12 Heterocycloalkyl, -NHC1-6 Alkyl, -N(C 1-6 Alkyl)2, or deuterated C 1-6 Alkoxy, the cycloalkyl, heterocycloalkyl is optionally substituted by 1 to 3 groups selected from deuterium, halogen, cyano, amino, hydroxyl, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy or deuterated C 1-6 In some embodiments, each R7' is independently H, D, amino, hydroxy, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, C 3-6 Cycloalkyl, C 3-12 Heterocycloalkyl, -NHC 1-4 Alkyl, -N(C 1-4 Alkyl)2, or deuterated C 1-4 Alkoxy, the cycloalkyl, heterocycloalkyl is optionally substituted by 1 to 3 groups selected from deuterium, halogen, cyano, amino, hydroxyl, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy or deuterated C 1-4 In some embodiments, each R7' is independently H, D, amino, hydroxy, C 1-4 Alkyl, halogenated C 1-2 Alkyl, deuterated C 1-2 Alkyl, C 1-2 Alkoxy, halogenated C 1-2 Alkoxy, C 3-6 Cycloalkyl, C 3-12 Heterocycloalkyl, -NHC 1-2 Alkyl, -N(C 1-2 Alkyl)2, or deuterated C 1-2 Alkoxy, the cycloalkyl, heterocycloalkyl is optionally substituted by 1 to 3 groups selected from deuterium, halogen, cyano, amino, hydroxyl, C 1-2 Alkyl, halogenated C 1-2 Alkyl, deuterated C 1-2 Alkyl, C 1-2 Alkoxy, halogenated C 1-2 Alkoxy or deuterated C 1-2 In some embodiments, each R7' is independently H, D, amino, hydroxy, C1-4 Alkyl; or

[0030] Each R7' is independently H, C 1-4 Alkyl or halogenated C 1-4 In some embodiments, each R7' is independently H or C 1-4 Alkyl; in some embodiments, each R7' is independently H;

[0031] Each R8' is independently H, D, C 1-6 Alkyl, C 1-4 Alkoxy, C 3-12 Carbocyclic ring, 4-12 membered heterocyclic ring containing 1-3 heteroatoms selected from N, O, S, wherein the alkyl and alkoxy groups are optionally substituted by 1-3 groups selected from halogen, =O, D, OH, NH2, CN, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 Alkyl, -C(O)C 1-4 Alkyl, -C(O)NHC 1-4 Alkyl, -C(O)NH2, -NHC(O)C 1-4 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy and deuterated C 1-6 The carbocyclic or heterocyclic ring is optionally substituted by 1-5 selected from R8 "; in some embodiments, each R8 'is independently H, C 1-4 Alkoxy, C 3-6 Cycloalkyl, phenyl, 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, or 5-6 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, wherein the cycloalkyl, phenyl, heterocycloalkyl or heteroaryl is optionally substituted by 1-3 halogen, =O, D, OH, NH2, CN, C 1-4 Alkyl, halogenated C 1-4 Alkyl, and C 1-4 In some embodiments, each R8' is independently H, C 1-4 Alkoxy, C 3-6 Cycloalkyl, phenyl, 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, or 5-6 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, wherein the cycloalkyl, phenyl, heterocycloalkyl or heteroaryl is optionally substituted by 1-3 halogen, =O, D, OH, NH2, CN, C 1-4 Alkyl, halogenated C 1-4 Alkyl, and C 1-4 In some embodiments, R8' is independently C 1-4Alkoxy, cyclopropyl, cyclobutyl, cyclopentyl, phenyl, tetrahydropyrrolyl, azetidinyl, piperidinyl, piperazinyl, morpholinyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, Thiphenyl, thiazolyl, pyrrolyl, imidazolyl, oxazolyl, pyridinyl, pyrazinyl, pyrimidinyl or pyridazinyl, optionally substituted by 1-3 groups selected from F, Cl, Br, I, =O, D, OH, NH2, CN, C 1-4 Alkyl, halogenated C 1-2 Alkyl, and C 1-2 Alkoxy group substitution; or

[0032] Each R8' is independently H, D, C 1-6 Alkyl, C 1-4 Alkoxy, C 3-12 Carbocyclic ring, 4-12 membered heterocyclic ring containing 1-3 heteroatoms selected from N, O, S, wherein the alkyl and alkoxy groups are optionally substituted by 1-3 groups selected from halogen, =O, D, OH, NH2, CN, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 Alkyl, -C(O)C 1-4 Alkyl, -C(O)NHC 1-4 Alkyl, -C(O)NH2, -NHC(O)C 1-4 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy and deuterated C 1-6 The carbocyclic or heterocyclic ring is optionally substituted by 1-5 selected from R8 "; in some embodiments, each R8 'is independently H, D, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-12 Carbocyclic ring, 4-12 membered heterocyclic ring containing 1-3 heteroatoms selected from N, O, S, wherein the alkyl and alkoxy groups are optionally substituted by 1-3 groups selected from halogen, =O, D, OH, NH2, CN, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, -C(O)C 1-2 Alkyl, -C(O)NHC 1-2 Alkyl, -C(O)NH2, -NHC(O)C 1-2 Alkyl, C 1-2 Alkoxy, halogenated C 1-2 Alkoxy and deuterated C 1-2 The carbocyclic or heterocyclic ring is optionally substituted by 1-5 groups selected from R8"; or

[0033] Each R8' is independently H, C 1-4Alkoxy, C 3-9 Carbocyclic ring, 4-10 membered heterocyclic ring containing 1-3 heteroatoms selected from N, O, S, wherein the carbocyclic ring or heterocyclic ring is optionally substituted by 1-3 heteroatoms selected from halogen, =O, D, OH, NH2, CN, C 1-4 Alkyl, -C(O)C 1-4 Alkyl, -C(O)NHC 1-4 Alkyl, -C(O)NH2, -NHC(O)C 1-4 Alkyl and C 1-4 In some embodiments, each R8' is independently H, C 1-4 Alkoxy, C 3-6 Cycloalkyl, phenyl, 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, or 5-6 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, wherein the cycloalkyl, phenyl, heterocycloalkyl or heteroaryl is optionally substituted by 1-3 halogen, =O, D, OH, NH2, CN, C 1-4 Alkyl, and C 1-4 In some embodiments, each R8' is independently H, C 1-4 Alkoxy, C 3-6 Cycloalkyl, phenyl, 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, or 5-6 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, wherein the cycloalkyl, phenyl, heterocycloalkyl or heteroaryl is optionally substituted by 1-3 halogen, =O, D, OH, NH2, CN, C 1-4 Alkyl, and C 1-4 In some embodiments, R8' is independently C 1-4 Alkoxy, cyclopropyl, cyclobutyl, cyclopentyl, phenyl, tetrahydropyrrolyl, azetidinyl, piperidinyl, piperazinyl, morpholinyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, Thiphenyl, thiazolyl, pyrrolyl, imidazolyl, oxazolyl, pyridinyl, pyrazinyl, pyrimidinyl or pyridazinyl, optionally substituted by 1-3 groups selected from F, Cl, Br, I, =O, D, OH, NH2, CN, C 1-4 Alkyl, and C 1-4 Alkoxy group substitution;

[0034] R8" is selected from halogen, D, OH, NH2, CN, C 1-6 Alkyl and C 1-6 Alkoxy, the alkyl or alkoxy group is optionally further substituted by 1 to 5 groups selected from halogen, deuterium, cyano, amino, hydroxyl, C 1-6 Alkyl or C 1-6In some embodiments, R8 is selected from halogen, D, OH, NH2, CN, C 1-4 Alkyl and C 1-4 Alkoxy, the alkyl or alkoxy group is optionally further substituted by 1 to 3 groups selected from halogen, deuterium, cyano, amino, hydroxyl, C 1-2 Alkyl or C 1-2 In some embodiments, R8" is selected from deuterium, F, Cl, methyl, ethyl, propyl, methoxy, ethoxy or propoxy, and the methyl, ethyl, propyl, methoxy, ethoxy or propoxy group is optionally further substituted with 1 to 3 groups selected from F, Cl, deuterium, hydroxy, methyl, ethyl, propyl, methoxy, ethoxy or propoxy;

[0035] Each r is independently 0, 1, 2, or 3; in some embodiments, each r is independently 0, 1, or 2;

[0036] p is 0, 1 or 2; in some embodiments, p is 0 or 1;

[0037] n and m are independently integers from 0 to 5; in some embodiments, n is 0, 1, 2, or 3; in some embodiments, m is 0, 1, 2, 3, or 4;

[0038] The compound of the present invention,

[0039] Among them, 1. Group no

[0040] 2. When yes R3 is H, C 1-4 Alkyl, C 1-4 Haloalkyl, R5 is H, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 When halogenated alkyl and R3 and R5 are not H at the same time, the group for

[0041] As a more specific first technical solution of the present invention, it relates to a compound represented by formula (Ia) or (Ib), its stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt,

[0042] in,

[0043] D is selected from:

[0044] D1 is selected from:

[0045] Ld is selected from C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl;

[0046] Rd1 is selected from 5-membered heteroaryl, wherein the heteroaryl is optionally substituted by 1-3 selected from halogen, D, CN, OH, C 1-4 Alkyl, C 1-4 Alkoxy and NH2 group substitution;

[0047] X is selected from CR1 or N;

[0048] Y, Z and V are independently CR5, C(R5)2, NR6, N, O or S, and the total number of O and S is not greater than 1;

[0049] Ring A is phenyl, a 5-7 membered heterocycloalkyl group containing 1-3 heteroatoms selected from N, O, and S, or a 5-6 membered heteroaryl group containing 1-3 heteroatoms selected from N, O, and S;

[0050] Ring B is C 3-12 Carbocyclic ring or 4-12 membered heterocyclic ring containing 1-3 heteroatoms selected from N, O, S;

[0051] Ring C is C 3-12 Carbocyclic ring or 4-12 membered heterocyclic ring containing 1-3 heteroatoms selected from N, O, S;

[0052] R1 is H, D, C 1-4 Alkyl, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, halogen, CN, OH, NH2 or COOH, wherein the alkyl, alkoxy, alkenyl, alkynyl is optionally substituted with 1-3 groups selected from halogen, D, CN, OH and NH2;

[0053] R2 is OH, NH2, -CH2NH2 or -NH-OH;

[0054] R3 and R4 are independently H, D, halogen, C 1-4 Alkyl, CN, OH, NH2, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl or C 3-6 Cycloalkyl; the alkyl, alkoxy, alkenyl, alkynyl or cycloalkyl group is optionally substituted by 1-3 groups selected from halogen, D, CN, OH and NH2;

[0055] Alternatively, R3 and R4 together form =NH;

[0056] Each R5 is independently H, D, =O, OH, C 1-4 Alkyl, CN, halogen, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, 5-6 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S or C 3-6 Cycloalkyl, wherein the alkyl, alkoxy, alkenyl, alkynyl, heteroaryl or cycloalkyl is optionally substituted by 1-3 groups selected from halogen, D, CN, OH, C 1-4 Alkyl, C 1-4 Alkoxy and NH2 group substitution;

[0057] R6 is H, D, C 1-4 Alkyl or C 3-6 Cycloalkyl; said alkyl or cycloalkyl is optionally substituted by 1-3 groups selected from halogen, D, CN, OH and NH2;

[0058] Each R7 is independently H, =O, D, OH, CN, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, N(R7')2, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-6 Cycloalkyl or 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl or heterocycloalkyl is optionally substituted by 1-3 halogen, D, OH, NH2, CN, C 1-4 Alkyl, -C(O)C 1-4 Alkyl, -C(O)NHC 1-4 Alkyl, -C(O)NH2, -NHC(O)C 1-4 Alkyl and C 1-4 Alkoxy group substitution;

[0059] Each R8 is independently H, D, nitro, amino, C 1-6 Alkoxy, =O, OH, CN, halogen, C 2-6 Alkenyl, C 2-6 Alkynyl, -SF5, N3, C 1-6 Alkyl, -COC 1-4 Alkyl, -N(R7')2, -C(=O)N(R7')2, -NR7'C(=O)-R7', -C(=O)-R7', -(CH2) r -O-(CH2) r -R8'、-SC 1-6 Alkyl, -S(O)-C 1-6 Alkyl, -S(O)2-C1-6 Alkyl, -S-(CH2) r -R8'、-NR7'-(CH2) r -R8' or -(CH2) r -R8', the alkyl, alkenyl, alkynyl group is optionally substituted by 1-5 groups selected from R8";

[0060] Each R7' is independently H, D, amino, hydroxyl, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, C 3-6 Cycloalkyl, C 3-12 Heterocycloalkyl, -NHC 1-6 Alkyl, -N(C 1-6 Alkyl)2, or deuterated C 1-6 Alkoxy, the cycloalkyl, heterocycloalkyl is optionally substituted by 1 to 3 groups selected from deuterium, halogen, cyano, amino, hydroxyl, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy or deuterated C 1-6 Alkoxy group substitution;

[0061] Each R8' is independently H, D, C 1-6 Alkyl, C 1-4 Alkoxy, C 3-12 Carbocyclic ring, 4-12 membered heterocyclic ring containing 1-3 heteroatoms selected from N, O, S, wherein the alkyl and alkoxy groups are optionally substituted by 1-3 groups selected from halogen, =O, D, OH, NH2, CN, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 Alkyl, -C(O)C 1-4 Alkyl, -C(O)NHC 1-4 Alkyl, -C(O)NH2, -NHC(O)C 1-4 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy and deuterated C 1-6 The carbocyclic or heterocyclic ring is optionally substituted by 1-5 groups selected from R8";

[0062] R8" is selected from halogen, D, OH, NH2, CN, C 1-6 Alkyl and C 1-6 Alkoxy, the alkyl or alkoxy group is optionally further substituted by 1 to 5 groups selected from halogen, deuterium, cyano, amino, hydroxyl, C1-6 Alkyl or C 1-6 Alkoxy group substitution;

[0063] Each r is independently 0, 1, 2 or 3;

[0064] p is 0, 1, or 2;

[0065] n and m are independently integers from 0 to 5;

[0066] The condition is,

[0067] 1. Group no

[0068] 2. When yes R3 is H, C 1-4 Alkyl, C 1-4 Haloalkyl, R5 is H, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 When halogenated alkyl and R3 and R5 are not H at the same time, the group for

[0069] As a more specific second technical solution of the present invention, there is provided a compound represented by formula (Ia), its stereoisomers, deuterated substances, solvates, or pharmaceutically acceptable salts,

[0070] in,

[0071] D is selected from:

[0072] X is selected from CR1 or N;

[0073] Y, Z and V are independently CR5, C(R5)2, NR6, N, O or S, and the total number of O and S is not greater than 1;

[0074] Ring A is phenyl, a 5-7 membered heterocycloalkyl group containing 1-3 heteroatoms selected from N, O, and S, or a 5-6 membered heteroaryl group containing 1-3 heteroatoms selected from N, O, and S;

[0075] Ring B is C 3-12 Carbocyclic ring or 4-12 membered heterocyclic ring containing 1-3 heteroatoms selected from N, O, S;

[0076] Ring C is C 3-12 Carbocyclic ring or 4-12 membered heterocyclic ring containing 1-3 heteroatoms selected from N, O, S;

[0077] R1 is H, D, C1-4 Alkyl, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, halogen, CN, OH, NH2 or COOH, wherein the alkyl, alkoxy, alkenyl, alkynyl is optionally substituted with 1-3 groups selected from halogen, D, CN, OH and NH2;

[0078] R2 is OH, NH2, -CH2NH2 or -NH-OH;

[0079] R3 and R4 are independently H, D, halogen, C 1-4 Alkyl, CN, OH, NH2, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl or C 3-6 Cycloalkyl; the alkyl, alkoxy, alkenyl, alkynyl or cycloalkyl group is optionally substituted by 1-3 groups selected from halogen, D, CN, OH and NH2;

[0080] Alternatively, R3 and R4 together form =NH;

[0081] Each R5 is independently H, D, =O, OH, C 1-4 Alkyl, CN, halogen, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, 5-6 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S or C 3-6 Cycloalkyl, wherein the alkyl, alkoxy, alkenyl, alkynyl, heteroaryl or cycloalkyl is optionally substituted by 1-3 groups selected from halogen, D, CN, OH, C 1-4 Alkyl, C 1-4 Alkoxy and NH2 group substitution;

[0082] R6 is H, D, C 1-4 Alkyl or C 3-6 Cycloalkyl; said alkyl or cycloalkyl is optionally substituted by 1-3 groups selected from halogen, D, CN, OH and NH2;

[0083] Each R7 is independently H, =O, D, OH, CN, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, N(R7')2, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-6Cycloalkyl or 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl or heterocycloalkyl is optionally substituted by 1-3 halogen, D, OH, NH2, CN, C 1-4 Alkyl, -C(O)C 1-4 Alkyl, -C(O)NHC 1-4 Alkyl, -C(O)NH2, -NHC(O)C 1-4 Alkyl and C 1-4 Alkoxy group substitution;

[0084] Each R8 is independently H, D, nitro, amino, C 1-6 Alkoxy, =O, OH, CN, halogen, C 2-6 Alkenyl, C 2-6 Alkynyl, -SF5, C 1-6 Alkyl, -COC 1-4 Alkyl, -N(R7')2, -C(=O)N(R7')2, -NR7'C(=O)-R7', -C(=O)-R7', -(CH2) r -O-(CH2) r -R8'、-SC 1-6 Alkyl, -S(O)-C 1-6 Alkyl, -S(O)2-C 1-6 Alkyl, -S-(CH2) r -R8'、-NR7'-(CH2) r -R8'、-(CH2) r -R8' or N3, the alkyl, alkenyl, alkynyl group is optionally substituted by 1-5 groups selected from R8";

[0085] Each R7' is independently H, D, amino, hydroxyl, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, C 3-6 Cycloalkyl, C 3-12 Heterocycloalkyl, -NHC 1-6 Alkyl, -N(C 1-6 Alkyl)2, or deuterated C 1-6 Alkoxy, the cycloalkyl, heterocycloalkyl is optionally substituted by 1 to 3 groups selected from deuterium, halogen, cyano, amino, hydroxyl, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy or deuterated C 1-6Alkoxy group substitution;

[0086] Each R8' is independently H, D, C 1-6 Alkyl, C 1-4 Alkoxy, C 3-12 Carbocyclic ring, 4-12 membered heterocyclic ring containing 1-3 heteroatoms selected from N, O, S, wherein the alkyl and alkoxy groups are optionally substituted by 1-3 groups selected from halogen, =O, D, OH, NH2, CN, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 Alkyl, -C(O)C 1-4 Alkyl, -C(O)NHC 1-4 Alkyl, -C(O)NH2, -NHC(O)C 1-4 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy and deuterated C 1-6 The carbocyclic or heterocyclic ring is optionally substituted by 1-5 groups selected from R8";

[0087] R8" is selected from halogen, D, OH, NH2, CN, C 1-6 Alkyl and C 1-6 Alkoxy, the alkyl or alkoxy group is optionally further substituted by 1 to 5 groups selected from halogen, deuterium, cyano, amino, hydroxyl, C 1-6 Alkyl or C 1-6 Alkoxy group substitution;

[0088] Each r is independently 0, 1, 2 or 3;

[0089] p is 0, 1, or 2;

[0090] n and m are independently integers from 0 to 5;

[0091] The condition is,

[0092] 1. Group no

[0093] 2. When yes R3 is H, C 1-4 Alkyl, C 1-4 Haloalkyl, R5 is H, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 When halogenated alkyl and R3 and R5 are not H at the same time, the group for

[0094] As a more specific third technical solution of the present invention, the compound of formula (Ia), its stereoisomers, deuterated substances, solvates, or pharmaceutically acceptable salts have the following structure:

[0095] The definitions of other groups are consistent with those of any of the above technical solutions.

[0096] As a more specific fourth technical solution of the present invention, there is provided a compound represented by formula (I), a stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt thereof,

[0097] wherein X is selected from CR1 or N;

[0098] Y, Z and V are independently CR5, C(R5)2, NR6, N, O or S, and the total number of O and S is not greater than 1;

[0099] Ring A is phenyl, a 5-7 membered heterocycloalkyl group containing 1-3 heteroatoms selected from N, O, and S, or a 5-6 membered heteroaryl group containing 1-3 heteroatoms selected from N, O, and S;

[0100] Ring B is C 3-12 Carbocyclic ring or 4-12 membered heterocyclic ring containing 1-3 heteroatoms selected from N, O, S;

[0101] Ring C is C 3-12 Carbocyclic ring or 4-12 membered heterocyclic ring containing 1-3 heteroatoms selected from N, O, S;

[0102] R1 is H, D, C 1-4 Alkyl, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, halogen, CN, OH, NH2 or COOH, wherein the alkyl, alkoxy, alkenyl, alkynyl is optionally substituted with 1-3 groups selected from halogen, D, CN, OH and NH2;

[0103] R2 is OH, NH2, -CH2NH2 or -NH-OH; or R2 is OH, NH2 or -NH-OH;

[0104] R3 and R4 are independently H, D, halogen, C 1-4 Alkyl, CN, OH, NH2, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl or C 3-6 Cycloalkyl; the alkyl, alkoxy, alkenyl, alkynyl or cycloalkyl group is optionally substituted by 1-3 groups selected from halogen, D, CN, OH and NH2;

[0105] Each R5 is independently H, D, =O, OH, C 1-4 Alkyl, CN, halogen, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl or C 3-6 Cycloalkyl, wherein the alkyl, alkoxy, alkenyl, alkynyl or cycloalkyl is optionally substituted with 1-3 groups selected from halogen, D, CN, OH and NH2;

[0106] R6 is H, D, C 1-4 Alkyl or C 3-6 Cycloalkyl; said alkyl or cycloalkyl is optionally substituted by 1-3 groups selected from halogen, D, CN, OH and NH2;

[0107] Each R7 is independently H, =O, D, OH, CN, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, N(R7')2, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-6 Cycloalkyl or 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl or heterocycloalkyl is optionally substituted by 1-3 halogen, D, OH, NH2, CN, C 1-4 Alkyl, -C(O)C 1-4 Alkyl, -C(O)NHC 1-4 Alkyl, -C(O)NH2, -NHC(O)C 1-4 Alkyl and C 1-4 Alkoxy group substitution;

[0108] Each R8 is independently H, D, =O, OH, CN, halogen, C 2-4 Alkenyl, C 2-4 Alkynyl, -SF5, C 1-4 Alkyl, -COC 1-4 Alkyl, -CONR7'C 1-4 Alkyl, -NR7'COC 1-4 Alkyl, N(R7')2, -O-(CH2)r-R8', -S-(CH2)r-R8', -NR7'-(CH2)r-R8' or -(CH2)r-R8', wherein the alkyl, alkenyl, alkynyl is optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-4 Alkyl and C 1-4 Alkoxy group substitution;

[0109] Each R7' is independently H, C 1-4 Alkyl or halogenated C 1-4 alkyl;

[0110] Each R8' is independently H, C 1-4 Alkoxy, C 3-9 Carbocyclic ring, 4-10 membered heterocyclic ring containing 1-3 heteroatoms selected from N, O, S, wherein the carbocyclic ring or heterocyclic ring is optionally substituted by 1-3 heteroatoms selected from halogen, =O, D, OH, NH2, CN, C 1-4 Alkyl, -C(O)C 1-4 Alkyl, -C(O)NHC 1-4 Alkyl, -C(O)NH2, -NHC(O)C 1-4 Alkyl and C 1-4 Alkoxy group substitution;

[0111] Each r is independently 0, 1, 2 or 3;

[0112] p is 0, 1, or 2;

[0113] n and m are independently integers from 0 to 5;

[0114] The condition is,

[0115] 1. Group no

[0116] 2. When yes R3 is H, C 1-4 Alkyl, C 1-4 Haloalkyl, R5 is H, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 When halogenated alkyl and R3 and R5 are not H at the same time, the group for

[0117] As a more specific fifth technical solution of the present invention, the compound represented by formula (Ia), formula (Ib), formula (I), formula (II-a) or formula (II), its stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt, wherein

[0118] D is selected from Further, D is selected from Further, D is selected from further,

[0119] D is selected from further,

[0120] D is selected from

[0121] The definitions of other groups are consistent with those of any of the above technical solutions.

[0122] As a more specific sixth technical solution of the present invention, the compound represented by formula (Ia), formula (Ib), formula (I), formula (II-a) or formula (II), its stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt, wherein

[0123] D is selected from

[0124] The definitions of other groups are consistent with those of any of the above technical solutions.

[0125] As a more specific seventh technical solution of the present invention, the compound represented by formula (Ia), formula (Ib), formula (I), its stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt,

[0126] wherein X is selected from CR1 or N;

[0127] Y, Z and V are independently CR5, N, O or S, and the total number of O and S is not greater than 1;

[0128] Ring A is phenyl, or a 5-6 membered heteroaryl group containing 1-3 heteroatoms selected from N, O, and S;

[0129] Ring B is phenyl, C 3-6 cycloalkyl, 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, and S, or 5-6 membered aryl containing 1-3 heteroatoms selected from N, O, and S;

[0130] Ring C is phenyl, C 3-6 cycloalkyl, 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, and S, or 5-6 membered aryl containing 1-3 heteroatoms selected from N, O, and S;

[0131] R1 is H, D, C 1-4 Alkyl, halogen, CN, wherein the alkyl is optionally substituted with 1 to 3 groups selected from halogen, D, CN and OH;

[0132] R2 is OH, NH2, -CH2NH2 or -NH-OH; or R2 is OH, NH2 or -NH-OH;

[0133] R3 and R4 are independently H, D, halogen, C 1-4Alkyl, CN, OH or NH2; the alkyl group is optionally substituted with 1-3 groups selected from halogen, D, CN, OH and NH2;

[0134] Each R5 is independently H, D, OH, C 1-4 Alkyl, CN, halogen, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, 5-membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, 4-5-membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S or C 3-4 Cycloalkyl, the alkyl, alkoxy, alkenyl, alkynyl, heteroaryl, heterocycloalkyl, cycloalkyl is optionally substituted by 1-3 groups selected from halogen, D, CN, OH, C 1-4 Alkyl, C 1-4 In certain embodiments, each R5 is independently H, D, OH, C 1-4 Alkyl, CN, halogen, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, 5-membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, 4-5-membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S or C 3-4 Cycloalkyl, the alkyl, alkoxy, alkenyl, alkynyl, heteroaryl, heterocycloalkyl, cycloalkyl is optionally substituted by 1-3 groups selected from halogen, D, CN, OH and NH2; or each R5 is independently H, D, OH, C 1-4 Alkyl, CN, halogen, C 1-4 Alkoxy, C 2-4 Alkenyl, or C 2-4 Alkynyl, wherein the alkyl, alkoxy, alkenyl or alkynyl group is optionally substituted with 1-3 groups selected from halogen, D, CN, OH and NH2;

[0135] Each R7 is independently H, =O, D, OH, CN, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, N(R7')2, C 2-4 Alkenyl, or C 2-4 Alkynyl, the alkyl, alkoxy, alkenyl, alkynyl is optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-4 Alkyl, -C(O)C 1-4 Alkyl, -C(O)NHC 1-4 Alkyl, -C(O)NH2, -NHC(O)C 1-4 Alkyl and C 1-4 Alkoxy group substitution;

[0136] Each R8 is independently H, D, =O, OH, CN, halogen, C 2-4 Alkenyl, C 2-4 Alkynyl, N3, C 1-4 Alkyl, -COC 1-4 Alkyl, -CONR7'C 1-4 Alkyl, -NR7'COC 1-4 Alkyl, N(R7')2, -O-(CH2)r-R8', or -(CH2)r-R8', wherein the alkyl, alkenyl, alkynyl is optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-4 Alkyl and C 1-4 or each R8 is independently H, D, =O, OH, CN, halogen, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkyl, -COC 1-4 Alkyl, -CONR7'C 1-4 Alkyl, -NR7'COC 1-4 Alkyl, N(R7')2, -O-(CH2)r-R8', or -(CH2)r-R8', wherein the alkyl, alkenyl, alkynyl is optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-4 Alkyl and C 1-4 Alkoxy group substitution;

[0137] Each R7' is independently H, C 1-4 Alkyl or halogenated C 1-4 alkyl;

[0138] Each R8' is independently H, C 1-4 Alkoxy, C 3-6 Cycloalkyl, phenyl, 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, or 5-6 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, wherein the cycloalkyl, phenyl, heterocycloalkyl or heteroaryl is optionally substituted by 1-3 halogen, =O, D, OH, NH2, CN, C 1-4 Alkyl, halogenated C 1-4 Alkyl, and C 1-4 or, each R8' is independently H, C 1-4 Alkoxy, C 3-6 Cycloalkyl, phenyl, 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, or 5-6 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, wherein the cycloalkyl, phenyl, heterocycloalkyl or heteroaryl is optionally substituted by 1-3 halogen, =O, D, OH, NH2, CN, C 1-4 Alkyl, and C 1-4Alkoxy group substitution;

[0139] Each r is independently 0, 1, or 2;

[0140] p is 0 or 1;

[0141] n and m are independently 0, 1, 2, 3 or 4;

[0142] The remaining groups are as described in any one of the first, second, third, fourth, fifth and sixth technical solutions.

[0143] As a more specific eighth technical solution of the present invention, the compound of the present invention, its stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt, the B ring is selected from

[0144] As a more specific ninth technical solution of the present invention, the compound represented by formula (Ia), formula (I), its stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt, has the structure of (II-a), formula (II):

[0145] The remaining groups are as described in the first, second, third, fourth, fifth, sixth, seventh and eighth technical solutions.

[0146] As a more specific tenth technical solution of the present invention, the compound represented by formula (Ia), formula (Ib), formula (I), formula (II-a) or formula (II), its stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt,

[0147] in,

[0148] X is selected from CR1 or N;

[0149] Y, Z and V are independently CR5, N, O or S, and the total number of O and S is not greater than 1;

[0150] Ring A is phenyl, thienyl, thiazolyl, pyrrolyl, imidazolyl, oxazolyl, pyridyl, pyrazinyl, pyrimidinyl or pyridazinyl;

[0151] D is selected from Or D is selected from

[0152] Alternatively, D is selected from

[0153] R1 is H, D, C 1-4 Alkyl, F, Cl, Br, I or CN, wherein the alkyl group is optionally substituted with 1-3 groups selected from F, Cl, Br, I, D, CN and OH;

[0154] R2 is OH, NH2, -CH2NH2 or -NH-OH;

[0155] R3 and R4 are independently H, D, F, Cl, Br, I, C 1-4 Alkyl or CN; the alkyl group is optionally substituted by 1-3 groups selected from F, Cl, Br, I, D, CN, OH and NH2;

[0156] Each R5 is independently H, D, OH, C 1-4 Alkyl, CN, F, Cl, Br, I, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, 5-membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, 4-5-membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S or C 3-4 Cycloalkyl, wherein the alkyl, alkoxy, alkenyl, alkynyl, heteroaryl, heterocycloalkyl, cycloalkyl is optionally substituted with 1-3 groups selected from F, Cl, Br, I, D, CN, OH, methoxy, and NH2; in certain embodiments, each R5 is independently H, D, OH, C 1-4 Alkyl, CN, F, Cl, Br, I, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, 5-membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, 4-5-membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S or C 3-4 Cycloalkyl, wherein the alkyl, alkoxy, alkenyl, alkynyl, heteroaryl, heterocycloalkyl, cycloalkyl is optionally substituted by 1-3 groups selected from F, Cl, Br, I, D, CN, OH and NH2; or, each R5 is independently H, D, OH, C 1-4 Alkyl, CN, F, Cl, Br, I, C 1-4 Alkoxy, C 2-4 Alkenyl, or C 2-4 Alkynyl, wherein the alkyl, alkoxy, alkenyl or alkynyl group is optionally substituted with 1-3 groups selected from F, Cl, Br, I, D, CN, OH and NH2;

[0157] Each R7 is independently H, D, OH, CN, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, N(R7')2, C 2-4 Alkenyl, or C 2-4 Alkynyl, the alkyl, alkoxy, alkenyl, alkynyl is optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-4 Alkyl and C 1-4 Alkoxy group substitution;

[0158] Each R8 is independently H, D, OH, CN, F, Cl, Br, I, C 2-4 Alkenyl, C 2-4 Alkynyl, N3, C 1-4 Alkyl, -COC 1-4 Alkyl, -CONR7'C 1-4 Alkyl, -NR7'COC 1-4 Alkyl, N(R7')2, -O-(CH2)r-R8', or -(CH2)r-R8', wherein the alkyl, alkenyl, alkynyl is optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-4 Alkyl and C 1-4 or each R8 is independently H, D, OH, CN, F, Cl, Br, I, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkyl, -COC 1-4 Alkyl, -CONR7'C 1-4 Alkyl, -NR7'COC 1-4 Alkyl, N(R7')2, -O-(CH2)r-R8', or -(CH2)r-R8', wherein the alkyl, alkenyl, alkynyl is optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-4 Alkyl and C 1-4 Alkoxy group substitution;

[0159] Each R7' is independently H, C 1-4 Alkyl or halogenated C 1-4 alkyl;

[0160] Each R8' is independently H, C 1-4 Alkoxy, C 3-6 Cycloalkyl, phenyl, 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, or 5-6 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, wherein the cycloalkyl, phenyl, heterocycloalkyl or heteroaryl is optionally substituted by 1-3 halogen, =O, D, OH, NH2, CN, C 1-4 Alkyl, halogenated C 1-4 Alkyl, and C 1-4 or, each R8' is independently H, C 1-4 Alkoxy, C 3-6Cycloalkyl, phenyl, 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, or 5-6 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, wherein the cycloalkyl, phenyl, heterocycloalkyl or heteroaryl is optionally substituted by 1-3 halogen, =O, D, OH, NH2, CN, C 1-4 Alkyl, and C 1-4 Alkoxy group substitution;

[0161] Each r is independently 0, 1, or 2;

[0162] p is 0 or 1;

[0163] n is 0, 1, 2, or 3;

[0164] m is 0, 1, 2, 3 or 4;

[0165] The remaining groups are as described in the first, second, third, fourth, fifth, sixth, seventh, eighth and ninth technical solutions.

[0166] As a more specific eleventh technical solution of the present invention, the compound represented by formula (Ia), formula (Ib), formula (I), formula (II-a) or formula (II), its stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt, wherein,

[0167] X is selected from CR1 or N;

[0168] Y, Z and V are independently CR5, N, O or S, and the total number of O and S is not greater than 1;

[0169] Ring A is phenyl, thienyl, thiazolyl, pyrrolyl, imidazolyl, oxazolyl, pyridyl, pyrazinyl, pyrimidinyl or pyridazinyl;

[0170] D is selected from

[0171] R1 is H, D, C 1-4 Alkyl, F, Cl, Br, I or CN, wherein the alkyl group is optionally substituted with 1-3 groups selected from F, Cl, Br, I, D, CN and OH;

[0172] R2 is OH, NH2, -CH2NH2 or -NH-OH;

[0173] R3 and R4 are independently H, D, F, Cl, Br, I, C 1-4 Alkyl or CN; the alkyl group is optionally substituted by 1-3 groups selected from F, Cl, Br, I, D, CN, OH and NH2;

[0174] Each R5 is independently H, D, OH, C 1-4Alkyl, CN, F, Cl, Br, I, C 1-4 Alkoxy, C 2-4 Alkenyl, or C 2-4 Alkynyl, wherein the alkyl, alkoxy, alkenyl or alkynyl group is optionally substituted with 1-3 groups selected from F, Cl, Br, I, D, CN, OH and NH2;

[0175] Each R7 is independently H, D, OH, CN, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, N(R7')2, C 2-4 Alkenyl, or C 2-4 Alkynyl, the alkyl, alkoxy, alkenyl, alkynyl is optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-4 Alkyl and C 1-4 Alkoxy group substitution;

[0176] Each R8 is independently H, D, OH, CN, F, Cl, Br, I, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkyl, -COC 1-4 Alkyl, -CONR7'C 1-4 Alkyl, -NR7'COC 1-4 Alkyl, N(R7')2, -O-(CH2)r-R8', or -(CH2)r-R8', wherein the alkyl, alkenyl, alkynyl is optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-4 Alkyl and C 1-4 Alkoxy group substitution;

[0177] Each R7' is independently H, C 1-4 Alkyl or halogenated C 1-4 alkyl;

[0178] Each R8' is independently H, C 1-4 Alkoxy, C 3-6 Cycloalkyl, phenyl, 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, or 5-6 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, wherein the cycloalkyl, phenyl, heterocycloalkyl or heteroaryl is optionally substituted by 1-3 halogen, =O, D, OH, NH2, CN, C 1-4 Alkyl, and C 1-4 Alkoxy group substitution;

[0179] Each r is independently 0, 1, or 2;

[0180] p is 0 or 1;

[0181] n is 0, 1, 2, or 3;

[0182] m is 0, 1, 2, 3 or 4;

[0183] The remaining groups are as described in any of the above technical solutions.

[0184] As a more specific twelfth technical solution of the present invention, the compound represented by formula (Ia), formula (Ib), formula (I), formula (II-a) or formula (II), its stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt,

[0185] X is selected from CR1 or N;

[0186] Y, Z and V are independently CR5, N, O or S, and the total number of O and S is not greater than 1;

[0187] Ring A is phenyl, thienyl, thiazolyl, pyrrolyl, imidazolyl, oxazolyl, pyridyl, pyrazinyl, pyrimidinyl or pyridazinyl;

[0188] R1 is H, D, C 1-4 Alkyl, F, Cl, Br, I or CN, wherein the alkyl group is optionally substituted with 1-3 groups selected from F, Cl, Br, I, D, CN and OH;

[0189] R2 is OH, NH2 or -NH-OH;

[0190] R3 and R4 are independently H, D, F, Cl, Br, I, C 1-4 Alkyl or CN; the alkyl group is optionally substituted by 1-3 groups selected from F, Cl, Br, I, D, CN, OH and NH2;

[0191] Each R5 is independently H, D, OH, C 1-4 Alkyl, CN, F, Cl, Br, I, C 1-4 Alkoxy, C 2-4 Alkenyl, or C 2-4 Alkynyl, wherein the alkyl, alkoxy, alkenyl or alkynyl group is optionally substituted with 1-3 groups selected from F, Cl, Br, I, D, CN, OH and NH2;

[0192] Each R7 is independently H, D, OH, CN, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, N(R7')2, C 2-4 Alkenyl, or C 2-4 Alkynyl, the alkyl, alkoxy, alkenyl, alkynyl is optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-4 Alkyl and C 1-4 Alkoxy group substitution;

[0193] Each R8 is independently H, D, OH, CN, F, Cl, Br, I, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkyl, -COC 1-4 Alkyl, -CONR7'C 1-4 Alkyl, -NR7'COC 1-4 Alkyl, N(R7')2, -O-(CH2)r-R8', or -(CH2)r-R8', wherein the alkyl, alkenyl, alkynyl is optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-4 Alkyl and C 1-4 Alkoxy group substitution;

[0194] Each R7' is independently H, C 1-4 Alkyl or halogenated C 1-4 alkyl;

[0195] Each R8' is independently H, C 1-4 Alkoxy, C 3-6 Cycloalkyl, phenyl, 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, or 5-6 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, wherein the cycloalkyl, phenyl, heterocycloalkyl or heteroaryl is optionally substituted by 1-3 halogen, =O, D, OH, NH2, CN, C 1-4 Alkyl, and C 1-4 Alkoxy group substitution;

[0196] Each r is independently 0, 1, or 2;

[0197] p is 0 or 1;

[0198] n is 0, 1, 2, or 3;

[0199] m is 0, 1, 2, 3 or 4;

[0200] The remaining groups are as described in any of the above technical solutions.

[0201] As a more specific thirteenth technical solution of the present invention, the compound represented by formula (Ia), formula (Ib), formula (I), formula (II-a) or formula (II), its stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt,

[0202] Each R8 is independently D, CN, F, Cl, Br, C 2-4 Alkenyl, C 2-4 Alkynyl, N3, C 1-4Alkyl, -O-(CH2)r-R8', or -(CH2)r-R8', wherein the alkyl, alkenyl, alkynyl is optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-2 Alkyl and C 1-2 Alkoxy group substitution;

[0203] The definitions of other groups are the same as those in any of the above technical solutions.

[0204] As a more specific fourteenth technical solution of the present invention, the compound represented by formula (Ia), formula (Ib), formula (I), formula (II-a) or formula (II), its stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt, wherein,

[0205] R8' is independently C 1-4 Alkoxy, cyclopropyl, cyclobutyl, cyclopentyl, phenyl, tetrahydropyrrolyl, azetidinyl, piperidinyl, piperazinyl, morpholinyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, Thiphenyl, thiazolyl, pyrrolyl, imidazolyl, oxazolyl, pyridinyl, pyrazinyl, pyrimidinyl or pyridazinyl, optionally substituted by 1-3 groups selected from F, Cl, Br, I, =O, D, OH, NH2, CN, C 1-4 Alkyl, halogenated C 1-2 Alkyl, and C 1-2 Alkoxy group substitution; or,

[0206] R8' is independently C 1-4 Alkoxy, cyclopropyl, cyclobutyl, cyclopentyl, phenyl, tetrahydropyrrolyl, azetidinyl, piperidinyl, piperazinyl, morpholinyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, Thiphenyl, thiazolyl, pyrrolyl, imidazolyl, oxazolyl, pyridinyl, pyrazinyl, pyrimidinyl or pyridazinyl, optionally substituted by 1-3 groups selected from F, Cl, Br, I, =O, D, OH, NH2, CN, C 1-4 Alkyl, and C 1-4 Alkoxy group substitution;

[0207] The remaining groups are as described in any of the above technical solutions.

[0208] As a more specific fifteenth technical solution of the present invention, the compound represented by formula (Ia), formula (Ib), formula (I), formula (II-a) or formula (II), its stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt, wherein D is selected from

[0209] D is selected from

[0210] The definitions of other groups are consistent with any of the above technical solutions.

[0211] As a more specific sixteenth technical solution of the present invention, the compound represented by formula (Ia), formula (Ib), formula (I), formula (II-a) or formula (II), its stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt, wherein,

[0212] R8 is selected from -O-(CH2) r -R 8’ , 3-12 membered heterocycloalkyl, C 3-12 Cycloalkyl, -O-(CH2) r -C 3-12 Cycloalkyl, -O-(CH2) r -(3-12 membered heterocycloalkyl) or -N(R 7’ ) 2, the CH2, cycloalkyl, heterocycloalkyl is optionally further substituted by 1-3 R8 ", preferably the cycloalkyl is further substituted by 1-3 R8";

[0213] Each R 7’ Each independently selected from hydrogen, deuterium, amino, hydroxyl, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy or deuterated C 1-4 alkoxy;

[0214] R 8’ Selected from C 1-4 Alkyl or C 1-4 Alkoxy, said alkyl or alkoxy optionally further substituted by 1 to 5 groups selected from deuterium, halogen, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy or deuterated C 1-4 Alkoxy group substitution;

[0215] Each R8" is independently selected from deuterium, =O, halogen, C 1-4 Alkyl or C 1-4 Alkoxy, the alkyl or alkoxy group is optionally further substituted by 1 to 3 groups selected from halogen, deuterium, hydroxyl, C 1-4 Alkyl or C 1-4 Alkoxy group substitution;

[0216] The definitions of other groups are the same as above.

[0217] As a more specific seventeenth technical solution of the present invention, the compound represented by formula (Ia), formula (Ib), formula (I), formula (II-a) or formula (II), its stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt, wherein,

[0218] R8 is selected from -O-(CH2) r -O(CH2) r CH3, 4-6 membered monocyclic heterocycloalkyl, 5-8 membered cyclic heterocycloalkyl, 5-11 membered spirocyclic heterocycloalkyl, 5-8 membered bridged heterocycloalkyl, -O-(CH2) r -C 3-8 Monocyclic cycloalkyl, -O-(CH2) r -4-6 membered monocyclic heterocycloalkyl, -O-(CH2) r -5-8 membered heterocyclic alkyl, -O-(CH2) r -5-8 membered spirocyclic heterocycloalkyl, -O-(CH2) r -5-8 membered bridged heterocycloalkyl or -N(CH3)2, wherein the heterocycloalkyl or cycloalkyl group is optionally further substituted with 1-3 R8", and preferably the cycloalkyl group is further substituted with 1-3 R8";

[0219] R8" are each independently selected from deuterium, =0, F, Cl, methyl, ethyl, propyl, methoxy, ethoxy or propoxy, said methyl, ethyl, propyl, methoxy, ethoxy or propoxy being optionally further substituted with 1 to 3 groups selected from F, Cl, deuterium, hydroxy, methyl, ethyl, propyl, methoxy, ethoxy or propoxy;

[0220] The definitions of other groups are the same as above.

[0221] As a more specific eighteenth technical solution of the present invention, the compound represented by formula (Ia), formula (Ib), formula (I), formula (II-a) or formula (II), its stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt, wherein,

[0222] Each R8 is independently CN, F, Cl, N3, vinyl, fluorovinyl, -O-CH2CH2-OCH3, -N(CH3)2, further

[0223] R8 is selected from -O-CH2CH2-OCH3, -N(CH3)2,

[0224] The definitions of other groups are the same as above.

[0225] As a more specific nineteenth technical solution of the present invention, the compound of formula (Ib), its stereoisomers, deuterated substances, solvates, or pharmaceutically acceptable salts, wherein,

[0226] D1 is selected from:

[0227] Ld is selected from C 2-4 Alkenyl, in certain embodiments, Ld is selected from

[0228] Rd1 is selected from 5-membered heteroaryl, wherein the heteroaryl is optionally substituted by 1-2 groups selected from NH2. In certain embodiments, Rd1 is selected from

[0229] As a more specific twentieth technical solution of the present invention, the compound of the present invention, its stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt is selected from the structure of the following Table 1.

[0230] Table 1

[0231] As a more specific twenty-first technical solution of the present invention, a pharmaceutical composition is provided, which contains the compound described in any one of the first to twentieth technical solutions, its stereoisomers, deuterated substances, solvates, or pharmaceutically acceptable salts, and pharmaceutically acceptable carriers and / or excipients.

[0232] As a more specific twenty-second technical solution of the present invention, a use is provided, namely, use of the compound described in any one of the first to twentieth technical solutions, its stereoisomers, deuterated substances, solvates, or pharmaceutically acceptable salts, or the composition described in the twenty-first technical solution in the preparation of a medicament for treating / preventing PRMT5·MTA-mediated diseases.

[0233] Furthermore, the PRMT5·MTA-mediated disease is selected from liver cancer, breast cancer, skin cancer, bladder cancer, pancreatic cancer or head and neck cancer.

[0234] The present invention also provides a composition or pharmaceutical formulation comprising a compound according to any of the foregoing embodiments, a stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier and / or excipient. The pharmaceutical composition may be in unit dosage form (a unit dosage is also referred to as a "dose strength").

[0235] Furthermore, the composition or pharmaceutical preparation of the present invention contains 1-1500 mg of the compound described in any one of the aforementioned schemes, its stereoisomers, deuterated substances, solvates, or pharmaceutically acceptable salts, and a pharmaceutically acceptable carrier and / or excipient.

[0236] The present invention also provides the use of a compound according to any of the preceding embodiments, or a stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt thereof, in the preparation of a medicament for treating or preventing a PRMT5·MTA-mediated disease. Furthermore, the PRMT5·MTA-mediated disease is liver cancer, breast cancer, skin cancer, bladder cancer, pancreatic cancer, or head and neck cancer.

[0237] The present invention also provides a method for treating a disease in a mammal, comprising administering to a subject a therapeutically effective amount of a compound of any of the foregoing schemes, a stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt thereof, wherein the disease is preferably liver cancer, breast cancer, skin cancer, bladder cancer, pancreatic cancer, or head and neck cancer, and wherein the therapeutically effective amount is preferably 1-1500 mg. In some embodiments, the mammal of the present invention includes a human.

[0238] As used herein, an "effective amount" or "therapeutically effective amount" refers to the administration of a sufficient amount of a compound disclosed herein to alleviate, to some extent, one or more symptoms of the disease or condition being treated. In some embodiments, the result is a reduction and / or alleviation of the signs, symptoms, or causes of a disease, or any other desired alteration of a biological system. For example, an "effective amount" for therapeutic uses is the amount of a composition comprising a compound disclosed herein required to provide a clinically significant reduction in disease symptoms. Examples of therapeutically effective amounts include, but are not limited to, 1-1500 mg, 1-1400 mg, 1-1300 mg, 1-1200 mg, 1-1000 mg, 1-900 mg, 1-800 mg, 1-700 mg, 1-600 mg, 1-500 mg, 1-400 mg, 1-300 mg, 1-250 mg, 1-200 mg, 1-150 mg, 1-125 mg, 1-100 mg, 1-80 mg, 1-60 mg, 1-50 mg, 1-40 mg, 1-25 mg, 1-20 mg, 5-1500 mg, 5-1000 mg, 5-900 mg, 5-800 mg, 5-700 mg, 5-6 00mg, 5-500mg, 5-400mg, 5-300mg, 5-250mg, 5-200mg, 5-150mg, 5-125mg, 5-100mg, 5-90mg, 5-70mg, 5-80mg, 5-60mg, 5-50mg, 5-40mg, 5-30mg, 5-2 5mg, 5-20mg, 10-1500mg, 10-1000mg, 10-900mg, 10-800mg, 10-700mg, 10-600mg, 10-500mg, 10-450mg, 10-400mg, 10-300mg, 10-250mg, 10-200mg, 10-150mg, 10-125mg, 10-100mg, 10-90mg, 10-80mg, 10-70mg, 10-60mg, 10-50mg, 10-4 0mg, 10-30mg, 10-20mg; 20-1500mg, 20-1000mg, 20-900mg, 20-800mg, 20-700mg, 20-6 00mg, 20-500mg, 20-400mg, 20-350mg, 20-300mg, 20-250mg, 20-200mg, 20-150mg, 20- 125mg, 20-100mg, 20-90mg, 20-80mg, 20-70mg, 20-60mg, 20-50mg, 20-40mg, 20-30mg;50-1500mg, 50-1000mg, 50-900mg, 50-800mg, 50-700mg, 50-600mg, 50-500mg, 50-400mg, 50-300mg, 50-250mg, 50-200mg, 50-150mg, 50-125mg, 5 0-100mg; 100-1500mg, 100-1000mg, 100-900mg, 100-800mg, 100-700mg, 100-600mg, 100-500mg, 100-400mg, 100-300mg, 100-250mg, 100-200mg;

[0239] In some embodiments, the pharmaceutical composition or formulation of the present invention contains the above-mentioned therapeutically effective amount of the compound of the present invention or its stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt;

[0240] The present invention relates to a pharmaceutical composition or pharmaceutical preparation comprising a therapeutically effective amount of a compound of the present invention, or a stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt thereof, and a carrier and / or excipient. The pharmaceutical composition may be in the form of a unit dosage form (the amount of the active ingredient in a unit dosage form is also referred to as the "drug strength"). In some embodiments, the pharmaceutical composition includes but is not limited to 1 mg, 1.25 mg, 2.5 mg, 5 mg, 10 mg, 12.5 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 110 mg, 120 mg, 125 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg g, 250 mg, 275 mg, 300 mg, 325 mg, 350 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 525 mg, 550 mg, 575 mg, 600 mg, 625 mg, 650 mg, 675 mg, 700 mg, 725 mg, 750 mg, 775 mg, 800 mg, 850 mg, 900 mg, 950 mg, 1000 mg, 1100 mg, 1200 mg, 1300 mg, 1400 mg, 1500 mg of a compound of the present invention or a stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt thereof.

[0241] A method for treating a disease in a mammal, comprising administering to a subject a therapeutically effective amount of a compound of the present invention, a stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier and / or excipient, wherein the therapeutically effective amount is preferably 1-1500 mg, and the disease is preferably liver cancer, breast cancer, skin cancer, bladder cancer, pancreatic cancer, or head and neck cancer.

[0242] A method for treating a disease in a mammal, comprising administering a compound of the present invention, a stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier and / or excipient to a subject at a daily dose of 1-1500 mg / day, wherein the daily dose can be a single dose or divided doses. In some embodiments, the daily dose includes but is not limited to 10-1500 mg / day, 20-1500 mg / day, 25-1500 mg / day, 50-1500 mg / day, 75-1500 mg / day, 100-1500 mg / day, 200-1500 mg / day, 10-1000 mg / day, 20-1000 mg / day, 25-1000 mg / day, 50-1000 mg / day, 75-1000 mg / day, In some embodiments, the daily dose includes but is not limited to 1 mg / day, 5 mg / day, 10 mg / day, 20 mg / day, 25 mg / day, 50 mg / day, 75 mg / day, 100 mg / day, 125 mg / day, 150 mg / day, 200 mg / day, 400 mg / day, 600 mg / day, 800 mg / day, 1000 mg / day, 1200 mg / day, 1400 mg / day, 1500 mg / day.

[0243] The present invention relates to a kit, which may include a composition in single-dose or multi-dose form, wherein the kit contains a compound of the present invention or a stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt thereof, and the amount of the compound of the present invention or its stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt is the same as that in the above-mentioned pharmaceutical composition.

[0244] The amount of the compound of the present invention or its stereoisomer, deuterated form, solvate or pharmaceutically acceptable salt in the present invention is in each case calculated based on the free base form.

[0245] "Preparation specifications" refers to the weight of the main drug contained in each vial, tablet or other unit preparation.

[0246] Synthesis route

[0247] Those skilled in the art can prepare the compounds of the present invention by combining known organic synthesis techniques, using commercially available chemicals and / or compounds described in the chemical literature as starting materials. "Commercially available chemicals" are obtained from reputable commercial sources, including suppliers such as Titan Technology, Anage Chemical, Shanghai Demo, Chengdu Kelon Chemical, Shaoyuan Chemical Technology, Nanjing Yaoshi, WuXi AppTec, and J&K Technology.

[0248] Specific and similar reactants can be selectively identified by indexes of known chemical substances prepared by the American Chemical Society's Chemical Abstracts Service, which are available in most public and university libraries and online. Chemicals that are known but not commercially available in the catalog are optionally prepared by custom chemical synthesis facilities, many of which standard chemical supply facilities (e.g., those listed above) offer custom synthesis services.

[0249] the term

[0250] Unless otherwise specified in the present invention, the terms of the present invention have the following meanings:

[0251] The carbon, hydrogen, oxygen, sulfur, nitrogen or halogen involved in the groups and compounds of the present invention include their isotopes, and the carbon, hydrogen, oxygen, sulfur, nitrogen or halogen involved in the groups and compounds of the present invention are optionally further replaced by one or more of their corresponding isotopes, wherein the isotopes of carbon include 12 C. 13 C and 14 C, hydrogen isotopes include protium (H), deuterium (deuterium, also known as heavy hydrogen), tritium (T, also known as super tritium), oxygen isotopes include 16 O. 17 O and 18 O, sulfur isotopes include 32 S. 33 S. 34 S and 36 S, nitrogen isotopes include 14 N and 15 N, an isotope of fluorine 19 F, chlorine isotopes include 35 Cl and 37 Isotopes of Cl, bromine include 79 Br and 81 Br.

[0252] "Halogen" herein refers to F, Cl, Br, I, or isotopes thereof.

[0253] "Halo" or "halogen-substituted" refers to substitution with one or more halogens selected from F, Cl, Br, I, or isotopes thereof. The upper limit of the number of halogen substituents is equal to the sum of the number of hydrogen atoms that can be replaced by the substituted group. Unless otherwise specified, the number of halogen substituents is any integer between 1 and the upper limit. When the number of halogen substituents is greater than 1, they may be the same or different halogens. Typical examples include 1-5 halogen substitutions, 1-3 halogen substitutions, 1-2 halogen substitutions, and 1 halogen substitution.

[0254] "Deuterium" refers to the isotope of hydrogen (H) and is synonymous with "D".

[0255] "Deuterated" or "deuterated compound" refers to a situation where a hydrogen atom on an alkyl, cycloalkyl, alkylene, aryl, heteroaryl, thiol, heterocycloalkyl, alkenyl, alkynyl or other group is replaced by at least one deuterium atom. The upper limit of the number of deuterated groups is equal to the sum of the number of replaceable hydrogen atoms in the substituted group. Unless otherwise specified, the number of deuterated groups is any integer between 1 and the upper limit, for example, 1-20 deuterium atoms, 1-10 deuterium atoms, 1-6 deuterium atoms, 1-3 deuterium atoms, 1-2 deuterium atoms or 1 deuterium atom.

[0256] “C x-y " group refers to a group containing x to y carbon atoms, such as "C 1-6 "Alkyl" refers to an alkyl group containing 1 to 6 carbon atoms.

[0257] "Alkyl" refers to a monovalent straight-chain or branched saturated aliphatic hydrocarbon group. Typically, it is an alkyl group having 1 to 20 carbon atoms, or an alkyl group having 1 to 8 carbon atoms, or an alkyl group having 1 to 6 carbon atoms, or an alkyl group having 1 to 4 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, neobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, and the like. The alkyl group may be further substituted with a substituent.

[0258] "Alkylene" refers to a divalent straight-chain or branched saturated alkyl group. Examples of alkylene groups include, but are not limited to, methylene, ethylene, and the like.

[0259] "Haloalkyl" refers to a situation where one or more hydrogen atoms in an alkyl group are replaced by one or more halogen atoms (such as fluorine, chlorine, bromine, iodine, or their isotopes). The upper limit of the number of halogen substituents is equal to the sum of the number of replaceable hydrogen atoms in the alkyl group. Unless otherwise specified, the number of halogen substituents is any integer between 1 and the upper limit. Typically, an alkyl group is substituted with 1-5 halogens, or 1-3 halogens, or 1-2 halogens, or 1 halogen. When the number of halogen substituents is greater than 1, they can be the same or different halogens. Specific examples include, but are not limited to, -CF3, -CH2Cl, -CH2CF3, -CCl2, CF3, etc.

[0260] "Alkoxy" or "alkyloxy" refers to an -O-alkyl group. For example, -OC 1-8 Alkyl, -OC 1-6 Alkyl, -OC 1-4 Alkyl or -OC 1-2 Specific non-limiting examples include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, n-hexyloxy, cyclopropyloxy, and cyclobutyloxy, and the like; the alkoxy groups may be optionally substituted with substituents.

[0261] "Haloalkoxy" refers to an -O-haloalkyl group. For example, -O-haloC 1-8 Alkyl, -O-halogenated C 1-6 Alkyl, -O-halogenated C 1-4 Alkyl or -O-halogenated C 1-2 Alkyl; the upper limit of the number of halogen substituents is equal to the sum of the number of hydrogen atoms that can be substituted by the substituted group. Unless otherwise specified, the number of halogen substituents is any integer between 1 and the upper limit, preferably 1-5 halogen substitutions, 1-3 halogen substitutions, 1-2 halogen substitutions, or 1 halogen substitution; when the number of halogen substituents is greater than 1, they may be the same or different halogen substitutions; non-limiting examples include monofluoromethoxy, difluoromethoxy, trifluoromethoxy, difluoroethyloxy, and the like.

[0262] "Alkenyl" refers to a straight or branched chain hydrocarbon group containing at least one carbon-carbon double bond (C=C), typically containing 2 to 18 carbon atoms, such as 2 to 8 carbon atoms, further such as 2 to 6 carbon atoms, and further such as 2 to 4 carbon atoms, examples of which include but are not limited to vinyl, allyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-methyl-1-butenyl, 2-methyl-1-butenyl, 2-methyl-2-butenyl, 2-methyl-3-butenyl, 2-methyl-4-butenyl, 2-methyl-5-butenyl, 2-methyl-6-butenyl, 2-methyl-7-butenyl, 2-methyl-8-butenyl, 2-methyl-9-butenyl, 2-methyl-1-butenyl, 2-methyl-2-butenyl, 2-methyl-3 ... -methyl-3-butenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 1-methyl-1-pentenyl, 2-methyl-1-pentenyl, 1-heptenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 1-octenyl, 3-octenyl, 1-nonenyl, 3-nonenyl, 1-decenyl, 4-decenyl, 1,3-butadiene, 1,3-pentadiene, 1,4-pentadiene and 1,4-hexadiene, etc.; the alkenyl group may be optionally further substituted with a substituent.

[0263] "Alkenylene" refers to a straight or branched divalent unsaturated hydrocarbon group containing at least one carbon-carbon double bond (C=C), typically containing 2 to 18 carbon atoms, such as 2 to 8 carbon atoms, further such as 2 to 6 carbon atoms, and further such as 2 to 4 carbon atoms. Non-limiting examples include ethynylene, and the alkenylene group may be optionally substituted with a substituent.

[0264] "Alkynyl" refers to a straight-chain or branched hydrocarbon group containing at least one carbon-carbon triple bond (C≡C), typically containing 2 to 18 carbon atoms, further containing 2 to 8 carbon atoms, further containing 2 to 6 carbon atoms, and further containing 2 to 4 carbon atoms. Examples include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, butynyl, 2-butynyl, 3-butynyl, 1-methyl-2-propynyl, 4-pentynyl, 3-pentynyl, 1-methyl-2-butynyl, 2-hexynyl, 3-hexynyl, 2-heptynyl, 3-heptynyl, 4-heptynyl, 3-octynyl, 3-nonynyl, and 4-decynyl. The alkynyl group may be optionally substituted with a substituent.

[0265] "Alkynylene" refers to a straight or branched divalent unsaturated hydrocarbon group containing a carbon-carbon triple bond (C≡C), typically containing 2 to 18 carbon atoms, further containing 2 to 8 carbon atoms, further containing 2 to 6 carbon atoms, further containing 2-4 carbon atoms, non-limiting examples include ethynylene, propynylene, butynylene, and the alkynylene group may be optionally substituted with a substituent.

[0266] "Cycloalkyl" refers to a saturated or partially unsaturated, non-aromatic carbocyclic hydrocarbon group containing no ring heteroatoms. Cycloalkyl can be monocyclic, bicyclic or polycyclic. Bicyclic or polycyclic rings can be cyclic, spirocyclic, bridged or a combination thereof. Bicyclic or polycyclic rings can include one or more aromatic rings, but the ring system as a whole does not have aromaticity. The attachment site can be on the aromatic ring or on the non-aromatic ring. Usually, the cycloalkyl group contains 3 to 20 carbon atoms, further contains 3-8 carbon atoms, and further contains 3-6 carbon atoms; when it is a monocyclic cycloalkyl group, it contains 3-15 carbon atoms, or 3-10 carbon atoms, or 3-8 carbon atoms, or 3-6 carbon atoms; when it is a bicyclic or polycyclic cycloalkyl group, it contains 5-12 carbon atoms, or 5-11 carbon atoms, or 6-10 carbon atoms; non-limiting examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, butenyl, cyclopentenyl, cyclohexenyl, The cycloalkyl group may be optionally substituted with a substituent.

[0267] "Cycloalkylene" refers to a divalent radical of a cycloalkyl group.

[0268] "Aryl" refers to a carbocyclic ring having aromaticity and not containing heteroatoms, including monocyclic aryl and condensed ring aryl. It generally contains 6 to 14 carbon atoms, and further contains 6 to 10 carbon atoms. Non-limiting examples include phenyl, naphthyl, anthracenyl, and phenanthrenyl. Aryl can optionally be substituted with a substituent.

[0269] "Carbocycle" or "carbocyclyl" refers to a saturated, partially unsaturated, or aromatic carbocycle, including aryl and cycloalkyl. Carbocycles can be monocyclic, bicyclic, or polycyclic, including bridged, fused, and spirocyclic rings and combinations thereof. Carbocycles typically have 3 to 12 carbon atoms, or 3-10 carbon atoms, or 3-6 carbon atoms. In non-limiting examples, monocyclic carbocycles include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or phenyl, and bicyclic bridged rings include etc., double ring and ring include etc., bicyclic spiro ring includes The carbocyclic ring may be optionally substituted with a substituent.

[0270] "Heterocycloalkyl" refers to a saturated or partially unsaturated non-aromatic carbocyclic ring containing 1, 2, 3, or 4 heteroatoms selected from N, S, and O. Heterocycloalkyl can be monocyclic, bicyclic, or polycyclic. Bicyclic or polycyclic rings can be bridged, fused, spirocyclic, or a combination thereof. Bicyclic or polycyclic rings can include one or more aromatic or heteroaromatic rings, but the ring system as a whole is not aromatic. The attachment point can be on the aromatic ring or on the non-aromatic ring. Heterocycloalkyl groups are usually 3 to 20-membered rings. When they are monocyclic heterocycloalkyl groups, they are usually 3 to 15-membered rings, or 3-10-membered rings, or 3-8-membered rings, or 3-6-membered rings; when they are bicyclic or polycyclic heterocycloalkyl groups, they are usually 5-12-membered rings, or 5-11-membered rings, or 6-9-membered rings. The heteroatoms N and S include their oxidation states. Non-limiting examples of heterocycloalkyl groups include azetidinyl, morpholinyl, piperazinyl, piperidinyl, tetrahydropyranyl, oxetanyl, pyranyl, azolidinyl, azolinyl, oxolanyl, oxekinyl, and the like. Heterocycloalkyl groups may be optionally substituted with substituents.

[0271] "Heteroaromatic ring" or "heteroaryl" unless otherwise specified refers to a ring containing 1 to 4 heteroatoms selected from N, O or S and their oxidation states and having aromatic properties, which may be monocyclic, bicyclic or polycyclic, and the bicyclic or polycyclic rings may be bridged, parallel, spirocyclic or combinations thereof; when bicyclic or polycyclic, it may be a fusion of a heteroaryl and an aryl, or a fusion of a heteroaryl and a heteroaryl, wherein either the heteroaryl or the aryl may be a connection site. Non-limiting examples include furanyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, indolyl, purinyl, The heteroaryl group may be optionally substituted by a substituent.

[0272] "Heterocycle" or "heterocyclyl" refers to a saturated or unsaturated, aromatic or non-aromatic ring containing 1 to 4 heteroatoms selected from N, O, or S and their oxidation states, and includes heteroaryl and heterocycloalkyl. Heterocycles include monocyclic heterocycles, bicyclic bridged heterocycles, bicyclic heterocycles, and bicyclic spiro heterocycles, or combinations thereof. Heterocycles are typically 3- to 12-membered heterocycles, 5- to 12-membered heterocycles, or 5- to 7-membered heterocycles. The heterocyclic group may be attached to a heteroatom or a carbon atom, and non-limiting examples include oxirane, aziridine, oxetanyl, azetidinyl, 1,3-dioxolanyl, 1,4-dioxolanyl, 1,3-dioxanyl, piperazinyl, azepanyl, pyridinyl, furanyl, thienyl, pyranyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, pyrazolyl, pyridazinyl, imidazolyl, piperidinyl, piperidinyl, morpholinyl, thiomorpholinyl, 1,3-dithianyl , dihydrofuranyl, dihydropyranyl, dithiolanyl, tetrahydrofuranyl, tetrahydropyrrolyl, tetrahydroimidazolyl, oxazolyl, dihydrooxazolyl, tetrahydrooxazolyl, tetrahydrothiazolyl, tetrahydropyranyl, benzimidazolyl, benzopyridinyl, pyrrolopyridinyl, benzodihydrofuranyl, azabicyclo[3.2.1]octyl, azabicyclo[5.2.0]nonyl, oxatricyclo[5.3.1.1]dodecyl, azaadamantyl and oxaspiro[3.3]heptanyl, The heterocyclic ring may be optionally substituted with a substituent.

[0273] "Heterocyclylene" refers to a substituted or unsubstituted, saturated or unsaturated, aromatic or non-aromatic divalent heterocyclic group. Non-limiting examples include wait.

[0274] "Spiro" refers to a polycyclic group that shares a carbon atom (called a spiro atom) between the rings, which may contain 0 or more double bonds or triple bonds, and may contain 0 to 5 heteroatoms selected from N, O, S, P, Si and their oxidation states. Typically, the spiro ring is a 6- to 14-membered ring, or a 6- to 12-membered ring, or a 6- to 10-membered ring. Typically, the spiro ring is a trispirotri (representing a three-membered ring spirotricycle), a trispirotetra, a trispiropenta, a trispirohexa, a tetraspirotetra, a tetraspiropenta, a tetraspirohexa, a pentaspiropenta or a pentaspirohexa. Non-limiting examples of spiro rings include The spiro ring may be optionally substituted with a substituent.

[0275] "Parallel ring" or "condensed ring" refers to a polycyclic group in which the rings share two adjacent ring atoms and a chemical bond, and may contain one or more double bonds or triple bonds, and the rings may contain 0 to 5 heteroatoms selected from N, S, O, P, Si and their oxidation states. Usually, the cyclic ring is a 5- to 20-membered ring, or a 5- to 14-membered ring, or a 5- to 12-membered ring, or a 5- to 10-membered ring. Usually, the cyclic ring is a three- to four-membered ring (representing a cyclic ring formed by a three-membered ring and a four-membered ring. According to the IUPC naming rules, it is possible to have a three-membered ring as the basic ring or a four-membered ring as the basic ring. The same applies below), a three- to five-ring ring, a three- to six-ring ring, a four- to four-ring ring, a four- to five-ring ring, a four- to six-ring ring, a five- to five-ring ring, a five- to six-ring ring, and a six- to six-ring ring. Non-limiting examples of cyclic rings include purine, quinoline, isoquinoline, benzopyran, benzofuran, benzothiophene, The cyclic ring may be aromatic or non-aromatic and may be optionally substituted with a substituent.

[0276] "Bridged ring" means two rings that share two non-adjacent ring atoms and may contain one or more double or triple bonds. A bridged ring may contain 0 to 5 heteroatoms selected from N, S, O, P, Si and their oxidation states. Typically, the number of ring atoms in a bridged ring is 5 to 20, or 5 to 14, or 5 to 12, or 5 to 10. Non-limiting examples of bridged rings include adamantane,

[0277] "Substitution" or "substituent" unless otherwise specified refers to any substitution at a position permitted by chemical theory, and the number of substituents complies with the chemical bond rules. Exemplary substituents include but are not limited to: C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-8 Heteroalkyl, C 5-12 Aryl, 5-12 membered heteroaryl, hydroxyl, C 1-6 Alkoxy, C 5-12 Aryloxy, thiol, C 1-6 Alkylthio, cyano, halogen, C 1-6 Alkylthiocarbonyl, C 1-6 Alkylcarbamoyl, N-carbamoyl, nitro, silyl, sulfinyl, sulfonyl, sulfoxide, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy, amino, phosphonic acid, -CO2(C 1-6 alkyl), -OC(=O)(C 1-6 alkyl), -OCO2(C 1-6 alkyl), -C(=O)NH2, -C(=O)N(C 1-6 alkyl)2,-OC(=O)NH(C 1-6 alkyl), -NHC(=O)(C1-6 alkyl), -N(C 1-6 alkyl)C(=O)(C 1-6 Alkyl), -NHCO2(C 1-6 alkyl), -NHC(=O)N(C 1-6 alkyl)2,-HC(=O)NH(C 1-6 alkyl), -NHC(=O)NH2, -NHSO2(C 1-6 alkyl), -SO2N(C 1-6 Alkyl)2,-SO2NH(C 1-6 alkyl), -SO2NH2, -SO2C 1-6 Alkyl, etc.

[0278] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and the description includes instances where the event or circumstance occurs and instances where it does not. For example, "alkyl optionally substituted with F" means that the alkyl group may but need not be substituted with F, and the description includes instances where the alkyl group is substituted with F and instances where the alkyl group is not substituted with F.

[0279] "Pharmaceutically acceptable salts" refer to salts of the compounds of the present invention that retain the biological effectiveness and properties of the free acids or free bases, and the free acids are reacted with non-toxic inorganic or organic bases, or the free bases are reacted with non-toxic inorganic or organic acids.

[0280] A "pharmaceutical composition" refers to a mixture of one or more compounds described herein, or stereoisomers, deuterated forms, solvates, pharmaceutically acceptable salts, or cocrystals thereof, with other ingredients, wherein the other ingredients include physiologically / pharmaceutically acceptable carriers and / or excipients.

[0281] "Carrier" refers to a system that does not cause significant irritation to the organism and does not eliminate the biological activity and properties of the administered compound, and can change the way the drug enters the human body and its distribution in the body, control the release rate of the drug and deliver the drug to the target organ. Non-limiting examples include microcapsules and microspheres, nanoparticles, liposomes, etc.

[0282] "Excipient" refers to a substance that is not itself a therapeutic agent but serves as a diluent, excipient, binder, and / or vehicle that is added to a pharmaceutical composition to improve its handling or storage properties or to allow or facilitate the formation of a compound or pharmaceutical composition into a unit dosage form for administration. As known to those skilled in the art, pharmaceutical excipients can serve a variety of functions and can be described as wetting agents, buffers, suspending agents, lubricants, emulsifiers, disintegrants, absorbents, preservatives, surfactants, colorants, flavoring agents, and sweeteners. Examples of pharmaceutical excipients include, but are not limited to: (1) sugars such as lactose, glucose, and sucrose; (2) starches such as corn starch and potato starch; (3) cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, cellulose acetate, hydroxypropyl methylcellulose, hydroxypropyl cellulose, microcrystalline cellulose, and cross-linked carboxymethylcellulose (e.g., cross-linked sodium carboxymethylcellulose); (4) tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients such as cocoa butter and suppository waxes; (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn starch, and maltodextrin. oil and soybean oil; (10) glycols such as propylene glycol; (11) polyols such as glycerol, sorbitol, mannitol and polyethylene glycol; (12) esters such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) pH buffer solutions; (21) polyesters, polycarbonates and / or polyanhydrides; and (22) other non-toxic compatible substances used in pharmaceutical preparations.

[0283] "Stereoisomers" refer to isomers resulting from different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers, and conformational isomers.

[0284] The compounds of the present invention also include their tautomers. For example, when the present invention describes a compound on the left side in which the pyrimidine ring is substituted with OH, it also includes the tautomer compound on the right side.

[0285] "Solvate" refers to a substance formed by a compound of the present invention or a salt thereof and a stoichiometric or non-stoichiometric amount of a solvent bound to the compound or salt thereof by non-covalent forces between the molecules. When the solvent is water, the solvate is a hydrate.

[0286] A "cocrystal" is a crystal formed by the active pharmaceutical ingredient (API) and cocrystal former (CCF) bound together by hydrogen bonds or other non-covalent bonds. Both the API and CCF are solid in their pure form at room temperature, and the components exist in a fixed stoichiometric ratio. A cocrystal is a multi-component crystal, encompassing both binary cocrystals formed between two neutral solids and multi-component cocrystals formed between a neutral solid and a salt or solvate. BRIEF DESCRIPTION OF THE DRAWINGS

[0287] FIG1 shows the in vivo efficacy results of compound 30 (tumor volume (a in the figure) and mouse body weight (b in the figure)).

[0288] FIG2 shows the in vivo efficacy results of compound 30 (tumor weight). DETAILED DESCRIPTION

[0289] The technical solutions of the present invention are described in detail below with reference to the accompanying drawings and embodiments, but the protection scope of the present invention includes but is not limited to them.

[0290] Test Method

[0291] The structures of the compounds were determined by nuclear magnetic resonance (NMR) or / and mass spectrometry (MS). NMR shifts (δ) were expressed in 10 -6 The unit of (ppm) is given. NMR measurements were performed using a Bruker Avance III 400 and Bruker Avance 300 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD). The internal standard was tetramethylsilane (TMS).

[0292] MS was determined using (Agilent 6120B (ESI) and Agilent 6120B (APCI));

[0293] HPLC determination was performed using an Agilent 1260DAD high pressure liquid chromatograph (Zorbax SB-C 18 100 × 4.6 mm, 3.5 μM);

[0294] Thin layer chromatography silica gel plates used were Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates. The specifications of the silica gel plates used for thin layer chromatography (TLC) were 0.15 mm to 0.20 mm, and the specifications used for thin layer chromatography separation and purification products were 0.4 mm to 0.5 mm.

[0295] Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as the carrier.

[0296] Example 1

[0297] 2-(4-(4-(Aminomethyl)-1-oxo-1,2-dihydroisoquinolin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropyloxy-3-fluorobenzonitrile (Compound 1)

[0298] 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydroisoquinolin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropoxy-3-fluorobenzonitrile(compound 1)

[0299] Step 1: 1-Hydroxy-6-chloroisoquinoline (Compound 1A) (5.00 g, 27.84 mmol, CAS No.: 131002-09-0, Supplier: Shanghai Titan Technology Co., Ltd.) was added to a mixed solution of tetrahydrofuran (120 mL) and dichloromethane (80 mL). After stirring for 15 minutes, N-bromosuccinimide (5.45 g, 30.62 mmol) was slowly added at a temperature below 30°C. After the addition was completed, the mixture was reacted at room temperature for 8 hours. After the reaction was completed, the reaction solution was concentrated to dryness, dichloromethane (50 mL) was added, stirred for 1 hour, and then filtered to obtain Compound 1B (6.20 g, yield: 86.11%), which was directly used in the next reaction.

[0300] LC-MS (ESI): m / z = 260.0 [M+H] + .

[0301] Step 2: Compound 1B (5.00 g, 19.34 mmol), zinc cyanide (1.14 g, 9.67 mmol), and 1,1'-bis(diphenylphosphino)ferrocene (1.08 g, 1.93 mmol) were added to N,N-dimethylformamide (150 mL). After nitrogen purge three times, tris(dibenzylideneacetone)dipalladium (0.89 g, 0.97 mmol) was added. The atmosphere was again purge three times with nitrogen, and the temperature was raised to 100°C for 2 hours. After completion of the reaction, the reaction solution was concentrated to dryness, added with dichloromethane (50 mL), and concentrated again to dryness. Dichloromethane (30 mL) was added, stirred for 1 hour, and filtered to obtain compound 1C (3.3 g, yield: 83.39%).

[0302] LC-MS (ESI): m / z = 203.1 [MH] - .

[0303] Step 3: Compound 1C (1.00 g, 4.84 mmol) was placed in a methanol solution (100 mL), and cobalt chloride (0.63 g, 4.84 mmol) was added. After stirring for 15 minutes, sodium borohydride (0.46 g, 12.10 mmol) was slowly added and stirred at room temperature for 2 hours. Di-tert-butyl dicarbonate (1.27 g, 5.81 mmol) was added and the mixture was allowed to react at room temperature overnight. After completion of the reaction, the mixture was filtered, the filtrate was washed with methanol, and the filtrate was concentrated to dryness. The residue was separated by silica gel column chromatography (dichloromethane:methanol (v:v) = 100%-90%) to obtain compound 1D (600 mg, yield: 39.89%).

[0304] LC-MS (ESI): m / z = 309.1 [M+H] + .

[0305] Step 4: Compound 1D (700 mg, 2.27 mmol), pinacol diboronate (1.15 g, 4.54 mmol), and potassium acetate (1.11 g, 11.35 mmol) were added sequentially to 1,4-dioxane (150 mL). The atmosphere was purged with nitrogen three times. Chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (0.18 g, 0.23 mmol) was added, and the atmosphere was purged with nitrogen again three times. The temperature was raised to 105°C and the reaction mixture was reacted for 1 hour. After cooling, the reaction mixture was concentrated to dryness, and the residue was separated by silica gel column chromatography (dichloromethane:methanol (v:v) = 100%-90%) to obtain compound 1E (800 mg, yield: 88.04%).

[0306] LC-MS (ESI): m / z = 309.1 [M+H] + .

[0307] Step 5: Compound 1F (100 mg, 0.27 mmol) (synthesis method reference patent: WO2021050915A1), compound 1E (110 mg, 0.27 mmol), and sodium bicarbonate (45 mg, 0.54 mmol) were added sequentially to a solution of 1,4-dioxane (9 mL) and water (3 mL). After nitrogen displacement three times, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (22 mg, 0.027 mmol) was added. The nitrogen atmosphere was again replaced three times, and the temperature was raised to 60°C and reacted for 1 hour. After completion of the reaction, the reaction solution was concentrated to dryness, and the residue was separated by silica gel column chromatography (dichloromethane:methanol (v:v) = 100%-90%) to obtain compound 1G (90 mg, yield: 59.10%).

[0308] LC-MS (ESI): m / z = 564.2 [M+H] +.

[0309] Step 6: Compound 1G (90 mg, 0.16 mmol) was added to a 4 mol / L methanolic hydrochloric acid solution (10 mL). After stirring at room temperature for 2 hours, the reaction solution was concentrated to dryness. The residue was separated by reverse-phase column chromatography (Biotage Isolera One, Agela C18 reverse-phase column, mobile phase: acetonitrile:water (v:v) = 98%-30%) to obtain compound 1 (6 mg, yield: 8.08%).

[0310] 1 H NMR(400MHz,DMSO-d6)δ11.06(s,1H),8.18(s,1H),8.13(d,1H),7.99(d,1H),7.45–7.3 8(m,2H),7.04(s,1H),4.21-4.18(m,1H),3.77(s,3H),3.52(s,2H),0.96–0.76(m,4H);

[0311] LC-MS (ESI): m / z = 462.1 [MH] - .

[0312] Example 2

[0313] 4-Chloro-6-cyclopropyloxy-3-fluoro-2-(4-(4-(hydroxymethyl)-7-oxo-6,7-dihydrothieno[2,3-d]pyridazin-2-yl)-1-methyl-1H-pyrazol-5-yl)benzonitrile (Compound 2)

[0314] 4-chloro-6-cyclopropoxy-3-fluoro-2-(4-(4-(hydroxymethyl)-7-oxo-6,7-dihydrothieno[2,3-d]pyridazin-2-yl)-1-methyl-1H-pyrazol-5-yl)benzonitrile(compound 2)

[0315] Step 1: Dissolve compound 1F (2.0 g, 4.8 mmol), pinacol borane (896.0 mg, 7.0 mmol), tetrakis(triphenylphosphine)palladium (554.0 mg, 0.48 mmol), and triethylamine (960.0 mg, 9.6 mmol) in 1,4-dioxane (20.0 mL). The atmosphere was purged with nitrogen three times and heated to 90°C overnight. The mixture was cooled to room temperature and passed through a silica gel column, eluting with ethyl acetate (50.0 mL). The filtrate was collected and concentrated, and the resulting residue was purified by column chromatography (petroleum ether:ethyl acetate (v:v) = 10:1) to provide compound 2B (1.5 g, yield: 75%).

[0316] LC-MS (ESI): m / z = 418.1 [M+H] + .

[0317] Step 2: Methyl 3-bromothiophene-2-carboxylate (Compound 2C) (10.1 g, 45.7 mmol) was dissolved in 1,4-dioxane (150 mL) and water (10 mL). [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (1.7 g, 2.3 mmol), cesium carbonate (29.3 g, 90.0 mmol), and potassium vinyl trifluoroborate (6.7 g, 50.0 mmol) were added. The atmosphere was purged with nitrogen three times and heated to 90°C for 2 h. The mixture was cooled to room temperature and passed through a short silica gel column, eluting with ethyl acetate. The filtrate was collected and concentrated. The resulting residue was purified by column chromatography (petroleum ether:ethyl acetate (v:v) = 20:1) to provide Compound 2D (6.1 g, yield: 80%).

[0318] LC-MS (ESI): m / z = 168.1 [M+H] + .

[0319] Step 3: Compound 2D (6.1 g, 36.5 mmol) was dissolved in tetrahydrofuran (250 mL). Lithium bis(trimethylsilyl)amide (40 mL, 1.0 M solution in tetrahydrofuran, 40.0 mmol) was slowly added under ice-cooling. The reaction was continued for 0.5 h, followed by the addition of iodine (10.1 g, 40.0 mmol). The temperature was slowly warmed to room temperature and allowed to react overnight. The reaction was quenched by the addition of aqueous sodium thiosulfate (100 mL). The mixture was extracted with ethyl acetate (100 mL x 3). The organic phase was dried over anhydrous sodium sulfate and concentrated to obtain a residue which was purified by column chromatography (petroleum ether:ethyl acetate (v:v) = 15:1) to afford compound 2E (8.5 g, yield: 81%).

[0320] LC-MS (ESI): m / z = 294.9 [M+H] + .

[0321] Step 4: Compound 2E (8.5 g, 29.5 mmol) was dissolved in acetone (50 mL), followed by potassium osmate dihydrate (0.9 g, 2.9 mmol) and 4-methylmorpholine-N-oxide (7.0 g, 60.0 mmol) dissolved in water (10 mL), and then added to the mixture. The mixture was allowed to react at room temperature for 2 h. The reaction was quenched by addition of saturated aqueous sodium sulfite solution, concentrated to remove the acetone, and extracted with ethyl acetate (100 mL x 3). The organic phase was dried over anhydrous sodium sulfate, and the resulting residue was purified by column chromatography (dichloromethane:methanol (v:v) = 10:1) to afford compound 2F (7.2 g, yield: 74%).

[0322] LC-MS (ESI): m / z = 328.9 [M+H] + .

[0323] Step 5: Dissolve compound 2F (7.2 g, 21.9 mmol) in N,N-dimethylformamide (100 mL), add imidazole (3.0 g, 44.0 mmol), and then add tert-butyldimethylsilyl chloride (3.3 g, 22.0 mmol). The mixture is allowed to react at room temperature for 2 h. The reaction is quenched by adding water (50 mL) and extracted with ethyl acetate (100 mL x 3). The organic phase is dried over anhydrous sodium sulfate and concentrated to obtain compound 2G (7.4 g crude product), which is used directly in the next reaction.

[0324] LC-MS (ESI): m / z = 443.0 [M+H] + .

[0325] Step 6: Dissolve compound 2G (7.4 g, 16.7 mmol) in ethyl acetate (100 mL), add 2-iodoacylbenzoic acid (9.8 g, 35.0 mmol), and heat to 80°C overnight. Cool to room temperature, filter to remove solids, and concentrate the filtrate to obtain compound 2H (6.4 g crude product), which is used directly in the next reaction.

[0326] LC-MS (ESI): m / z = 441.0 [M+H] + .

[0327] Step 7: Dissolve compound 2H (6.4 g, 14.5 mmol) in ethanol (20 mL) and slowly add hydrazine hydrate (6.0 mL). A solid appears after approximately 5 minutes. Stirring is continued for 10 minutes. Filter, rinse the filter cake with ethanol (5.0 mL), and collect and concentrate the filtrate to obtain compound 2I (4.5 g crude product), which is used directly in the next reaction.

[0328] LC-MS (ESI): m / z = 423.0 [M+H] + .

[0329] Step 8: Compound 2I (590.0 mg, 1.4 mmol), compound 2B (600.0 mg, 1.4 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (91.0 mg, 0.14 mmol), and sodium bicarbonate (403.2 mg, 4.8 mmol) were dissolved in 1,4-dioxane (10.0 mL) and water (3.0 mL). The atmosphere was purged with nitrogen three times and heated to 80°C for 2 h. After cooling to room temperature, the reaction mixture was filtered through a silica gel short column. The filter cake was rinsed with ethyl acetate (50.0 mL). The filtrate was collected and concentrated. The resulting residue was purified by column chromatography (petroleum ether:ethyl acetate (v:v) = 1:1) to provide compound 2J (690.0 mg, yield: 84%).

[0330] LC-MS (ESI): m / z = 586.1 [M+H] + .

[0331] Step 9: Dissolve compound 2J (690.0 mg, 1.1 mmol) in tetrahydrofuran (20 mL), then slowly add tetrabutylammonium fluoride (1.0 M in tetrahydrofuran, 1.5 mL). React at room temperature for 1 h. The resulting residue was directly concentrated and purified by column chromatography (dichloromethane:methanol (v:v) = 10:1) to afford compound 2 (480.0 mg, 99% yield).

[0332] 1 H NMR(400MHz,DMSO-d6)δ12.70(s,1H),8.28(s,1H),8.09(d,1H),7.69(s,1H),5.44(s,1 H),4.57(d,2H),4.38–4.09(m,1H),3.78(s,3H),0.95–0.92(m,2H),0.88–0.74(m,2H);

[0333] LC-MS (ESI): m / z = 472.0 [M+H] + .

[0334] Example 3

[0335] 2-(4-(4-(Aminomethyl)-7-oxo-6,7-dihydrothieno[2,3-d]pyridazin-2-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropyloxy-3-fluorobenzonitrile (Compound 3)

[0336] 2-(4-(4-(aminomethyl)-7-oxo-6,7-dihydrothieno[2,3-d]pyridazin-2-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropoxy-3-fluorobenzonitrile(compound 3)

[0337] Step 1: Dissolve compound 2 (420.0 mg, 0.9 mmol) in ethyl acetate (10 mL), add 2-iodoacylbenzoic acid (504.0 mg, 1.8 mmol), and heat under reflux overnight. Cool to room temperature, filter, rinse the filter cake with ethyl acetate, collect the filtrate, and concentrate. The resulting residue is purified by column chromatography (dichloromethane:methanol (v:v) = 10:1) to provide compound 3A (400.0 mg, yield: 95%).

[0338] LC-MS (ESI): m / z = 470.0 [M+H] + .

[0339] Step 2: Compound 3A (400.0 mg, 0.8 mmol) was dissolved in methanol (10 mL), and ammonium acetate (246.0 mg, 3.2 mmol) was added. The mixture was heated under reflux for 1 h, and then sodium cyanoborohydride (201.0 mg, 3.2 mmol) was added and reacted for 4 h. The mixture was cooled to room temperature and extracted with ethyl acetate (20 mL x 3). The organic phase was dried over anhydrous sodium sulfate and concentrated. The resulting crude product was purified by column chromatography (dichloromethane:methanol (v:v) = 10:1) to afford compound 3 (70.0 mg, yield: 17%).

[0340] 1 H NMR(400MHz,DMSO-d6)δ12.66(s,1H),8.30(s,1H),8.09(d,1H),7.85(s,1H),4. 42–4.15(m,3H),3.87(s,2H),3.77(s,3H),0.95–0.93(m,2H),0.90–0.73(m,2H);

[0341] LC-MS (ESI): m / z = 471.0 [M+H] + .

[0342] Example 4

[0343] 2-(4-(5-(aminomethyl)-8-oxo-7,8-dihydropyrido[2,3-d]pyridazin-3-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropyloxy-3-fluorobenzonitrile (Compound 4)

[0344] 2-(4-(5-(aminomethyl)-8-oxo-7,8-dihydropyrido[2,3-d]pyridazin-3-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropoxy-3-fluorobenzonitrile(compound 4)

[0345] Step 1: Compound 4A (15 g, 59.89 mmol), isopropenylboronic acid pinacol ester (10.57 g, 62.88 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (2.19 g, 2.99 mmol), and potassium carbonate (20.69 g, 149.72 mmol) were dissolved in 1,4-dioxane (125 mL) and water (25 mL). The atmosphere was then replaced with nitrogen three times and the reaction was carried out at 80°C for 2 h. The mixture was quenched by the addition of water (150 mL) and ethyl acetate (100 mL), filtered, and the filtrate was extracted with ethyl acetate (150 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting residue was purified by column chromatography (petroleum ether:ethyl acetate (v:v) = 5:1) to afford compound 4B (12.0 g, yield: 94.67%).

[0346] LCMS (ESI): m / z = 212.1 [M+H] + .

[0347] Step 2: Compound 4B (12 g, 56.70 mmol) was dissolved in tetrahydrofuran (120 mL) and water (120 mL), and potassium osmate dihydrate (420 mg, 1.13 mmol) and sodium periodate (42.45 g, 198.45 mmol) were added. The mixture was allowed to react overnight at room temperature. The mixture was filtered, and the filtrate was quenched with sodium thiosulfate solution (200 mL). The mixture was then extracted with ethyl acetate (200 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography (petroleum ether:ethyl acetate (v:v) = 2:1) to afford compound 4C (8.8 g, yield: 72.65%).

[0348] LCMS (ESI): m / z = 214.1 [M+H] + .

[0349] Step 3: Compound 4C (8.8 g, 41.19 mmol) was dissolved in ethanol (130 mL), and hydrazine hydrate (3.75 mL, 61.73 mmol) was added and refluxed at 80°C for 2 hours. The mixture was cooled to room temperature and filtered. The filter cake was rinsed once with ethanol, collected, and dried to obtain compound 4D (6.5 g, yield: 80.67%).

[0350] LCMS (ESI): m / z = 196.0 [M+H] + .

[0351] Step 4: Compound 4D (2.0 g, 10.22 mmol) was dissolved in acetonitrile (25 mL), and phosphorus oxychloride (7.84 g, 51.06 mmol) was added. The mixture was reacted at 80°C for 1.5 hours. After cooling, the mixture was concentrated under reduced pressure to remove excess phosphorus oxychloride and acetonitrile. The residue was dissolved in dichloromethane (200 mL) and washed with saturated sodium bicarbonate (200 mL). The organic phase was separated and dried over anhydrous sodium sulfate, filtered, and concentrated to afford compound 4E (2.0 g crude product), which was used directly in the next reaction.

[0352] Step 5: Compound 4E (2.0 g, 9.34 mmol) was dissolved in acetonitrile (25 mL), and azobisisobutyronitrile (0.31 g, 1.89 mmol) and N-bromosuccinimide (2.49 g, 14.04 mmol) were added. The reaction was carried out at 80°C for 16 hours. After cooling, the mixture was concentrated under reduced pressure to remove the acetonitrile. The residue was dissolved in dichloromethane (20 mL) and purified by column chromatography to obtain compound 4F (0.73 g, yield: 26.68%).

[0353] LCMS (ESI): m / z = 291.9 [M+H] + .

[0354] Step 6: Compound 4F (0.73 g, 2.49 mmol) was dissolved in N,N-dimethylformamide (6 mL). Potassium phthalimide (0.48 g, 2.61 mmol) was added portionwise at room temperature and allowed to react for 1 hour. After completion of the reaction, water (60 mL) was added, followed by extraction with ethyl acetate (50 mL x 3). The organic phase was washed with water (40 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated to afford compound 4G (0.80 g crude product), which was used directly in the next reaction.

[0355] LCMS (ESI): m / z = 359.0 [M+H] + .

[0356] Step 7: Compound 4G (0.8 g, 2.23 mmol) was dissolved in acetic acid (8 mL), and potassium acetate hydrate (1.3 g, 11.19 mmol) was added. The mixture was reacted at 100°C for 1 hour. After cooling, the acetic acid was removed by concentration under reduced pressure. Water (50 mL) was added to the residue, which was then extracted with ethyl acetate (50 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to afford compound 4H (0.80 g crude product), which was used directly in the next reaction.

[0357] LCMS (ESI): m / z = 341.0 [M+H] + .

[0358] Step 8: Compound 4H (0.8 g) was dissolved in ethanol (15 mL), and hydrazine hydrate (0.18 g, 3.6 mmol) was added and reacted at 80°C for 2 hours. After cooling, the mixture was concentrated under reduced pressure to obtain compound 4I (0.6 g crude product), which was used directly in the next reaction.

[0359] LCMS (ESI): m / z = 211.1 [M+H] + .

[0360] Step 9: Dissolve compound 4I (0.8 g, 2.85 mmol) in dichloromethane (12 mL), then add triethylamine (0.87 g, 8.61 mmol) and di-tert-butyl dicarbonate (1.24 g, 5.7 mmol). Incubate at room temperature for 2 hours. Filter the reaction mixture and collect the filter cake to obtain compound 4J (0.55 g crude product), which is used directly in the next reaction.

[0361] LCMS (ESI): m / z = 311.1 [M+H] + .

[0362] Step 10: Compound 4J (0.25 g, 0.80 mmol), compound 2B (0.37 g, 0.89 mmol), potassium acetate (0.39 g, 3.98 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (0.065 g, 0.079 mmol) were dissolved in acetonitrile (3 mL) and water (1 mL). The atmosphere was then replaced with nitrogen three times and the reaction was carried out at 130°C for 1 h. The mixture was directly concentrated, and dichloromethane (10 mL) was added. The filtrate was filtered and concentrated. The resulting residue was purified by column chromatography (dichloromethane:methanol (v:v) = 25:1) to afford compound 4K (0.15 g, yield: 33.13%).

[0363] LCMS (ESI): m / z = 566.1 [M+H] + .

[0364] Step 11: Add compound 4K (0.15 g, 0.27 mmol) and dichloromethane (4 mL) to a 25 mL single-necked flask. Add trifluoroacetic acid (1 mL) dropwise and stir at room temperature for 1.5 hours. Concentrate the mixture at 35°C, add dichloromethane (10 mL) and triethylamine (0.3 g), stir for five minutes, and concentrate the reaction mixture. The resulting crude product is purified by preparative HPLC. Preparation method: 1. Apparatus: Waters 2767 Preparative Liquid Chromatography; Column: SunFire@PrepC18 (19 mm × 250 mm). 2. Dissolve the sample in methanol and filter through a 0.45 μm filter to prepare a sample solution. 3. Preparative Chromatography Conditions: a. Mobile Phase A, B Composition: Mobile Phase A: Acetonitrile; Mobile Phase B: Water (containing 0.1% NH3·H2O); b. Gradient elution, Mobile Phase A content from 20% to 70%; c. Flow rate: 20 mL / min; d. Elution time: 15 min. Retention time: 3.10 min. Compound 4 (0.032 g, yield: 25.44%) was obtained.

[0365] 1 H NMR(400MHz,DMSO-d6)δ12.66(s,1H),8.90(d,1H),8.39(s,1H),8.22(d,1H),8.00 (d,1H),4.24–4.15(m,1H),3.84(d,2H),3.81(s,3H),0.90(dd,2H),0.81(dd,2H);

[0366] LC-MS (ESI): m / z = 466.1 [M+H] + .

[0367] Example 5

[0368] 2-(4-(2-(Aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropyloxy-3-fluorobenzonitrile (Compound 5)

[0369] 2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropoxy-3-fluorobenzonitrile(compound 5)

[0370] Step 1: Compound 5A (4 g, 16.7 mmol, CAS number: 403850-89-5, supplier: Shanghai Bid Pharmaceutical Technology Co., Ltd.) and 1-bromopyrrolidine-2,5-dione (2.96 g, 16.7 mmol) were dissolved in N,N-dimethylformamide (60 mL) and stirred at 40°C for 12 hours. After completion of the reaction, water (20 mL) was added and the mixture was extracted with ethyl acetate (20 mL × 3). The organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. The resulting residue was concentrated and purified by column chromatography (dichloromethane:ethyl acetate (v:v) = 17:100) to obtain compound 5B (1.9 g, yield: 36%).

[0371] 1 H NMR (400MHz, DMSO-d6) δ12.72(s,1H),8.03(d,1H),7.91-7.88(m,1H),7.71-7.69(m,1H),4.54(s,2H).

[0372] Step 2: Dissolve compound 5B (0.75 g, 2.36 mmol), (2,4-dimethoxyphenyl)methanamine (0.51 g, 3.05 mmol), and potassium carbonate (0.65 g, 4.72 mmol) in N,N-dimethylformamide (20 mL). Stir and react at room temperature for 1 hour. Add water (10 mL) and extract with ethyl acetate (20 mL x 3). The organic phase is washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a residue that is purified by preparative HPLC. Purification method: 1. Instrument: Waters 2767 Preparative HPLC; Column: SunFire@PrepC18 (19 mm x 250 mm). 2. Dissolve the sample in methanol and filter through a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Mobile phase A, B composition: Mobile phase A: acetonitrile; Mobile phase B: water; b. Gradient elution, mobile phase A content ranging from 10% to 40%; c. Flow rate 80 mL / min; d. Elution time 20 min. Retention time 10.50 min. Compound 5C (0.27 g, yield: 28%) was obtained.

[0373] LC-MS (ESI): m / z = 404.2 [M+H] + .

[0374] Step 3: Compound 5C (0.2 g, 0.49 mmol), compound 2B (0.37 g, 0.89 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (0.036 g, 0.049 mmol), and potassium acetate (0.096 g, 0.98 mmol) were dissolved in a mixed solvent of acetonitrile (9 mL) and water (3 mL). The atmosphere was replaced with nitrogen three times and the mixture was heated to 130°C and stirred for 1 hour in a microwave oven. Water (10 mL) was added and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. The resulting residue was concentrated and purified by column chromatography (dichloromethane:methanol (v:v) = 100:8) to afford compound 5D (0.13 g, yield: 43%).

[0375] LC-MS (ESI): m / z = 615.1 [M+H] + .

[0376] Step 4: Compound 5D (0.1 g, 0.16 mmol) was dissolved in a mixture of trifluoroacetic acid (4 mL) and concentrated hydrochloric acid (1 mL) and reacted at 50°C for 3 hours. After completion of the reaction, the resulting residue was directly concentrated and purified by preparative HPLC. Preparation method: 1. Apparatus: Waters 2767 preparative liquid chromatography; Chromatographic column: SunFire@PrepC18 (19 mm × 250 mm). 2. The sample was dissolved in methanol and filtered through a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Mobile phase A and B composition: Mobile phase A: acetonitrile; Mobile phase B: Water (containing 0.1% NH3 / H2O); b. Gradient elution, mobile phase A content ranging from 5% to 50%; c. Flow rate: 20 mL / min; d. Elution time: 20 min. Retention time: 15 min. Compound 5 (0.0015 g, yield: 2%) was obtained.

[0377] 1 H NMR(400MHz,DMSO-d6)δ12.31(s,1H),8.13(s,1H),8.05–7.97(m,2H),7.43(d,1H),7.27-7.2 4(m,1H),4.20-4.16(m,1H),4.04(s,2H),3.76(s,3H),0.95-0.90(m,2H),0.84–0.76(m,2H);

[0378] LC-MS (ESI): m / z = 465.1 [M+H] + .

[0379] Example 6

[0380] 7-(5-(3-chloro-6-cyano-5-cyclopropyloxy-2-fluorophenyl)-1-methyl-1H-pyrazol-4-yl)-4-oxo-3,4-dihydrophthalazine-1-carboximidamide (Compound 6)

[0381] 7-(5-(3-chloro-6-cyano-5-cyclopropoxy-2-fluorophenyl)-1-methyl-1H-pyrazol-4-yl)-4-oxo-3,4-dihydrophthalazine-1-carboximidamide(compound 6)

[0382] Step 1: Add compound 6A (1.3 g, 4.59 mmol, synthesis reference: US202178994) to a sealed tube, add ammonia solution (20 mL, 7 M in methanol), and heat to 100°C for 16 hours. The reaction solution was cooled and filtered to obtain compound 6B (1.1 g crude product), which was used directly in the next reaction.

[0383] LC-MS (ESI): m / z = 265.9 [MH] - .

[0384] Step 2: Compound 6B (1.1 g crude product, 4.10 mmol) and dioxane (20 mL) were added to a flask, followed by phosphorus oxychloride (1.87 mL, 20.46 mmol) and allowed to react at 120°C for 6 hours. The reaction solution was concentrated and diluted with dichloromethane. 30 mL of water and saturated sodium carbonate solution were added to adjust the pH to approximately 8. The solution was extracted with dichloromethane, and the organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. The resulting residue was concentrated and separated by column chromatography (petroleum ether:ethyl acetate (v:v) = 3:1) to afford compound 6C (690 mg, yield: 67.3%).

[0385] LC-MS (ESI): m / z = 248.0 [MH] - .

[0386] Step 3: Ammonium chloride (612 mg, 11.45 mmol) and xylene (10 mL) were added to a flask. Trimethylaluminum (5.4 mL, 10.80 mmol, 2 M in hexane) was added dropwise under an ice bath and allowed to react at room temperature for 1 hour. Compound 6C (540 mg, 2.16 mmol) was then added and the temperature was raised to 120°C for 24 hours. Diethylene glycol dimethyl ether (10 mL) was then added and the temperature was raised to 160°C for 8 hours. After cooling, methanol (5 mL) was added and stirred for 1 hour. The mixture was filtered and the filter cake was washed with ethyl acetate / methanol (10 mL / 10 mL). The filtrate was collected and dried over anhydrous sodium sulfate and concentrated to give compound 6D (3 g crude product), which was used directly in the next reaction.

[0387] LC-MS (ESI): m / z = 266.9 [M+H] + .

[0388] Step 4: Compound 6D (3 g crude product, 11.23 mmol), methanol (30 mL), and water (10 mL) were added to a flask. Potassium carbonate (4.66 g, 33.69 mmol) and di-tert-butyl dicarbonate (4.9 g, 22.46 mmol) were added and reacted at room temperature for 4 hours. Ethyl acetate (50 mL) and saturated sodium chloride solution (50 mL) were added to the system, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. The resulting residue was concentrated and purified by column chromatography (petroleum ether:ethyl acetate (v:v) = 1:1) to obtain compound 6E (380 mg, two-step yield: 47.91%).

[0389] LCMS (ESI): m / z = 367.0 [M+H] + .

[0390] Step 5: Compound 6E (180 mg, 0.49 mmol), compound 2B (250 mg, 0.60 mmol), and sodium bicarbonate (164 mg, 1.96 mmol) were dissolved in 1,4-dioxane (9 mL) and water (3 mL) under nitrogen protection. [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (18 mg, 0.025 mmol) was added and reacted at 80°C under nitrogen protection for 2 hours. After cooling, the product was directly concentrated, and the resulting residue was separated by column chromatography (petroleum ether:ethyl acetate (v:v) = 1:4) to provide compound 6F (80 mg, yield: 28.25%).

[0391] LCMS (ESI): m / z = 578.2 [M+H] + .

[0392] Step 6: Compound 6F (80 mg, 0.14 mmol) and dichloromethane (6 mL) were added to a flask, followed by trifluoroacetic acid (1 mL). The mixture was reacted at 35°C for 2 hours. The resulting residue was directly concentrated and purified by preparative HPLC. Preparation method: 1. Apparatus: Waters 2767 preparative liquid chromatography; Chromatographic column: SunFire@PrepC18 (19 mm × 250 mm). 2. The sample was dissolved in DMF and filtered through a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Mobile phase A and B composition: Mobile phase A: acetonitrile; Mobile phase B: Water (containing 0.1% NH3 / H2O); b. Gradient elution, mobile phase A content ranging from 10% to 40%; c. Flow rate: 20 mL / min; d. Elution time: 10 min. Retention time: 3.28 min. Compound 6 (23 mg, yield: 34%) was obtained.

[0393] 1 H NMR(400MHz,CD3OD)δ8.33(d,1H),8.10(s,1H),7.89(d,1H),7.85(s,1H),7.80 (t,1H),4.09-4.07(m,1H),3.81(s,3H),0.94-0.91(m,2H),0.87-0.84(m,2H);

[0394] LC-MS (ESI): m / z = 478.4 [M+H] + .

[0395] Example 7

[0396] 2-(4-(4-(aminomethyl)-8-(1-methyl-1H-pyrazol-4-yl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropyloxy-3-fluorobenzonitrile (Compound 7)

[0397] 2-(4-(4-(aminomethyl)-8-(1-methyl-1H-pyrazol-4-yl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropoxy-3-fluorobenzonitrile(compound 7)

[0398] Step 1: Dissolve compound 7A (100 g, 410 mmol) in acetonitrile (1.1 L), then add triethylamine (110 mL, 820 mmol), followed by di-tert-butyl dicarbonate (119 g, 533 mmol) and 4-dimethylaminopyridine (5.0 g, 41 mmol). Heat to 50°C and stir for 2 hours. After cooling, concentrate directly, add water (1 L), and extract with ethyl acetate (1 L x 3). The organic phases are combined, dried, and concentrated to give compound 7B (123 g, yield: 87.23%).

[0399] 1 H NMR (400MHz, DMSO-d6) δ8.88(s,1H),7.74(t,1H),7.63(d,1H),3.84(s,3H),2.27(s,3H),1.48(s,9H).

[0400] Step 2: Compound 7B (123 g, 358 mmol) and lithium hydroxide monohydrate (45 g, 1.07 mol) were dissolved in tetrahydrofuran (800 mL) and water (250 mL) and stirred at room temperature for 18 hours. The mixture was concentrated directly, and the resulting residue was diluted with water (300 mL). The pH was adjusted to 2-3 with 2 mol / L dilute hydrochloric acid, and the aqueous phase was extracted with ethyl acetate (1 L x 3). The organic phase was collected, dried, and concentrated to obtain compound 7C (102 g, yield: 86.45%).

[0401] 1 H NMR (400MHz, DMSO-d6) δ8.82(s,1H),7.65(d,1H),7.57(d,1H),2.27(s,3H),1.47(s,9H).

[0402] Step 3: Compound 7C (102 g, 308.9 mmol) was dissolved in N,N-dimethylformamide (1.0 L). N,N-diisopropylethylamine (102 mL, 620 mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) (153 g, 403 mmol) were added at room temperature. The mixture was stirred at room temperature for 30 min. Finally, dimethylhydroxylamine hydrochloride (45.6 g, 465 mmol) was added and the mixture was reacted at room temperature for 2 h. The reaction solution was poured into 2 L of water and extracted three times with ethyl acetate (1 L x 3). The organic phase was collected, dried, and concentrated. The resulting residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (v:v) = 3:1) to obtain compound 7D (82 g, yield: 71.12%).

[0403] 1H NMR (400MHz, DMSO-d6) δ8.78(s,1H),7.63(s,1H),7.24(d,1H),3.43(s,3H),3.26(s,3H),2.04(s,3H),1.47(s,9H).

[0404] Step 4: Compound 7D (82 g, 220 mmol) was dissolved in tetrahydrofuran (800 mL). Methylmagnesium bromide (220 mL, 660 mmol, 3 M in diethyl ether) was added at 0°C and the mixture was slowly warmed to room temperature for 16 hours. Saturated ammonium chloride solution (1 L) was added to the reaction system, stirred for 10 minutes, and then extracted with ethyl acetate (500 mL x 3). The organic phase was collected, dried, and concentrated. The resulting residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (v:v) = 3:1) to obtain compound 7E (70 g, yield: 97.08%).

[0405] 1 H NMR (400MHz, DMSO-d6) δ8.84(s,1H),7.67(d,1H),7.62(d,1H),2.53(s,3H),2.13(s,3H),1.47(s,9H).

[0406] Step 5: Compound 7E (53 g, 160 mmol) was dissolved in tert-butyl alcohol (500 mL) and water (500 mL), anhydrous potassium carbonate (44 g, 320 mmol) was added, and the mixture was heated to 70°C with stirring. Finally, potassium permanganate (179 g, 1.13 mol) was added to the reaction system in six batches at 30-min intervals. After the addition was complete, the reaction was continued at 70°C for 1 hour. The reaction solution was cooled to room temperature, filtered through celite, and the filtrate was collected. The pH was then adjusted to 2-3 with 2 mol / L dilute hydrochloric acid, and then extracted with a mixed solvent of ethyl acetate and tetrahydrofuran (v:v = 10:1) (700 mL x 3). The organic phase was collected, dried, and concentrated to obtain compound 7F (39 g crude product), which was used directly in the next reaction.

[0407] LC-MS (ESI): m / z = 388.1 [M+H] + .

[0408] Step 6: Compound 7F (39 g, 100 mmol) was dissolved in ethanol (200 mL), followed by the addition of hydrazine hydrate (7.24 mL, 120 mmol). The mixture was heated to 75°C for 15 hours. The mixture was cooled to room temperature, filtered, and the filter cake was washed with ethanol (100 mL). The filter cake was collected and dried to afford compound 7G (17 g, yield: 43.81%).

[0409] 1H NMR (400MHz, DMSO-d6) δ12.00(s,1H),8.63(s,1H),8.05(s,1H),1.50(s,9H).

[0410] Step 7: Compound 7G (17 g, 44.2 mmol) was dissolved in methanol (150 mL), and methanolic hydrochloric acid (4 mol / L, 240 mL) was added. The mixture was heated to 70°C for 2 hours. After cooling, the mixture was concentrated directly. Water (200 mL) was added to the residue, and the pH was adjusted to 8 with sodium bicarbonate. The residue was filtered, and the filter cake was washed with water (100 mL). The filter cake was collected and dried to obtain compound 7H (13 g, yield: 98.56%).

[0411] 1 H NMR (400MHz, DMSO-d6) δ12.87(s,1H),7.58(s,2H),7.13(s,1H),3.87(s,3H);

[0412] LC-MS (ESI): m / z = 298.0 [M+H] + .

[0413] Step 8: Compound 7H (13 g, 43.6 mmol) and p-toluenesulfonic acid (33 g, 174.4 mmol) were dissolved in acetonitrile (2.0 L) and replaced with nitrogen twice. Sodium nitrite (7.5 g, 109 mmol) was then dissolved in water (20 mL) and added dropwise to the above system. The reaction was maintained at this temperature for 1 hour. Finally, potassium iodide (21.7 g, 130.8 mmol) was dissolved in water (20 mL) and added dropwise to the above reaction solution. After completion, the mixture was warmed to room temperature and reacted for 2 hours. Saturated sodium thiosulfate solution (200 mL) was added to the system and stirred for 10 minutes. The organic phase was then concentrated to remove the organic phase and filtered. The filter cake was washed with clean water (300 mL). The filter cake was collected and dried to obtain compound 7I (8.2 g, yield: 45.98%).

[0414] 1 H NMR (400MHz, DMSO-d6) δ13.24(s,1H),8.73(d,1H),8.61(d,1H),3.90(s,3H);

[0415] LC-MS (ESI): m / z = 409.1 [M+H] + .

[0416] Step 9: Compound 7I (8.2 g, 20 mmol) was dissolved in a mixture of tetrahydrofuran (240 mL) and ethanol (240 mL). Anhydrous calcium chloride (2.5 g, 2.4 mmol) was added at 0°C. Then, sodium borohydride (1.47 g, 40 mmol) was added to the reaction system in four batches at 10-min intervals. The reaction was allowed to proceed for 10 min. Saturated ammonium chloride solution (100 mL) was added to the system, and the mixture was stirred at room temperature for 10 min. The mixture was filtered, and the filter cake was washed with water (100 mL). The filter cake was collected and dried. The solid was then mixed with methanol (200 mL), stirred for three hours, filtered, and the filter cake was collected and dried to obtain compound 7J (4.8 g, yield: 62.84%).

[0417] 1 H NMR (400MHz, DMSO-d6) δ12.61(s,1H),8.55(d,1H),8.28(d,1H),5.55(t,1H),4.61(d,2H);

[0418] LC-MS (ESI): m / z = 381.1 [M+H] + .

[0419] Step 10: Dissolve compound 7J (0.95 g, 2.5 mmol) in thionyl chloride (10 mL) and heat to 70°C for 2 hours. After cooling, concentrate the mixture. Add petroleum ether (20 mL) to the resulting residue, stir for 20 minutes, filter, and wash the filter cake with petroleum ether (30 mL). Collect and dry the filter cake to obtain compound 7K (0.92 g, yield: 89.93%).

[0420] 1 H NMR (400MHz, DMSO-d6) δ12.86(s,1H),8.60(d,1H),8.27(d,1H),5.01(s,2H).

[0421] Step 11: Compound 7K (0.92 g, 2.24 mmol) was dissolved in N,N-dimethylformamide (20 mL), and potassium phthalimide (0.42 g, 2.24 mmol) was added. The mixture was allowed to react at room temperature for 2 hours. The reaction mixture was poured into water (200 mL), stirred for 10 minutes, and then filtered. The filter cake was washed with water (50 mL), collected, and dried to obtain compound 7L (1.1 g, yield: 96.32%).

[0422] 1H NMR (400MHz, DMSO-d6) δ12.54(s,1H),8.61(d,1H),8.40(d,1H),7.96–7.94(m,2H),7.90–7.88(m,2H),5.12(s,2H);

[0423] LC-MS (ESI): m / z = 510.3 [M+H] + .

[0424] Step 12: Compound 7L (300 mg, 0.59 mmol), 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (110 mg, 0.53 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (43 mg, 0.059 mmol), and sodium bicarbonate (200 mg, 2.36 mmol) were dissolved in a mixed solvent of N,N-dimethylformamide and water (v:v = 2:1, 12 mL). The atmosphere was replaced with nitrogen three times and heated to 70°C for 5 hours. The mixture was cooled to room temperature and concentrated directly. The resulting residue was purified by silica gel column chromatography (dichloromethane:methanol (v:v) = 20:1 to 10:1) to obtain compound 7M (180 mg, yield: 65.72%).

[0425] LC-MS (ESI): m / z = 466.1 [M+H] + .

[0426] Step 13: Compound 7M (180 mg, 0.38 mmol), pinacol diboron (190 mg, 0.76 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (27 mg, 0.038 mmol), and potassium acetate (88 mg, 1.8 mmol) were dissolved in 1,4-dioxane (10 mL). The atmosphere was replaced with nitrogen three times and heated to 80°C for 16 hours. After cooling, the mixture was concentrated directly. The resulting residue was mixed with a mixed solvent of petroleum ether and ethyl acetate (v:v = 10:1, 20 mL), stirred at room temperature for 1 hour, and filtered. The filter cake was washed once with a mixed solvent of petroleum ether and ethyl acetate (v:v = 10:1, 20 mL). The filter cake was collected and dried to obtain compound 7N (300 mg crude product), which was used directly in the next reaction.

[0427] LC-MS (ESI): m / z = 430.1 [M+H] + .

[0428] Step 14: Dissolve compound 7N (300 mg, 0.58 mmol), compound 1F (242 mg, 0.58 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (42 mg, 0.058 mmol), and sodium bicarbonate (101 mg, 1.2 mmol) in a mixed solvent of 1,4-dioxane and water (v:v = 5:1, 18 mL). Replace the atmosphere with nitrogen three times and heat to 80°C for 5 hours. After cooling, concentrate the mixture to obtain compound 7O (600 mg crude product), which is used directly in the next step.

[0429] LC-MS (ESI): m / z = 676.1 [M+H] + .

[0430] Step 15: Dissolve compound 7O (600 mg) in ethanol (30 mL), then add hydrazine hydrate (0.1 mL) and react at 80°C for 1 hour. Cool and concentrate directly, and the resulting residue is purified by preparative HPLC. Preparation method: 1. Apparatus: Waters 2767 preparative liquid chromatography; Chromatographic column: SunFire@PrepC18 (19 mm × 250 mm). 2. Dissolve the sample in methanol and filter through a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Mobile phase A and B composition: Mobile phase A: acetonitrile; Mobile phase B: Water (containing 0.1% NH3·H2O); b. Gradient elution, mobile phase A content ranging from 20% to 60%; c. Flow rate: 20 mL / min; d. Elution time: 20 min. Retention time: 3.26 min. Compound 7 (26 mg, yield: 12.58%) was obtained.

[0431] 1 H NMR(400MHz,DMSO-d6)δ12.18(s,1H),8.35(s,1H),8.05(d,1H),7.84(s,1H),7.59(s,1H),7.50(s,1 H),7.40(s,1H),4.21(s,1H),3.86(s,3H),3.79(s,3H),3.75(s,2H),0.92(d,2H),0.84–0.74(m,2H);

[0432] LC-MS (ESI): m / z = 545.1 [M+H] + .

[0433] Example 8 and Example 9

[0434] (2S)-2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydroisoquinolin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(3-methylazetidin-1-yl)benzonitrile and (2R)-2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydroisoquinolin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(3-methylazetidin-1-yl)benzonitrile (Compound 8, Compound 9)

[0435] (2S)-2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydroisoquinolin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(3-methylazetidin-1-yl)benzonitrile and(2R)-2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydroisoquinolin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(3-methylazetidin-1-yl)benzonitrile(compound 8, compound 9)

[0436] Step 1: Compound 8A (30 g, 118.84 mmol, synthesis method reference: US202178994), 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (40 g, 190.2 mmol), cesium fluoride (30 g, 202 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (8.7 g, 11.9 mmol) were weighed into a three-necked flask. Cyclopentyl methyl ether was added, and nitrogen was protected. The reaction was carried out at 90°C for 3 hours. A mixed solvent of petroleum ether and ethyl acetate (v:v = 1:1, 250 mL) was added, stirred, filtered, and the filtrate was collected. The filter cake was washed twice with a mixed solvent of petroleum ether and ethyl acetate (v:v = 1:1, 250 mL). The combined filtrates were concentrated, and the resulting residue was separated by column chromatography (petroleum ether:ethyl acetate (v:v) = 10:1) to give compound 8B (18.80 g, yield: 62%).

[0437] LC-MS (ESI): m / z = 254.0 [M+H] + .

[0438] Step 2: Compound 8B (3.00 g, 11.83 mmol) was dissolved in acetonitrile (80 mL), and N-iodosuccinimide (5.32 g, 23.66 mmol) and trifluoroacetic acid (0.67 g, 5.92 mmol) were added, followed by stirring at 50°C for 4 h. The mixture was directly concentrated, and the resulting residue was separated by column chromatography (petroleum ether:ethyl acetate (v:v) = 1:5) to provide compound 8C (4.30 g, yield: 95%).

[0439] LC-MS (ESI): m / z = 379.9 [M+H] + .

[0440] Step 3: Compound 8C (1.14 g, 3.00 mmol), compound 1E (1.00 g, 2.50 mmol), and sodium bicarbonate (0.63 g, 7.50 mmol) were added sequentially to a solution of 1,4-dioxane (75 mL) and water (25 mL). The atmosphere was purged with nitrogen three times, and then [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (0.18 g, 0.25 mmol) was added. The atmosphere was purged with nitrogen three more times, and the temperature was raised to 60°C for 1 hour. After cooling, the mixture was directly concentrated, and the resulting residue was separated by silica gel column chromatography (dichloromethane:methanol (v:v) = 100%-90%) to obtain compound 8D (680 mg, yield: 51.72%).

[0441] LC-MS (ESI): m / z = 526.2 [M+H] + .

[0442] Step 4: Compound 8D (400 mg, 0.76 mmol), potassium carbonate (0.32 g, 2.32 mmol), and 3-methylazetidine (65 mg, 0.91 mmol) were added sequentially to N,N-dimethylformamide (20 mL) and the mixture was heated to 80°C for 1 hour. After cooling, the mixture was filtered. Ethyl acetate (50 mL) and water (50 mL) were added to the filtrate for separation. The organic phase was collected, dried, and concentrated. The resulting residue was purified by silica gel column chromatography (dichloromethane:methanol (v:v) = 100%-90%) to obtain compound 8E (79 mg, yield: 18.01%).

[0443] LC-MS (ESI): m / z = 575.0 [MH] - .

[0444] Step 5: Compound 8E (79 mg, 0.13 mmol) was separated by SFC to yield P1 (retention time 1.404 min, designated as compound 8E-1) and P2 (retention time 2.049 min, designated as compound 8E-2). Separation conditions were: Waters 150 preparative SFC (SFC-26), ChiralCel OX column, 250×30 mm ID, 5 μm, mobile phase: A: CO2, B: isopropanol and acetonitrile (containing 0.1% ammonia), gradient: B 50%, flow rate: 120 mL / min, column pressure: 100 bar. Column temperature: 25°C; wavelength: 220 nm; cycle time: 4.5 min. Sample preparation: Dissolve the compound in acetonitrile. Injection: 3 mL per injection. Workup: After separation, concentrate the product by rotary evaporation at 35°C, then dry the solvent in a lyophilizer at -80°C. Compound 8E-1 (38 mg, 48.1%) and compound 8E-2 (40 mg, 50.6%) were obtained.

[0445] Compound 8E-1: LC-MS (ESI): m / z=577.5 [M+H] + .

[0446] Compound 8E-2: LC-MS (ESI): m / z = 577.5 [M+H] + .

[0447] Step 6: Dissolve compound 8E-1 (38 mg, 0.066 mmol) in dichloromethane (2 mL), then add trifluoroacetic acid (0.5 mL) and stir at room temperature for 1 hour. Concentrate the resulting residue and purify it by preparative HPLC. Separation method: 1. Apparatus: Waters 2767 preparative liquid chromatography; Column: SunFire@PrepC18 (19 mm × 250 mm). 2. Filter the sample through a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Mobile phase A and B composition: Mobile phase A: acetonitrile; Mobile phase B: Water (containing 0.1% NH3·H2O); b. Gradient elution, mobile phase A content ranging from 10% to 70%; c. Flow rate: 20 mL / min; d. Elution time: 15 min. Retention time: 11.0 min. Compound 8 (14 mg, yield: 44%) was obtained.

[0448] Compound 8: 1H NMR(400MHz, CDCl3)δ8.23(s,1H),7.92(s,1H),7.55(s,1H),7.14(s,2H),6.56(d,1H),4.31-4.23(m,3H), 3.81(s,4H),3.77-3.73(m,1H),3.70-3.66(m,1H),2.79(s,1H),1.26(s,3H); LC-MS(ESI):m / z=475.0[MH] - .

[0449] Compound 8E-2 (40 mg, 0.069 mmol) was dissolved in dichloromethane (2 mL), followed by the addition of trifluoroacetic acid (0.5 mL). The reaction was stirred at room temperature for 1 hour. The resulting residue was directly concentrated and purified by preparative HPLC. Separation method: 1. Apparatus: Waters 2767 preparative HPLC; Column: SunFire@PrepC18 (19 mm × 250 mm). 2. The sample was filtered through a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Mobile phase A and B composition: Mobile phase A: acetonitrile; Mobile phase B: Water (containing 0.1% NH3·H2O); b. Gradient elution, mobile phase A content from 10% to 70%; c. Flow rate: 20 mL / min; d. Elution time: 15 min. Retention time: 12.5 min. Compound 9 (14.6 mg, yield: 44%) was obtained.

[0450] Compound 9: 1 H NMR(400MHz, CDCl3)δ8.23(s,1H),7.92(s,1H),7.54(s,1H),7.15(s,2H),6.56(d,1H),4.30-4.23(m,3H), 3.82(s,4H),3.77-3.73(m,1H),3.70-3.66(m,1H),2.79(s,1H),1.25(s,3H); LC-MS(ESI):m / z=475.0[MH] - .

[0451] Example 10

[0452] 2-(4-(2-aminoethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropyloxy-3-fluorobenzonitrile (Compound 10)

[0453] 2-(4-(4-(2-aminoethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropoxy-3-fluorobenzonitrile(compound 10)

[0454] Step 1: Compound 10A (2.98 g, 11 mmol, synthesis method reference: Journal of Medicinal Chemistry, 2022, vol. 65, #3, p. 1749-1766) was dissolved in anhydrous acetonitrile (30 mL). Trimethylsilyl cyanide (2.75 mL, 22 mmol) was added. After nitrogen displacement, tetrabutylammonium fluoride (11.51 g, 44 mmol) was added and heated to 80°C with stirring for 3 h. After cooling to room temperature, the reaction was quenched with water (100 mL) and extracted with ethyl acetate (200 mL x 3). The organic phase was dried over anhydrous sodium sulfate and concentrated. The resulting residue was purified by slurrying with a mixed solvent of petroleum ether and dichloromethane (v:v = 10:1) to obtain compound 10B (2.6 g, yield: 90%).

[0455] 1 H NMR (400MHz, DMSO-d6) δ12.86(s,1H),8.17(dd,2H),8.06(dd,1H),4.49(s,2H);

[0456] LC-MS (ESI): m / z = 264.0 [M+H] + .

[0457] Step 2: Dissolve compound 10B (1.0 g, 3.8 mmol) in ethanol (15 mL), add nickel chloride (0.41 g, 3.1 mmol) and sodium borohydride (11.51 g, 30.4 mmol), and heat to 45°C with stirring for 3 hours to obtain an ethanol solution of compound 10C, which was used directly in the next reaction.

[0458] LC-MS (ESI): m / z = 268.0 [M+H] + .

[0459] Step 3: To a solution of compound 10C in ethanol (15 mL) were slowly added triethylamine (0.16 g, 15.2 mmol) and di-tert-butyl dicarbonate (6.63 g, 30.4 mmol) dropwise. The mixture was allowed to react at room temperature overnight. Water (100 mL) was added, and the mixture was extracted with ethyl acetate (200 mL x 3). The organic phase was dried over anhydrous sodium sulfate and concentrated. The resulting residue was purified by column chromatography (petroleum ether:ethyl acetate (v / v) = 5:1) to afford compound 10D (0.068 g, 4.9% yield).

[0460] LC-MS (ESI): m / z = 368.2 [M+H] + .

[0461] Step 4: Compound 10D (68 mg, 0.18 mmol), compound 2B (150 mg, 0.36 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (29 mg, 0.036 mmol), and sodium bicarbonate (53 mg, 0.63 mmol) were dissolved in 1,4-dioxane (10 mL) and water (2 mL). The atmosphere was replaced with nitrogen three times, and the mixture was heated to 80°C and stirred for 2 hours. After cooling, water (50 mL) was added, and the mixture was extracted with ethyl acetate (100 mL x 3). The organic phase was dried over anhydrous sodium sulfate, and the resulting residue was concentrated and separated by column chromatography (petroleum ether:ethyl acetate (v / v) = 1:5) to obtain compound 10E (87 mg, yield: 83%).

[0462] LC-MS (ESI): m / z = 579.3 [M+H] + .

[0463] Step 5: Dissolve compound 10E (87 mg, 0.15 mmol) in dichloromethane (4 mL), add trifluoroacetic acid (1 mL), and stir at room temperature for 1 hour. Concentrate the resulting residue and purify it by preparative HPLC. Preparation method: 1. Apparatus: Waters 2767 preparative liquid chromatography; Chromatographic column: SunFire@PrepC18 (19 mm × 250 mm). 2. Dissolve the sample in methanol and filter through a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Mobile phase A, B composition: Mobile phase A: acetonitrile; Mobile phase B: water; b. Gradient elution, mobile phase A content ranging from 10% to 40%; c. Flow rate: 20 mL / min; d. Elution time: 10 min. Compound 10 (5 mg, yield: 7%) was obtained.

[0464] 1H NMR(400MHz,DMSO-d6)δ12.44(s,1H),8.26(s,1H),8.16(t,1H),8.02(t,1H),7.67(d,1H ),7.56(s,1H),4.24–4.13(m,1H),3.77(d,3H),2.80(dd,4H),0.90(t,2H),0.83(d,2H);

[0465] LC-MS (ESI): m / z = 479.50 [M+H] + .

[0466] Example 11 and Example 12

[0467] (2S)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropyloxy-3-fluorobenzonitrile and (2R)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropyloxy-3-fluorobenzonitrile (Compound 11, Compound 12)

[0468] (2S)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropoxy-3-fluorobenzonitrile and(2R)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropoxy-3-fluorobenzonitrile(compound 11, compound 12)

[0469] Step 1: Dissolve compound 5C (4 g, 9.89 mmol) in trifluoroacetic acid (20 mL), heat to 50°C, and stir for 1 hour. Concentrate directly, add saturated aqueous sodium carbonate (50 mL) to the resulting residue, stir for half an hour, filter, collect the filter cake, wash it twice with water, and dry it to obtain compound 11A (2.4 g crude product), which is used directly in the next reaction.

[0470] LC-MS (ESI): m / z = 254.0 [M+H] + .

[0471] Step 2: Dissolve compound 11A (2.5 g, 9.84 mmol) in methanol (20 mL) and dichloromethane (20 mL), then add di-tert-butyl dicarbonate (6.5 g, 29.51 mmol) and stir at room temperature for 1 hour. Concentrate directly, add water (50 mL), stir evenly, filter, collect the filter cake, wash it three times with petroleum ether, and dry it to obtain compound 11B (1.6 g crude product), which is used directly in the next reaction.

[0472] LC-MS (ESI): m / z = 354.0 [M+H] + .

[0473] Step 3: Compound 11B (0.1 g, 0.24 mmol), compound 2B (0.16 g, 0.24 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (0.018 g, 0.024 mmol), and sodium bicarbonate (0.06 g, 0.72 mmol) were dissolved in a mixed solvent of 1,4-dioxane (5 mL) and water (1 mL). The atmosphere was replaced with nitrogen three times, and the mixture was heated to 80°C and stirred for 1 hour. After cooling, water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phase was collected, washed with saturated brine, and dried over anhydrous sodium sulfate. The resulting residue was concentrated and purified by column chromatography (dichloromethane:methanol (v:v) = 100:8) to obtain compound 11C (0.08 g, yield: 59.00%).

[0474] LC-MS (ESI): m / z = 565.2 [M+H] + .

[0475] Step 4: Compound 11C (320 mg) was separated by chiral SFC to afford P1 (retention time: 1.891 min, designated as compound 11C-1) and P2 (retention time: 2.139 min, designated as compound 11C-2). Separation method: Instrument: Waters 150 SFC; Column: Chiralcel OD-Column; Mobile phases: A for CO2 and B for EtOH (0.1% NH3·H2O); Gradient: 35% phase B isocratic elution; Flow rate: 80 mL / min; Column pressure: 100 bar; Column temperature: 25°C; Absorption wavelength: 220 nm, Cycle time: 3 min. Sample preparation: The compound was dissolved in acetonitrile at a concentration of 5 mg / mL; injection: 3 mL was injected each time; treatment: after separation, it was concentrated by rotary evaporation at 35°C, and then the solvent was dried by freeze dryer at -80°C to obtain compounds 11C-1 (93.6 mg, 29.25%) and 11C-2 (100 mg, 31.25%).

[0476] Step 5: Compound 11C-1 (93.6 mg, 0.17 mmol) was dissolved in a solution of hydrogen chloride in dioxane (5 mL, 4 M) and stirred at room temperature for 2 hours. The reaction system was concentrated and diluted with dichloromethane, followed by sodium bicarbonate solution to make the solution alkaline. The liquid was extracted with dichloromethane, and the organic phase was collected, dried, and concentrated. The resulting residue was separated by reverse-phase column chromatography (water:acetonitrile (v / v) = 67:33) to provide compound 11 (38 mg, yield: 48.08%).

[0477] Compound 11: 1 H NMR (400MHz, DMSO-d6) δ8.16(s,1H),8.00(dd,2H),7.37(d,1H),7.21(dd,1H),4.25–4.15(m,1H),3.76(s,3H),3.63(s,2H),0.94–0.75(m,4H);

[0478] LC-MS (ESI): m / z = 465.4 [M+H] + .

[0479] Compound 11C-2 (100 mg, 0.18 mmol) was dissolved in a solution of hydrogen chloride in dioxane (5 mL, 4 M) and stirred at room temperature for 2 hours. The reaction system was concentrated and diluted with dichloromethane, followed by sodium bicarbonate solution to make the solution alkaline. The liquid was extracted with dichloromethane, and the organic phase was collected, dried, and concentrated. The crude product was separated by reverse-phase column chromatography (water:acetonitrile (v / v) = 67:33) to afford compound 12 (40 mg, yield: 47.80%).

[0480] Compound 12: 1 H NMR (400MHz, DMSO-d6) δ8.16(s,1H),8.00(dd,2H),7.37(d,1H),7.21(dd,1H),4.26–4.16(m,1H),3.76(s,3H),3.65(s,2H),0.97–0.74(m,4H);

[0481] LC-MS (ESI): m / z = 465.5 [M+H] + .

[0482] Example 13

[0483] 1-(Aminomethyl)-7-(5-(3-chloro-6-cyano-5-cyclopropyloxy-2-fluorophenyl)-1-methyl-1H-pyrazol-4-yl)-4-oxo-3,4-dihydrophthalazine-5-carbonitrile (Compound 13)

[0484] 1-(aminomethyl)-7-(5-(3-chloro-6-cyano-5-cyclopropoxy-2-fluorophenyl)-1-methyl-1H-pyrazol-4-yl)-4-oxo-3,4-dihydrophthalazine-5-carbonitrile(compound 13)

[0485] Step 1: Dissolve compound 7L (300 mg, 0.59 mmol), zinc cyanide (70 mg, 0.53 mmol), and tetrakis(triphenylphosphine)palladium (68 mg, 0.059 mmol) in N,N-dimethylformamide (8 mL). The atmosphere was purged with nitrogen three times and heated to 80°C for 4 hours. After cooling, the mixture was concentrated directly, and the resulting residue was purified by column chromatography (petroleum ether:ethyl acetate (v:v) = 1:1 to 1:2) to afford compound 13A (130 mg, 53.85% yield).

[0486] 1 H NMR (400MHz, DMSO-d6) δ12.86(s,1H),8.73(d,1H),8.65(d,1H),7.95(dd,2H),7.90(dd,2H),5.21(s,2H);

[0487] LC-MS (ESI): m / z = 409.0 [M+H] + .

[0488] Step 2: Compound 13A (130 mg, 0.32 mmol), pinacol diboron (163 mg, 0.64 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (23 mg, 0.032 mmol), and potassium acetate (76 mg, 0.8 mmol) were dissolved in 1,4-dioxane (6 mL). The atmosphere was purged with nitrogen three times and heated to 80°C for 16 hours. After cooling, the mixture was concentrated directly. The resulting residue was added to a mixture of petroleum ether and ethyl acetate (v:v = 10:1, 20 mL) and stirred for 1 hour. The residue was filtered and the filter cake was washed once with a mixture of petroleum ether and ethyl acetate (v:v = 10:1, 20 mL). The filter cake was collected and dried to obtain compound 13B (210 mg crude product), which was used directly in the next reaction.

[0489] LC-MS (ESI): m / z = 375.30 [M+H] + .

[0490] Step 3: Compound 13B (210 mg, 0.46 mmol), compound 1F (191 mg, 0.46 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (33 mg, 0.046 mmol), and sodium bicarbonate (77 mg, 0.92 mmol) were dissolved in a mixture of 1,4-dioxane and water (v:v = 5:1, 12 mL). The atmosphere was replaced with nitrogen three times and heated to 80°C for 5 hours. After cooling, the mixture was concentrated to obtain compound 13C (400 mg crude product), which was used directly in the next reaction.

[0491] Step 4: Dissolve compound 13C (400 mg) in ethanol (20 mL), then add hydrazine hydrate (0.1 mL) and heat to 80°C for 1 hour. After cooling, concentrate directly, and the resulting residue is purified by preparative HPLC. Preparation method: 1. Apparatus: Waters 2767 preparative liquid chromatography; Chromatographic column: SunFire@PrepC18 (19 mm × 250 mm). 2. Dissolve the sample in methanol and filter through a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Mobile phase A and B composition: Mobile phase A: acetonitrile; Mobile phase B: Water (containing 0.1% NH3·H2O); b. Gradient elution, mobile phase A content from 50% to 90%; c. Flow rate: 20 mL / min; d. Elution time: 20 min. Retention time: 4.91 min. Compound 13 (18 mg, yield: 11.48%) was obtained.

[0492] 1 H NMR(400MHz,DMSO-d6)δ12.79(s,1H),8.42(s,1H),8.35(s,1H),8.03(d,1H), 7.91(s,1H),4.21(s,1H),3.80(s,3H),3.73(s,2H),0.91(d,2H),0.80(d,2H);

[0493] LC-MS (ESI): m / z = 490.50 [M+H] + .

[0494] Example 14 and Example 15

[0495] (2S)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(1-methylcyclopropyloxy)benzonitrile and (2R)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(1-methylcyclopropyloxy)benzonitrile (Compound 14, Compound 15)

[0496] (2S)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(1-methylcyclopropoxy)benzonitrile and(2R)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(1-methylcyclopropoxy)benzonitrile(compound 14, compound 15)

[0497] Step 1: Compound 11B (0.1 g, 0.28 mmol), pinacol diboron (0.078 g, 0.31 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (0.02 g, 0.028 mmol), and potassium acetate (0.082 g, 0.84 mmol) were dissolved in 1,4-dioxane (5 mL). The atmosphere was purged with nitrogen three times and heated to 80°C for 16 hours. After cooling, the mixture was concentrated directly. The resulting residue was added to a mixture of petroleum ether and ethyl acetate (v:v = 10:1, 20 mL) and stirred for 1 hour. The residue was filtered and the filter cake was washed once with a mixture of petroleum ether and ethyl acetate (v:v = 10:1, 20 mL). The filter cake was collected and dried to afford compound 14A (0.2 g crude product), which was used directly in the next step.

[0498] LC-MS (ESI): m / z = 402.2 [M+H] + .

[0499] Step 2: Dissolve 1-methylcyclopropanol (5.7 g, 79.05 mmol) in tetrahydrofuran (100 mL) and add sodium hydride (3.15 g, 79.05 mmol) portionwise. Heat to 40°C and stir for 2 h. Cool to room temperature, then add compound 8C (10 g, 26.35 mmol) and react at room temperature for 8 h. The reaction is quenched by adding methanol and concentrated directly. The resulting residue is purified by silica gel column chromatography (ethyl acetate:petroleum ether (v:v) = 1:5) to afford compound 14B (2.5 g, yield: 21.98%).

[0500] LC-MS (ESI): m / z = 432.3 [M+H] + .

[0501] Step 3: Compound 14B (0.5 g, 1.16 mmol), compound 14A (0.47 g, 1.16 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (0.085 g, 0.12 mmol), and sodium bicarbonate (0.29 g, 3.48 mmol) were dissolved in a mixed solvent of 1,4-dioxane (10 mL) and water (2 mL). The atmosphere was replaced with nitrogen three times, and then heated to 80°C and stirred for 2 hours. Water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phase was collected, washed, dried, and concentrated. The resulting residue was purified by silica gel column chromatography (dichloromethane:methanol (v:v) = 100:8) to obtain compound 14C (0.3 g, yield: 44.67%).

[0502] LC-MS (ESI): m / z = 579.6 [M+H] + .

[0503] Step 4: Compound 14C (100 mg) was separated by chiral SFC to afford P1 (retention time: 1.754 min, designated as compound 14C-1) and P2 (retention time: 2.035 min, designated as compound 14C-2). Separation method: Instrument: Waters 150 SFC; Column: Chiralcel OD-Column; Mobile phases: A for CO2 and B for MeOH (0.1% NH3·H2O); Gradient: 35% phase B isocratic elution; Flow rate: 100 mL / min; Column pressure: 100 bar; Column temperature: 25°C; Absorption wavelength: 220 nm; Cycle time: 2 min. Sample preparation: The compound was dissolved in acetonitrile at a concentration of 2 mg / mL; injection: 2.5 mL was injected each time; treatment: after separation, it was concentrated by rotary evaporation at 35°C, and then the solvent was dried by freeze dryer at -80°C to obtain compounds 14C-1 (24 mg, 23.51%) and 14C-2 (22 mg, 22.12%).

[0504] Step 5: Compound 14C-1 (20 mg, 0.035 mmol) was dissolved in 4 M hydrogen chloride in dioxane (2 mL) and reacted at 25°C for 2 hours. The reaction system was concentrated and diluted with dichloromethane, followed by alkalinity with sodium bicarbonate solution. The liquid was extracted with dichloromethane, and the organic phase was collected, dried, and concentrated. The resulting residue was purified by reverse-phase column chromatography (water:acetonitrile (v / v) = 67:33) to afford compound 14 (8 mg, yield: 47.73%).

[0505] Compound 14: 1H NMR(400MHz,DMSO-d6)δ8.15(s,1H),7.98(d,1H),7.82(d,1H),7.38(d,1H),7.20(dd,1H),3.76 (s,3H),3.71(s,2H),1.58(s,3H),1.05-1.02(m,2H),0.90(t,2H); LC-MS(ESI):m / z=479.4[M+H] + .

[0506] Compound 14C-2 (20 mg, 0.035 mmol) was dissolved in 4 M hydrogen chloride in dioxane (2 mL) and reacted at 25°C for 2 hours. The reaction system was concentrated and diluted with dichloromethane, followed by alkalinity with sodium bicarbonate solution. The liquid was extracted with dichloromethane, and the organic phase was collected, dried, and concentrated. The resulting residue was purified by reverse-phase column chromatography (water:acetonitrile (v / v) = 67:33) to afford compound 15 (8 mg, yield: 47.73%).

[0507] Compound 15: 1 H NMR(400MHz,DMSO-d6)δ8.16(s,1H),7.98(d,1H),7.82(d,1H),7.37(d,1H),7.20(dd,1H),3.76 (s,3H),3.66(s,2H),1.58(s,3H),1.05-1.02(m,2H),0.90(t,2H); LC-MS(ESI):m / z=479.5[M+H] + .

[0508] Example 16 and Example 17

[0509] (2S)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(3-methylazetidin-1-yl)benzonitrile and (2R)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(3-methylazetidin-1-yl)benzonitrile (Compound 16, Compound 17)

[0510] 2S)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(3-methylazetidin-1-yl)benzonitrile and(2R)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(3-methylazetidin-1-yl)benzonitrile(compound 16, compound 17)

[0511] Step 1: Compound 8C (10 g, 26.35 mmol), 14A (12.7 g, 31.62 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (1.93 g, 2.64 mmol), and potassium acetate (6.64 g, 79.05 mmol) were dissolved in 1,4-dioxane (100 mL) and water (20 mL). The atmosphere was replaced with nitrogen three times and the mixture was heated to 80°C for 4 hours. After cooling, the mixture was concentrated directly. The resulting residue was added to a mixture of petroleum ether and ethyl acetate (v:v = 10:1, 20 mL) and stirred for 1 hour. The filter cake was collected by filtration and purified by column chromatography (methanol:dichloromethane (v:v) = 1:20) to provide compound 16A (9 g, yield: 64.82%).

[0512] LC-MS (ESI): m / z = 527.1 [M+H] + .

[0513] Step 2: 3-(Methoxymethyl)azetidine hydrochloride (0.31 g, 2.89 mmol), compound 16A (1 g, 1.90 mmol), and potassium carbonate (0.78 g, 5.63 mmol) were dissolved in N,N-dimethylformamide (20 mL), heated to 80°C, and stirred for 2 h. The mixture was cooled to room temperature and concentrated. The resulting residue was purified by silica gel column chromatography (methanol:dichloromethane (v:v) = 1:20) to provide compound 16B (0.8 g, yield: 72.84%).

[0514] LC-MS (ESI): m / z = 578.6 [M+H] + .

[0515] Step 3: Compound 16B (600 mg) was separated by chiral SFC to afford P1 (retention time: 2.025 min, designated as compound 16B-1) and P2 (retention time: 2.288 min, designated as compound 16B-2). Separation method: Instrument: Waters 150 SFC; Column: Chiralcel OJ-Column; Mobile phases: A for CO2 and B for MeOH (0.1% NH3·H2O); Gradient: 45% phase B isocratic elution; Flow rate: 120 mL / min; Column pressure: 100 bar; Column temperature: 25°C; Absorption wavelength: 220 nm; Cycle time: 3.5 min. Sample preparation: The compound was dissolved in methanol at a concentration of 10 mg / mL; injection: 3 mL was injected each time; treatment: after separation, it was concentrated by rotary evaporation at 35°C, and then the solvent was dried by freeze dryer at -80°C to obtain compounds 16B-1 (80 mg, 13.33%) and 16B-2 (80 mg, 13.33%).

[0516] Step 5: Dissolve 16B-1 (80 mg, 0.14 mmol) in 4 M hydrogen chloride in dioxane (2 mL) and react at 25°C for 2 hours. The reaction system was concentrated and diluted with dichloromethane, followed by alkalinity with sodium bicarbonate solution. The liquid was extracted with dichloromethane, and the organic phase was collected, dried, and concentrated. The resulting residue was purified by reverse-phase column chromatography (water:acetonitrile (v / v) = 67:33) to afford compound 16 (40 mg, yield: 59.78%).

[0517] Compound 16: 1 H NMR(400MHz,DMSO-d6)δ8.14(d,1H),7.99(d,1H),7.38–7.35(m,1H),7.34–7.25(m,2H), 6.64-6.61(m,1H),3.75(s,3H),3.72–3.57(m,4H),3.15–3.12(m,2H),1.10-0.95(m,4H);

[0518] LC-MS(ESI):m / z=514.5[M+HCl+H] + .

[0519] 16B-2 (80 mg, 0.14 mmol) was dissolved in 4 M hydrogen chloride in dioxane (2 mL) and reacted at 25°C for 2 hours. The reaction system was concentrated and diluted with dichloromethane, followed by sodium bicarbonate solution to make the solution alkaline. The mixture was extracted with dichloromethane, and the organic phase was collected, dried, and concentrated. The resulting residue was purified by reverse-phase column chromatography (water:acetonitrile (v / v) = 67:33) to afford compound 17 (41 mg, yield: 60.05%).

[0520] Compound 17: 1 H NMR(400MHz,DMSO-d6)δ8.14(d,1H),7.99(d,1H),7.37–7.33(m,1H),7.33–7.27(m,2H), 6.64-6.61(m,1H),3.75(s,3H),3.72–3.57(m,4H),3.18–3.09(m,2H),1.10-0.95(m,4H);

[0521] LC-MS(ESI):m / z=514.5[M+HCl+H] + .

[0522] Example 18 and Example 19

[0523] (2S)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-6-azido-4-chloro-3-fluorobenzonitrile and (2R)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-6-azido-4-chloro-3-fluorobenzonitrile (Compound 18, Compound 19)

[0524] (2S)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-6-azido-4-chloro-3-fluorobenzonitrile and(2R)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-6-azido-4-chloro-3-fluorobenzonitrile(compound 18, compound 19)

[0525] Step 1: Dissolve compound 16A (0.5 g, 0.95 mmol) in N,N-dimethylformamide (10 mL), cool to -20°C, add sodium azide (0.068 g, 1.04 mmol), and slowly warm to room temperature for 5 hours. The mixture was concentrated, and the resulting residue was purified by silica gel column chromatography (methanol:dichloromethane (v:v) = 1:20) to afford compound 18A (0.4 g, yield: 76.56%).

[0526] LC-MS (ESI): m / z = 550.1 [M+H] + .

[0527] Step 2: Compound 18A (400 mg) was separated by chiral SFC to afford P1 (retention time: 1.156 min, designated as compound 18A-1) and P2 (retention time: 1.479 min, designated as compound 18A-2). Separation method: Instrument: Waters 150 SFC; Column: Chiralcel IC-Column; Mobile phases: A for CO2 and B for EtOH (0.1% NH3·H2O); Gradient: 45% phase B isocratic elution; Flow rate: 70 mL / min; Column pressure: 100 bar; Column temperature: 25°C; Absorption wavelength: 220 nm; Cycle time: 9 min. Sample preparation: The compound was dissolved in acetonitrile at a concentration of 2 mg / mL; injection: 2.5 mL was injected each time; treatment: after separation, it was concentrated by rotary evaporation at 35°C, and then the solvent was dried by freeze dryer at -80°C to obtain compounds 18A-1 (128 mg, 32.00%) and 18A-2 (126 mg, 31.5%).

[0528] Step 3: Compound 18A-1 (128 mg, 0.23 mmol) was dissolved in 4 M hydrogen chloride in dioxane (2 mL) and reacted at 25°C for 2 hours. The reaction system was concentrated and diluted with dichloromethane, followed by alkalinity with sodium bicarbonate solution. The liquid was extracted with dichloromethane, and the organic phase was collected, dried, and concentrated. The resulting residue was purified by reverse-phase column chromatography (water:acetonitrile (v / v) = 67:33) to afford compound 18 (50 mg, yield: 48.33%).

[0529] Compound 18: 1 H NMR (400MHz, DMSO-d6) δ8.21(d,1H),8.16(s,1H),7.97(d,1H),7.39(d,1H),7.23–7.16(m,1H),3.76(s,3H),3.61(s,2H);

[0530] LC-MS (ESI): m / z = 450.5 [M+H] + .

[0531] Compound 18A-2 (126 mg, 0.32 mmol) was dissolved in 4M hydrogen chloride in dioxane (2 mL) and reacted at 25°C for 2 hours. The reaction system was concentrated and diluted with dichloromethane, followed by alkalinity with sodium bicarbonate solution. The liquid was extracted with dichloromethane, and the organic phase was collected, dried, and concentrated. The resulting residue was purified by reverse-phase column chromatography (water:acetonitrile (v / v) = 67:33) to afford compound 19 (50 mg, yield: 48.32%).

[0532] Compound 19: 1 H NMR (400MHz, DMSO-d6) δ8.20(d,1H),8.16(s,1H),7.97(d,1H),7.39(d,1H),7.23–7.16(m,1H),3.76(s,3H),3.60(s,2H);

[0533] LC-MS (ESI): m / z = 450.5 [M+H] + .

[0534] Example 20

[0535] 2-(4-(4-(Aminomethyl)-1-oxo-8-vinyl-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropyloxy-3-fluorobenzonitrile (Compound 20)

[0536] 2-(4-(4-(aminomethyl)-1-oxo-8-vinyl-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropoxy-3-fluorobenzonitrile(compound 20)

[0537] Step 1: Compound 7L (5.0 g, 9.8 mmol) was added to a reaction flask and mixed with anhydrous ethanol (25 mL). Hydrazine hydrate (3 mL, 39.2 mmol) was then added and reacted at 80°C for 1 hour. After the reaction, the mixture was cooled to room temperature and filtered. The filter cake was washed once with anhydrous ethanol (25 mL) and once with water (50 mL). The filter cake was collected and dried to obtain compound 20A (3.5 g, yield: 94.59%).

[0538] LC-MS (ESI): m / z = 382.0 [M+H] + .

[0539] Step 2: Compound 20A (3.5 g, 9.2 mmol) and triethylamine (3.8 mL, 27.6 mmol) were added to a reaction flask and dissolved with dichloromethane (60 mL). Di-tert-butyl dicarbonate (4.0 g, 18.4 mmol) was then added and allowed to react at room temperature for 3 hours. After completion of the reaction, the mixture was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (v:v) = 2:1 to 1:2) to obtain compound 20B (1.0 g, yield: 22.73%).

[0540] 1 H NMR (400MHz, DMSO-d6) δ12.64(s,1H),8.56(d,1H),8.23(s,1H),7.40(s,1H),4.35(d,2H),1.40(s,9H);

[0541] LC-MS (ESI): m / z = 480.3 [M+H] + .

[0542] Step 3: Compound 20B (400 mg, 0.83 mmol), bistriphenylphosphine palladium dichloride (57 mg, 0.083 mmol), and tetraethylammonium chloride (270 mg, 1.63 mmol) were added to a reaction flask, dissolved with N,N-dimethylformamide (15 mL), and the atmosphere was replaced with nitrogen three times. Tributyl vinyl tin (260 mg, 0.83 mmol) was then added, and the mixture was reacted at 110°C for 2 hours. After cooling, the reaction solution was poured into water (100 mL) and extracted with ethyl acetate (50 mL × 3). The organic phases were combined, washed once with saturated brine, and dried over anhydrous sodium sulfate. The filtrate was filtered and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (v:v) = 2:1 to 0:1) to obtain compound 20C (200 mg, yield: 63.49%).

[0543] 1 H NMR(400MHz,DMSO-d6)δ12.57(s,1H),8.15(s,1H),8.13–8.07(m,1H),8.06(d ,1H),7.36–7.30(m,1H),5.83(d,1H),5.45(d,1H),4.38(t,2H),1.40(s,9H);

[0544] LC-MS (ESI): m / z = 382.1 [M+H] + .

[0545] Step 4: Compound 20C (100 mg, 0.26 mmol), compound 2B (109 mg, 0.26 mmol), chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (21 mg, 0.026 mmol), and sodium bicarbonate (45 mg, 0.53 mmol) were added to a reaction flask. 1,4-dioxane and water (v:v = 10:1, 10 mL) were then added. The atmosphere was purged with nitrogen three times and the reaction was carried out at 90°C for 2 hours. After cooling, the mixture was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (v:v) = 4:1 to 1:1) to afford compound 20D (56 mg, yield: 36.51%).

[0546] LC-MS (ESI): m / z = 591.3 [M+H] + .

[0547] Step 5: Compound 20D (56 mg, 0.095 mmol) was added to a reaction flask and dissolved in dichloromethane (2 mL). Trifluoroacetic acid (0.6 mL) was then added and allowed to react at room temperature for 30 min. The organic solvent was removed by direct concentration under reduced pressure, and the residue was purified by preparative HPLC. Method: 1. Apparatus: Waters 2767 Preparative HPLC; Column: SunFire@PrepC18 (19 mm × 250 mm). 2. The sample was dissolved in methanol and filtered through a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatographic conditions: a. Mobile phase A and B composition: Mobile phase A: acetonitrile; Mobile phase B: Water (containing 0.1% TFA); b. Gradient elution, mobile phase A content ranging from 20% to 60%; c. Flow rate: 20 mL / min. d. Elution time: 20 min; Retention time: 3.45 min. Compound 20 (19 mg, yield: 33.04%) was obtained.

[0548] 1 H NMR(400MHz,DMSO-d6)δ12.81(s,1H),8.43(s,1H),8.14–8.04(m,2H),7.79(s,1H),7.38(s,1H),5.30( d,1H),5.11(d,1H),4.38(d,2H),4.26–4.20(m,1H),3.81(s,3H),0.94–0.93(m,2H),0.85–0.72(m,2H);

[0549] LC-MS (ESI): m / z = 491.5 [M+H] + .

[0550] Example 21

[0551] 2-(4-(4-(Aminomethyl)-8-ethynyl-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropyloxy-3-fluorobenzonitrile (Compound 21)

[0552] 2-(4-(4-(aminomethyl)-8-ethynyl-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropoxy-3-fluorobenzonitrile(compound 21)

[0553] Step 1: Compound 20B (0.1 g, 0.21 mmol), trimethylethynylsilane (41 mg, 0.42 mmol), cuprous iodide (8 mg, 0.42 mmol), and triethylamine (42 mg, 0.42 mmol) were dissolved in N,N-dimethylformamide (5 mL). Nitrogen was bubbled through the mixture for 1 minute, followed by the addition of bistriphenylphosphine palladium dichloride (15 mg, 0.021 mmol). Nitrogen was bubbled through the mixture for another half minute, and the mixture was stirred on ice for 1 hour. The mixture was then filtered, and the filtrate was diluted with water and extracted with ethyl acetate. The organic phase was washed sequentially with sodium bicarbonate solution and saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a residue which was purified by silica gel column chromatography (petroleum ether:ethyl acetate (v:v) = 3:2) to afford compound 21A (30 mg, yield: 32%).

[0554] LCMS (ESI): m / z = 450.1 [M+H] + .

[0555] Step 2: Compound 21A (80 mg, 0.18 mmol), compound 2B (110 mg, 0.27 mmol), sodium bicarbonate (30 mg, 0.36 mmol), and chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (14 mg, 0.018 mmol) were dissolved in 1,4-dioxane (5 mL) and water (0.5 mL). The mixture was heated to 90°C and stirred for 2 h under nitrogen. After cooling, the mixture was filtered and the filtrate was concentrated. The resulting residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (v / v) = 1:0 to 1:1) to afford compound 21B (40 mg, yield: 34%).

[0556] LCMS (ESI): m / z = 661.2 [M+H] + .

[0557] Step 3: Compound 21B (40 mg, 0.06 mmol) was dissolved in dichloromethane (2 mL), followed by the addition of trifluoroacetic acid (0.6 mL) and stirring at room temperature for 1 hour. The reaction mixture was concentrated to dryness, and the crude solid was added to anhydrous methanol (3 mL), followed by the addition of potassium carbonate (17 mg, 0.12 mmol) and stirring at room temperature for 2 hours. The mixture was filtered, and the filtrate was concentrated to dryness. The resulting residue was purified by silica gel column chromatography (dichloromethane:methanol (v / v) = 1:0 to 9:1) to afford compound 21 (8 mg, yield: 27%).

[0558] 1 H NMR(400MHz,CD3OD)δ8.18(s,1H),7.95(d,1H),7.72(d,1H),7.68(dd,1H),4.15-4.11 (m,1H),3.95(s,1H),3.93(d,2H),3.85(s,3H),0.97-0.93(m,2H),0.89-0.85(m,2H);

[0559] LCMS (ESI): m / z = 489.1 [M+H] + .

[0560] Example 22

[0561] 2-(4-(4-(Aminomethyl)-8-(azetidin-1-yl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropyloxy-3-fluorobenzonitrile (Compound 22)

[0562] 2-(4-(4-(aminomethyl)-8-(azetidin-1-yl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropoxy-3-fluorobenzonitrile(compound 22)

[0563] Step 1: Compound 7L (0.25 g, 0.49 mmol), anhydrous potassium phosphate (310 mg, 1.47 mmol), cuprous iodide (47 mg, 0.24 mmol), and azetidine (56 mg, 0.98 mmol) were dissolved in N,N-dimethylformamide (5 mL). Nitrogen was bubbled through the mixture for 1 minute, then heated to 90°C and stirred for 2 hours. After cooling, the mixture was filtered. The filtrate was diluted with water and extracted with ethyl acetate. The organic phase was collected, washed with sodium bicarbonate solution and saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a residue which was purified by silica gel column chromatography (petroleum ether:ethyl acetate (v:v) = 1:1) to afford compound 22A (30 mg, yield: 14%).

[0564] LCMS (ESI): m / z = 439.0 [M+H] + .

[0565] Step 2: Compound 22A (30 mg, 0.068 mmol), compound 2B (43 mg, 0.10 mmol), sodium bicarbonate (17 mg, 0.20 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (5 mg, 0.0068 mmol) were dissolved in 1,4-dioxane (3 mL) and water (0.3 mL). The mixture was heated to 90°C and stirred for 2 h under nitrogen. After cooling, the mixture was filtered and the filtrate was concentrated. The resulting residue was purified by silica gel column chromatography (dichloromethane:methanol (v / v) = 10:1) to afford compound 22B (40 mg, 94% yield).

[0566] LCMS (ESI): m / z = 650.2 [M+H] + .

[0567] Step 3: Compound 22B (40 mg, 0.062 mmol) was dissolved in anhydrous methanol (3 mL), followed by the addition of aqueous methylamine (0.3 mL). The mixture was heated to 45°C for 3 hours. After cooling, the mixture was concentrated to dryness. The resulting residue was purified by silica gel column chromatography (dichloromethane:methanol (v / v) = 1:0 to 9:1) to afford compound 22 (10 mg, 31% yield).

[0568] 1 H NMR(400MHz,CD3OD)δ8.01(s,1H),7.82(d,1H),6.92(s,1H),6.27(s,1H),4.02-3.97(m,3H ),3.84-3.78(m,4H),3.71(s,3H),2.21-2.15(m,2H),0.85-0.80(m,2H),0.79-0.73(m,2H);

[0569] LCMS (ESI): m / z = 520.2 [M+H] + .

[0570] Examples 23 and 24

[0571] (2S)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-vinylbenzonitrile and (2R)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-vinylbenzonitrile (Compound 23, Compound 24)

[0572] (2S)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-vinylbenzonitrile and(2R)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-vinylbenzonitrile(compound 23, compound 24)

[0573] Step 1: Dissolve compound 14A (2.00 g, 3.80 mmol) in tetrahydrofuran (10 mL), add aqueous ammonia (40%, 50 mL), and heat to 100°C in a sealed container with stirring for 6 hours. After cooling, add ethyl acetate (200 mL) and water (200 mL) to separate the liquids. The organic phase is collected, washed with saturated brine (200 mL x 3), dried over anhydrous sodium sulfate, and concentrated to obtain compound 23A (1.86 g crude product), which is used directly in the next reaction.

[0574] LC-MS (ESI): m / z = 525.2 [M+H] + .

[0575] Step 2: Compound 23A (1.86 g, 3.55 mmol) was dissolved in acetonitrile (50 mL), and copper bromide (0.87 g, 3.91 mmol) was added. The mixture was cooled to -15°C to -10°C under nitrogen. Tert-butyl nitrite (0.50 g, 4.85 mmol) was slowly added, and the temperature was naturally raised to 30°C with stirring for 1 h. Ethyl acetate (200 mL) and water (200 mL) were added for separation. The organic phase was washed with saturated brine (200 mL x 3), dried over anhydrous sodium sulfate, and concentrated to obtain compound 23B (2.00 g crude product), which was used directly in the next reaction.

[0576] LC-MS (ESI): m / z = 587.2 [M+H] + .

[0577] Step 3: Compound 23B (2.00 g, 3.40 mmol), potassium ethylene trifluoroborate (0.68 g, 5.10 mmol), and potassium carbonate (2.82 g, 20.40 mmol) were added to a mixed solution of 1,4-dioxane (100 mL) and water (20 mL). After nitrogen displacement, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (0.12 g, 0.17 mmol) was added. After nitrogen displacement again, the temperature was raised to 90°C and the reaction was allowed to react for 2 hours. After cooling and filtration, ethyl acetate (200 mL) and water (200 mL) were added to the filtrate for separation. The organic phase was washed with saturated brine (200 mL × 3), dried over anhydrous sodium sulfate, and concentrated to obtain compound 23C (1.50 g crude product), which was used directly in the next reaction.

[0578] LC-MS (ESI): m / z = 535.1 [M+H] + .

[0579] Step 4: Compound 23C (1.50 g) was separated by chiral SFC to afford P1 (retention time: 1.927 min, designated as compound 23C-1) and P2 (retention time: 2.093 min, designated as compound 23C-2). Separation method: Instrument: Waters 150 SFC; Column: Chiralcel OD-Column; Mobile phases: A for CO2 and B for MeOH (0.1% NH3·H2O); Gradient: 35% phase B isocratic elution; Flow rate: 100 mL / min; Column pressure: 100 bar; Column temperature: 25°C; Absorption wavelength: 220 nm; Cycle time: 5 min. Sample preparation: The compound was dissolved in methanol at a concentration of 10 mg / mL; injection: 3.5 mL was injected each time; treatment: after separation, it was concentrated by rotary evaporator at 35°C, and then the solvent was dried by freeze dryer at -80°C to obtain compound 23C-1 (211 mg, 14.1%) and compound 23C-2 (121 mg, 8.1%).

[0580] Step 5: Dissolve compound 23C-1 (211 mg) in 20% trifluoroacetic acid in dichloromethane (5 mL) and stir at room temperature for 2 hours. Concentrate directly to dryness, add methanol solution (15 mL) and solid sodium bicarbonate, and adjust the pH to alkaline. The reaction mixture is directly separated by reverse-phase column chromatography (acetonitrile:water (v / v) = 5% to 50%) to obtain compound 23 (123 mg, yield: 76.5%).

[0581] Compound 23: 1 H NMR(400MHz, CDCl3)δ8.14(d,1H),7.90(s,1H),7.85(d,1H),7.46(s,1H),7.15(d,1H),6.98-6 .88(m,1H),5.94(d,1H),5.63(d,1H),3.93(s,2H),3.82(s,3H); LC-MS(ESI):m / z=435.5[M+H] + .

[0582] Compound 23C-2 (121 mg) was dissolved in 20% trifluoroacetic acid in dichloromethane (5 mL) and stirred at room temperature for 2 hours. The mixture was concentrated to dryness, and methanol (15 mL) and solid sodium bicarbonate were added to adjust the pH to alkaline. The reaction mixture was directly separated by reverse-phase column chromatography (acetonitrile:water (v / v) = 5% to 50%) to obtain compound 24 (19 mg, yield: 19.9%).

[0583] Compound 24: 1H NMR(400MHz, CDCl3)δ8.14(d,1H),7.91(s,1H),7.85(d,1H),7.46(s,1H),7.15(d,1H),6.96-6 .92(m,1H),5.94(d,1H),5.63(d,1H),3.94(s,2H),3.83(s,3H); LC-MS(ESI):m / z=435.5[M+H] + .

[0584] Examples 25 and 26

[0585] (2S)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-(3,3-difluoroazetidin-1-yl)-3-fluorobenzonitrile and (2R)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-(3,3-difluoroazetidin-1-yl)-3-fluorobenzonitrile (Compound 25, Compound 26)

[0586] (2S)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-(3,3-difluoroazetidin-1-yl)-3-fluorobenzonitrile and(2R)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydroquinazolin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-(3,3-difluoroazetidin-1-yl)-3-fluorobenzonitrile(compound 25, compound 26)

[0587] Step 1: Dissolve compound 8C (3.8 g, 10.01 mmol), 3,3-difluoroazetidine hydrochloride (1.56 g, 12 mmol), and potassium carbonate (2.77 g, 20 mmol) in dimethyl sulfoxide (40 mL) and stir at 80°C for 4 hours. After cooling, extract with ethyl acetate (100 mL) and water (100 mL). The organic phase is collected and concentrated to dryness. The residue is separated by silica gel column chromatography (ethyl acetate:petroleum ether (v:v) = 0-25%) to obtain compound 25A (2.5 g, yield: 55.2%).

[0588] LC-MS (ESI): m / z = 453.2 [M+H] + .

[0589] Step 2: Compound 14A (0.64 g, 1.77 mmol), compound 25A (800 mg, 1.77 mmol), and sodium bicarbonate (0.45 g, 5.31 mmol) were added to a mixed solution of 1,4-dioxane (90 mL) and water (30 mL). After nitrogen replacement three times, [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (0.13 g, 0.18 mmol) was added. After nitrogen replacement again, the temperature was raised to 60°C and the reaction was allowed to react for 2 hours. After cooling, the mixture was directly filtered. Ethyl acetate (300 mL) and water (300 mL) were added to the filtrate for separation. The organic phase was washed with saturated brine (300 mL × 3), dried over anhydrous sodium sulfate, and concentrated to dryness to obtain compound 25B (389 mg crude product), which was used directly in the next reaction.

[0590] LC-MS (ESI): m / z = 600.2 [M+H] + .

[0591] Step 3: Compound 25B (380 mg) was separated by chiral SFC to afford P1 (retention time: 1.284 min, designated as compound 25B-1) and P2 (retention time: 1.335 min, designated as compound 25B-2). Separation method: Instrument: Waters 150 SFC; Column: Chiralpak AS Column; Mobile phases: A for CO2 and B for EtOH (0.1% NH3·H2O); Gradient: 15% phase B isocratic elution; Flow rate: 100 mL / min; Column pressure: 100 bar; Column temperature: 25°C; Absorption wavelength: 220 nm; Cycle time: 2.5 min. Sample preparation: The compound was dissolved in ethanol at a concentration of 8.0 mg / mL; injection: 2 mL was injected each time; treatment: after separation, it was concentrated by rotary evaporation at 35°C, and then the solvent was dried by freeze dryer at -80°C to obtain compound 25B-1 (49 mg, 12.9%) and compound 25B-2 (41 mg, 10.8%).

[0592] Step 4: Dissolve compound 25B-1 (49 mg) in 20% trifluoroacetic acid in dichloromethane (3 mL) and stir at room temperature for 2 hours. Concentrate directly to dryness, add methanol (15 mL) and solid sodium bicarbonate to adjust the pH to alkaline. The reaction mixture is directly separated by reverse-phase column chromatography (acetonitrile:water (v / v) = 5% to 50%) to obtain compound 25 (11 mg, yield: 26.9%).

[0593] Compound 25: 1 H NMR(400MHz,DMSO-d6)δ8.16(s,1H),7.99(d,1H),7.40(d,1H),7.32–7.23(m,2 H),4.77-4.55(m,4H),3.76(s,3H),3.60(s,2H); LC-MS(ESI):m / z=500.5[M+H] + .

[0594] Compound 25B-2 (41 mg) was dissolved in 20% trifluoroacetic acid in dichloromethane (3 mL) and stirred at room temperature for 2 hours. The mixture was concentrated to dryness, and a methanol solution (15 mL) was added. Solid sodium bicarbonate was added to adjust the pH to alkaline. The reaction mixture was directly separated by reverse-phase column chromatography (acetonitrile:water (v / v) = 5% to 50%) to provide compound 26 (13 mg, yield: 19.9%).

[0595] Compound 26: 1 H NMR(400MHz,DMSO-d6)δ8.16(s,1H),7.99(d,1H),7.40(d,1H),7.32–7.25(m,2 H),4.71-4.55(m,5H),3.76(s,4H),3.60(s,2H); LC-MS(ESI):m / z=500.5[M+H] + .

[0596] Example 27

[0597] 6-(5-(3-chloro-6-cyano-5-cyclopropyloxy-2-fluorophenyl)-1-methyl-1H-pyrazol-4-yl)quinoline-3-carboxamide (Compound 27)

[0598] 6-(5-(3-chloro-6-cyano-5-cyclopropoxy-2-fluorophenyl)-1-methyl-1H-pyrazol-4-yl)quinoline-3-carboxamide(compound 27)

[0599] Step 1: Dissolve 6-bromoquinoline-3-carboxamide (250.0 mg, 1.0 mmol), compound 2B (583.0 mg, 1.4 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (73.0 mg, 0.1 mmol), and sodium bicarbonate (385.2 mg, 4.0 mmol) in 1,4-dioxane (10.0 mL) and water (3.0 mL). The atmosphere was purged with nitrogen three times and heated to 80°C for 2 h. The mixture was cooled to room temperature and filtered through a funnel filled with silica gel. The filter cake was rinsed with ethyl acetate (50.0 mL). The filtrate was collected and concentrated. The resulting residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (v:v) = 1:1) to provide compound 27 (690.0 mg, yield: 84%).

[0600] 1 H NMR(400MHz,DMSO-d6)δ9.23(d,1H),8.70(d,1H),8.27(s,1H),8.15(s,1H),8.07–7.94(m,2H),7.8 1(d,1H),7.75–7.59(m,2H),4.26–4.11(m,1H),3.78(s,3H),0.91–0.89(m,2H),0.85–0.74(m,2H);

[0601] LC-MS (ESI): m / z = 462.1 [M+H] + .

[0602] Example 28

[0603] 2-(4-(2-(Aminomethyl)-4-oxo-3,4-dihydropyrido[2,3-d]pyrimidin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(1-methylcyclopropyloxy)benzonitrile (Compound 28)

[0604] 2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydropyrido[2,3-d]pyrimidin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(1-methylcyclopropoxy)benzonitrile(compound 28)

[0605] Step 1: Compound 14B (2 g, 4.63 mmol), pinacol diboronate (2.96 g, 23.17 mmol), triethylamine (2.81 g, 27.77 mmol), and tetrakis(triphenylphosphine)palladium (0.54 g, 0.46 mmol) were dissolved in 1,4-dioxane (25 mL). The mixture was then purged with nitrogen three times and maintained under a nitrogen atmosphere. The mixture was heated to 90°C and stirred for 12 hours. After cooling, the reaction mixture was diluted with water and extracted three times with ethyl acetate. The organic phase was collected, dried, and concentrated. The resulting residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether (v:v) = 10-13%) to provide compound 28A (1.25 g, yield: 63%).

[0606] LC-MS (ESI): m / z = 432.20 [M+H] + .

[0607] Step 2: Sodium metal (0.27 g, 11.7 mmol) was added portionwise to methanol (50 mL). After complete dissolution, 2-chloroacetonitrile (13.1 g, 173.5 mmol) was added, followed by stirring at room temperature for 40 minutes. 2-Amino-6-chloropyridine-3-carboxylic acid (Compound 28B) (10 g, 57.9 mmol) was then dissolved in methanol (50 mL) and added to the reaction system. The mixture was reacted at room temperature for 2 hours. The reaction solution was directly concentrated, and the resulting residue was purified by silica gel column chromatography (methanol:dichloromethane (v:v) = 3%) to afford Compound 28C (3.6 g, yield: 27%).

[0608] LC-MS (ESI): m / z = 230.1 [M+H] + .

[0609] Step 3: Compound 28C (2.2 g, 9.56 mmol) was dissolved in aqueous ammonia (20 mL) and stirred at room temperature for 2 hours until dissolved. The reaction solution was directly concentrated, and the resulting crude product was dissolved in methanol (15 mL). Triethylamine (3.87 g, 38.24 mmol) and di-tert-butyl dicarbonate (3.13 g, 14.3 mmol) were then added, and stirring was continued at room temperature for 3 hours. The reaction solution was directly concentrated, and the resulting residue was purified by silica gel column chromatography (methanol:dichloromethane (v:v) = 3-6%) to obtain compound 28D (2.8 g, yield: 94%).

[0610] LC-MS (ESI): m / z = 311.3 [M+H] + .

[0611] Step 4: Compound 28A (0.4 g, 0.93 mmol), compound 28D (0.40 g, 1.30 mmol), potassium carbonate (0.32 g, 2.33 mmol), and [1,1-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (0.068 g, 0.093 mmol) were dissolved in 1,4-dioxane (10 mL) and water (2.5 mL). The mixture was then replaced with nitrogen three times and maintained under a nitrogen atmosphere. The mixture was heated to 100°C and stirred for 2 hours. After cooling, the reaction mixture was diluted with water and extracted three times with ethyl acetate. The organic phase was collected, dried, and concentrated. The resulting residue was purified by silica gel column chromatography (methanol:dichloromethane (v:v) = 10-12%) to provide compound 28E (0.52 g, yield: 96%).

[0612] LC-MS (ESI): m / z = 580.2 [M+H] + .

[0613] Step 5: Dissolve compound 28E (0.15 g, 0.26 mmol) in dichloromethane (4 mL), then add trifluoroacetic acid (1 mL) and stir at room temperature for 2 hours. Concentrate directly, and the resulting residue is purified by preparative HPLC. Method: 1. Apparatus: Waters 2767 preparative liquid chromatography; Column: SunFire@PrepC18 (19 mm × 250 mm). 2. Filter the sample through a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Mobile phase A and B composition: Mobile phase A: acetonitrile; Mobile phase B: water (containing 0.1% ammonium acetate); b. Gradient elution, mobile phase A content from 30% to 80%; c. Flow rate: 15 mL / min; d. Elution time: 40 min. Retention time: 30 min. Compound 28 (14 mg, yield: 11%) was obtained.

[0614] 1 H NMR (400MHz, DMSO-d6) δ8.43(s,1H),8.36(d,1H),7.78(d,1H),7.67(d,1H),3.78(s,3H),3.58(s,2H),1.62(s,3H),1.03(dd,2H),0.92(s,2H).

[0615] LC-MS (ESI): m / z = 480.1 [M+H] + .

[0616] Example 29 and Example 30

[0617] (2S)-2-(4-(4-(aminomethyl)-1-oxo-8-vinyl-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(1-methylcyclopropyloxy)benzonitrile and (2R)-2-(4-(4-(aminomethyl)-1-oxo-8-vinyl-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(1-methylcyclopropyloxy)benzonitrile (Compound 29, Compound 30)

[0618] (2S)-2-(4-(4-(aminomethyl)-1-oxo-8-vinyl-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(1-methylcyclopropoxy)benzonitrile and(2R)-2-(4-(4-(aminomethyl)-1-oxo-8-vinyl-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(1-methylcyclopropoxy)benzonitrile(compound 29, compound 30)

[0619] Step 1: Compound 20C (1.0 g, 2.63 mmol), compound 28A (1.13 g, 2.63 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (190 mg, 0.26 mmol), and potassium carbonate (1.09 g, 7.89 mmol) were dissolved in a mixture of 1,4-dioxane and water (v:v = 6:1, 70 mL). The atmosphere was replaced with nitrogen three times and the mixture was reacted at 85°C for 6 hours. After cooling, the mixture was directly concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (v:v) = 4:1 to 1:1) to obtain compound 29A (690 mg, yield: 43.40%).

[0620] LC-MS (ESI): m / z = 605.7 [M+H] + .

[0621] Step 2: Compound 29A (0.69 g, 1.14 mmol) was separated by SFC to afford P1 (retention time 1.142 min, designated as compound 29A-1) and P2 (retention time 1.195 min, designated as compound 29A-2). Method: Instrument: Waters 150 SFC, Column: Chiralcel AS Column (250 × 30 mm × 10 μm), Mobile Phase: A for CO2; B for 0.1% NH3·H2O in MeOH, Gradient: B 15%, Flow Rate: 100 mL / min, Column Pressure: 100 bar, Column Temperature: 25°C, Absorption Wavelength: 220 nm, Cycle Time: 6.3 min. Sample preparation: The compound was dissolved in acetonitrile at a concentration of 10 mg / mL; injection: 3 mL was injected each time; treatment: after separation, it was concentrated by rotary evaporation at 35°C, and then the solvent was dried by freeze dryer at -80°C to obtain compounds 29A-1 (92.8 mg, 13.45%) and 29A-2 (66.9 mg, 9.96%).

[0622] Compound 29A-1: LC-MS (ESI): m / z=605.7 [M+H] + .

[0623] Compound 29A-2: LC-MS (ESI): m / z=605.7 [M+H] + .

[0624] Step 3: Compound 29A-1 (92.8 mg, 0.15 mmol) was dissolved in dichloromethane (8 mL), followed by the addition of trifluoroacetic acid (1.6 mL) and the reaction mixture was allowed to react at room temperature for 2 hours. The reaction mixture was then concentrated, and the residue was dissolved in dichloromethane (50 mL), then adjusted to alkalinity with saturated sodium bicarbonate solution (50 mL). The mixture was allowed to stand for separation, and the dichloromethane phase was separated. The aqueous phase was extracted with dichloromethane (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and the filtrate was collected by filtration and concentrated. The resulting residue was purified by silica gel column chromatography (dichloromethane:methanol (v:v) = 20:1) to afford compound 29 (75 mg, yield: 96.87%).

[0625] Compound 29: 1 H NMR(400MHz,DMSO-d6)δ12.36(s,1H),8.39(s,1H),8.13(dd,1H),7.85(d,1H),7.71(s,1H),7.61(s, 1H),5.38(d,1H),5.31(d,1H),3.82–3.77(m,5H),1.59(s,3H),1.05–1.00(m,2H),0.95–0.90(m,2H);

[0626] LC-MS (ESI): m / z = 505.9 [M+H] + .

[0627] Compound 29A-2 (66.9 mg, 0.11 mmol) was dissolved in dichloromethane (6 mL), followed by the addition of trifluoroacetic acid (1.2 mL) and the reaction was allowed to react at room temperature for 2 hours. The reaction system was then concentrated, and the residue was dissolved in dichloromethane (50 mL) and adjusted to alkalinity with saturated sodium bicarbonate solution (50 mL). The mixture was allowed to stand for separation, and the dichloromethane phase was separated. The aqueous phase was extracted with dichloromethane (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and the filtrate was collected by filtration and concentrated. The resulting residue was purified by silica gel column chromatography (dichloromethane:methanol (v:v) = 20:1) to obtain compound 30 (43 mg, yield: 77.25%).

[0628] Compound 30: 1 H NMR(400MHz,DMSO-d6)δ12.35(s,1H),8.39(s,1H),8.13(dd,1H),7.85(d,1H),7.71(s,1H),7.61(s,1 H),5.38(d,1H),5.31(d,1H),3.81–3.77(m,5H),1.59(s,3H),0.94–0.91(m,,2H),0.87–0.83(m,2H);

[0629] LC-MS (ESI): m / z = 505.9 [M+H] + .

[0630] Example 31 and Example 32

[0631] (2S)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydropyrido[2,3-d]pyrimidin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(1-methylcyclopropyloxy)benzonitrile and (2R)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydropyrido[2,3-d]pyrimidin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(1-methylcyclopropyloxy)benzonitrile (Compound 31, Compound 32)

[0632] (2S)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydropyrido[2,3-d]pyrimidin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(1-methylcyclopropoxy)benzonitrile and(2R)-2-(4-(2-(aminomethyl)-4-oxo-3,4-dihydropyrido[2,3-d]pyrimidin-7-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(1-methylcyclopropoxy)benzonitrile(compound 31, compound 32)

[0633] Step 1: Compound 28E (1.6 g, 2.76 mmol) was separated by chiral SFC to afford P1 (retention time: 2.590 min, designated as compound 28E-1) and P2 (retention time: 3.949 min, designated as compound 28E-2). Separation method: Instrument: MGⅡ preparative SFC (SFC-14); Column: ChiralPak IE, 250×30 mm ID, 10 μm; Mobile phase: A for CO2 and B for EtOH; Gradient: 30% phase B; Flow rate: 80 mL / min; Column pressure: 100 bar; Column temperature: 38°C; Absorption wavelength: 220 nm; Cycle time: 2.5 min. Sample preparation: The compound was dissolved in ethanol and injected at a rate of 2 mL each time. Treatment: After separation, the mixture was concentrated by rotary evaporation at 35°C, and then the solvent was dried by freeze drying at -80°C to obtain compound 28E-1 (0.45 g, 28%) and compound 28E-2 (0.53 g, 33%).

[0634] Step 2: Dissolve compound 28E-1 (0.45 g, 0.78 mmol) in dichloromethane (8 mL), then add trifluoroacetic acid (2 mL) and stir at room temperature for 2 hours. Concentrate the resulting residue and purify it by preparative HPLC. Method: 1. Apparatus: Waters 2767 preparative HPLC; Column: SunFire@PrepC18 (19 mm × 250 mm). 2. Filter the sample through a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Mobile phase A and B composition: Mobile phase A: acetonitrile; Mobile phase B: Water (containing 0.5% NH3 / H2O); b. Gradient elution, mobile phase A content ranging from 5% to 60%; c. Flow rate: 20 mL / min; d. Elution time: 15 min. Retention time: 10.0 min. Compound 31 (19 mg, yield: 5%) was obtained.

[0635] Compound 31: 1 H NMR(400MHz,DMSO-d6)δ8.41(s,1H),8.36(d,1H),7.78(d,1H),7.64(d,1H), 3.78(s,3H),3.64(s,2H),1.62(s,3H),1.11-1.00(m,2H),0.98-0.89(m,2H);

[0636] LC-MS (ESI): m / z = 480.5 [M+H] + .

[0637] Compound 28E-2 (0.53 g, 0.91 mmol) was dissolved in dichloromethane (8 mL), followed by the addition of trifluoroacetic acid (2 mL) and stirring at room temperature for 2 hours. The resulting residue was directly concentrated and purified by preparative HPLC. Method: 1. Apparatus: Waters 2767 preparative HPLC; Column: SunFire@PrepC18 (19 mm × 250 mm). 2. The sample was filtered through a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Mobile phase A and B composition: Mobile phase A: acetonitrile; Mobile phase B: Water (containing 0.5% NH3 / H2O); b. Gradient elution, mobile phase A content from 5% to 60%; c. Flow rate: 20 mL / min; d. Elution time: 15 min. Retention time: 10.0 min. Compound 32 (51 mg, yield: 11%) was obtained.

[0638] Compound 32: 1H NMR(400MHz,DMSO-d6)δ8.42(s,1H),8.36(d,1H),7.78(d,1H),7.65(d,1H), 3.78(s,3H),3.63(s,2H),1.62(s,3H),1.11–1.01(m,2H),0.96-0.90(m,2H);

[0639] LC-MS (ESI): m / z = 480.5 [M+H] + .

[0640] Example 33

[0641] 2-(4-(4-Aminomethyl)-8-vinyl-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(1-fluorocyclobutyl)benzonitrile (Compound 33)

[0642] 2-(4-(4-(aminomethyl)-8-ethenyl-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(1-fluorocyclobutyl)benzonitrile(compound 33)

[0643] Step 1: Compound 8C (7 g, 27.6 mmol) was weighed into a flask, and N,N-dimethylformamide (70 mL) and diethyl malonate (8.84 g, 55.20 mmol) were added. Then, sodium hydride (3.31 g, 82.81 mmol) was slowly added. The mixture was heated to 80°C for 15 hours. The reaction mixture was extracted with ethyl acetate, and the organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated. The resulting residue was purified by column chromatography (petroleum ether:ethyl acetate (v / v) = 10:1) to provide compound 33A (4.1 g, yield: 37.7%).

[0644] LC-MS (ESI): m / z = 394.2 [M+H] + .

[0645] Step 2: Compound 33A (4.1 g, 10.43 mmol) was dissolved in dimethyl sulfoxide (40 mL), and lithium chloride (1.33 g, 31.28 mmol) was added. The mixture was heated to 100°C for 15 hours. The reaction solution was extracted with ethyl acetate, and the organic phase was collected, washed with saturated brine, and dried over anhydrous sodium sulfate. The filtrate was collected by filtration and concentrated. The resulting residue was purified by column chromatography (petroleum ether:ethyl acetate (v / v) = 5:1) to provide compound 33B (2 g, yield: 59.6%).

[0646] LC-MS (ESI): m / z = 322.3 [M+H] + .

[0647] Step 3: Compound 33B (1.7 g, 5.28 mmol) was dissolved in dimethyl sulfoxide (10 mL), and diphenyl(vinyl)sulfonium trifluoromethanesulfonate (2.3 g, 6.34 mmol) was added. The mixture was reacted at 25°C for 5 minutes. 1,8-diazabicyclo[5.4.0]undec-7-ene (2.4 g, 15.84 mmol) was then added and the mixture was reacted at 25°C for 2 hours. The reaction solution was extracted with ethyl acetate, and the organic phase was collected, washed with saturated brine, and dried over anhydrous sodium sulfate. The filtrate was collected by filtration and concentrated. The resulting residue was purified by column chromatography (petroleum ether:ethyl acetate (v / v) = 5:1) to afford compound 33C (1.6 g, yield: 87%).

[0648] LC-MS (ESI): m / z = 348.2 [M+H] + .

[0649] Step 4: Compound 33C (1.1 g, 3.16 mmol) was weighed into a flask, tetrahydrofuran (10 mL) was added, and lithium borohydride (137.8 mg, 6.33 mmol) was added. The mixture was stirred at 25°C, and then water (1 mL) was slowly added. The system was reacted at 25°C for 15 hours. The reaction solution was extracted with ethyl acetate, and the organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the resulting residue was purified by column chromatography (petroleum ether:ethyl acetate (v / v) = 5:1) to obtain compound 33D (890 mg, yield: 92%).

[0650] LC-MS (ESI): m / z = 306.1 [M+H] + .

[0651] Step 5: Compound 33D (300 mg, 0.98 mmol) was dissolved in dichloromethane (10 mL), cooled to -78°C, and diethylaminosulfur trifluoride (315.9 mg, 1.96 mmol) was added with stirring. The mixture was slowly warmed to room temperature and stirred for 2 hours. The reaction solution was slowly added to saturated aqueous sodium bicarbonate (20 mL) and extracted with dichloromethane. The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The resulting residue was purified by column chromatography (petroleum ether:ethyl acetate (v / v) = 8:1) to provide compound 33E (240 mg, yield: 79.7%).

[0652] LC-MS (ESI): m / z = 308.2 [M+H] + .

[0653] Step 6: Compound 33E (0.22 g, 0.71 mmol), N-iodosuccinimide (0.24 g, 1.06 mmol), and acetonitrile (5 mL) were added to a single-necked flask. Trifluoroacetic acid (16 mg, 0.14 mmol) was added, and the mixture was heated to 50°C and stirred for 3 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography (petroleum ether:ethyl acetate (v:v) = 5:1) to provide compound 33F (0.24 g, yield: 78.0%).

[0654] LC-MS (ESI): m / z=434.0[M+H]+.

[0655] Step 7: Compound 33F (0.2 g, 0.46 mmol), pinacol borane (0.35 g, 2.77 mmol), triethylamine (0.28 g, 2.77 mmol), and tetrakis(triphenylphosphine)palladium (0.05 g, 0.05 mmol) were weighed into a flask. 1,4-dioxane (3 mL) was added and the mixture was heated to 90°C under nitrogen for 16 hours. The reaction solution was extracted with ethyl acetate, and the organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the resulting residue was purified by column chromatography (petroleum ether:ethyl acetate (v:v) = 5:1) to provide compound 33G (130 mg, 65.0% yield).

[0656] LC-MS (ESI): m / z = 434.1 [M+H] + .

[0657] Step 8: Compound 33G (0.13 g, 0.30 mmol), compound 20C (0.11 g, 0.30 mmol), sodium bicarbonate (0.06 g, 0.75 mmol), and chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (0.02 g, 0.03 mmol) were dissolved in 1,4-dioxane (3 mL) and water (0.5 mL). The mixture was heated to 90°C under nitrogen and stirred for 12 hours. The reaction mixture was filtered and concentrated under reduced pressure. The residue was purified by column chromatography (petroleum ether:ethyl acetate (v:v) = 1:1) to afford compound 33H (0.07 g, yield: 38.5%).

[0658] LC-MS (ESI): m / z = 607.6 [M+H] + .

[0659] Step 9: Dissolve compound 33H (0.03, 0.05 mmol) in dichloromethane (3 mL), add trifluoroacetic acid (1 mL), and react at room temperature for 1 hour. The reaction solution was directly concentrated, and the resulting residue was purified by preparative HPLC. Method: 1. Apparatus: Waters 2767 preparative liquid chromatography; Chromatographic column: SunFire@PrepC18 (19 mm × 250 mm). 2. Dissolve the sample in methanol and filter through a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Mobile phase A and B composition: Mobile phase A: acetonitrile; Mobile phase B: Water (containing 0.1% TFA); b. Gradient elution, mobile phase A content from 20% to 60%; c. Flow rate: 20 mL / min; d. Elution time: 20 min. Retention time: 6.94 min. Compound 33 (8 mg, yield: 31.9%) was obtained.

[0660] LC-MS (ESI): m / z = 507.5 [M+H] + .

[0661] Biological testing

[0662] 1. PRMT5 and PRMT5·MTA binding assay

[0663] 1.1 PRMT5 enzyme activity test method

[0664] Prepare 1× assay buffer (10 mM Tris, 1 mM DTT, 0.01% BSA, 0.01% Tween-20, adjusted to pH 8.0). Dissolve the test compound in DMSO to a 10 mM stock solution and dilute to a 100-fold final concentration. Add 100 nL of compound to each well in the treatment group, and 100 nL of DMSO to the blank and negative control wells. Then, add 5 μL of PRMT5 / MEP50 (BPS, Cat#31921) enzyme solution (final concentration 1 nM) to the treatment and negative control wells, and 5 μL of 1× assay buffer to the blank wells. Incubate at room temperature for 15 minutes. Add 5 μL of a mixture of H4(1-21)S1ac (0.035 μM; GL Biochem, customized) and SAM (0.434 μM; Sigma, Cat#A7007) to each well and incubate at room temperature for 1 hour. Prepare 1× Epigenetics buffer (PerkinElmer, Cat#AL008F) to dilute the acceptor and donor beads. Add 15 μL of acceptor beads (final concentration 10 μg / mL; PerkinElmer, Cat#AL150C) and donor beads (final concentration 10 μg / mL; PerkinElmer, Cat#AS106M) and incubate at room temperature for 1 hour in the dark. Detection was performed using the Alpha module of an Enspire microplate reader. The resulting data were used to calculate the inhibition rate in Excel: Inhibition rate (%) = (maximum negative control - detection signal value) / (maximum negative control - minimum blank control) × 100. IC values ​​were fitted using XL-Fit. 50 value.

[0665] 1.2 PRMT5·MTA enzyme activity assay

[0666] Prepare 1× assay buffer (10 mM Tris, 1 mM DTT, 0.01% BSA, 0.01% Tween-20, adjusted to pH 8.0). Dissolve the test compound in DMSO to a 10 mM stock solution and dilute to a 100-fold final concentration. Add 100 nL of compound to each well in the treatment group, and 100 nL of DMSO to the blank and negative control wells. Then, add 5 μL of a solution of PRMT5 / MEP50 (BPS, Cat#31921) (final concentration 1 nM) and MTA (final concentration 1 μM) to the treatment and negative control wells, and 5 μL of 1× assay buffer to the blank wells. Incubate at room temperature for 15 minutes. Add 5 μL of a mixture of H4(1-21)S1ac (0.035 μM; GL Biochem, customized) and SAM (0.434 μM; Sigma, Cat#A7007) to each well and incubate at room temperature for 1 hour. Prepare 1× Epigenetics buffer (PerkinElmer, Cat# AL008F) to dilute the acceptor and donor beads. Add 15 μL of acceptor beads (final concentration 10 μg / mL; PerkinElmer, #AL150C) and donor beads (final concentration 10 μg / mL; PerkinElmer, Cat# AS106M) and incubate at room temperature for 1 hour in the dark. Detection is then performed using the Alpha module of an Enspire microplate reader.

[0667] The obtained data were used to calculate the inhibition rate in Excel: inhibition rate (%) = (maximum negative control - detection signal value) / (maximum negative control - minimum blank control) × 100, and IC was fitted using XL-Fit. 50 value.

[0668] The compounds of the present invention have an IC<100 nM for PRMT5·MTA 50 IC values ​​of some compounds 50 The values ​​are shown in Table 2, where AA represents IC 50 <1nM, A represents 1nM≤IC 50 <10nM, B represents 10nM≤IC 50 <50nM, C represents 50nM≤IC 50 <100nM.

[0669] Table 2 Inhibitory activity of compounds against PRMT5·MTA

[0670] Conclusion: The compounds of the present invention, such as the example compounds, have significant inhibitory activity against PRMT5·MTA. Some preferred compounds, such as compound 30, have an IC 50At the same time, in the absence of MTA, the inhibitory activity against PRMT5 is relatively weak, that is, the compounds of the present invention have selective inhibition against PRMT5·MTA.

[0671] 2. MIAPACA-2 cell proliferation inhibition experiment

[0672] MIAPACA-2 cells were cultured in DMEM (ATCC, Cat#30-2002; supplemented with 10% FBS and 1% double-antibody) until the confluence reached approximately 85%, and then plated. The culture medium was discarded, and the cells were rinsed with 1× PBS and digested with trypsin (Gibco, Cat#15400-054). When the cells became round and began to detach, culture medium was added to terminate the digestion. The cells were pipetted off and transferred to a sterile centrifuge tube and centrifuged at 1000 rpm for 3 minutes. After centrifugation, the cells were removed and the supernatant discarded. Culture medium was added to the centrifuge tube to resuspend the cells and count them. Based on the count results, the cell suspension was adjusted to the appropriate concentration and poured into the sample reservoir. The cells were added to the first 96-well cell plate (Corning, Cat#3903). At the same time, 10 Day 0 wells were plated in the second 96-well cell plate. The compounds were dissolved in DMSO to 10 mM and stored for later use. During the experiment, the compound stock solution was serially diluted 5-fold. 24 hours after plating, add the diluted compound to the first 96-well cell plate, set up a DMSO vehicle control group, and continue incubation at 37°C, 5% CO2 for 5 days. After 5 days of incubation, slowly and carefully aspirate the supernatant from the first 96-well cell plate using a multichannel pipette and discard it. Then add TrypLE (Gibco, Cat#12604-013) to each well and place in a 37°C incubator for 5 minutes until the cells are completely detached. Then, add culture medium to terminate the digestion and pipette evenly. Take a third 96-well cell plate (Corning, Cat#3903), add DMEM medium and the digested cell suspension to each well, and mix thoroughly with a pipette. Add the diluted compound and set up a DMSO vehicle control group. Continue incubation at 37°C, 5% CO2 for 5 days. 24 hours after plating, CELL VIABILITY reagent (Vazyme, Cat# DD1101-03) was added to each well of the second 96-well cell plate. CELL VIABILITY reagent was added to each well of the first 96-well cell plate on the 5th day after administration. CELL VIABILITY reagent was added to each well of the third 96-well cell plate on the 10th day after the first administration. The cells were incubated at room temperature for 10 minutes. After the incubation, the cells were gently shaken 5 times, and then the chemiluminescence readings were measured using a microplate reader. The cell proliferation inhibition rate was calculated according to the following formula: [1-(25×T test-T0 / 25×T control-T0)]×100. The GI of the compound for inhibiting cell proliferation was then calculated using the DoseResp function using Origin 9.2 software. 50value.

[0673] Effect of the compounds of the present invention on the GI function of MIAPACA-2 cells 50 Less than 1000nM, some compounds GI 50 Less than 100nM, some compounds GI 50 Less than 50nM.

[0674] The test results of some embodiments are shown in Table 3, where A<50nM, 50nM≤B<100nM, and 100nM≤C<500nM:

[0675] Table 3 Antiproliferation activity of compounds on MIAPACA-2 cells

[0676] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good inhibitory activity against MIAPACA-2 cells.

[0677] 3. HCT-116 cell proliferation inhibition experiment

[0678] HCT-116 cells were cultured in RPMI 1640 medium (ATCC, Cat#30-2001; supplemented with 10% FBS and 1% double-antibody) until the confluence reached approximately 85% before plating. The medium was discarded, and the cells were washed with 1× PBS and then digested with trypsin (Gibco, Cat#15400-054). When the cells became round and began to detach, the medium was added to terminate the digestion. The cells were pipetted off and transferred to a sterile centrifuge tube and centrifuged at 1000 rpm for 3 minutes. After centrifugation, the cells were removed and the supernatant discarded. The centrifuge tube was resuspended in medium and counted. According to the counting results, the cell suspension was adjusted to the appropriate concentration and poured into the sample tank. It was added to the first 96-well cell plate (Corning, Cat#3903). At the same time, 10 Day 0 wells were plated in the second 96-well cell plate. The compound was dissolved in DMSO to 10 mM and stored for later use. During the experiment, the compound stock solution was diluted 5-fold in sequence. 24 hours after plating, add the diluted compound to the first 96-well plate, set up a DMSO vehicle control group, and continue incubation at 37°C, 5% CO2 for 5 days. After 5 days of incubation, slowly and carefully aspirate the supernatant from the first 96-well plate using a multichannel pipette and discard it. Then, add TrypLE (Gibco, Cat#12604-013) to each well and place in a 37°C incubator for 5 minutes until the cells are completely detached. Then, add culture medium to terminate the digestion and pipette evenly. Take a third 96-well plate (Corning, Cat#3903), add RPMI-1640 medium and the digested cell suspension to each well, and mix thoroughly with a pipette. Add the diluted compound and set up a DMSO vehicle control group. Continue incubation at 37°C, 5% CO2 for 5 days. 24 hours after plating, CELL VIABILITY reagent (Vazyme, Cat# DD1101-03) was added to each well of the second 96-well cell plate. CELL VIABILITY reagent was added to each well of the first 96-well cell plate on the 5th day after administration. CELL VIABILITY reagent was added to each well of the third 96-well cell plate on the 10th day after the first administration. The cells were incubated at room temperature for 10 minutes. After the incubation, the cells were gently shaken 5 times, and then the chemiluminescence readings were measured using a microplate reader. The cell proliferation inhibition rate was calculated according to the following formula: [1-(25×T test-T0 / 25×T control-T0)]×100. The GI of the compound for inhibiting cell proliferation was then calculated using the DoseResp function using Origin 9.2 software. 50 value.

[0679] The test results of some embodiments are shown in Table 4, where A>500nM, 100nM≤B<500nM, and C<100nM:

[0680] Table 4 Inhibitory activity of compounds on HCT-116 cell proliferation

[0681] Conclusion: The compounds of the present invention, such as the compounds in the examples, have weak inhibitory activity against HCT-116 cells.

[0682] 4. HCT-116MTAP- / - cell proliferation inhibition experiment

[0683] HCT-116MTAP - / -Cells were cultured in RPMI 1640 medium (ATCC, Cat#30-2001; supplemented with 10% FBS and 1% double antibody) until the confluence reached about 85% before plating. The culture medium was discarded, and the cells were washed with 1×PBS and then digested with trypsin (Gibco, Cat#15400-054). When the cells became round and began to fall off, the culture medium was added to terminate the digestion. The cells were blown off and transferred to a sterile centrifuge tube and centrifuged at 1000 rpm for 3 minutes. After centrifugation, the cells were removed and the supernatant was discarded. The culture medium was added to the centrifuge tube to resuspend the cells and count them. According to the counting results, the cell suspension was adjusted to the appropriate concentration and poured into the sample tank. It was added to the first 96-well cell plate (Corning, Cat#3903). At the same time, 10 Day 0 wells were plated in the second 96-well cell plate. The compound was dissolved in DMSO to 10 mM and stored for later use. During the experiment, the compound stock solution was diluted 5-fold in sequence. 24 hours after plating, add the diluted compound to the first 96-well plate, set up a DMSO vehicle control group, and continue incubation at 37°C, 5% CO2 for 5 days. After 5 days of incubation, slowly and carefully aspirate the supernatant from the first 96-well plate using a multichannel pipette and discard. Add TrypLE (Gibco, Cat#12604-013) to each well and place in a 37°C incubator for 5 minutes until the cells are completely detached. Then, add culture medium to terminate digestion and pipette evenly. Take a third 96-well plate (Corning, Cat#3903), add RPMI-1640 medium and the digested original cell suspension to each well, and mix thoroughly with a pipette. Add the diluted compound, set up a DMSO vehicle control group, and continue incubation at 37°C, 5% CO2 for 5 days. 24 hours after plating, CELL VIABILITY reagent (Vazyme, Cat# DD1101-03) was added to each well of the second 96-well cell plate. CELL VIABILITY reagent was added to each well of the first 96-well cell plate on the 5th day after administration. CELL VIABILITY reagent was added to each well of the third 96-well cell plate on the 10th day after the first administration. The cells were incubated at room temperature for 10 minutes. After the incubation, the cells were gently shaken 5 times, and then the chemiluminescence readings were measured using a microplate reader. The cell proliferation inhibition rate was calculated according to the following formula: [1-(25×T test-T0 / 25×T control-T0)]×100. The GI of the compound for inhibiting cell proliferation was then calculated using the DoseResp function using Origin 9.2 software. 50 value.

[0684] Table 5 Proliferation inhibition activity of compounds on HCT-116MTAP- / - cells

[0685] Conclusion: The compounds of the present invention, such as the compounds in the examples, have an effect on HCT-116MTAP.- / - Strong cell inhibitory activity.

[0686] 5. Cell sDMA detection test

[0687] 5.1. Detection of sDMA in MIAPACA-2 cells

[0688] MIAPACA-2 cells were cultured in DMEM (ATCC, Cat#30-2002; supplemented with 10% FBS and 1% double-antibody) until confluence reached approximately 85% before plating. The cells were discarded, rinsed with 1× PBS, and then digested with trypsin (Gibco, Cat#15400-054). When the cells became round and began to detach, culture medium was added to terminate digestion. The cells were pipetted off and transferred to a sterile centrifuge tube. Centrifuged at 1000 rpm for 3 minutes. After centrifugation, the supernatant was removed and discarded. Culture medium was added to the centrifuge tube to resuspend the cells and count them. Based on the count results, the cell suspension was adjusted to the appropriate concentration and poured into the sample well. 100 μL was added to each well of a 96-well plate. The compound was dissolved in DMSO to 10 mM and stored until ready for use. During the experiment, the compound was serially diluted 5-fold in culture medium. 100 μL of the diluted compound was added to the 96-well plate with the cells. A DMSO vehicle control group was also set up. The cells were cultured at 37°C and 5% CO2 for 3 days. After 3 days, each well was washed twice with cold PBS. Protease and phosphatase inhibitor cocktail (CST, Cat#5872S) and RIPA lysis buffer were mixed in the required volume and added to a 96-well plate at 50 μL / well. The plates were placed on ice for 10 minutes, with repeated pipetting until the cells were completely lysed. The plates were then centrifuged at 2500 rpm at 4°C for 35 minutes, and the supernatant protein samples were collected. Protein detection was performed using a BCA kit (Beyotime, Cat#P0011). Based on the assay results, the protein concentration of the lysate was adjusted to the desired concentration using PBS carbonate-bicarbonate buffer, pH 9.6 (Sigma, Cat#C3041). 100 μL was then added to each well of a black high-binding 96-well plate (Nunc Maxisorp, Cat#437111) and incubated at room temperature for 2 hours. The lysate was removed and the plate was washed four times with PBST (Cell Signaling, Cat#9809S). Blocking was performed with 5% BSA blocking buffer (solarbio, Cat#SW3015) for 1 hour, followed by washing four times with PBST. 100 μL of either sDMA antibody (CST, Cat#13222S) or SMD3 antibody (Abgent, Cat#AP12451A) was added to each well and shaken overnight at 4°C. After removing the antibody, the plate was washed four times with PBST. 100 μL of rabbit secondary antibody (Cell Signaling Technology, Cat#7074) was added to each well and incubated at room temperature for 1 hour. After removing the secondary antibody, the plate was washed four times with PBST. 100 μL of Immobilon Forte Western HRP substrate (Millipore, Cat#WBLUF0500) was added to each well and incubated with shaking for 15 minutes before detection using chemiluminescence on a BMG microplate reader.After data normalization (SDMA reading / SMD3 reading), Graphpad Prism 8.3.0 software was used to plot the data and calculate the IC. 50 value.

[0689] 5.2. SDMA Detection in HCT-116 Cells

[0690] HCT-116 cells were cultured in RPMI 1640 medium (ATCC, Cat#30-2001, supplemented with 10% FBS and 1% double antibody) until the confluence reached about 85% and then plated. Discard the medium, rinse with 1×PBS, and then add trypsin (Gibco, Cat#15400-054) for digestion. When the cells become round and begin to fall off, add medium to terminate digestion. The cells were blown down and transferred to a sterile centrifuge tube and centrifuged at 1000 rpm for 3 minutes. After centrifugation, the cells were removed and the supernatant was discarded. The centrifuge tube was added with medium to resuspend the cells and counted. According to the counting results, the cell suspension was adjusted to the appropriate concentration and poured into the sample tank. 100 μL was added to each well of the 96-well plate. The compound was dissolved in DMSO to 10 mM and stored for later use. During the experiment, the compound was diluted 5-fold in sequence with culture medium. Add 100 μL of diluted compound to a 96-well plate containing cells. A DMSO vehicle control group was also established and cultured at 37°C, 5% CO2 for 3 days. After 3 days, each well was washed twice with cold PBS. Protease and phosphatase inhibitor cocktail (CST, Cat#5872S) and RIPA lysis buffer were mixed to the required volume and added 50 μL / well to the 96-well plate. The plate was incubated on ice for 10 minutes, with repeated pipetting until the cells were completely lysed. The supernatant was then centrifuged at 2500 rpm at 4°C for 35 minutes, and the supernatant protein sample was collected. Protein detection was performed using a BCA kit (Beyotime, Cat#P0011). Based on the assay results, the protein concentration of the lysate was adjusted to the desired concentration using PBS carbonate-bicarbonate buffer, pH 9.6 (Sigma, Cat#C3041). 100 μL was then added to each well of a black high-binding 96-well plate (Nunc Maxisorp, Cat#437111) and incubated at room temperature for 2 hours. The lysate was removed and the plate was washed four times with PBST (Cell Signaling, Cat#9809S). Blocking was performed with 5% BSA blocking buffer (solarbio, Cat#SW3015) for 1 hour, followed by washing four times with PBST. 100 μL of either sDMA antibody (CST, Cat#13222S) or SMD3 antibody (Abgent, Cat#AP12451A) was added to each well and shaken overnight at 4°C. After removing the antibody, the plate was washed four times with PBST. 100 μL of rabbit secondary antibody (Cell Signaling Technology, Cat#7074) was added to each well and incubated at room temperature for 1 hour. After removing the secondary antibody, the plate was washed four times with PBST. 100 μL of Immobilon Forte Western HRP substrate (Millipore, Cat#WBLUF0500) was added to each well and incubated with shaking for 15 minutes before detection using chemiluminescence on a BMG microplate reader.After data normalization (SDMA reading / SMD3 reading), Graphpad Prism 8.3.0 software was used to plot the data and calculate the IC. 50 value.

[0691] Table 6 Inhibitory activity of compounds on SDMA production in HCT-116 cells

[0692] Conclusion: The compounds of the present invention, such as the compounds in the examples, have weak inhibitory activity on SDMA production in HCT-116 cells.

[0693] 5.3 HCT-116MTAP - / - SDMA detection in cells

[0694] HCT-116MTAP- / - cells were cultured with RPMI 1640 medium (ATCC, #30-2001, supplemented with 10% FBS and 1% double antibody) until the confluence reached about 85% and then plated. Discard the culture medium, rinse with 1×PBS, and then add trypsin (Gibco, Cat#15400-054) for digestion. When the cells become round and begin to fall off, add culture medium to terminate digestion, blow down the cells and transfer them to a sterile centrifuge tube, centrifuge at 1000rpm for 3 minutes, remove the cells after centrifugation, and discard the supernatant. Add culture medium to the centrifuge tube to resuspend the cells and count them. According to the counting results, adjust the cell suspension to the appropriate concentration and pour it into the sample tank. Add 100 μL to each well of the 96-well plate. Dissolve the compound to 10mM with DMSO and save it for later use. During the experiment, dilute the compound in a 5-fold gradient with culture medium. Add 100 μL of diluted compound to a 96-well plate containing cells. A DMSO vehicle control group was also established and cultured at 37°C, 5% CO2 for 3 days. After 3 days, each well was washed twice with cold PBS. Protease and phosphatase inhibitor cocktail (CST, Cat#5872S) and RIPA lysis buffer were mixed to the required volume and added 50 μL / well to the 96-well plate. The plate was incubated on ice for 10 minutes, with repeated pipetting until the cells were completely lysed. The supernatant was then centrifuged at 2500 rpm at 4°C for 35 minutes, and the supernatant protein sample was collected. Protein detection was performed using a BCA kit (Beyotime, Cat#P0011). Based on the assay results, the protein concentration of the lysate was adjusted to the desired concentration using PBS carbonate-bicarbonate buffer, pH 9.6 (Sigma, Cat#C3041). 100 μL was then added to each well of a black high-binding 96-well plate (Nunc Maxisorp, Cat#437111) and incubated at room temperature for 2 hours. The lysate was removed and the plate was washed four times with PBST (Cell Signaling, Cat#9809S). Blocking was performed with 5% BSA blocking buffer (solarbio, Cat#SW3015) for 1 hour, followed by washing four times with PBST. 100 μL of either sDMA antibody (CST, Cat#13222S) or SMD3 antibody (Abgent, Cat#AP12451A) was added to each well and shaken overnight at 4°C. After removing the antibody, the plate was washed four times with PBST. 100 μL of rabbit secondary antibody (Cell Signaling Technology, Cat#7074) was added to each well and incubated at room temperature for 1 hour. After removing the secondary antibody, the plate was washed four times with PBST. 100 μL of Immobilon Forte Western HRP substrate (Millipore, Cat#WBLUF0500) was added to each well and incubated with shaking for 15 minutes before detection using chemiluminescence on a BMG microplate reader.After data normalization (SDMA reading / SMD3 reading), Graphpad Prism 8.3.0 software was used to plot the data and calculate the IC. 50 value.

[0695] Table 7 Inhibitory activity of compounds on SDMA production in HCT-116MTAP- / - cells

[0696] Conclusion: The compounds of the present invention, such as the compounds in the examples, have an effect on HCT-116MTAP. - / - The cell-generated SDMA has a strong inhibitory activity.

[0697] 6. hERG potassium channel effect test

[0698] Experimental platform: electrophysiology manual patch clamp system

[0699] Cell line: Chinese hamster ovary (CHO) cell line stably expressing hERG potassium channel

[0700] Experimental methods: CHO (Chinese Hamster Ovary) cells stably expressing hERG potassium channels were used to record hERG potassium channel currents using the whole-cell patch clamp technique at room temperature. Glass microelectrodes were pulled from glass electrode blanks (BF150-86-10, Sutter) using a puller. The tip resistance after perfusing the electrode liquid was about 2-5 MΩ. The glass microelectrode was inserted into the amplifier probe to connect to the patch clamp amplifier. The clamping voltage and data recording were controlled and recorded by a computer using pClamp 10 software, with a sampling frequency of 10 kHz and a filter frequency of 2 kHz. After obtaining the whole-cell recording, the cell was clamped at -80 mV to induce the hERG potassium current (I hERG ) was administered with a 2-second depolarization step from -80 mV to +20 mV, followed by repolarization to -50 mV for 1 second before returning to -80 mV. This voltage stimulus was administered every 10 seconds, and administration began after confirming that the hERG potassium current was stable (for at least 1 minute). Compounds were administered for at least 1 minute at each test concentration, and at least two cells (n ≥ 2) were tested for each concentration.

[0701] Data processing: Data analysis was performed using pClamp 10, GraphPad Prism 5, and Excel software. The degree of inhibition of hERG potassium current (peak hERG tail current induced at -50 mV) by different compound concentrations was calculated using the following formula: Inhibition% = [1–(I / Io)] × 100%

[0702] Wherein, Inhibition% represents the inhibition percentage of the compound on hERG potassium current, and I and Io represent the amplitude of hERG potassium current before and after drug addition, respectively.

[0703] Compound IC 50 Calculated using GraphPad Prism 5 software by fitting the following equation: Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC50-X)×HillSlope))

[0704] Among them, X is the Log value of the test sample concentration, Y is the inhibition percentage at the corresponding concentration, and Bottom

[0705] and Top are the minimum and maximum inhibition percentages, respectively.

[0706] 7. CYP3A4 enzyme inhibition test

[0707] The purpose of this study was to evaluate the effects of test substances on the activity of the CYP3A4 enzyme of cytochrome P450 (CYP) in human liver microsomes using an in vitro test system. A specific probe substrate for the CYP3A4 enzyme was incubated with human liver microsomes and different concentrations of the test substance. Reduced nicotinamide adenine dinucleotide phosphate (NADPH) was added to initiate the reaction. After the reaction, the sample was processed and the metabolites produced by the specific substrate were quantitatively detected by liquid chromatography-tandem mass spectrometry (LC-MS / MS). Changes in CYP enzyme activity were measured and the IC values ​​were calculated. 50 The inhibitory potential of the test substance on each CYP3A4 enzyme was evaluated by the incubation concentration of 0 to 30 μM under the test conditions.

[0708] Conclusion: The compounds of the present invention, such as the compounds in the examples, have weak inhibition on the CYP3A4 enzyme.

[0709] 8. Pharmacokinetic test in mice

[0710] 8.1. Experimental Animals: Male C57 mice, 22-25 g, 6 mice per compound, purchased from Chengdu Dashuo Experimental Animal Co., Ltd.

[0711] 8.2 Experimental Design: On the day of the experiment, (6 × number of test compounds n) C57 mice were randomly divided into groups according to body weight. They were fasted but not watered for 12-14 hours before administration and fed 4 hours after administration.

[0712] Before and after drug administration, 0.06 mL of blood was collected via the orbital cavity under isoflurane anesthesia. The blood was placed in an EDTAK2 centrifuge tube and centrifuged at 5000 rpm at 4°C for 10 minutes to collect plasma. Blood was collected at 0, 5, 15, 30 minutes, 1, 2, 4, 7, and 24 hours for both the intravenous and oral gavage groups. All samples were stored at -80°C prior to analysis and quantitative analysis was performed using LC-MS / MS.

[0713] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good pharmacokinetic characteristics in mice.

[0714] 9. Pharmacokinetic test in rats

[0715] 9.1. Experimental Animals: Male SD rats, approximately 220 g, 6-8 weeks old, 6 rats per compound, purchased from Chengdu Dashuo Experimental Animal Co., Ltd.

[0716] 9.2 Experimental Design: On the day of the experiment, (6 × number of test compounds n) SD rats were randomly divided into groups according to body weight. They were fasted but not watered for 12-14 hours before administration and fed 4 hours after administration.

[0717] Before and after drug administration, 0.15 mL of blood was collected via the orbital cavity under isoflurane anesthesia. The blood was placed in an EDTAK2 centrifuge tube and centrifuged at 5000 rpm at 4°C for 10 minutes to collect plasma. Blood was collected from both the intravenous and oral gavage groups at 0, 5, 15, 30 minutes, and 1, 2, 4, 6, 8, and 24 hours. All samples were stored at -80°C prior to analysis and quantitative analysis was performed using LC-MS / MS.

[0718] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good pharmacokinetic characteristics in rats.

[0719] 10. Beagle dog pharmacokinetic test

[0720] 10.1. Experimental Animals: Male beagle dogs, weighing approximately 8-11 kg, 6 per compound, purchased from Beijing Mas Biotechnology Co., Ltd.

[0721] 10.2 Test Methods: On the day of the test, (6 × number of test compounds) beagle dogs were randomly divided into groups based on body weight. They were fasted (but not watered) for 12-14 hours prior to dosing and fed 4 hours after dosing. Dosing was performed according to Table 8.

[0722] Table 8 Dosage information Note: Intravenous administration solvent: 5% DMA + 5% Solutol + 90% Saline; Oral administration solvent: 0.5% MC

[0723] (DMA: dimethylacetamide; Solutol: polyethylene glycol-15-hydroxystearate; Saline: normal saline; MC: methylcellulose solution)

[0724] Before and after dosing, 1 mL of blood was collected from the jugular vein or limb vein into an EDTAK2 centrifuge tube. The samples were centrifuged at 5000 rpm at 4°C for 10 minutes, and plasma was collected. Blood was collected at the following time points for both the intravenous and oral administration groups: 0, 5, 15, 30 minutes, and 1, 2, 4, 6, 8, 10, 12, and 24 hours. All samples were stored at -80°C prior to analysis and quantitative analysis was performed using LC-MS / MS.

[0725] Table 9 Pharmacokinetic parameters of test compounds in beagle dog plasma -:not applicable.

[0726] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good pharmacokinetic characteristics in beagle dogs.

[0727] 11. Monkey pharmacokinetic test

[0728] 11.1. Experimental Animals: Male cynomolgus monkeys, 3-5 kg, 3-6 years old, 4 per compound. Purchased from Suzhou Xishan Biotechnology Co., Ltd.

[0729] 11.2 Test Method: On the day of the test, (4 × the number of test compounds n) monkeys were randomly divided into groups according to body weight. They were fasted but not watered for 14-18 hours before administration and fed 4 hours after administration.

[0730] Before and after dosing, 1.0 mL of blood was collected from a limb vein and placed in an EDTAK2 centrifuge tube. The samples were centrifuged at 5000 rpm at 4°C for 10 minutes, and plasma was collected. For both the intravenous and oral gavage groups, blood was collected at the following time points: 0, 5, 15, 30 minutes, and 1, 2, 4, 6, 8, 10, 12, 24, 48, and 72 hours. All samples were stored at -80°C prior to analysis and quantitative analysis was performed using LC-MS / MS.

[0731] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good pharmacokinetic characteristics in monkeys.

[0732] 12. In vivo efficacy experiments

[0733] HCT-116MTAP- / - cells were cultured in RPMI-1640 medium at 37°C in a 5% CO2 incubator. Cells were passaged every 2 to 3 days. When the required number of cells was reached, the cells were harvested, counted, and adjusted to an appropriate concentration with PBS. The cells were then diluted 1:1 with Matrigel to a concentration of 2.5×107 cells / ml. A volume of 5×106 cells / 200 μL was administered to each mouse (purchased from Weitong Lihua Laboratory Animal Technology Co., Ltd., Balb / c nude, female) and inoculated into the right forelimb subaxillary region. Tumors were sorted by tumor size and then grouped for dosing (the day of group dosing was designated Day 0). Tumors and animal body weights were measured three times weekly during dosing. After the experiment, multi-point PK analysis was performed, tumor samples were collected, and tumor weights were recorded and photographed. The in vivo efficacy results of compound 30 are shown in FIG1 (tumor volume (a in the figure) and mouse body weight (b in the figure)), and the in vivo efficacy results of compound 30 (tumor weight) are shown in FIG2 .

[0734] The experimental data were expressed as mean ± standard error (SEM). The data were statistically analyzed using one-way ANOVA plus Dunnett's multiple comparison test in Graphpad Prism. P < 0.05 was considered to be significantly different.

[0735] (MRTX1719 structure: )

[0736] Conclusion: The compounds of the present invention, such as the compounds in the examples, are effective in mice CDX (HCT-116MTAP - / - It has a good tumor-suppressing effect in the cell model.

Claims

1. A compound represented by formula (Ia) or (Ib), a stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt thereof, in, D is selected from: D1 is selected from: Ld is selected from C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl; Rd1 is selected from 5-membered heteroaryl, wherein the heteroaryl is optionally substituted by 1-3 selected from halogen, D, CN, OH, C 1-4 Alkyl, C 1-4 Alkoxy and NH2 group substitution; X is selected from CR1 or N; Y, Z and V are independently CR5, C(R5)2, NR6, N, O or S, and the total number of O and S is not greater than 1; Ring A is phenyl, a 5-7 membered heterocycloalkyl group containing 1-3 heteroatoms selected from N, O, and S, or a 5-6 membered heteroaryl group containing 1-3 heteroatoms selected from N, O, and S; Ring B is C 3-12 Carbocyclic ring or 4-12 membered heterocyclic ring containing 1-3 heteroatoms selected from N, O, S; Ring C is C 3-12 Carbocyclic ring or 4-12 membered heterocyclic ring containing 1-3 heteroatoms selected from N, O, S; R1 is H, D, C 1-4 Alkyl, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, halogen, CN, OH, NH2 or COOH, wherein the alkyl, alkoxy, alkenyl, alkynyl is optionally substituted with 1-3 groups selected from halogen, D, CN, OH and NH2; R2 is OH, NH2, -CH2NH2 or -NH-OH; R3 and R4 are independently H, D, halogen, C 1-4 Alkyl, CN, OH, NH2, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl or C 3-6 Cycloalkyl; the alkyl, alkoxy, alkenyl, alkynyl or cycloalkyl group is optionally substituted by 1-3 groups selected from halogen, D, CN, OH and NH2; Alternatively, R3 and R4 together form =NH; Each R5 is independently H, D, =O, OH, C 1-4 Alkyl, CN, halogen, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, 5-6 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S or C 3-6 Cycloalkyl, wherein the alkyl, alkoxy, alkenyl, alkynyl, heteroaryl or cycloalkyl is optionally substituted by 1-3 groups selected from halogen, D, CN, OH, C 1-4 Alkyl, C 1-4 Alkoxy and NH2 group substitution; R6 is H, D, C 1-4 Alkyl or C 3-6 Cycloalkyl; said alkyl or cycloalkyl is optionally substituted by 1-3 groups selected from halogen, D, CN, OH and NH2; Each R7 is independently H, =O, D, OH, CN, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, N(R7')2, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-6 Cycloalkyl or 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl or heterocycloalkyl is optionally substituted by 1-3 halogen, D, OH, NH2, CN, C 1-4 Alkyl, -C(O)C 1-4 Alkyl, -C(O)NHC 1-4 Alkyl, -C(O)NH2, -NHC(O)C 1-4 Alkyl and C 1-4 Alkoxy group substitution; Each R8 is independently H, D, nitro, amino, C 1-6 Alkoxy, =O, OH, CN, halogen, C 2-6 Alkenyl, C 2-6 Alkynyl, -SF5, N3, C 1-6 Alkyl, -COC 1-4 Alkyl, -N(R7')2, -C(=O)N(R7')2, -NR7'C(=O)-R7', -C(=O)-R7', -(CH2) r -O-(CH2) r -R8'、-SC 1-6 Alkyl, -S(O)-C 1-6 Alkyl, -S(O)2-C 1-6 Alkyl, -S-(CH2) r -R8'、-NR7'-(CH2) r -R8' or -(CH2) r -R8', the alkyl, alkenyl, alkynyl group is optionally substituted by 1-5 groups selected from R 8” replace; Each R7' is independently H, D, amino, hydroxyl, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, C 3-6 Cycloalkyl, C 3-12 Heterocycloalkyl, -NHC 1-6 Alkyl, -N(C 1-6 Alkyl)2, or deuterated C 1-6 Alkoxy, the cycloalkyl, heterocycloalkyl is optionally substituted by 1 to 3 groups selected from deuterium, halogen, cyano, amino, hydroxyl, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy or deuterated C 1-6 Alkoxy group substitution; Each R8' is independently H, D, C 1-6 Alkyl, C 1-4 Alkoxy, C 3-12 Carbocyclic ring, 4-12 membered heterocyclic ring containing 1-3 heteroatoms selected from N, O, S, wherein the alkyl and alkoxy groups are optionally substituted by 1-3 groups selected from halogen, =O, D, OH, NH2, CN, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 Alkyl, -C(O)C 1-4 Alkyl, -C(O)NHC 1-4 Alkyl, -C(O)NH2, -NHC(O)C 1-4 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy and deuterated C 1-6 The carbocyclic or heterocyclic ring is optionally substituted by 1-5 groups selected from R 8” replace; R 8” Selected from halogen, D, OH, NH2, CN, C 1-6 Alkyl and C 1-6 Alkoxy, the alkyl or alkoxy group is optionally further substituted by 1 to 5 groups selected from halogen, deuterium, cyano, amino, hydroxyl, C 1-6 Alkyl or C 1-6 Alkoxy group substitution; Each r is independently 0, 1, 2 or 3; p is 0, 1, or 2; n and m are independently integers from 0 to 5; The condition is, 1. Group no 2. When yes R3 is H, C 1-4 Alkyl, C 1-4 Haloalkyl, R5 is H, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 When halogenated alkyl and R3 and R5 are not H at the same time, the group for 2. The compound of formula (Ia) according to claim 1, its stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt, has the following structure:

3. The compound according to claim 1, its stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt, D is selected from 4. The compound according to any one of claims 1 to 3, its stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt, wherein: X is selected from CR1 or N; Y, Z and V are independently CR5, N, O or S, and the total number of O and S is not greater than 1; Ring A is phenyl, or a 5-6 membered heteroaryl group containing 1-3 heteroatoms selected from N, O, and S; Ring B is phenyl, C 3-6 cycloalkyl, 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, and S, or 5-6 membered heteroaryl containing 1-3 heteroatoms selected from N, O, and S; Ring C is phenyl, C 3-6 cycloalkyl, 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, and S, or 5-6 membered heteroaryl containing 1-3 heteroatoms selected from N, O, and S; R1 is H, D, C 1-4 Alkyl, halogen, CN, wherein the alkyl is optionally substituted with 1 to 3 groups selected from halogen, D, CN and OH; R2 is OH, NH2, -CH2NH2 or -NH-OH; R3 and R4 are independently H, D, halogen, C 1-4 Alkyl, CN, OH or NH2, wherein the alkyl group is optionally substituted with 1 to 3 groups selected from halogen, D, CN, OH and NH2; Each R5 is independently H, D, OH, C 1-4 Alkyl, CN, halogen, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, 5-membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, 4-5-membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S or C 3-4 Cycloalkyl, the alkyl, alkoxy, alkenyl, alkynyl, heteroaryl, heterocycloalkyl, cycloalkyl is optionally substituted by 1-3 groups selected from halogen, D, CN, OH, C 1-4 Alkyl, C 1-4 Alkoxy, and NH2 group substitution; Each R7 is independently H, =O, D, OH, CN, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, N(R7')2, C 2-4 Alkenyl, or C 2-4 Alkynyl, the alkyl, alkoxy, alkenyl, alkynyl is optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-4 Alkyl, -C(O)C 1-4 Alkyl, -C(O)NHC 1-4 Alkyl, -C(O)NH2, -NHC(O)C 1-4 Alkyl and C 1-4 Alkoxy group substitution; Each R8 is independently H, D, =O, OH, CN, halogen, C 2-4 Alkenyl, C 2-4 Alkynyl, N3, C 1-4 Alkyl, -COC 1-4 Alkyl, -CONR7'C 1-4 Alkyl, -NR7'COC 1-4 Alkyl, N(R7')2, -O-(CH2)r-R8', or -(CH2)r-R8', wherein the alkyl, alkenyl, alkynyl is optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-4 Alkyl and C 1-4 Alkoxy group substitution; Each R7' is independently H, C 1-4 Alkyl or halogenated C 1-4 alkyl; Each R8' is independently H, C 1-4 Alkoxy, C 3-6 Cycloalkyl, phenyl, 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, or 5-6 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, wherein the cycloalkyl, phenyl, heterocycloalkyl or Heteroaryl is optionally substituted by 1-3 groups selected from halogen, =O, D, OH, NH2, CN, C 1-4 Alkyl, halogenated C 1-4 Alkyl, and C 1-4 Alkoxy group substitution; Each r is independently 0, 1, or 2; p is 0 or 1; n and m are independently 0, 1, 2, 3 or 4.

5. The compound according to claim 1, its stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt, wherein: Ring B is selected from 6. The compound according to claim 1, its stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt, wherein: The compound has the structure of formula (II-a), (II):

7. The compound according to claim 6, its stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt, wherein: X is selected from CR1 or N; Y, Z and V are independently CR5, N, O or S, and the total number of O and S is not greater than 1; Ring A is phenyl, thienyl, thiazolyl, pyrrolyl, imidazolyl, oxazolyl, pyridyl, pyrazinyl, pyrimidinyl or pyridazinyl; D is selected from R1 is H, D, C 1-4 Alkyl, F, Cl, Br, I or CN, wherein the alkyl group is optionally substituted with 1-3 groups selected from F, Cl, Br, I, D, CN and OH; R2 is OH, NH2, -CH2NH2 or -NH-OH; R3 and R4 are independently H, D, F, Cl, Br, I, C 1-4 Alkyl or CN; the alkyl group is optionally substituted by 1-3 groups selected from F, Cl, Br, I, D, CN, OH and NH2; Each R5 is independently H, D, OH, C 1-4 Alkyl, CN, F, Cl, Br, I, C 1-4 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, 5-membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, 4-5-membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S or C 3-4 Cycloalkyl, wherein the alkyl, alkoxy, alkenyl, alkynyl, heteroaryl, heterocycloalkyl, cycloalkyl is optionally substituted with 1-3 groups selected from F, Cl, Br, I, D, CN, OH, methoxy, and NH2; Each R7 is independently H, D, OH, CN, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, N(R7')2, C 2-4 Alkenyl, or C 2-4 Alkynyl, the alkyl, alkoxy, alkenyl, alkynyl is optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-4 Alkyl and C 1-4 Alkoxy group substitution; Each R8 is independently H, D, OH, CN, F, Cl, Br, I, C 2-4 Alkenyl, C 2-4 Alkynyl, N3, C 1-4 Alkyl, -COC 1-4 Alkyl, -CONR7'C 1-4 Alkyl, -NR7'COC 1-4 Alkyl, N(R7')2, -O-(CH2)r-R8', or -(CH2)r-R8', wherein the alkyl, alkenyl, alkynyl is optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-4 Alkyl and C 1-4 Alkoxy group substitution; Each R7' is independently H, C 1-4 Alkyl or halogenated C 1-4 alkyl; Each R8' is independently H, C 1-4 Alkoxy, C 3-6 Cycloalkyl, phenyl, 4-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, or 5-6 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, wherein the cycloalkyl, phenyl, heterocycloalkyl or heteroaryl is optionally substituted by 1-3 halogen, =O, D, OH, NH2, CN, C 1-4 Alkyl, halogenated C 1-4 Alkyl, and C 1-4 Alkoxy group substitution; Each r is independently 0, 1, or 2; p is 0 or 1; n is 0, 1, 2, or 3; m is 0, 1, 2, 3 or 4.

8. The compound according to claim 7, its stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt, wherein: Each R8 is independently D, CN, F, Cl, Br, C 2-4 Alkenyl, C 2-4 Alkynyl, N3, C 1-4 Alkyl, -O-(CH2)r-R8', or -(CH2)r-R8', wherein the alkyl, alkenyl, alkynyl is optionally substituted by 1-3 groups selected from halogen, D, OH, NH2, CN, C 1-2 Alkyl and C 1-2 Alkoxy groups are substituted.

9. The compound according to claim 6, its stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt, wherein: R8' is independently C 1-4 Alkoxy, cyclopropyl, cyclobutyl, cyclopentyl, phenyl, tetrahydropyrrolyl, azetidinyl, piperidinyl, piperazinyl, morpholinyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, Thiphenyl, thiazolyl, pyrrolyl, imidazolyl, oxazolyl, pyridinyl, pyrazinyl, pyrimidinyl or pyridazinyl, optionally substituted by 1-3 groups selected from F, Cl, Br, I, =O, D, OH, NH2, CN, C 1-4 Alkyl, halogenated C 1-2 Alkyl, and C 1-2 Alkoxy groups are substituted.

10. The compound according to claim 1, its stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt, wherein: D1 is selected from: Ld is selected from C 2-4 Alkenyl, preferably Rd1 is selected from 5-membered heteroaryl, wherein the heteroaryl is optionally substituted by 1-2 groups selected from NH2, and Rd1 is preferably 11. The compound according to claim 1, its stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt, wherein the compound is selected from the structure in Table 1.

12. A pharmaceutical composition comprising the compound according to any one of claims 1 to 11, or a stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier and / or excipient.

13. Use of the compound according to any one of claims 1 to 11, its stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt, or the composition according to claim 12, in the preparation of a medicament for treating / preventing a disease mediated by PRMT5·MTA. The use according to claim 13, wherein the PRMT5·MTA-mediated disease is selected from liver cancer, breast cancer, skin cancer, bladder cancer, pancreatic cancer or head and neck cancer.

15. A pharmaceutical composition or pharmaceutical formulation comprising 1-1500 mg of a compound according to any one of claims 1-11 or a stereoisomer, deuterated form, solvate, prodrug, metabolite, pharmaceutically acceptable salt or cocrystal thereof and a pharmaceutically acceptable excipient.

16. A method for treating a disease in a mammal, the method comprising administering to a subject a therapeutically effective amount of a compound according to any one of claims 1 to 11, or a stereoisomer, deuterated form, solvate, prodrug, metabolite, pharmaceutically acceptable salt, or cocrystal thereof, preferably 1 to 1500 mg, wherein the disease is preferably liver cancer, breast cancer, skin cancer, bladder cancer, pancreatic cancer, or head and neck cancer.

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