Heterocyclic alkynyl-substituted amide derivative as well as preparation method and application thereof

Abstract

The invention relates to a heterocyclic alkynyl-substituted amide derivative as shown in a formula (I), and a preparation method and application of the heterocyclic alkynyl-substituted amide derivative. The invention also relates to a pharmaceutical composition containing the compound as an active ingredient and application of the compound or the pharmaceutical composition in treating and / or preventing related diseases mediated by PRMT5.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, specifically to a class of heterocyclic alkyne-substituted amide derivatives, their preparation methods, and their applications. Background Technology

[0002] Epigenetics-regulated gene expression plays a vital biological role in protein maturation and cell differentiation, and is crucial in many human diseases. Arginine guanidinomethylation catalyzed by protein arginine methyltransferase (PRMT) is a common post-translational modification in eukaryotic cells, affecting various biological processes such as cell signaling, gene transcription, mRNA translation, DNA recombination, and repair (Cell Mol. Life Sci. 2015.72(11):2041-2059).

[0003] PRMT5 is a member of the PRMT family, and its mediated methylation plays an important role in maintaining normal intracellular homeostasis. However, more and more studies have shown that abnormal expression of PRMT5 is associated with the occurrence of various tumors. It is overexpressed in various tumors and the mechanisms of occurrence differ in different tumors (CellMol.Life Sci.2015.72(11):2041-2059).

[0004] Homozygous deletion of tumor suppressor genes is a key driver of tumorigenesis. The deletion of the tumor suppressor gene CDKN2A, located on human chromosome 9p21, is one of the most frequently mutated genes in tumors, occurring in 15% of cases. Due to its close resemblance to CDKN2A, the methionine phosphorylase gene (MTAP) is frequently deleted in tumors, playing a crucial role in the methionine and adenine rescue pathway (Cell Reports, 2016, 15:574–587). MTAP deletion leads to the accumulation of its substrate, methylthioadenosine (MTA). Because MTA is structurally similar to S-adenosylmethionine (SAM), it selectively competes with SAM for binding to PRMT5, partially inhibiting PRMT5 activity and sensitizing further PRMT5 inhibition, i.e., synthetic lethality (Science, 2016, VOL 351ISSUE 6278:1214-1217).

[0005] However, PRMT5 is a known essential gene. Knockout or siRNA silencing of PRMT5 in normal tissues leads to abnormal physiological functions, such as reduced blood cell counts, infertility, skeletal muscle loss, and myocardial hypertrophy (Journal of Clinical Investigation, 2015, 125(9):3532-44). Currently, none of the PRMT5 inhibitors in the clinical stage can induce synthetic lethality due to MTAP deficiency. Therefore, new strategies are needed to exploit the metabolic vulnerability caused by MTAP deficiency.

[0006] Developing small molecule inhibitors targeting PRMT5·MTA can preferentially act on MTAP-deficient tumor cells. Since normal cells do not lack MTAP and have low MTA concentrations, they do not have a significant inhibitory effect on normal cells, thereby increasing the therapeutic index (AACR Annual Meeting, 2021, Abstract LB003), providing a new strategy for tumor treatment. Summary of the Invention

[0007] The problem the invention aims to solve:

[0008] Although several patent applications for PRMT5 inhibitors have been published, the huge market demand from MTAP- / - cancer patients, coupled with the less-than-ideal clinical efficacy of existing PRMT5 inhibitors, necessitates further development of new compounds. Through continuous efforts, the inventors of this application have designed compounds with the structure shown in general formula (I) and discovered that compounds with this structure exhibit excellent PRMT5 inhibitory effects and have great application potential.

[0009] Solution for solving the problem:

[0010] In order to solve the above problems, the inventors of this application have conducted in-depth research and discovered that a class of heterocyclic alkyne-substituted amide derivatives can achieve the desired purpose, resulting in the completion of this invention.

[0011] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0012] The compounds represented by Formula I, their tautomers, stereoisomers, pharmaceutically acceptable salts, or deuterated compounds thereof:

[0013]

[0014] in,

[0015] Selected from single or double bonds, or It does not exist;

[0016] when When X is a single bond or a double bond, X, Y, and Z are independently selected from CR. 4 R 5 NR 6 O, N, S, CR 4 ;

[0017] when When Y does not exist, X is hydrogen and Z is CR. 4 R 5 ;

[0018] W is selected from CR 7 Or N;

[0019] A is selected from CR 8 R 9 NR 8 Or O;

[0020] The ring M is selected from 5-10 aryl or 5-10 heteroaryl, wherein the 5-10 heteroaryl contains 1-4 heteroatoms selected from N, O or S;

[0021] R 3 R 4 R 5 R 6 R 7 R 8 R 9 Independently selected from hydrogen, deuterium, halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 cycloalkyl, C 3-6 Cycloalkoxy; wherein, the C 1-6 Alkyl, C 1-6 The alkoxy group is unsubstituted or surrounded by one or more R groups. i Replace; the C 3-6 cycloalkyl, C 3-6 The cycloalkoxy group is unsubstituted or surrounded by one or more R groups. j replace;

[0022] R 1 Selected from C 1-6 Alkyl, C 3-10 cycloalkyl-(C 1-6 alkyl) m -, 4-10 membered heterocyclic group-(C 1-6 alkyl) n -, 5-10 quinone heteroaryl-(C 1-6 alkyl) p -, 5-10 aryl-(C 1-6 alkyl) q-; where n = 0 or 1, m = 0 or 1, p = 0 or 1, q = 0 or 1; the heterocyclic group and the heteroaryl group each independently contain 1-4 heterocyclic atoms selected from O, N or S; wherein, the C 1-6 The alkyl group is unsubstituted or contains one or more R groups. i Replace; the C 3-10 The cycloalkyl group is unsubstituted or contains one or more R groups. j Substitution; the 4-10 membered heterocyclic group is unsubstituted or replaced by one or more R groups. k Substitution; the 5-10 aryl and 5-10 heteroaryl groups are unsubstituted or independently substituted by one or more R groups. l replace;

[0023] R 2 Selected from: 5-10-membered heteroaryl groups and 4-10-membered heterocyclic groups, wherein each of the 5-10-membered heteroaryl group and the 4-10-membered heterocyclic group independently contains 1-4 heteroatoms selected from N, O, or S; wherein the 5-10-membered heteroaryl group and the 4-10-membered heterocyclic group are unsubstituted or converted by one or more identical or different R atoms. b replace;

[0024] n1 and n2 are each independently 0, 1, and 2;

[0025] Each R i R j R k R l Each C is independently selected from hydroxyl, deuterium, halogen, cyano, and optionally substituted by one or more C groups selected from halogen, cyano, or hydroxyl. 1-6 Alkyl group, optionally surrounded by one or more groups selected from halogen, cyano, hydroxyl, -NR'R", C 1-6 alkoxy-substituted C 1-6 Alkoxy, C 3-10 cycloalkyl, optionally with one or more C 1-6 Alkyl-substituted 5-10-membered heteroaryl groups, wherein the 5-10-membered heteroaryl group contains 1-4 heteroatoms selected from N, O, and S; wherein R' and R" are each independently selected from H and C. 1-6 Alkyl, halogenated C 1-6 Alkyl groups, or R' and R" together with the nitrogen atom they are attached to, form 4-8 membered heterocyclic groups, wherein the 4-8 membered heterocyclic groups contain 1-4 heteroatoms selected from N, O, and S;

[0026] Each R b Each is independently selected from hydroxyl, deuterium, cyano, halogen, -NR'R", C1-C6 alkyl, C 1-6Alkoxy, C3-C6 cycloalkyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, and C3-C6 cycloalkyl are unsubstituted or each is independently substituted by one or more substituents selected from halogen, hydroxyl, cyano, and C1-C6 alkoxy; wherein in -NR'R”, R' and R” are each independently selected from hydrogen, C 1-6 Alkyl, Halogenated C 1-6 Alkyl groups, or R' and R" together with the nitrogen atom to which they are attached, form 4-8 membered heterocyclic groups, which contain 1-4 heteroatoms selected from N, O, and S.

[0027] In some specific implementations, cyclic M is selected from 5-6 aryl or 5-6 heteroaryl.

[0028] In this invention, unless otherwise specified, each occurrence of a 5-10 heteroaryl group preferably refers to each containing 1-4 heteroatoms selected from N, O, and S; each occurrence of a 5-6 heteroaryl group preferably refers to each containing 1-3 heteroatoms selected from N, O, and S.

[0029] In some specific embodiments, ring M is selected from phenyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrazolyl, imidazolyl, triazolyl, pyrroleyl, furanyl, oxazolyl, isoxazolyl, and thiazolyl.

[0030] In some specific embodiments, ring M is selected from phenyl or pyridyl.

[0031] In some specific implementation plans, It is a single or double bond, and X, Y, and Z are independently selected.

[0032] CR 4 R 5 NR 6 O, N, S, CR 4 .

[0033] In some specific implementation plans, It does not exist, Y does not exist, X is hydrogen, Z is CR. 4 R 5 .

[0034] In some specific implementation schemes, A is selected from O.

[0035] In some specific implementations, n1 and n2 are each independently 0 or 1.

[0036] In some specific implementation schemes, the group Selected from

[0037] In some specific implementation schemes, the group Selected from

[0038] In some specific implementation schemes, the group Selected from:

[0039] In some specific implementation schemes, the group Selected from:

[0040] In some specific implementation schemes, R 2 The compounds are selected from 5-6-membered heteroaryl groups and 4-6-membered heterocyclic groups; each of the 5-6-membered heteroaryl group and the 4-6-membered heterocyclic group independently contains 1-3 heteroatoms selected from N, O, or S; the 5-6-membered heteroaryl group and the 4-6-membered heterocyclic group are unsubstituted or independently converted by one or more identical or different R atoms. b replace.

[0041] In some specific implementation schemes, R 2 The group is selected from 5-6-membered heteroaryl, 4-6-membered heterocyclic, and 5-6-membered heteroaryl-5-6-membered heterocyclic groups; each of the 5-6-membered heteroaryl, 4-6-membered heterocyclic, and 5-6-membered heterocyclic groups independently contains 1, 2, or 3 heteroatoms selected from N, O, or S; the 5-6-membered heteroaryl, 4-6-membered heterocyclic, and 5-6-membered heteroaryl-5-6-membered heterocyclic groups are unsubstituted or independently converted by one or more identical or different R atoms. b replace.

[0042] In some specific implementation schemes, R 2 The aryl group is selected from 5-6-membered heteroaryl groups, wherein the 5-6-membered heteroaryl group contains 1-3 heteroatoms selected from N, O, or S; the 5-6-membered heteroaryl group is unsubstituted or converted by one or more identical or different R atoms. b replace.

[0043] In some specific implementation schemes, R 2 The group is selected from pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyridyl, pyrazinyl, pyrimidinyl, and pyridazinyl, and is either unsubstituted or independently bound by one, two, or three R groups. b replace.

[0044] In some specific implementation schemes, R 2 Selected from 5-6-membered heteroaryl groups, wherein the 5-6-membered heteroaryl group contains one, two, or three N heteroatoms; the 5-6-membered heteroaryl group is unsubstituted or converted by one or more identical or different R atoms. b replace.

[0045] In some specific implementation schemes, R 2 Selected from pyrazolyl, imidazole, and pyridyl groups, which are unsubstituted or independently bound by one, two, or three R groups. b replace.

[0046] In some specific implementation schemes, R 2 Selected from It is either unreplaced or independently controlled by one, two, or three Rs. b replace.

[0047] In some specific implementation schemes, R 2 The 5-membered heteroaryl group is selected from 5-membered heteroaryl groups, wherein the 5-membered heteroaryl group contains 1-3 heteroatoms selected from N, O, or S, and the 5-membered heteroaryl group is unsubstituted or surrounded by 1, 2, or 3 R atoms. b replace.

[0048] In some specific implementation schemes, R 2 Selected from pyrazolyl, imidazole, thiazolyl, isothiazolyl, oxazolyl, and isoxazolyl, which are unsubstituted or surrounded by one, two, or three R groups. b replace.

[0049] In some specific implementation schemes, R 2 Selected from 5-membered heteroaryl groups, wherein the 5-membered heteroaryl group contains 1-2 heteroatoms selected from N or S, and wherein the 5-membered heteroaryl group is unsubstituted or converted by 1, 2 or 3 R atoms. b replace.

[0050] In some specific implementation schemes, R 2 Selected from pyrazolyl, imidazole, thiazolyl, and isothiazolyl groups, which are unsubstituted or independently bound by one, two, or three R groups. b replace.

[0051] In some specific implementation schemes, R 2 Selected from 5-membered heteroaryl groups, wherein the 5-membered heteroaryl group contains 1, 2, or 3 N heteroatoms, and wherein the 5-membered heteroaryl group is unsubstituted or converted by 1, 2, or 3 identical or different R atoms. b replace.

[0052] In some specific implementation schemes, R 2 Selected from pyrazolyl or imidazole groups, which are unsubstituted or independently bound by one, two, or three R groups. b replace.

[0053] In some specific implementation schemes, R 2 Selected from pyrazolyl groups, which are unsubstituted or independently composed of one, two, or three R groups. b replace.

[0054] In some specific implementation schemes, R 2 Selected from It is either unreplaced or independently controlled by one, two, or three Rs. b replace.

[0055] In some specific implementation schemes, R 2 Selected from

[0056]

[0057] In some specific implementation schemes, R 2 The group is selected from 6-membered heteroaryl groups, wherein the 6-membered heteroaryl group contains one, two, or three heteroatoms selected from N, O, or S; the 6-membered heteroaryl group is unsubstituted or substituted with one or more identical or different R atoms. b replace.

[0058] In some specific implementation schemes, R 2 Selected from 6-membered heteroaryl groups, wherein the 6-membered heteroaryl group contains 1, 2, or 3 N heteroatoms; wherein the 6-membered heteroaryl group is unsubstituted or converted by 1, 2, or 3 identical or different R atoms. b replace.

[0059] In some specific implementation schemes, R 2 Selected from pyridinyl, wherein the pyridinyl group is unsubstituted or converted by one, two, or three identical or different R groups. b replace.

[0060] In some specific implementation schemes, R 2 Selected from It is unsubstituted or replaced by one, two, or three identical or different Rs. b replace.

[0061] In some specific implementation schemes, R 2 Selected from

[0062]

[0063] In some specific implementation schemes, R 2 The compounds are selected from 5-6-membered heteroaryl and 5-6-membered heterocyclic groups, wherein each of the 5-6-membered heteroaryl and 5-6-membered heterocyclic groups independently contains 1, 2, or 3 heteroatoms selected from N, O, or S; the 5-6-membered heteroaryl and 5-6-membered heterocyclic groups are unsubstituted or modified by one or more identical or different R atoms. b replace.

[0064] In some specific implementation schemes, R 2 Selected from 5-6-membered heteroaryl and 5-6-membered heterocyclic groups, wherein each of the 5-6-membered heteroaryl and 5-6-membered heterocyclic group independently contains 1, 2, or 3 N heteroatoms; wherein the 5-6-membered heteroaryl and 5-6-membered heterocyclic group is unsubstituted or converted by one or more identical or different R atoms. b replace.

[0065] In some specific implementation schemes, R 2Selected from 5-membered heteroaryl and 6-membered heterocyclic groups, wherein each of the 5-membered heteroaryl and 6-membered heterocyclic group independently contains 1, 2, or 3 N heteroatoms; wherein the 5-membered heteroaryl and 6-membered heterocyclic group is unsubstituted or converted by one or more identical or different R atoms. b replace.

[0066] In some specific implementation schemes, R 2 Selected from imidazolidenepiperazinyl; said imidazolidenepiperazinyl group is unsubstituted or converted by one, two, or three identical or different R groups. b replace.

[0067] In some specific implementation schemes, R 2 Selected from

[0068] In some specific implementation schemes, R 2 The compounds are selected from 5-6-membered heteroaryl and 5-6-membered heterocyclic groups, 5-6-membered heteroaryl and 5-6-membered heteroaryl groups, and 5-6-membered heteroaryl phenyl groups, wherein each of the 5-6-membered heteroaryl and 5-6-membered heterocyclic groups independently contains 1, 2, or 3 heteroatoms selected from N, O, or S; and wherein the 5-6-membered heteroaryl and 5-6-membered heterocyclic groups, 5-6-membered heteroaryl and 5-6-membered heteroaryl phenyl groups are unsubstituted or converted by one or more identical or different R atoms. b replace.

[0069] In some specific implementation schemes, R 2 The molecule is selected from 5-6-membered heteroaryl and 5-6-membered heterocyclic groups, 5-6-membered heteroaryl and 5-6-membered heteroaryl groups, and 5-6-membered heteroaryl phenyl groups, wherein the 5-6-membered heteroaryl group is selected from imidazolyl, pyrazolyl, and pyridinyl, and the 5-6-membered heterocyclic group is selected from pyrrolidinyl, piperidinyl, and piperazinyl; the 5-6-membered heteroaryl and 5-6-membered heterocyclic group, 5-6-membered heteroaryl and 5-6-membered heteroaryl phenyl groups are unsubstituted or substituted with one or more identical or different R groups. b replace.

[0070] In some specific implementation schemes, R 2 The compound is selected from imidazopyrrolidinyl, imidazopiperidinyl, imidazopiperazinyl, pyrazolophenyl, and pyrazolopyridinyl, wherein the imidazopyrrolidinyl, imidazopiperidinyl, imidazopiperazinyl, pyrazolophenyl, and pyrazolopyridinyl are unsubstituted or substituted with one or more identical or different R groups. b replace.

[0071] In some specific implementation schemes, R 2 Selected from

[0072] In some specific implementation schemes, R 2The group is selected from 4-6 membered heterocyclic groups; the 4-6 membered heterocyclic group contains 1-3 heteroatoms selected from N, O or S; the 4-6 membered heterocyclic group is unsubstituted or converted by 1, 2 or 3 identical or different R atoms. b replace.

[0073] In some specific implementation schemes, R 2 Selected from 4-6-membered heterocyclic alkyl groups and 4-6-membered heterocyclic alkenyl groups; each of the 4-6-membered heterocyclic alkyl groups and 4-6-membered heterocyclic alkenyl groups independently contains 1-3 heteroatoms selected from N, O, or S; the 4-6-membered heterocyclic alkyl groups and 4-6-membered heterocyclic alkenyl groups are unsubstituted or independently separated by 1, 2, or 3 identical or different R atoms. b replace.

[0074] In some specific implementation schemes, R 2 Selected from 4-6-membered heterocyclic alkyl groups and 4-6-membered heterocyclic alkenyl groups; each of the 4-6-membered heterocyclic alkyl groups and 4-6-membered heterocyclic alkenyl groups independently contains 1-2 heteroatoms selected from N or O; the 4-6-membered heterocyclic alkyl groups and 4-6-membered heterocyclic alkenyl groups are unsubstituted or independently separated by 1, 2 or 3 identical or different R atoms. b replace.

[0075] In some specific implementation schemes, R 2 Selected from tetrahydro-2H-pyranyl, oxetyl, 1,2,3,6-tetrahydropyridyl, and 3,6-dihydro-2H-pyranyl; wherein the tetrahydro-2H-pyranyl, oxetyl, 1,2,3,6-tetrahydropyridyl, and 3,6-dihydro-2H-pyranyl are unsubstituted or independently converted by one, two, or three identical or different R groups. b replace.

[0076] In some specific implementation schemes, R 2 Selected from The It is either unsubstituted or independently replaced by one, two, or three identical or different Rs. b replace.

[0077] In some specific implementation schemes, R 2 Selected from Where R 2a With R 2b Together with the carbon atoms they are attached to, they form a 4-6 membered heterocyclic group, which contains 1-3 heteroatoms selected from N, O, or S; the 4-6 membered heterocyclic group is unsubstituted or independently bound by 1, 2, or 3 identical or different R atoms. b replace.

[0078] In some specific implementation schemes, R 2 Selected from Where R 2a With R 2b Together with the carbon atoms they are connected to, they form 4-6 membered heterocyclic alkyl groups or 4-6 membered heterocyclic alkenyl groups; each of the 4-6 membered heterocyclic alkyl groups and 4-6 membered heterocyclic alkenyl groups independently contains 1-3 heteroatoms selected from N, O, or S; the 4-6 membered heterocyclic alkyl groups and 4-6 membered heterocyclic alkenyl groups are unsubstituted or independently bound by 1, 2, or 3 identical or different R atoms. b replace.

[0079] In some specific implementation schemes, R 2 Selected from Where R 2a With R 2b Together with the carbon atoms they are connected to, they form 4-6 membered heterocyclic alkyl groups or 4-6 membered heterocyclic alkenyl groups; each of the 4-6 membered heterocyclic alkyl groups and 4-6 membered heterocyclic alkenyl groups independently contains 1-2 heteroatoms selected from N or O; the 4-6 membered heterocyclic alkyl groups and 4-6 membered heterocyclic alkenyl groups are unsubstituted or independently bound by 1, 2 or 3 identical or different R atoms. b replace.

[0080] In some specific implementation schemes, R 2 Selected from Where R 2a With R 2b Together with the carbon atoms they are linked to, they form tetrahydro-2H-pyranyl, oxetyl, 1,2,3,6-tetrahydropyridyl, and 3,6-dihydro-2H-pyranyl; said tetrahydro-2H-pyranyl, oxetyl, 1,2,3,6-tetrahydropyridyl, and 3,6-dihydro-2H-pyranyl are unsubstituted or independently bound by one, two, or three identical or different R atoms. b replace.

[0081] In some specific implementation schemes, R 2 Selected from Among them, groups It is either unsubstituted or independently replaced by one, two, or three identical or different Rs. b replace.

[0082] In some specific implementation schemes, each R b Each of the following is independently selected from H, C1-C6 alkyl, C1-C6 alkoxy, and C3-C6 cycloalkyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, and C3-C6 cycloalkyl are unsubstituted or are independently substituted by one, two, three, or four halogens.

[0083] In some specific implementation schemes, each R bEach of the following is independently selected from H, C1-C3 alkyl, C1-C3 alkoxy, and C3-C5 cycloalkyl, wherein the C1-C3 alkyl, C1-C3 alkoxy, and C3-C5 cycloalkyl are unsubstituted or are independently substituted by one, two, three, or four substituents selected from F, Cl, or Br.

[0084] In some specific implementation schemes, each R b Each is independently selected from H, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0085] In some specific implementation schemes, each R b Each is independently selected from methyl, methoxy, and cyclopropyl.

[0086] In some specific implementation schemes, each R b Each is independently selected from cyclopropyl.

[0087] In some specific implementation schemes, each R b Each is independently selected from methoxy groups.

[0088] In some specific implementation schemes, each R b Each is independently selected from methyl groups.

[0089] In some specific implementation schemes, R 2 Selected from

[0090]

[0091] In some specific implementation schemes, R 2 Selected from

[0092] In some specific implementation schemes, each R b Each of the following is independently selected from H, deuterium, halogen, C1-C6 alkyl, C1-C6 alkoxy, and C3-C6 cycloalkyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, and C3-C6 cycloalkyl are unsubstituted or are independently substituted by one, two, three, or four halogens.

[0093] In some specific implementation schemes, each R b Each of the following is independently selected from H, deuterium, halogen, C1-C3 alkyl, C1-C3 alkoxy, and C3-C5 cycloalkyl, wherein the C1-C3 alkyl, C1-C3 alkoxy, and C3-C5 cycloalkyl are unsubstituted or are independently substituted by one, two, three, or four halogens.

[0094] In some specific implementation schemes, each R bEach of the following groups is independently selected from H, deuterium, F, Cl, Br, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl, wherein the methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl groups are unsubstituted or independently substituted by one, two, three, or four substituents selected from F, Cl, and Br.

[0095] In some specific implementation schemes, each R b Each is independently selected from H, deuterium, F, Cl, Br, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -CHF2, -CF3.

[0096] In some specific implementation schemes, each R b Each is independently selected from H, deuterium, F, Cl, methyl, ethyl, isopropyl, methoxy, cyclopropyl, -CHF2, -CF3.

[0097] In some specific implementations, the R described herein b It can be further replaced by deuterium.

[0098] In some specific implementation schemes, each R b Each is independently selected from H, deuterium, F, Cl, methyl, ethyl, isopropyl, methoxy, cyclopropyl, -CHF2, -CF3, -CD3.

[0099] In some specific implementation schemes, R 2 Selected from;

[0100]

[0101] In some specific implementation schemes, R 2 Selected from

[0102]

[0103] In some specific implementation schemes, R 2 Selected from

[0104]

[0105] In some specific implementation schemes, R 3 Selected from hydrogen, halogens, C 1-6 Alkyl, C 3-6 Cycloalkyl.

[0106] In some specific implementation schemes, R 3 Selected from hydrogen, F, Cl, Br, C 1-4 Alkyl, C5-6 Alkyl, C 3-5 Cycloalkyl.

[0107] In some specific implementation schemes, R 3 Selected from hydrogen, F, Cl, Br, C 1-4 Alkyl, C 3-5 Cycloalkyl.

[0108] In some specific implementation schemes, R 3 Selected from hydrogen, F, Cl, Br, methyl, ethyl, propyl, isopropyl, butyl, cyclopropyl, cyclobutyl, and cyclopentyl.

[0109] In some specific implementation schemes, R 3 Selected from hydrogen, F, Cl, methyl, ethyl, and cyclopropyl.

[0110] In some specific implementation schemes, R 3 Selected from hydrogen, F, Cl, Br, C 1-4 alkyl.

[0111] In some specific implementation schemes, R 3 Selected from hydrogen, F, Cl, methyl, ethyl;

[0112] In some specific implementation schemes, R 3 Selected from hydrogen, F, and methyl.

[0113] In some specific implementation schemes, the group Selected from

[0114] In some specific implementation schemes, the group Selected from In some specific implementation schemes, the group Selected from:

[0115]

[0116] In some specific implementation schemes, the group Selected from:

[0117] In some specific implementation schemes, the group Selected from:

[0118]

[0119] In some specific implementation schemes, the group Selected from

[0120] In some specific implementation schemes, the group Selected from

[0121] Where W is selected from N or CR 7 .

[0122] In some specific implementation schemes, W is selected from CR 7 .

[0123] In some specific implementation schemes, W is selected from N.

[0124] In some specific implementation schemes, R 4 R 5 R 6 R 7 Each is independently selected from hydrogen, halogen, and C. 1-6 alkyl.

[0125] In some specific implementation schemes, R 4 R 5 R 6 R 7 Each is independently selected from hydrogen, halogen, and C. 1-4 Alkyl, C 5-6 alkyl.

[0126] In some specific implementation schemes, R 4 R 5 R 6 R 7 Each is independently selected from hydrogen, halogen, and C. 1-4 alkyl.

[0127] In some specific implementation schemes, R 4 R 5 R 6 R 7 Each is independently selected from hydrogen, F, Cl, Br, methyl, ethyl, propyl, and isopropyl.

[0128] In some specific implementation schemes, R 4 R 5 R 6 R 7 Each is independently selected from hydrogen, F, Cl, and methyl.

[0129] In some specific implementations, the R described herein 4 R 5 R 6 R 7 It can be further replaced by deuterium.

[0130] In some specific implementation schemes, R 4 R 5 R 6 R 7Each is independently selected from hydrogen, F, Cl, methyl, and -CD3.

[0131] In some specific implementation schemes, the group Selected from

[0132] Wherein, W is selected from N or CR 7 .

[0133] In some specific implementation schemes, the group Selected from

[0134] Wherein, W is selected from N or CR 7 R 7 Selected from H, F, Cl, and methyl.

[0135] In some specific implementation schemes, the group Selected from:

[0136]

[0137] In some specific implementation schemes, the group Selected from:

[0138] In some specific implementation schemes, the group Selected from:

[0139]

[0140] In some specific implementation schemes, the group Selected from:

[0141]

[0142] In some specific implementation schemes, the group Selected from:

[0143] In some specific implementation schemes, R 1 Selected from: C 1-6 Alkyl, C 3-10 cycloalkyl, C 3-10 cycloalkyl-C 1-6 Alkyl-, 5-10-membered heteroaryl-; wherein the C1-C6 alkyl group is unsubstituted or converted by one or more R i Replacement; the C3-C 10 The cycloalkyl group is unsubstituted or contains one or more R groups. j Substitution; the 5-10 membered heteroaryl group is unsubstituted or replaced by one or more R groups. l replace.

[0144] In some specific implementation schemes, R 1 Selected from C 1-4 Alkyl, C 3-6 Monocycloalkyl, C 5-8 Bridged cycloalkyl, C 5-8 Spirocycloalkyl, C 3-6 Monocycloalkyl-C 1-4 Alkyl-, C 5-8 Bridged cycloalkyl-C 1-4 Alkyl-, C 5-8 Spirocycloalkyl-C 1-4 Alkyl-, 5-6-membered heteroaryl; wherein, the C 1-4 The alkyl group is unsubstituted or contains one or more R groups. i Replace; the C 3-6 Monocycloalkyl, C 5-8 Bridged cycloalkyl, C 5-8 Spirocycloalkyl is unsubstituted or converted by one or more R j Substitution; the 5-10 membered heteroaryl group is unsubstituted or replaced by one or more R groups. l replace.

[0145] In some specific implementation schemes, R 1 Selected from methyl, ethyl, propyl, butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, Pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrazolyl, imidazoleyl, triazolyl, pyrroleyl, furanyl, oxazolyl, isoxazolyl, thiazolyl; wherein the methyl, ethyl, propyl, and butyl groups are unsubstituted or substituted with one or more R groups. i Substitution; the cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, It is unreplaced or replaced by one or more R j Substitution; the pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrazolyl, imidazoleyl, triazolyl, pyrroleyl, furanyl, oxazolyl, isoxazolyl, and thiazolyl groups are unsubstituted or substituted by one or more R groups. l replace.

[0146] In some specific implementation schemes, R 1 Selected from methyl, ethyl, cyclopropyl, cyclobutyl, cyclopentyl, Pyrazolyl, imidazole, triazolyl, pyrroleyl; wherein the methyl or ethyl group is unsubstituted or is surrounded by one, two, or three R groups. i Substitution; the cyclopropyl, cyclobutyl, cyclopentyl, Is it unreplaced or replaced by 1, 2 or 3 Rs? jSubstitution; the pyrazolyl, imidazole, triazolyl, and pyrroleyl groups are unsubstituted or replaced by one, two, or three R groups. l replace.

[0147] In some specific implementation schemes, R 1 Selected from methyl, ethyl, cyclopropyl, Pyrazolyl; wherein the methyl or ethyl group is unsubstituted or is occupied by one, two, or three R groups. i Substitution; the cyclopropyl, Is it unreplaced or replaced by 1, 2 or 3 Rs? j Substitution; the pyrazol group is unsubstituted or replaced by one, two, or three R groups. l replace.

[0148] It should be understood that the substituent R in this invention 1 In the definition: R i R always substitutes for alkyl or alkylene groups. j Substitution always occurs on a cycloalkyl group, including monocycloalkyl, bridged cycloalkyl, and spirocycloalkyl; R k Always substituted on the heterocyclic group; R l It always substitutes for the aryl or heteroaryl group. For example, If the substituent R appears in the middle, j Then, R j Substitution sites are located on cyclopropyl or superior.

[0149] In some specific implementation schemes, R 1 Selected from methyl, ethyl, cyclopropyl, Pyrazolyl; wherein the methyl, ethyl, cyclopropyl, It is unsubstituted; the pyrazol group is unsubstituted or surrounded by one, two, or three R groups. l replace.

[0150] In some specific implementation schemes, each R i R j R k R l Each is independently selected from halogens and halogenated C. 1-6 Alkyl, C 1-6 Alkyl, C 3-6 Cycloalkyl.

[0151] In some specific implementation schemes, each R i R j R k R l Each is independently selected from halogens and halogenated C. 1-4 Alkyl, C 1-4 Alkyl, C 3-6 Cycloalkyl.

[0152] In some specific implementation schemes, each R i R j R k R l Each is independently selected from halogens, C 1-4 Alkyl, C 3-6 Cycloalkyl.

[0153] In some specific implementation schemes, each R i R j R k R l Each is independently selected from F, Cl, Br, methyl, ethyl, propyl, isopropyl, cyclopropyl, and cyclobutyl.

[0154] In some specific implementation schemes, each R i R j R k R l Each is independently selected from F, Cl, methyl, and ethyl.

[0155] In some specific implementation schemes, each R i R j R k R l Each of the substances is independently selected from methyl groups.

[0156] In some specific implementation schemes, each R l Each is independently selected from halogens and halogenated C. 1-6 Alkyl, C 1-6 Alkyl, C 3-6 Cycloalkyl.

[0157] In some specific implementation schemes, each R l Each is independently selected from halogens and halogenated C. 1-4 Alkyl, C 1-4 Alkyl, C 3-6 Cycloalkyl.

[0158] In some specific implementation schemes, each R l Each is independently selected from halogens, C 1-4 Alkyl, C 3-6 Cycloalkyl.

[0159] In some specific implementation schemes, each R l Each is independently selected from F, Cl, Br, methyl, ethyl, propyl, isopropyl, cyclopropyl, and cyclobutyl.

[0160] In some specific implementation schemes, each R l Each is independently selected from F, Cl, methyl, and ethyl.

[0161] In some specific implementation schemes, each R l Each of the substances is independently selected from methyl groups.

[0162] In some specific implementation schemes, each R l Each is independently selected from D, F, Cl, methyl, and ethyl.

[0163] In some specific implementation schemes, R 1 Selected from methyl, ethyl, cyclopropyl,

[0164] In some specific implementation schemes, R 1 Selected from methyl, ethyl, and cyclopropyl.

[0165] In some specific implementation schemes, R 1 Selected from methyl.

[0166] In some specific implementation schemes, R 1 Selected from -CD3.

[0167] In some specific implementations, equation (I) has the structure described in equation IA:

[0168] Among them, R 1 R 2 R 3 ,A,n1,n2,ring M,W,X,Y,Z, Each is defined as described above.

[0169] Alternatively, formula (I) has the structure described in formula IB:

[0170] Among them, R 1 R 2 R 3 ,A,n1,n2,ring M,W,X,Y,Z, Each is defined as described above.

[0171] As an exemplary compound of the present invention, the compound shown in formula (I) is selected from any of the following specific compounds:

[0172]

[0173]

[0174]

[0175]

[0176]

[0177]

[0178]

[0179] Furthermore, the present invention also provides the S-configuration of the above exemplary compounds:

[0180]

[0181]

[0182]

[0183]

[0184]

[0185]

[0186]

[0187] The term "pharmaceutically acceptable salt" refers to the salt of the compounds of this invention, prepared by reacting a compound having specific substituents discovered in this invention with a relatively non-toxic acid or base. When the compounds of this invention contain relatively acidic functional groups, base addition salts can be obtained by contacting such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. When the compounds of this invention contain relatively basic functional groups, acid addition salts can be obtained by contacting such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent.

[0188] The term "deuterated compound" refers to a compound of the present invention comprising at least one deuterium atom, specifically meaning that one or more hydrogen atoms in the compound of the present invention can be replaced or substituted by a deuterium atom. In some embodiments, the compound comprises two or more deuterium atoms. In some embodiments, the compound comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 deuterium atoms. Methods for synthesizing isotopes into organic compounds are known in the art.

[0189] Preparation method:

[0190] This invention also provides a method for preparing the compound. The preparation of the compound of formula (I) can be accomplished by the following exemplary methods and examples, but these methods and examples should not be considered in any way as limiting the scope of the invention. The compound of this invention can also be synthesized by synthetic techniques known to those skilled in the art, or by combining synthetic methods known in the art with the method described in this invention. The product obtained from each reaction step is obtained using separation techniques known in the art, including but not limited to extraction, filtration, distillation, crystallization, chromatographic separation, etc. The starting materials and chemical reagents required for synthesis can be conventionally synthesized according to literature (such as those provided by Scifinder) or purchased.

[0191] Synthesis process route:

[0192] Route 1:

[0193]

[0194] Step 1: Treating the compound shown in Formula I-1 with di-tert-butyl dicarbonate yields the compound shown in Formula I-2;

[0195] Step 2: Under alkaline conditions well known in the art, use R 1 -Q 2 The compound shown in Formula I-2 can be treated with the compound shown to obtain the compound shown in Formula I-3;

[0196] Step 3: Under coupling conditions well-known in the art, use The compound shown in Formula I-3 can be treated with the compound shown to obtain the compound shown in Formula I-4;

[0197] Step 4: The compound shown in Formula I-4 is deprotected under acidic conditions to obtain the compound shown in Formula I-5.

[0198] Step 5: Under condensation conditions well known in the art, the compound shown in formula I-5 is treated with the compound shown in I-6 to obtain the compound shown in formula I.

[0199] Among them, Q 1 Q 2 Halogens, X, Y, Z, W, A, R 1 R 2 R 3 The definitions of n1, n2, and ring M are as defined in equation (I) of this paper.

[0200] Route 2:

[0201]

[0202] Step 1: Under condensation conditions well known in the art, the compound shown in formula II-1 is treated with the compound shown in I-6 to obtain the compound shown in formula II-2;

[0203] Step 2: Under coupling conditions well-known in the art, using The compound shown in Formula II-2 can be treated with the compound shown to obtain the compound shown in Formula II-3;

[0204] Step 3: The compound shown in Formula II-3 is treated with tetrabutylammonium fluoride to obtain the compound shown in Formula II-4;

[0205] Step 4: Under coupling conditions well known in the art, the compound shown in formula II-4 is treated with the compound shown in R2-Q2 to obtain the compound shown in formula I;

[0206] or,

[0207] Step 2: Under coupling conditions well-known in the art, using The compound shown in Formula II-2 can be treated with the compound shown to obtain the compound shown in Formula I;

[0208] Among them, Q 1 Q 2 Halogens, X, Y, Z, W, A, R 1 R 2 R 3 The definitions of n1, n2, and ring M are as defined in equation (I) of this paper.

[0209] Route 3:

[0210]

[0211] Step 1: Under condensation conditions well known in the art, treating the compound shown in formula III-1 with the compound shown in I-6 yields the compound shown in formula II-4.

[0212] Step 2: Under coupling conditions well known in the art, the compound shown in formula II-4 is treated with the compound shown in R2-Q2 to obtain the compound shown in formula I;

[0213] Among them, Q 2 Halogens, X, Y, Z, W, A, R 1 R 2 R 3 The definitions of n1, n2, and ring M are as defined in equation (I) of this paper.

[0214] Pharmaceutical composition:

[0215] The present invention also provides a pharmaceutical composition for treating and / or preventing diseases associated with abnormal PRMT5 expression, the pharmaceutical composition comprising a therapeutic and / or preventive effective amount of the compound as described above or its tautomers, stereoisomers, pharmaceutically acceptable salts or deuterated compounds thereof, and optionally pharmaceutical excipients.

[0216] In some implementations, the disease associated with abnormal PRMT5 expression is a tumor or cancer.

[0217] In some implementations, the disease associated with abnormal PRMT5 expression refers to a disease associated with abnormal PRMT5 expression and MTAP deficiency.

[0218] In some implementations, the disease associated with abnormal PRMT5 expression refers to tumors or cancers with MTAP deficiency.

[0219] Methods for preparing various pharmaceutical compositions containing a certain amount of active ingredient are known, or will be obvious to those skilled in the art according to the disclosure of the present invention. As described in REMINGTON'S PHARMACEUTICAL SCIENCES, Martin, EW, ed., Mack Publishing Company, 19th ed. (1995), the method for preparing the pharmaceutical composition includes incorporating appropriate pharmaceutical excipients, carriers, diluents, etc.

[0220] The present invention also provides a PRMT5 inhibitor comprising a therapeutic and / or preventative effective amount of the compound as described above or its tautomers, stereoisomers, pharmaceutically acceptable salts or deuterated compounds thereof.

[0221] Medical uses:

[0222] The present invention also provides the use of the compounds as described above, or their tautomers, stereoisomers, pharmaceutically acceptable salts or their deuterated compounds, or the pharmaceutical compositions as described above, in the preparation of PRMT5 inhibitors.

[0223] The present invention also provides the compounds as described above, or their tautomers, stereoisomers, pharmaceutically acceptable salts or their deuterated compounds, or the pharmaceutical compositions as described above, in the preparation of medicaments for the treatment and / or prevention of diseases associated with abnormal PRMT5 expression.

[0224] The present invention also provides the use of the compounds as described above, or their tautomers, stereoisomers, pharmaceutically acceptable salts or their deuterated compounds, or the pharmaceutical compositions as described above, in the treatment and / or prevention of diseases associated with abnormal PRMT5 expression.

[0225] The present invention also provides compounds as described above, or tautomers, stereoisomers, pharmaceutically acceptable salts thereof, or deuterated compounds thereof, or pharmaceutical compositions as described above, for the treatment and / or prevention of diseases associated with abnormal PRMT5 expression.

[0226] In some implementations, the disease associated with abnormal PRMT5 expression is a tumor or cancer.

[0227] In some implementations, the disease associated with abnormal PRMT5 expression refers to a disease with MTAP deficiency associated with abnormal PRMT5 expression.

[0228] In some implementations, the disease associated with abnormal PRMT5 expression refers to tumors or cancers with MTAP deficiency.

[0229] The present invention also provides a method for treating and / or preventing diseases associated with abnormal PRMT5 expression, comprising administering to an individual in need a therapeutic and / or preventative effective amount of the compound as described above or its tautomers, stereoisomers, pharmaceutically acceptable salts or deuterated compounds thereof, or the pharmaceutical composition as described above.

[0230] In some implementations, the disease associated with abnormal PRMT5 expression is a tumor or cancer.

[0231] In some implementations, the disease associated with abnormal PRMT5 expression refers to a disease associated with abnormal PRMT5 expression and MTAP deficiency.

[0232] In some implementations, the disease associated with abnormal PRMT5 expression refers to tumors or cancers with MTAP deficiency.

[0233] In this invention, "treatment" generally refers to achieving the desired pharmacological and / or physiological effect. This effect may be preventative based on the complete or partial prevention of the disease or its symptoms; and / or therapeutic based on the partial or complete stabilization or cure of the disease and / or side effects resulting from the disease. As used herein, "treatment" encompasses any treatment of a patient's disease, including: (a) preventing the disease or symptoms occurring in a patient who has not yet been diagnosed with the disease; (b) suppressing the symptoms of the disease, i.e., preventing its progression; or (c) alleviating the symptoms of the disease, i.e., causing the disease or symptoms to regress.

[0234] In this invention, "subject" and "individual in need" refer to vertebrates. In some embodiments, vertebrates refer to mammals. Mammals include, but are not limited to, livestock (such as cattle), pets (such as cats, dogs, and horses), primates, mice, and rats. In some embodiments, mammals refer to humans.

[0235] In this invention, "effective amount" refers to the amount that effectively achieves the desired therapeutic or preventative effect at the necessary dose and time. The "therapeutic effective amount" of the substance / molecule of this invention may vary depending on factors such as an individual's disease state, age, sex, weight, and the substance / molecule's ability to elicit the desired response in the individual. Therapeutic effective amount also encompasses the amount in which the beneficial therapeutic effect of the substance / molecule outweighs any toxic or harmful consequences. "Preventative effective amount" refers to the amount that effectively achieves the desired preventative effect at the necessary dose and time. Typically, but not necessarily, the preventative effective amount will be lower than the therapeutic effective amount because the preventative dose is administered to the subject before the onset of the disease or in its early stages. In the case of cancer, the therapeutically effective amount of the drug may reduce the number of cancer cells; shrink the tumor volume; inhibit (i.e., slow down, preferably stop) the infiltration of cancer cells into surrounding organs; inhibit (i.e., slow down, preferably stop) tumor metastasis; inhibit tumor growth to some extent; and / or alleviate one or more symptoms associated with cancer to some extent.

[0236] Terminology definition:

[0237] According to the conventions of the art, In the structural formula of this paper, the bond is used to describe the connection point between the part or substituent and the parent nucleus or main structure.

[0238] The hyphen "-" that does not appear between two letters or symbols is used to indicate the connection point of a substituent. For example, C 3-6 cycloalkyl-(C 1-6 alkyl) r -meaning through (C) 1-6 alkyl) r - Connected to the rest of the molecule.

[0239] As used in this article, “substituted” means that any one or more hydrogen atoms on a specified atom or group are selectively substituted by a specified group, provided that the substitution does not exceed the normal valence state of the specified atom.

[0240] In various parts of this specification, the substituents of the compounds disclosed herein are disclosed according to the type or scope of the groups. In particular, the invention includes every independent secondary combination of the various members of these group types and scopes. For example, the term "C..." 1-6 "alkyl" specifically refers to independently disclosed methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl, and C6 alkyl, or independently disclosed "C 1-4 "alkyl", or independently disclosed "C 1-3 alkyl".

[0241] The term "alkyl" refers to both branched and straight-chain saturated aliphatic hydrocarbon groups having a specified number of carbon atoms. For example, "C 1-6"Alkyl" refers to C1, C2, C3, C4, C5, and C6. Additionally, for example, "C..." 1-6 "Alkyl" refers to an alkyl group having 1 to 6 carbon atoms. An alkyl group can be unsubstituted or substituted, whereby one or more of its hydrogen atoms are replaced by another chemical group. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl (e.g., n-propyl and isopropyl), butyl (e.g., n-butyl, isobutyl, tert-butyl), pentyl (e.g., n-pentyl, isopentyl, neopentyl), etc. Those skilled in the art will understand that C is used herein. 1-6 Alkyl groups contain monovalent C 1-6 Alkyl, divalent C 1-6 Alkylenes, such as C3-C6 cycloalkyl-(C1-C6 alkyl) r The C1-C6 alkyl in - refers to C1-C6 alkylene.

[0242] The term "alkoxy" refers to any of the above-mentioned alkyl groups (e.g., C10, C20, C30, C40, C50, C6 ... 1-6 Alkyl, C 1-4 Alkyl, C 1-3 Alkyl groups, etc., which are attached to the rest of the molecule by oxygen atoms (-O-).

[0243] The term "halogenated C" 1-6 Alkyl or halogenated C 1-6 "Alkoxy" refers to an alkyl or alkoxy group in which one or more (e.g., two or three) hydrogen atoms are replaced by a halogen atom, such as fluorine, chlorine, or bromine. The definition of the alkyl or alkoxy group is as described above. In some embodiments, the term "halogenated C" is used... 1-6 "Alkyl" is preferably fluorinated, and can be, for example, -CF3, -CHF2, -CH2F, -CH2CH2F, -CH2CHF2, -CH2CF3, etc. In some embodiments, the term "halogenated C" is used. 1-6 The alkoxy group is preferably fluorinated, for example, it can be -OCF3, -OCHF2, -OCH2F, -OCH2CH2F, -OCH2CHF2, -OCH2CF3, etc.

[0244] The term "hydroxy-substituted C" 1-6 "Alkyl" refers to an alkyl group in which one hydrogen atom is replaced by a hydroxyl group, as defined above. As an example, the "hydroxyl-substituted C..." 1-6 "alkyl" can be hydroxymethyl.

[0245] The term "cycloalkyl" refers to a cyclic alkyl group, including monocyclic, bicyclic, or polycyclic systems. When a cycloalkyl group is bicyclic or polycyclic, each ring should be a saturated carbocyclic ring or carbocyclic residue. The possible connections between two rings in a bicyclic or polycyclic cycloalkyl group include bridging, fusion, or helicaling. For example, C 3-10 Cycloalkyl refers to compounds including C3, C4, C5, C6, C7, C8, C9, and C6.10 Cycloalkyl groups. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0246] The term "cycloalkenyl" refers to a cycloalkyl group as defined above, which has at least one carbon-carbon double bond, for example...

[0247] The term "cycloalkoxy" refers to any of the above-mentioned cycloalkyl groups (e.g., C10, C20, C30, C40, C50, C60, C7 ... 3-6 Cycloalkyl groups are alkyl groups that are attached to the rest of the molecule by an oxygen atom (-O-).

[0248] The term "carbocyclic ring" or "carbocyclic residue" refers to any stable 3-, 4-, 5-, 6-, or 7-membered monocyclic ring or 5-, 6-, 7-, 8-, 9-, 10-, 11-, 12-, 13-, or 14-membered bicyclic or polycyclic ring, wherein any ring can be saturated, partially saturated, unsaturated, or aromatic. The connection between each pair of bicyclic or polycyclic carbocyclic rings can include bridging, fusion, or helicaling. Examples of such carbocyclic rings include, but are not limited to, cyclopropyl, cyclobutyl, cyclobutenyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptenyl, cycloheptyl, adamantyl, cyclooctyl, phenyl, naphthyl, [2,2,2]bicyclooctane, etc. wait.

[0249] The term "aryl" refers to a monocyclic, bicyclic, or tricyclic aromatic hydrocarbon group having 5 to 14 carbon atoms in its ring moiety. When the "aryl" is bicyclic or tricyclic, each ring is an aromatic ring. The two rings of a bicyclic or tricyclic aryl group can be linked in various ways, including bridging, fusion, and helicaling. Examples include phenyl and naphthyl groups, each of which can be substituted.

[0250] The terms "heterocyclic," "heterocyclic," or "heterocyclic group" are used interchangeably and refer to substituted and unsubstituted 4-8 membered monocyclic or bicyclic groups, 8-10 membered bicyclic or tricyclic groups, and 10-14 membered tricyclic or polycyclic groups, wherein at least one ring has at least one heteroatom (O, S, or N), preferably having one, two, or three heteroatoms selected from O, S, and N. Each heteroatom-containing ring in the group may contain one or two oxygen or sulfur atoms and / or one to four nitrogen atoms, limited by the total number of heteroatoms in each ring being four or less, and further limited by the ring containing at least one carbon atom. In some preferred embodiments, the heteroatom refers only to N or O, and the total number of these does not exceed three, preferably only one or two heteroatoms. Carbon and sulfur atoms may optionally be oxidized, nitrogen atoms may optionally be quaternized, and ring atoms on the heterocycle may optionally be substituted with =O (oxo) when the valence allows. (e.g.: The fused rings completing the bicyclic and tricyclic groups may contain only carbon atoms and may be saturated, partially saturated, or completely unsaturated, aromatic or non-aromatic. Heterocyclic groups may be attached to any available nitrogen or carbon atom. As previously mentioned, heterocyclic groups include “spiroheterocyclic,” “heterobridged,” and “heterocyclic alkenyl,” etc., as described below. Exemplary heterocyclic groups include, but are not limited to, azirrobutyl, oxacyclobutyl, pyrrolidinyl, imidazolinyl, oxazolidinyl, isoxazolinyl, thiazolinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, 2-oxopiperidinyl, 2-oxopiperidinyl, 2-oxopiperidinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxaazirroheptatrienyl, 1-pyridoneyl, 4-piperidinoneyl, tetrahydropyranyl, morpholinyl, 1,3-dioxacyclopentyl, quininecycloyl, etc.

[0251] The term "saturated heterocyclic group" refers to the monocyclic, bicyclic, or tricyclic group in the aforementioned "heterocyclic group" where the group is in a completely saturated state. The "heterocyclic group" is as described above. As an example, the saturated heterocyclic group can be a morpholino group (e.g., ...). ), piperidinyl (e.g.) ), piperazine group etc.

[0252] The term "heterocyclic alkenyl" refers to a heterocyclic group that has at least one carbon-carbon double bond in a heterocycle as defined above, for example...

[0253] The term "heteroaryl" refers to the aforementioned aryl groups, both substituted and unsubstituted, having at least one heteroatom (O, N, or S) in at least one ring. These include aromatic 5-8 membered monocyclic groups, 8-10 membered bicyclic groups, and 10-14 membered tricyclic groups. The heteroatom-containing ring preferably has one, two, or three heterocyclic atoms selected from O, N, or S. Each heteroatom-containing ring of the heteroaryl group may contain one or two oxygen or sulfur atoms and / or one to four nitrogen atoms, with the limitation that the total number of heteroatoms in each ring is four or less and each ring has at least one carbon atom. For bicyclic or tricyclic heteroaryl groups, each ring is an aromatic ring.

[0254] The term "spirocyclic" refers to a bicyclic structure having a common ring atom, where each monocyclic ring is a saturated or unsaturated, aromatic or non-aromatic carbon ring having 3-7 carbon atoms. Exemplary spirocyclic alkyl groups include, but are not limited to: spirocyclic[4.5]decane, spirocyclic[3.4]octane, spirocyclic[2.3]hexane, etc. Etc. In this document, the spirocyclic group does not include the aryl group as defined above.

[0255] The terms "saturated spirocyclic" and "spirocyclic alkyl" refer to a "spirocyclic group" as defined above, in which each monocycle is fully saturated. Exemplary spirocyclic alkyl groups include, but are not limited to: spirocyclic[4.5]decane, spirocyclic[3.4]octane, and spirocyclic[2.3]hexane.

[0256] The term "spiroheterocyclic group" refers to a bicyclic structure having one shared ring atom or a tricyclic structure having two independent shared ring atoms, wherein each monocyclic ring is a saturated or unsaturated monocyclic group having 3-8 ring atoms, wherein at least one ring has 1 or 2 ring atoms selected from N, O or S(O). n The heteroatom is a carbon atom, where n is an integer from 0 to 2, and the remaining ring atoms are carbon atoms. Additionally, one or two ring carbon atoms in the heterocyclic group are optionally replaced by a -CO- group. Exemplary spiroheterocyclic groups include, but are not limited to, 5-azaspiro[2.3]hexane and 6-oxaspiro[3.4]-7-octanone. In this document, the spiroheterocyclic groups do not include heteroaryl groups as defined above.

[0257] The terms "saturated spiroheterocyclic" and "spiroheterocyclic alkyl" refer to a "spiroheterocyclic group" as defined above, where each monocycle is fully saturated. Exemplary spirocyclic alkyl groups include, but are not limited to:

[0258] The term "bridged ring group" refers to a 3- to 8-membered monocyclic carbocyclic group, whether saturated or unsaturated, aromatic or non-aromatic, in which two non-adjacent ring atoms are connected by a cross ring (CRR). n Group, C 2-6 alkenyl, C 2-6 Alkyne groups or bonds are linked, where n is an integer from 1 to 3, and each R is independently H or methyl (where (CRR) n Group, C 2-6 alkenyl, C 2-6 The alkynyl group or bond is also referred to herein as a bridging group. The bridging group is optionally substituted by one or two substituents independently selected from alkyl, halogen, alkoxy, hydroxy, or cyano groups, wherein the alkyl, alkoxy, alkenyl, and alkynyl groups are each defined as described above. Examples of bridging groups include, but are not limited to, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, etc. It should be understood herein that when two non-adjacent ring atoms of a monocyclic 4- to 7-membered hydrocarbon group are connected by a bond, the “bridging group” may be referred to as a “fused ring group” or “fused ring group.” In this document, the bridging group does not include aryl groups as defined above.

[0259] The terms "saturated bridged cycloalkanes" and "bridged cycloalkyl groups" refer to "bridged cycloalkanes" as defined above, where each ring is in a saturated state. Here, "saturated" should be understood as each carbon atom in the bridged cycloalkanes being in a saturated state (including the carbon atoms in the bridging groups).

[0260] The term "heterobridged cyclogroup" refers to a "bridged cyclogroup" as defined above, having one, two, three, or four cyclic carbon atoms (including carbon atoms in the bridging group) replaced by heteroatoms selected from N, O, or S(O)n, where n is an integer from 0 to 2. Examples of "heterobridged cyclogroups" include, but are not limited to, 2-azabicyclo[2.2.2]octane, quinine ring, 7-oxabicyclo[2.2.1]heptane, etc. In this document, the heterobridged cyclogroups do not include heteroaryl groups as defined above.

[0261] The terms "saturated heterobridged cyclogroup" and "heterobridged cycloalkyl" refer to "heterobridged cyclogroups" as defined above, where each ring is in a saturated state. Here, "saturated" should be understood as each ring atom in the bridged cyclogroup being in a saturated state (including carbon / heteroatoms in the bridging group).

[0262] Unless otherwise specified, when referring to a clearly named aryl (e.g., phenyl), cycloalkyl (e.g., cyclohexyl), heterocyclic (e.g., pyrrolidinyl, piperidinyl, morpholinyl) or heteroaryl (e.g., imidazolyl, pyrazolyl, triazolyl), the reference means a ring having 0 to 3, preferably 0 to 2, substituents selected as needed from the substituents described above for aryl, cycloalkyl, heterocyclic and / or heteroaryl.

[0263] The term "heteroatoms" should include oxygen, sulfur, and nitrogen.

[0264] The term "halogen" should include "F, Cl, Br, I".

[0265] When the term "unsaturated" is used in the text to refer to a ring or group, the ring or group may be completely unsaturated or partially unsaturated.

[0266] When the term "saturation" is used in the text to refer to a ring or group, unless otherwise specified, the ring or group should be completely saturated.

[0267] From all the above descriptions, it will be apparent to those skilled in the art that its name is any group of a compound name, such as "C". 3-10 "Cycloalkyl-C(O)-" should refer to the part conventionally derived from it, such as from the C-shaped part. 3-10 It is constructed by substituting a carbonyl group with a cycloalkyl group, wherein the cycloalkyl group is as defined above. Other similar compound names can be understood with reference to the foregoing.

[0268] The term "optional" means that it is optional. For example, "optional is selected by 1 to 3 Rs". d Replacement C 1-6 "alkyl" indicates that the C 1-6 Alkyl groups can be 1 to 3 R d It can be replaced, or it can be replaced by 1 to 3 Rs. dIt replaces [other definitions]. Other similar definitions can be understood by referring to the foregoing content.

[0269] The term "optional oxo-substitution of ring atoms in heterocyclic groups" refers to the optional substitution of ring atoms on a heterocyclic group with =O (oxo-substitution) (e.g.: The ring atom CH2 undergoes oxidation to yield a group. ).

[0270] Throughout the specification, groups and their substituents may be selected by those skilled in the art to provide stable moieties and compounds and compounds that can be used as pharmaceutically acceptable compounds and / or intermediate compounds that can be used to prepare pharmaceutically acceptable compounds.

[0271] In this document, unless otherwise explicitly stated, the descriptive phrase “…each independently selected” used throughout the document can mean either that the specific options expressed by the same or different symbols in different groups do not affect each other, or that the specific options expressed by the same or different symbols in the same group do not affect each other.

[0272] The term "XXX is substituted at any substituted position by one or more substituents selected from YYY" means that XXX can be substituted at any substituted position by one or more substituents selected from YYY. When XXX is substituted at any substituted position by multiple substituents selected from YYY, the multiple substituents can be the same or different. The multiple substituents can be two or more, preferably two, three, or four, more preferably two or three. For example, C 1-6 An alkyl group is substituted by one or more substituents selected from cyano and hydroxyl groups at any substituted position, which indicates that C 1-6 Alkyl groups can be substituted by one or more cyano groups at any substituted position, or by one or more hydroxyl groups at any substituted position, or by one or more cyano groups and hydroxyl groups (e.g., one cyano group and one hydroxyl group, or two cyano groups and one hydroxyl group, or two cyano groups and two hydroxyl groups, etc.) at any substituted position simultaneously.

[0273] In this article, when the structure and chemical name are inconsistent, the structure shall prevail.

[0274] Invention effects:

[0275] The compounds of this invention have excellent PRMT5 inhibitory activity, and compared with the MTAP wild-type (WT) HCT-116 cell line, the compounds of this invention have excellent selective inhibitory activity against the human MTAP-deficient HCT-116 cell line. They can be used to develop small molecule drugs targeting PRMT5·MTA and preferentially act on MTAP-deficient tumor cells.

[0276] Furthermore, experiments have shown that the compounds of this invention exhibit varying degrees of advantages in terms of stability in human liver microsomes, mouse pK, rat pK, and in vivo antitumor effects in constructed mouse tumor models.

[0277] Furthermore, the compounds of the present invention have a higher safety window than existing compounds and are significantly superior to existing technologies in terms of safety (such as inhibition of toxicity in hERG, CYP, etc.). Detailed Implementation

[0278] It should be understood that the terminology used herein is intended to describe specific embodiments and is not intended to be limiting. Furthermore, although any methods, apparatus, and materials similar to or equivalent to those described herein may be used to practice or test the invention, preferred methods, apparatus, and materials are described here.

[0279] The structure of the compound was determined by nuclear magnetic resonance (NMR) or mass spectrometry (MS). NMR measurements were performed using a Bruker ASCENA-400 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD). The internal standard was tetramethylsilane (TMS). Chemical shifts were expressed in 10⁻¹⁰ increments. -6 (ppm) is given as the unit.

[0280] Reaction monitoring and MS determination were performed using a Thermofisher ESQ (ESI) mass spectrometer.

[0281] The HPLC determination was performed using a Thermo Fisher Scientific U3000 DAD high-performance liquid chromatograph (GL Sciences ODS-HL HP 3μm 3.0*100mm column).

[0282] Thin-layer chromatography (TLC) used Qingdao Ocean GF254 silica gel plates. The silica gel plates used in TLC had a diameter of 0.15–0.2 mm, while the purified products were separated using high-performance thin-layer chromatography (HPLC) preparative plates with a diameter of 0.9–1.0 mm. Column chromatography used Qingdao Ocean 200–300 mesh silica gel as the carrier. The developing solvent systems were A: dichloromethane and methanol; and B: petroleum ether and ethyl acetate. The volume ratio of the solvents was adjusted according to the polarity of the compounds. For medium-pressure preparative liquid chromatography (PLC) purification, a Biotage Isera One preparative PL was used.

[0283] In the following embodiments, unless otherwise specified, all reaction materials can be purchased from suppliers in the SciFinder database. For example, some reagents in the embodiments of this invention were purchased from manufacturers such as Saen Chemical Technology (Shanghai) Co., Ltd., Shanghai Shaoyuan Reagent Co., Ltd., Nanjing Yaoshi Technology Co., Ltd., Jiangsu Aikon Biomedical R&D Co., Ltd., and Shanghai Bid Pharmaceutical Technology Co., Ltd. Furthermore, unless otherwise specified, all raw materials used in the embodiments of this invention are of analytical grade. Unless otherwise specified, all ratios of two liquid substances mentioned herein are volume ratios; all percentages of substances mentioned are mass percentages.

[0284] Example 1: 4-Amino-N,1-Dimethyl-N-(6-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 1)

[0285] Preparation of Compound 1c: Compound 1a (86 mg, 0.38 mmol) was dissolved in N,N-dimethylacetamide (3 mL), and compound 1b (94.2 mg, 0.39 mmol, preparation method referred to WO2022169948A1), N,N-diisopropylethylamine (205.17 μL, 1.24 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (235.5 mg, 0.62 mmol) were added. After addition, the mixture was stirred at 25 °C for 3 hours. The reaction solution was quenched with water, extracted with ethyl acetate (3 x 15 mL), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to normal-phase silica gel column chromatography (dichloromethane:methanol = 10:1) to obtain compound 1c (60 mg, 0.13 mmol), ESI-MS (m / z): 452.3 [M+H]. + .

[0286] Preparation of Compound 1

[0287] Under nitrogen protection, 4-ethynyl-1-methyl-1H-pyrazole (32 mg, 0.3 mmol), cuprous iodide (6 mg, 0.03 mmol), N,N-diisopropylethylamine (39 mg, 0.3 mmol), and tetrakis(triphenylphosphine)palladium (23 mg, 0.02 mmol) were added sequentially to a solution of compound 1c (66 mg, 0.15 mmol) in N-methylpyrrolidone (1 mL). After addition, the mixture was reacted in a microwave oven at 70 °C for 3 hours. Upon completion of the reaction, water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation. The mixture was purified by reversed-phase column chromatography (acetonitrile: 0.05% ammonium bicarbonate aqueous solution = 0:100%–50%:50%) to give compound 1 (19 mg). ESI-MS (m / z): 478.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.33(s,1H),8.28(s,1H),8.08(s,1H),7.69(s,1H),7.65–7.63(m2H),7.45(d,J=7.6Hz, 1H),7.16(s,2H),7.10(d,J=8.0Hz,1H),6.99(s,1H),4.81–4.68(m,2H),4.44(s,3H),3.87(s,3H),2.70(s,3H).

[0288] Example 2: 4-Amino-N-cyclopropyl-1-methyl-N-{6-[(1-methylpyrazol-4-yl)ethynyl]-2,3-dihydro-1-benzofuran-3-yl}pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 2)

[0289] Preparation of compound 2b: Cyclopropylamine (0.38 mL, 5.48 mmol) and acetic acid (0.09 mL, 1.49 mmol) were added to a solution of compound 2a (1.0 g, 4.98 mmol) in dichloromethane (10 mL), and the reaction was carried out overnight at 25 °C. After the reaction was completed, the solution was concentrated to give compound 2b (1.0 g). ESI-MS (m / z): 240.0 [M+H] + .

[0290] Preparation of Compound 2c: Potassium tert-butoxide (1.16 g, 10.41 mmol) was added to a solution of trimethyl sulfoxide (2.3 g, 10.41 mmol) in tetrahydrofuran (10 mL). The mixture was stirred at 25 °C for 30 minutes. Compound 2b (1.0 g, 4.16 mmol) was dissolved in tetrahydrofuran (5 mL) and added dropwise to the system. The reaction was allowed to proceed at room temperature for 1 hour, then heated to 50 °C and reacted for 3 hours. Potassium tert-butoxide (4.16 mmol, 467 mg) was then added, and the reaction was continued overnight at 25 °C. After the reaction was complete, the mixture was quenched with water (15 mL), extracted with ethyl acetate (20 x 3 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Normal-phase column chromatography (ethyl acetate / petroleum ether = 0:100%–50%:50%) yielded compound 2c (200.0 mg). ESI-MS (m / z): 254.2 [M+H] + .

[0291] Preparation of Compound 2d: Compound 1b (114.4 mg, 0.47 mmol), O-(IH-benzotriazol-1-yl)-N,N,N',N'-tetramethylisourea boron tetrafluoride (189.6 mg, 0.59 mmol), and N,N-diisopropylethylamine (0.20 mL, 1.18 mmol) were added to a solution of compound 2c (100 mg, 0.39 mmol) in N,N-dimethylacetamide (5 mL). The reaction was carried out at 25 °C for 3 hours. Water (20 mL) was added, and the mixture was extracted with ethyl acetate (20 x 3 mL). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and subjected to normal-phase column chromatography (methanol / dichloromethane = 0:100%–10%:90%) to give compound 2d (130.0 mg). ESI-MS (m / z): 478.2 [M+H] + .

[0292] Preparation of compound 2

[0293] Under nitrogen protection, 4-ethynyl-1-methyl-1H-pyrazole (21 mg, 0.2 mmol), cuprous iodide (4 mg, 0.02 mmol), N,N-diisopropylethylamine (26 mg, 0.2 mmol), and tetrakis(triphenylphosphine)palladium (12 mg, 0.01 mmol) were added sequentially to a solution of compound 2d (48 mg, 0.1 mmol) in N-methylpyrrolidone (1 mL). After addition, the mixture was microwaved at 70 °C for 3 hours. Upon completion of the reaction, water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation. The mixture was purified by reversed-phase column chromatography (acetonitrile: 0.05% ammonium bicarbonate aqueous solution = 0:100%–40%:60%) to give compound 2 (23 mg). ESI-MS (m / z): 503.7 [M+H]+ . 1 H NMR(400MHz,DMSO-d6)δ8.46(d,J=2.0Hz,1H),8.28(s,1H),8.08(s,1H),7.77(d d,J=8.0,2.0Hz,1H),7.70(d,J=0.8Hz,1H),7.60–7.57(m,2H),7.16(s,2H),7.0 9(dd,J=7.6,1.2Hz,1H),6.98(d,J=1.2Hz,1H),5.96(dd,J=9.2,4.0Hz,1H),4.8 5–4.67(m,2H),4.42(s,3H),3.87(s,3H),2.88–2.79(m,1H),0.46–0.06(m,4H).

[0294] Example 3: 4-Amino-N-methyl-N-(6-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1,3-dihydrofuran[3,4-c]quinoline-8-carboxamide (Compound 3)

[0295] Preparation of Compound 3b: Compound 3a (50.6 mg, 0.22 mmol, preparation method referred to WO2022169948 A1), N,N-diisopropylethylamine (108.93 μL, 0.66 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (208.4 mg, 0.55 mmol) were added to a solution of compound 1a (50 mg, 0.22 mmol) in N,N-dimethylformamide (2 mL). The reaction mixture was reacted overnight at 25 °C. The reaction solution was then diluted with water (50 mL), extracted with ethyl acetate (3 x 10 mL), and the organic phase was washed three times with saturated brine. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The filtrate was then subjected to normal-phase column chromatography (dichloromethane / methanol = 10:1) to obtain compound 3b (50 mg). ESI-MS (m / z): 439.92 [M+H] + .

[0296] Preparation of Compound 3: Cuprous iodide (4.3 mg, 0.02 mmol), tetrakis(triphenylphosphine)palladium (13.1 mg, 0.01 mmol), N,N-diisopropylethylamine (56.43 μL, 0.34 mmol), and 4-ethynyl-1-methylpyrazole (155.8 mg, 1.59 mmol) were added to a solution of 3b (50 mg, 0.11 mmol) in 2 mL of N-methylpyrrolidone. After addition, the mixture was microwaved at 70 °C for 3 hours under nitrogen protection. The reaction solution was filtered through diatomaceous earth, water (50 mL) was added, and the mixture was extracted with ethyl acetate (3 x 10 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude compound was obtained by normal-phase column chromatography (dichloromethane / methanol = 10:1), followed by reverse-phase column chromatography (acetonitrile / 0.05% ammonium bicarbonate solution = 0:100%–40%:60%) to give compound 3 (16 mg). ESI-MS (m / z): 466.1 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ8.06(s,1H),7.68–7.57(m,2H),7.39(d,J=7.6Hz,1H),7.08(d,J=7.6Hz,1H),6.96(s,1H), 6.69(s,2H),6.30–5.64(m,1H),5.40–5.33(m,2H),5.06–4.97(m,2H),4.72–4.60(m,2H),3.85(s,3H),2.62(s,3H).

[0297] Example 4: 4-Amino-N-(6-((1-cyclopropyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 4)

[0298]

[0299] To a solution of N-methylpyrrolidone (63 mg, 0.14 mmol) in 2 mL, 4 mg (0.02 mmol) of cuprous iodide, 32 mg (0.03 mmol) of tetrakis(triphenylphosphine) palladium, 90 mg (0.7 mmol) of N,N-diisopropylethylamine, and 55 mg (0.42 mmol) of 1-cyclopropyl-4-ethynyl-1H-pyrazole were added. The mixture was then microwaved at 80 °C for 3 hours under nitrogen protection. The reaction solution was filtered through diatomaceous earth, and water (15 mL) was added. Extraction was performed with ethyl acetate (3 x 10 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Reversed-phase column chromatography (acetonitrile / 0.05% ammonium bicarbonate solution = 0:100%–50%:50%) yielded compound 4 (15 mg). ESI-MS (m / z): 504.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.33(s,1H),8.28(s,1H),8.18(s,1H),7.71–7.62(m,3H),7.45(d,J=7.6Hz,1H),7.16–7.06(m, 3H),6.97(s,1H),6.39–5.84(m,1H),4.79–4.69(m,2H),4.44(s,3H),3.79–3.74(m,1H),2.70(s,3H),1.11–0.97(m,4H).

[0300] Compound 4 was separated by SFC (equipment: Waters 150Prep-SFC; column: Chiral AS column; mobile phase A: CO2, mobile phase B: methanol (containing 0.1% NH3·H2O); B%: 35%) to obtain compounds 4A and 4B. Compound 4A: SFC analytical method: equipment: SHIMADZU LC-20AD; column: Chiral AS column; mobile phase A: CO2, mobile phase B: methanol (containing 0.05% diethylamine); B%: 5-40%; retention time: 2.176 min, ee = 100%.

[0301] Compound 4B: SFC analysis method: Equipment: SHIMADZU LC-20AD; Column: Chiral AS column; Mobile phase A: CO2, Mobile phase B: Methanol (containing 0.05% diethylamine); B%: 5-40%; Retention time: 2.565 min, ee = 100%.

[0302] Example 5: 4-Amino-7-fluoro-N,1-dimethyl-N-(6-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazol[4,3-c]quinoline-8-carboxamide (Compound 5)

[0303]

[0304] Preparation of compound 5b

[0305] Compound 5a (57.2 mg, 0.22 mmol, preparation method according to WO2022169948 A1), N,N-diisopropylethylamine (108.93 μL, 0.66 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (208.4 mg, 0.55 mmol) were added to a solution of compound 1a (50 mg, 0.22 mmol) in N,N-dimethylformamide (2 mL). The reaction mixture was reacted overnight at 25 °C. The reaction mixture was then diluted with water (50 mL), extracted with ethyl acetate (3 x 10 mL), and washed three times with saturated brine. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and subjected to normal-phase column chromatography (dichloromethane / methanol = 10:1) to give compound 5b (55 mg). ESI-MS (m / z): 469.95 [M+H] + .

[0306] Preparation of Compound 5: Cuprous iodide (4.5 mg, 0.02 mmol), tetrakis(triphenylphosphine)palladium (13.5 mg, 0.01 mmol), N,N-diisopropylethylamine (58.11 μL, 0.35 mmol) and 4-ethynyl-1-methylpyrazole (62.0 mg, 0.58 mmol) were added to a 2 mL solution of N-methylpyrrolidone (55 mg, 0.12 mmol) of Compound 5b. The mixture was then microwaved at 70 °C for 3 hours under nitrogen protection. The reaction solution was filtered through diatomaceous earth, water (50 mL) was added, and the mixture was extracted with ethyl acetate (3 x 10 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Normal-phase column chromatography (methanol / dichloromethane = 10:1) was performed to give the crude compound, followed by reverse-phase column chromatography (acetonitrile / 0.05% ammonium bicarbonate solution = 0:100%–45%:55%) to give compound 5 (18 mg). ESI-MS (m / z): 496.0 [M+H] + . 1H NMR(400MHz,DMSO-d6)δ8.29–8.22(m,2H),8.05(s,1H),7.67(s,1H),7.39–7.27(m,4H),7.12–7.06(m,1H), 6.99–6.92(m,1H),6.48–5.56(m,1H),4.83–4.64(m,2H),4.41–4.39(m,3H),3.85(s,3H),2.69–2.57(m,3H).

[0307] Compound 5 was separated by SFC (equipment: Waters 150Prep-SFC; column: Chiral AS column; mobile phase A: CO2, mobile phase B: methanol (containing 0.1% NH3·H2O); B%: 35%) to obtain compounds 5A and 5B. Compound 5A: SFC analytical method: equipment: SHIMADZU LC-20AD; column: Chiral AS column; mobile phase A: CO2, mobile phase B: methanol (containing 0.05% diethylamine); B%: 5-40%; retention time: 1.997 min, ee = 99.7%. 1 H NMR(400MHz,DMSO-d6)δ8.29–8.24(m,2H),8.07(s,1H),7.67(s,1H),7.40–7.29(m,4H),7.12–7.07(m,1H), 6.99–6.94(m,1H),6.48–5.56(m,1H),4.83–4.64(m,2H),4.41–4.39(m,3H),3.85(s,3H),2.69–2.58(m,3H). 1 H NMR (400MHz, DMSO-d6, 90℃) δ8.25–8.23(m,2H),7.96(s,1H),7.60(s,1H),7.34–7.30(m,2H),7.07(dd,J= 7.6,1.4Hz,1H),6.98(s,2H),6.92(s,1H),4.69–4.58(m,2H),4.38(s,3H),3.84(s,3H),2.65(s,3H).[α] D 25 +201.4 (c 1 mg / mL, DMF).

[0308] Compound 5B: SFC analysis method: Equipment: SHIMADZU LC-20AD; Column: Chiral AS column; Mobile phase A: CO2, Mobile phase B: Methanol (containing 0.05% diethylamine); B%: 5-40%; Retention time: 2.237 min, ee = 100%.1 H NMR(400MHz,DMSO-d6)δ8.33–8.22(m,2H),8.05(s,1H),7.67(s,1H),7.40–7.27(m,4H),7.12–7.06(m,1H), 6.99–6.95(m,1H),6.46–5.56(m,1H),4.83–4.63(m,2H),4.41–4.39(m,3H),3.85(s,3H),2.69–2.57(m,3H). 1 H NMR (400MHz, DMSO-d6, 90℃) δ8.25–8.23(m,2H),7.96(s,1H),7.60(s,1H),7.34–7.31(m,2H),7.07(d, J=7.6Hz,1H),6.98(s,2H),6.92(s,1H),4.70–4.58(m,2H),4.38(s,3H),3.84(s,3H),2.65(s,3H).[α] D 25 -215.6 (c1mg / mL, DMF).

[0309] Example 5A: (S)-4-amino-7-fluoro-N,1-dimethyl-N-(6-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazol[4,3-c]quinoline-8-carboxamide (Compound 5A)

[0310]

[0311] Compound Int-1 was separated by SFC (equipment: Shimadzu Prep-SFC; column: Opti-ChiralA1 column (20*250mm, 5μm); mobile phase A: CO2, B: ethanol (containing 0.1% NH3·H2O); B%: 35%), yielding two components.

[0312] Compound Int-1A: SFC analysis method: Equipment: SHIMADZU LC-20AD; Column: Opti-Chiral A1-3 column (100*4.6mm, 3μm); Mobile phase A: CO2, Mobile phase B: Ethanol (containing 0.05% diethylamine); B%: 30%; Retention time: 1.924 min, ee = 100%.

[0313] The preparation process of compound Int-3A is referenced in WO2024131901 A1.

[0314] Confirmation of the absolute configuration of compound Int-2A: The crystal structure of Int-2A was determined by MicroED, and it belongs to the triclinic system, space group P1 (No. 1). The cell constants measured by MicroED are... α = 97.51(12)°, β = 89.49(15)°, γ = 99.60(18)°, cell volume The Z' of the system is 1, and the asymmetric unit of the crystal is composed of 1 API molecule.

[0315] Preparation of compound 1a-A

[0316] Compound Int-3A (310 mg, 0.95 mmol) was dissolved in tetrahydrofuran (2 mL), and 4 M dioxane hydrochloride (2 mL) was added. The reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was then evaporated to dryness, slurried with methyl tert-butyl ether (5 mL), and filtered to obtain 1a-A (180 mg). ESI-MS (m / z): 227.96 [M+H] +

[0317] Preparation of compound 5A: The synthetic process for preparing compound 5A from compound 1a-A is the same as that for preparing compound 5 from compound 1a. ESI-MS (m / z): 496.0 [M+H] + [α] D 25 +213.4 (c 1 mg / mL, DMF).

[0318] Example 6: 4-Amino-N,1-Dimethyl-N-(6-((1-methyl-1,2,3,6-tetrahydropyridin-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 6)

[0319]

[0320] Preparation of compound 6b

[0321] Compound 1b (48.4 mg, 0.20 mmol), triethylamine (0.07 mL, 0.50 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (156.0 mg, 0.30 mmol) were added to a solution of compound 6a (41.9 mg, 0.20 mmol, prepared according to WO2024131901 A1) in N,N-dimethylacetamide (3 mL). The mixture was reacted at 25 °C for 18 hours. The reaction solution was diluted with ethyl acetate (10 mL) and water (10 mL), separated, and the organic phase was concentrated and purified by high-performance liquid chromatography (HPLC) to obtain compound 6b (5 mg). ESI-MS (m / z): 398.17 [M+H] + .

[0322] Preparation of compound 6

[0323] Under nitrogen protection, 1-methyl-1,2,3,6-tetrahydropyridine-4-yltrifluoromethanesulfonate (64 mg, 0.26 mmol), cuprous iodide (9.90 mg, 0.05 mmol), N,N-diisopropylethylamine (134.16 mg, 1.04 mmol), and tetrakis(triphenylphosphine)palladium (30.1 mg, 0.03 mmol) were added sequentially to a solution of compound 6b (50 mg, 0.13 mmol) in N,N-dimethylformamide (2 mL). After the addition was complete, the reaction was carried out at 25 °C for 3 hours. The reaction mixture was diluted with water (10 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated. The crude product was obtained by column chromatography (methanol:dichloromethane = 1:20), followed by reverse-phase chromatography (acetonitrile / 0.05% ammonium bicarbonate aqueous solution = 0:100%–40%:60%) to give compound 6 (15 mg). ESI-MS (m / z): 493.38 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.33(s,1H),8.28(s,1H),7.67–7.62(m,2H),7.43(d,J=7.6Hz,1H),7.16(s,2H),7.05(d,J=7.6Hz,1H),6.9 4(s,1H),6.43–5.80(m,2H),4.75–4.70(m,2H),4.44(s,3H),2.99–2.96(m,2H),2.69(s,3H),2.50–2.47(m,2H),2.31–2.26(m,3H).

[0324] Example 7: 4-Amino-N-ethyl-1-methyl-N-(6-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 7)

[0325]

[0326] Preparation of compound 7b

[0327] Compound 1b (100 mg, 0.41 mol) was dissolved in N,N-dimethylformamide (10 mL), and N,N-diisopropylethylamine (0.36 mL, 2.1 mol) was added. The mixture was stirred in an ice bath for 5 min, followed by the addition of N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (236 mg, 0.62 mol). The reaction was continued for another 5 min, and finally 7a (110 mg, 0.45 mol) was added. The reaction was continued for one hour. The reaction solution was diluted with water (30 mL), extracted with ethyl acetate (20 mL * 3), and the organic phase was washed three times with saturated brine. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The filtrate was then subjected to column chromatography (methanol:dichloromethane = 1:20) to obtain compound 7b (120 mg). ESI-MS (m / z): 465.91 [M+H] + .

[0328] Preparation of compound 7

[0329] Under nitrogen protection, 4-ethynyl-1-methyl-1H-pyrazole (54.7 mg, 0.52 mmol), cuprous iodide (9.8 mg, 0.05 mmol), N,N-diisopropylethylamine (0.18 mL, 1.03 mmol), and tetrakis(triphenylphosphine)palladium (29.8 mg, 0.026 mmol) were added sequentially to a solution of compound 7b (120 mg, 0.26 mmol) in N-methylpyrrolidone (3 mL). After addition, the mixture was reacted in a microwave oven at 70 °C for three hours. The reaction solution was then diluted with water (50 mL), extracted with ethyl acetate (20 mL x 3), and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation. The solution was then subjected to column chromatography (methanol:dichloromethane = 1:20) to give compound 7 (60 mg). ESI-MS (m / z): 492.22 [M+H] + . 1H NMR (400MHz, DMSO) δ8.28(s,2H),8.08(s,1H),7.69–7.59(m,3H),7.48(d,J=7.6Hz,1H),7.16(s,2H),7.08(dd,J=7.6 1.2Hz,1H),6.99(s,1H),5.92(s,1H),4.80–4.63(m,2H),4.44(s,3H),3.87(s,3H),3.30–3.22(m,2H),0.99–0.89(m,3H).

[0330] Example 8: 4-Amino-7-chloro-N-(6-(((1-cyclopropyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 8)

[0331]

[0332] Preparation of compound 8b

[0333] Compound 8a (279 mg, 1.01 mmol, preparation method referred to WO2022169948 A1), triethylamine (255.1 mg, 2.52 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (576.3 mg, 1.51 mmol) were added to a solution of compound 1a (230 mg, 1.01 mmol) in N,N-dimethylformamide (10 mL). The reaction mixture was reacted overnight at 25 °C. The reaction solution was then diluted with water (50 mL), extracted with ethyl acetate (2 x 25 mL), and the organic phase was washed three times with saturated brine. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Normal-phase column chromatography (dichloromethane / methanol = 50:1) was performed to give compound 8b (280 mg). ESI-MS (m / z): 487.89 [M+H] + .

[0334] Preparation of compound 8c

[0335] To a solution of compound 8b (280 mg, 0.58 mmol) in N-methylpyrrolidone (3 mL), cuprous iodide (21.9 mg, 0.12 mmol), tetratetraphenylphosphine palladium (66.5 mg, 0.06 mmol), N,N-diisopropylethylamine (371.52 mg, 2.88 mmol), and trimethylsilylacetylene (113 mg, 1.15 mmol) were added. The mixture was then microwaved at 70 °C for 3 hours under nitrogen protection. The reaction solution was filtered through diatomaceous earth, and water (50 mL) was added. Extraction was performed with ethyl acetate (2 x 25 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Normal-phase column chromatography (dichloromethane / methanol = 20:1) was performed to give compound 8c (220 mg). ESI-MS (m / z): 504.3 [M+H] + .

[0336] Preparation of compound 8d

[0337] To a tetrahydrofuran (5 mL) solution of compound 8c (220 mg, 0.44 mmol), tetrabutylammonium fluoride (870 μL, 0.87 mmol) was added, and the reaction was carried out at 25 °C for 1 h. The reaction mixture was then extracted with water (15 mL) and ethyl acetate (2 x 25 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The filtrate was purified by preparative chromatography (dichloromethane / methanol = 10:1) to give compound 8d (160 mg). ESI-MS (m / z): 432.03 [M+H] + .

[0338] Preparation of compound 8

[0339] Under nitrogen protection, 1-cyclopropyl-4-iodopyrazole (46.6 mg, 0.20 mmol), cuprous iodide (7.60 mg, 0.04 mmol), N,N-diisopropylethylamine (51.6 mg, 0.4 mmol), and tetrakis(triphenylphosphine)palladium (23 mg, 0.02 mmol) were added sequentially to a solution of compound 8d (43 mg, 0.10 mmol) in N,N-dimethylformamide (2 mL). After the addition was complete, the reaction mixture was reacted at 25 °C for 3 hours. The reaction mixture was then diluted with water (10 mL) and extracted with ethyl acetate (10 mL * 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by evaporation. The crude product was obtained by column chromatography (methanol:dichloromethane = 1:20), and then subjected to reversed-phase chromatography (acetonitrile / 0.05% ammonium bicarbonate aqueous solution = 0:100%–40%:60%) to give compound 8 (28 mg). ESI-MS (m / z): 538.15 [M+H] + . 1HNMR(400MHz,DMSO-d6)δ8.41–8.21(m,2H),8.19–8.15(m,1H),7.72–7.61(m,2H),7.54–7.30(m,3H),7.15–6.94(m,2H),6.53–6.48and 5.46–5.42(m,1H),4.91–4.53(m,2H),4.48–4.38(m,3H),3.79–3.74(m,1H),2.73–2.50(m,3H),1.11–1.04(m,2H),1.03–0.97(m,2H).

[0340] Example 8A: (S)-4-amino-7-chloro-N-(6-((1-cyclopropyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 8A)

[0341]

[0342] The synthetic process for preparing compound 8A from compounds 1a-A is the same as that for preparing compound 8 from compound 1a. ESI-MS (m / z): 538.0 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.41–8.21(m,2H),8.19–8.15(m,1H),7.72–7.61(m,2H),7.54–7.38(m,1H),7.32–7.30(m,2H),7.15–6 .94(m,2H),6.53–5.42(m,1H),4.91–4.53(m,2H),4.48–4.38(m,3H),3.79–3.74(m,1H),2.73–2.50(m,3H),1.11–0.97(m,4H).

[0343] Example 9: 4-Amino-7-chloro-N,1-dimethyl-N-(6-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 9)

[0344] In a 10 mL microwave-safe tube, reactant 8b (48.6 mg, 0.10 mmol), cuprous iodide (3.8 mg, 0.02 mmol), tetrakis(triphenylphosphine)palladium (11.5 mg, 0.01 mmol), and N-methylpyrrolidone (2 mL) were added. The mixture was bubbled with nitrogen for 3 minutes, followed by the addition of N,N-diisopropylethylamine (52.94 μL, 0.32 mmol) and N-methyl-4-ynylpyrazole (21.2 mg, 0.20 mmol). The reaction mixture was stirred in a microwave at 70 °C for 3 hours. After the reaction was complete, the reaction was quenched with water, extracted three times with ethyl acetate (30 mL), and the organic phases were combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to column chromatography (methanol:dichloromethane = 1:20) to obtain the crude product. Compound 9 (15 mg) was then separated by reverse-phase column chromatography (acetonitrile:0.05% ammonium bicarbonate aqueous solution = 0:100%–35%:65%). ESI-MS (m / z): 511.90 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ8.41–8.21(m,2H),8.10–8.05(m,1H),7.72–7.61(m,2H),7.54–7.30(m,3H),7.15–6.91(m,2H),6.54–6.48and 5.47–5.42(m,1H),4.87–4.56(m,2H),4.47–4.41(m,3H),3.87–3.86(m,3H),2.73–2.50(m,3H).

[0345] Compound 9 was separated by SFC (equipment: Waters 150Prep-SFC; column: Chiral AS column; mobile phase A: CO2, mobile phase B: methanol (containing 0.1% NH3·H2O); B%: 45%) to obtain compounds 9A and 9B. Compound 9A: SFC analytical method: equipment: SHIMADZU LC-20AD; column: Chiral AS column; mobile phase A: CO2, mobile phase B: methanol (containing 0.05% diethylamine); B%: 5-40%; retention time: 2.187 min, ee = 100%. ESI-MS (m / z): 512.2 [M+H] + .

[0346] Compound 9B: SFC analysis method: Equipment: SHIMADZU LC-20AD; Column: Chiral AS column; Mobile phase A: CO2, Mobile phase B: Methanol (containing 0.05% diethylamine); B%: 5-40%; Retention time: 2.376 min, ee = 100%. ESI-MS (m / z): 512.3 [M+H] +.

[0347] Example 10: 4-Amino-N,1,7-Trimethyl-N-(6-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 10)

[0348]

[0349] Preparation of compound 10b

[0350] Compound 10a (256.3 mg, 0.92 mmol, preparation method according to WO2022169948 A1), triethylamine (232.9 mg, 2.30 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (526.2 mg, 1.38 mmol) were added to a solution of compound 1a (210 mg, 0.92 mmol) in N,N-dimethylformamide (10 mL). The reaction mixture was reacted overnight at 25 °C. The reaction solution was then diluted with water (50 mL), extracted with ethyl acetate (2 x 25 mL), and the organic phase was washed three times with saturated brine. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Normal-phase column chromatography (dichloromethane / methanol = 50:1) was performed to give compound 10b (360 mg). ESI-MS (m / z): 468.07 [M+H] + .

[0351] Preparation of compound 10

[0352] Add reactant 10b (46.6 mg, 0.10 mmol), cuprous iodide (3.8 mg, 0.02 mmol), tetrakis(triphenylphosphine)palladium (11.5 mg, 0.01 mmol), and solvent N-methylpyrrolidone (2 mL) to a 10 mL microwave-safe tube. Bubble under nitrogen for 3 minutes, then add N,N-diisopropylethylamine (52.94 μL, 0.32 mmol) and N-methyl-4-ynylpyrazole (21.2 mg, 0.20 mmol). Stir the reaction mixture in a microwave at 70 °C for 3 hours. After the reaction was completed, the reaction was quenched with water, and the organic phases were extracted three times with ethyl acetate (30 ml) and combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to column chromatography (methanol:dichloromethane = 1:20) to obtain the crude product. Compound 10 (15 mg) was then separated by reverse-phase column chromatography (acetonitrile:0.05% ammonium bicarbonate aqueous solution = 0:100%–35%:65%). ESI-MS (m / z): 491.96 [M+H] + . 1H NMR(400MHz,DMSO-d6)δ8.24(s,1H),8.07–8.04(m,2H),7.69(s,1H),7.52–7.43(m,2H),7.17–6.95(m,4H),6.55–6.48and 5.54–5.45(m,1H),4.91–4.54(m,2H),4.39(s,3H),3.87(s,3H),3.30(s,3H),2.72–2.37(m,3H).

[0353] Example 10A: (S)-4-amino-N,1,7-trimethyl-N-(6-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 10A)

[0354]

[0355] The synthetic process for preparing compound 10A from compound 1a-A is the same as the synthetic process for preparing compound 10 from compound 1a.

[0356] Compound 10A: ESI-MS (m / z): 492.01 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ8.24(s,1H),8.07–8.04(s,1H),7.69(s,1H),7.52–7.44(m,2H),7.14–6.95(m ,4H),6.55–5.45(m,1H),4.91–4.54(m,2H),4.39(s,3H),3.87(s,3H),3.30(s,3H),2.72–2.37(s,3H).

[0357] Example 11: 4-Amino-N-(6-((1-cyclopropyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1,7-trimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 11)

[0358]

[0359] Preparation of Compound 11b: Compound 6a (25 mg, 0.12 mmol), triethylamine (0.05 mL, 0.36 mmol), and O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethylisourea boron tetrafluoride (57.8 mg, 0.18 mmol) were added to a solution of compound 10a (30.8 mg, 0.12 mmol) in N,N-dimethylacetamide (5 mL). The reaction mixture was reacted overnight at 25 °C. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (10 x 3 mL). The organic phase was washed three times with saturated brine. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Reversed-phase column chromatography (acetonitrile / 0.05% ammonium bicarbonate aqueous solution = 0:100%-50%:50%) was performed to obtain compound 11a (7.0 mg), ESI-MS (m / z): 412.15.

[0360] Compound 11 was prepared under nitrogen protection by adding 1-cyclopropyl-4-iodopyrazole (56.9 mg, 0.24 mmol), cuprous iodide (9.90 mg, 0.05 mmol), N,N-diisopropylethylamine (134.16 mg, 1.04 mmol), and tetrakis(triphenylphosphine)palladium (30.1 mg, 0.03 mmol) sequentially to a solution of compound 11a (50 mg, 0.12 mmol) in N,N-dimethylformamide (2 mL). After addition, the reaction was carried out at 25 °C for 3 hours. The reaction mixture was diluted with water (10 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated. The crude product was obtained by column chromatography (methanol:dichloromethane = 1:20), and compound 11 (20 mg) was prepared by reverse-phase chromatography (acetonitrile / 0.05% ammonium bicarbonate aqueous solution = 0:100%–40%:60%). ESI-MS (m / z): 518.22 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ8.39(s,1H),8.30–8.10(m,2H),7.95–7.38(m,4H),7.26–6.94(m,3H),6.52–6.50and 5.50–5.42(m,1H),4.89–4.42(m,5H),3.80–3.74(m,1H),2.73and 2.49(s,3H),2.56and 2.39(s,3H),1.14–1.05(m,2H),1.05–0.95(m,2H).

[0361] Example 12: 4-Amino-N-(6-((1-cyclopropyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 12)

[0362] Preparation of compound 12a

[0363] Under nitrogen protection, trimethylsilylacetylene (338.5 mg, 3.45 mmol), cuprous iodide (65.6 mg, 0.34 mmol), N,N-diisopropylethylamine (890.9 mg, 6.89 mmol), and tetrakis(triphenylphosphine)palladium (199.1 mg, 0.17 mmol) were added sequentially to a solution of compound 5b (808 mg, 1.72 mmol) in N-methylpyrrolidone (10 mL). The mixture was then reacted in a microwave oven at 100 °C for three hours. The reaction solution was diluted with water (200 mL), extracted with ethyl acetate (30 mL x 3), and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation. The resulting solution was then subjected to column chromatography (methanol:dichloromethane = 1:20) to give compound 12a (595 mg). ESI–MS (m / z): 488.18 [M+H] + .

[0364] Preparation of compound 12b

[0365] A solution of tetrabutylammonium fluoride (2.44 mL, 1 M) was added to a tetrahydrofuran (10 mL) solution of compound 12a (595 mg, 1.22 mmol), and the mixture was allowed to react at room temperature for one hour. Water was added, and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and subjected to column chromatography (methanol:dichloromethane = 1:20) to give compound 12b (470 mg). ESI–MS (m / z): 416.12 [M+H] + .

[0366] Preparation of compound 12

[0367] Under nitrogen protection, 1-cyclopropyl-4-iodo-1H-pyrazole (70.2 mg, 0.3 mmol), cuprous iodide (5.7 mg, 0.03 mmol), N,N-diisopropylethylamine (77.5 mg, 0.6 mmol), and tetrakis(triphenylphosphine)palladium (17.3 mg, 0.015 mmol) were added sequentially to a solution of compound 12b (62.3 mg, 0.15 mmol) in N-methylpyrrolidone (1.5 mL). The reaction mixture was then reacted in an oil bath at 50 °C for three hours. Water (100 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated. The extract was then subjected to column chromatography (methanol:dichloromethane = 1:20) to give compound 12 (25 mg). ESI–MS (m / z): 521.97 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.32–8.24(m,2H),8.18(s,1H),7.68(s,1H),7.42–7.28(m,4H),7.14–7.05(m,1H),7.02–6.92(m, 1H),6.51–5.58(m,1H),4.88–4.66(m,2H),4.44–4.40(m,3H),3.81–3.73(m,1H),2.71–2.59(m,3H),1.09–0.95(m,4H).

[0368] Example 12A: (S)-4-amino-N-(6-((1-cyclopropyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 12A)

[0369]

[0370] The synthetic process for preparing compound 12A from compound 5b-A is the same as the synthetic process for preparing compound 12 from compound 5b.

[0371] Compound 12A: ESI–MS (m / z): 521.94 [M+H] + . 1H NMR (400MHz, DMSO) δ8.34–8.23(m,2H),8.18(s,1H),7.69(s,1H),7.45–7.29(m,4H),7.16–7.05(m,1H),7.02–6.92(m, 1H),6.50–5.56(m,1H),4.87–4.65(m,1H),4.44–4.40(m,3H),3.84–3.70(m,1H),2.71–2.60(m,1H),1.11–0.96(m,4H).

[0372] Example 13: 4-Amino-7-fluoro-N-(6-((1-isopropyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 13)

[0373]

[0374] Following the preparation procedure of compound 12, compound 1-cyclopropyl-4-iodo-1H-pyrazole was replaced with compound 4-iodo-1-isopropyl-1H-pyrazole (70.8 mg, 0.3 mmol), and the reaction was carried out at 50 °C for 3 hours under nitrogen protection to obtain compound 13 (28 mg). ESI–MS (m / z): 524.05 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.39–8.22(m,2H),8.17(s,1H),7.70(s,1H),7.44–7.28(m,4H),7.17–7.05(m,1H),7.02–6.93(m, 1H),6.51–5.52(m,1H),4.87–4.66(m,2H),4.57–4.48(m,1H),4.44–4.40(m,3H),2.71–2.60(m,3H),1.45–1.42(m,6H).

[0375] Example 14: 4-Amino-7-chloro-N-(6-(((1-isopropyl-1H-pyrazole-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazole[4,3-c]quinoline-8-carboxamide (Compound 14)

[0376]

[0377] Following the preparation procedure of compound 8, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 4-iodo-1-(prop-2-yl)pyrazole (21.9 mg, 0.09 mmol), and the reaction was carried out at 50 °C for 3 hours under nitrogen protection to obtain compound 14 (12 mg). ESI-MS (m / z): 540.17 [M+H] + . 1 H NMR(400MHz, DMSO+D2O)δ8.41–8.19(m,2H),8.14–8.08(m,1H),7.77–7.64(m,2H),7.49–7.25(m,1H),7.12–6.88(m,2H),6.51–6.42and 5.42–5.34(m,1H),4.84–4.46(m,3H),4.44and 4.38(s,3H),2.7–2.47(m,3H),1.45–1.34(m,6H).

[0378] Example 15: 4-Amino-7-chloro-N-(6-((1-ethyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazol[4,3-c]quinoline-8-carboxamide (Compound 15)

[0379]

[0380] Following the preparation procedure of compound 8, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 1-ethyl-4-iodopyrazole (30.8 mg, 0.14 mmol), and the reaction was carried out at 50 °C for 3 hours under nitrogen protection to obtain compound 15 (16.4 mg). ESI-MS (m / z): 526.0 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.43–8.11(m,3H),7.72–7.61(m,2H),7.54–7.29(m,3H),7.17–7.04(m,1H),7.04–6.91(m,1H),6.55–6.48and 5.48–5.38(m,1H),4.86–4.55(m,2H),4.47and 4.41(s,3H),4.21–4.09(m,2H),2.73–2.50(m,3H),1.44–1.34(m,3H).

[0381] Example 16: 4-Amino-N-(6-(((1-cyclopropyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N-methyl-1,3-dihydrofuran[3,4-c]quinoline-8-carboxamide (Compound 16)

[0382]

[0383] Preparation of Compound 16b: Compound 16a (109 mg, 0.44 mmol, preparation method referred to WO2022169948 A1), triethylamine (110.9 mg, 1.10 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (250.5 mg, 0.66 mmol) were added to a solution of compound 1a (100 mg, 0.44 mmol) in N,N-dimethylformamide (10 mL). The reaction mixture was reacted overnight at 25 °C. The reaction solution was then diluted with water (50 mL), extracted with ethyl acetate (2 x 25 mL), and the organic phase was washed three times with saturated brine. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and subjected to normal-phase column chromatography (dichloromethane / methanol = 50:1) to obtain compound 16b (118 mg). ESI-MS (m / z): 459.85 [M+H] + .

[0384] Preparation of compound 16c

[0385] To a solution of 16b (118 mg, 0.26 mmol) in 3 mL of N-methylpyrrolidone, cuprous iodide (9.8 mg, 0.05 mmol), tetratetraphenylphosphine palladium (29.8 mg, 0.03 mmol), triethylamine (104.2 mg, 2.88 mmol), and trimethylsilylacetylene (50.6 mg, 0.51 mmol) were added. The mixture was then microwaved at 70 °C for 3 hours under nitrogen protection. The reaction solution was filtered through diatomaceous earth, and water (50 mL) was added. Extraction was performed with ethyl acetate (2 x 25 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Normal-phase column chromatography (dichloromethane / methanol = 20:1) was performed to give compound 16c (105 mg). ESI-MS (m / z): 476.2 [M+H] + .

[0386] Preparation of Compound 16d: Tetrabutylammonium fluoride (440 μL, 0.44 mmol) was added to a tetrahydrofuran (5 mL) solution of compound 16c (105 mg, 0.22 mmol), and the reaction was carried out at 25 °C for 1 h. The reaction solution was then extracted with water (15 mL) and ethyl acetate (2 x 25 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by preparative chromatography (dichloromethane / methanol = 15:1) to obtain compound 16d (89.1 mg). ESI-MS (m / z): 404.09 [M+H] + .

[0387] Preparation of compound 16

[0388] Under nitrogen protection, 1-cyclopropyl-4-iodopyrazole (49.9 mg, 0.22 mmol), cuprous iodide (8.10 mg, 0.04 mmol), triethylamine (43.1 mg, 0.43 mmol), and tetrakis(triphenylphosphine)palladium (24.6 mg, 0.02 mmol) were added sequentially to a solution of compound 16d (43 mg, 0.11 mmol) in N,N-dimethylformamide (2 mL). After addition, the reaction mixture was reacted at 25 °C for 3 hours. The reaction mixture was then diluted with water (10 mL) and extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by evaporation. The crude product was obtained by column chromatography (methanol:dichloromethane = 1:20). Compound 16 (18 mg) was obtained by reverse-phase preparation (acetonitrile / 0.05% ammonium bicarbonate aqueous solution = 0:100%–40%:60%). ESI-MS (m / z): 510.18 [M+H] + . 1 HNMR(400MHz,DMSO-d6)δ8.19–8.18(m,1H),7.69–7.65(m,2H),7.38–7.31(m,2H),7.13–7.07(m,1H),7.00–6.95(m,1H),6.85(s,2H),6.45–6.42and 5.52–5.49(m,1H),5.38–5.32(m,2H),5.04–5.00(m,2H),4.82–4.59(m,2H),3.80–3.74(m,1H),2.68and 2.55(s,3H),1.11–0.97(m,4H).

[0389] Example 16A: (S)-4-amino-N-(6-(((1-cyclopropyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N-methyl-1,3-dihydrofuran[3,4-c]quinoline-8-carboxamide (Compound 16A)

[0390] The synthetic process for preparing compound 16A from compound 1a-A is the same as the synthetic process for preparing compound 16 from compound 1a.

[0391] Compound 16A: ESI-MS (m / z): 510.08 [M+H] + . 1H NMR (400MHz, DMSO) δ8.20–8.17(m,1H),7.71–7.62(m,2H),7.40–7.26(m,2H),7.14–7.05(m,1H),7.01–6.94(m,1H),6.85(s,2H),6.47–6.38and 5.53–5.46(m,1H),5.40–5.32(m,2H),5.04–5.00(m,2H),4.83–4.56(m,2H),3.82–3.72(m,1H),2.68and 2.55(s,3H),1.10–1.05(m,2H),1.02–0.97(m,2H).

[0392] Example 17: 4-Amino-N-(6-((1-ethyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 17)

[0393]

[0394] Following the preparation procedure of compound 12, compound 1-cyclopropyl-4-iodo-1H-pyrazole was replaced with compound 1-ethyl-4-iodo-1H-pyrazole (66.6 mg, 0.3 mmol) to obtain compound 17 (30 mg). ESI–MS (m / z): 510.05 [M+H] + . 1 HNMR (400MHz, DMSO) δ8.36–8.21(m,2H),8.13(s,1H),7.70(s,1H),7.43–7.28(m,4H),7.16–7.05(m,1H),7.03–6.94(m, 1H),6.50–5.54(m,1H),4.87–4.61(m,2H),4.45–4.40(m,3H),4.20–4.13(m,2H),2.71–2.60(m,3H),1.42–1.37(m,3H).

[0395] Example 18: 4-Amino-7-chloro-N,1-dimethyl-N-(6-((1-methyl-1H-pyrazol-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 18)

[0396]

[0397] Following the preparation procedure of compound 8, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 3-iodo-1-methylpyrazole (41.6 mg, 0.20 mmol) to obtain compound 18 (21 mg). ESI-MS (m / z): 511.97 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ8.41–8.21(m,2H),7.79–7.78(m,1H),7.71–7.61(m,1H),7 .57–7.28(m,3H),7.20–6.96(m,2H),6.53–5.43(m,2H),4.87–4.55(m,2H),4.46and 4.41(s,3H),3.88–3.87(m,3H),2.74–2.53(m,3H).

[0398] Example 19: 4-Amino-N-(6-((1-isopropyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 19)

[0399]

[0400] Under nitrogen protection, 4-iodo-1-isopropyl-1H-pyrazole (59.0 mg, 0.25 mmol), cuprous iodide (4.8 mg, 0.025 mmol), N,N-diisopropylethylamine (64.6 mg, 0.5 mmol), and tetrakis(triphenylphosphine)palladium (14.5 mg, 0.0125 mmol) were added sequentially to a solution of compound 6b (49.6 mg, 0.125 mmol) in N-methylpyrrolidone (1.5 mL). The reaction mixture was then reacted in an oil bath at 50 °C for three hours. Water (100 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated. The extract was then subjected to column chromatography (methanol:dichloromethane = 1:20) to give compound 19 (30 mg). ESI–MS (m / z): 506.05 [M+H] + . 1H NMR (400MHz, DMSO) δ8.34(s,1H),8.28(s,1H),8.17(s,1H),7.70(s,1H),7.68–7.62(m,2H),7.48–7.42(m,1H),7.18(s,2H),7.12 –7.07(m,1H),6.98(s,1H),6.30–5.87(m,1H),4.79–4.70(m,2H),4.57–4.49(m,1H),4.44(s,3H),2.70(s,3H),1.46–1.43(m,6H).

[0401] Example 19A: (S)-4-amino-N-(6-((1-isopropyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 19A)

[0402]

[0403] The preparation process of compound 6b-A is described in WO2024131901 A1. The synthetic process for preparing compound 19A from compound 6b-A is described in the synthetic process for preparing compound 19 from compound 6b.

[0404] Compound 19A: ESI–MS (m / z): 506.13 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.33(s,1H),8.28(s,1H),8.17(s,1H),7.70(s,1H),7.67–7.62(m,2H),7.48–7.42(m,1H),7.16(s, 2H),7.11–7.06(m,1H),6.97(s,1H),4.80–4.67(m,2H),4.57–4.48(m,1H),4.44(s,3H),2.69(s,3H),1.45–1.42(m,6H).

[0405] Example 20: 4-Amino-N-(6-((1-ethyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 20)

[0406]

[0407] Following the preparation procedure of compound 19, compound 4-iodo-1-isopropyl-1H-pyrazole was replaced with compound 1-ethyl-4-iodo-1H-pyrazole (55.5 mg, 0.25 mmol) to obtain compound 20 (15 mg). ESI–MS (m / z): 492.02 [M+H] + . 1 HNMR(400MHz,DMSO)δ8.36(s,2H),8.14(s,1H),7.70–7.39(m,6H),7.14–7.07(m,1H),6.99(s,1H),6 .36–5.82(m,1H),4.82–4.68(m,2H),4.45(s,3H),4.20–4.13(m,2H),2.70(s,3H),1.43–1.36(m,3H).

[0408] Example 20A: (S)-4-amino-N-(6-((1-ethyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 20A)

[0409]

[0410] The synthetic process for preparing compound 20A from compound 6b-A is the same as the synthetic process for preparing compound 20 from compound 6b.

[0411] Compound 20A: ESI–MS (m / z): 492.02 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.34(s,1H),8.28(s,1H),8.13(s,1H),7.70(s,1H),7.68–7.61(m,2H),7.48–7.42(m,1H),7.19(s, 2H),7.12–7.07(m,1H),6.98(s,1H),4.80–4.68(m,2H),4.44(s,3H),4.22–4.11(m,2H),2.69(s,3H),1.41–1.37(m,3H).

[0412] Example 21: 4-Amino-7-fluoro-N,1-dimethyl-N-(6-((1-methyl-1H-pyrazol-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 21)

[0413]

[0414] Following the preparation procedure of compound 12, compound 1-cyclopropyl-4-iodo-1H-pyrazole was replaced with compound 3-iodo-1-methylpyrazole (21 mg, 0.10 mmol), and the reaction was carried out at 25 °C for 3 hours to obtain compound 21 (20 mg). ESI-MS (m / z): 495.74 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ8.40–8.24(m,2H),7.81–7.78(m,1H),7.51–7.26(m, 4H),7.21–7.12(m,1H),7.09–7.01(m,1H),6.55–6.51(m,1H),6.50–6.45and 5.64–5.57(m,1H),4.86–4.65(m,2H),4.44–4.41(m,3H),3.89(s,3H),2.72and 2.61(s,3H).

[0415] Example 21A: (S)-4-amino-7-fluoro-N,1-dimethyl-N-(6-((1-methyl-1H-pyrazol-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 21A)

[0416]

[0417] The synthetic process for preparing compound 21A from compound 12b-A is the same as the synthetic process for preparing compound 21 from compound 12b.

[0418] Compound 21A: ESI-MS (m / z): 496.08 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.40–8.24(m,2H),7.81–7.78(m,1H),7.51–7.26(m,4H),7.21–7.12(m,1H),7.09–7.01(m, 1H),6.55–6.51(m,1H),6.51–5.57(m,1H),4.90–4.62(m,2H),4.44–4.41(s,3H),3.89(s,3H),2.72and2.61(s,3H).

[0419] Example 22: 4-Amino-7-chloro-N-(6-((1,5-dimethyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1Hpyrazolo[4,3-c]quinoline-8-carboxamide (Compound 22)

[0420]

[0421] Preparation of compound 22b: N-iodosuccinimide (2.5 g, 10.93 mmol) was added to a solution of compound 22a (1.0 g, 10.41 mmol) in N,N-dimethylformamide (10 mL). After addition, the reaction mixture was reacted at 25 °C for 1 hour. The reaction mixture was then diluted with water (10 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, the solvent was removed by evaporation, and the mixture was subjected to column chromatography (petroleum ether: ethyl acetate = 6:1) to obtain 22b (1.75 g).

[0422] Preparation of compound 22

[0423] Following the preparation procedure of compound 8, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 22b (41.1 mg, 0.19 mmol) to obtain compound 22 (15 mg). ESI-MS (m / z): 526.07 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.41–8.21(m,2H),7.72–7.26(m,5H),7.15–6.93(m,2H),6.55–6.48and 5.48–5.40(m,1H),4.86–4.57(m,2H),4.47and 4.41(s,3H),3.78–3.77(m,3H),2.76–2.41(m,3H),2.38–2.36(m,3H).

[0424] Example 22A: (S)-4-amino-7-chloro-N-(6-((1,5-dimethyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1Hpyrazolo[4,3-c]quinoline-8-carboxamide (Compound 22A)

[0425]

[0426] The synthetic process for preparing compound 22A from compound 22b and compound 8d-A is the same as the synthetic process for preparing compound 22 from compound 22b and compound 8d.

[0427] Compound 22A: ESI-MS (m / z): 525.95 [M+H] + .

[0428] Example 23: 4-Amino-N-(6-((1,5-dimethyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N,1-dimethyl-1Hpyrazolo[4,3-c]quinoline-8-carboxamide (Compound 23)

[0429] Following the preparation procedure of compound 12, compound 1-cyclopropyl-4-iodo-1H-pyrazole was replaced with compound 22b (44.4 mg, 0.20 mmol), and the reaction was carried out at room temperature for 3 hours to obtain compound 23 (15 mg). ESI-MS (m / z): 510.38 [M+H] + . 1 HNMR(400MHz,DMSO-d6)δ8.41–8.21(m,2H),7.59(s,1H),7.47–7.20(m,4H),7.17–7.05(m,1H),7.04–6.96(m,1H),6.49–6.43and 5.62–5.54(m,1H),4.87–4.65(m,2H),4.43–4.41(m,3H),3.78(s,3H),2.72and 2.60(s,3H),2.37(s,3H).

[0430] Compound 23 was separated by SFC (equipment: Waters 150Prep-SFC; column: Chiral AS column; mobile phase A: CO2, mobile phase B: methanol (containing 0.1% NH3·H2O); B%: 40%) to obtain compounds 23A and 23B. Compound 23A: SFC analytical method: equipment: SHIMADZU LC-20AD; column: Chiral AS column; mobile phase A: CO2, mobile phase B: methanol (containing 0.05% diethylamine); B%: 5-40%; retention time: 1.948 min, ee = 100%. [α] D 25 +247 (c 1 mg / mL, DMF).

[0431] Compound 23B: SFC analysis method: Equipment: SHIMADZU LC-20AD; Column: Chiral AS column; Mobile phase A: CO2, Mobile phase B: Methanol (containing 0.05% diethylamine); B%: 5-40%; Retention time: 2.155 min, ee = 100%. [α] D 25 -271 (c 1 mg / mL, DMF).

[0432] Example 24: 4-Amino-N-(6-(((3-chloro-1-methyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 24)

[0433] Preparation of Compound 24b: N-iodosuccinimide (235.2 mg, 1.05 mmol) was added to a solution of 3-chloro-1-methylpyrazole (116 mg, 1.0 mmol) in acetonitrile (3 mL). The reaction mixture was reacted at 25 °C for 1 hour. Water (10 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation. The solution was then subjected to column chromatography (petroleum ether:ethyl acetate = 6:1) to give compound 24b (210 mg). ESI-MS (m / z): 242.89 [M+H] + .

[0434] Preparation of compound 24

[0435] Following the preparation procedure of compound 12, compound 1-cyclopropyl-4-iodo-1H-pyrazole was replaced with compound 24b (24.2 mg, 0.10 mmol), and the reaction was carried out at 25 °C for 3 hours to obtain compound 24 (20 mg). ESI-MS (m / z): 530.17 [M+H] + . 1 HNMR(400MHz,DMSO-d6)δ8.42–8.15(m,2H),7.97–7.73(m,1H),7.49–7.28(m,4H),7.20–7.12(m,1H),7.04–6.97( m,1H),6.51–6.39and5.63–5.52(m,1H),4.86–4.62(m,2H),4.43–4.41(s,3H),3.85(s,3H),2.71and2.60(s,3H).

[0436] Compound 24 was separated by SFC (equipment: Waters 150Prep-SFC; column: Chiral AS column; mobile phase A: CO2, mobile phase B: methanol (containing 0.1% NH3·H2O); B%: 40%) to obtain compounds 24A and 24B. Compound 24A: SFC analytical method: equipment: SHIMADZU LC-20AD; column: Chiral AS column; mobile phase A: CO2, mobile phase B: methanol (containing 0.05% diethylamine); B%: 5-40%; retention time: 0.789 min, ee = 99.71%.

[0437] Compound 24B: SFC analysis method: Equipment: SHIMADZU LC-20AD; Column: Chiral AS column; Mobile phase A: CO2, Mobile phase B: Methanol (containing 0.05% diethylamine); B%: 5-40%; Retention time: 1.213 min, ee = 99.83%.

[0438] Example 25: 4-Amino-N-(6-((1,3-dimethyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N,1-dimethyl-1Hpyrazolo[4,3-c]quinoline-8-carboxamide (Compound 25)

[0439]

[0440] Following the preparation procedure of compound 12, compound 1-cyclopropyl-4-iodo-1H-pyrazole was replaced with compound 25a (22.2 mg, 0.10 mmol), and the reaction was carried out at 25 °C for 3 hours to obtain compound 25 (20 mg). ESI-MS (m / z): 510.16 [M+H] + . 1 HNMR(400MHz,DMSO-d6)δ8.40–8.25(m,2H),7.94(s,1H),7.41–7.30(m,2H),7.15–7.06(m,1H),7.03–6.95(m,1H),6.49–6.43and 5.62–5.55(m,1H),4.87–4.60(m,2H),4.43–4.41(m,3H),3.78(s,3H),2.71and 2.60(s,3H),2.24(s,3H).

[0441] Compound 25 was separated by SFC (equipment: Waters 150Prep-SFC; column: Chiral AS column; mobile phase A: CO2, mobile phase B: methanol (containing 0.1% NH3·H2O); B%: 40%) to obtain compounds 25A and 25B. Compound 25A: SFC analytical method: equipment: SHIMADZU LC-20AD; column: Chiral AS column; mobile phase A: CO2, mobile phase B: methanol (containing 0.05% diethylamine); B%: 5-40%; retention time: 1.832 min, ee = 99.94%. [α] D 25 +231.4 (c 1 mg / mL, DMF).

[0442] Compound 25B: SFC analysis method: Equipment: SHIMADZU LC-20AD; Column: Chiral AS column; Mobile phase A: CO2, Mobile phase B: Methanol (containing 0.05% diethylamine); B%: 5-40%; Retention time: 2.019 min, ee = 100%. [α] D 25 -253 (c 1mg / mL, DMF).

[0443] Example 26: 4-Amino-7-chloro-N-(6-((1,3-dimethyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1Hpyrazolo[4,3-c]quinoline-8-carboxamide (Compound 26)

[0444]

[0445] Following the preparation procedure of compound 8, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 25a (22.2 mg, 0.10 mmol), and the reaction was carried out at 25 °C for 3 hours to obtain compound 26 (20 mg). ESI-MS (m / z): 525.98 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ8.44–8.21(m,2H),7.95–7.93(m,1H),7.70–7.63(m,1H),7.52–7.27(m,3H),7.14–6.95(m,2H),6.52–6.50and 5.47–5.42(m,1H),4.89–4.55(m,2H),4.47–4.41(m,3H),3.79and 3.78(s,3H),2.74–2.51(m,3H),2.25–2.23(m,3H).

[0446] Example 26A: (S)-4-amino-7-chloro-N-(6-((1,3-dimethyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1Hpyrazolo[4,3-c]quinoline-8-carboxamide (Compound 26A)

[0447]

[0448] The synthetic process for preparing compound 26A from compound 8d-A is the same as the synthetic process for preparing compound 26 from compound 8d.

[0449] Compound 26A: ESI-MS (m / z): 525.97 [M+H] + .1 H NMR (400MHz, DMSO-d6) δ8.44–8.21(m,2H),7.95–7.93(m,1H),7.70–7.63(m,1H),7.52–7.27(m,3H),7.14–6.95(m,2H ),6.52–5.42(m,1H),4.89–4.55(m,2H),4.47–4.41(m,3H),3.79–3.78(m,3H),2.74–2.51(m,3H),2.25–2.23(m,3H).

[0450] Example 27: 4-Amino-N-ethyl-7-fluoro-1-methyl-N-(6-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 27)

[0451]

[0452] Preparation of compound 27a

[0453] Compound 5a (106 mg, 0.41 mol) was dissolved in N,N-dimethylformamide (10 mL), and N,N-diisopropylethylamine (0.36 mL, 2.04 mol) was added. The mixture was stirred in an ice bath for 5 min, followed by the addition of N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (232.53 mg, 0.61 mol). The reaction was continued for 5 min, and finally 7a (113.54 mg, 0.41 mol) was added. The reaction was continued for one hour. The reaction solution was diluted with water (100 mL), extracted with ethyl acetate (20 mL * 3), and the organic phase was washed three times with saturated brine. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The filtrate was then subjected to column chromatography (methanol:dichloromethane = 1:20) to obtain compound 27a (120 mg). ESI-MS (m / z): 484.11 [M+H] + .

[0454] Preparation of compound 27

[0455] Under nitrogen protection, 4-ethynyl-1-methyl-1H-pyrazole (53.0 mg, 0.50 mmol), cuprous iodide (9.5 mg, 0.05 mmol), N,N-diisopropylethylamine (0.17 mL, 0.99 mmol), and tetrakis(triphenylphosphine)palladium (28.7 mg, 0.025 mmol) were added sequentially to a solution of compound 27a (120 mg, 0.25 mmol) in N-methylpyrrolidone (3 mL). After addition, the mixture was microwaved at 100 °C for three hours. Water (50 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation. The solution was then subjected to column chromatography (methanol:dichloromethane = 1:20) to give compound 27 (15 mg). ESI-MS (m / z): 510.1 [M+H] + . 1 ¹H NMR (400 MHz, DMSO) δ 8.38–8.17 (m, 2H), 8.07 (s, 1H), 7.69 (s, 1H), 7.50–7.26 (m, 4H), 7.14–6.95 (m, 2H), 6.20–5.53 (m, 1H), 4.91–4.53 (m, 2H), 4.42 (s, 3H), 3.87 (s, 3H), 3.25–3.05 (m, 2H), 1.10–0.72 (m, 3H). Example 28: 4-amino-7-chloro-N-ethyl-1-methyl-N-(6-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 28)

[0456]

[0457] Preparation of compound 28a

[0458] Following the preparation procedure of compound 27a, compound 5a was replaced with compound 8a (120 mg, 0.43 mol) to obtain compound 28a (160 mg). ESI-MS (m / z): 499.94 [M+H] + .

[0459] Preparation of compound 28

[0460] Following the preparation procedure of compound 27, compound 27a was replaced with compound 28a (100 mg, 0.2 mmol) to obtain compound 28 (20 mg). ESI-MS (m / z): 526.2 [M+H] + . 1H NMR (400MHz, DMSO) δ8.40–8.15(m,2H),8.11–8.04(m,1H),7.72–7.62(m,2H),7.55–7.36(m,1H),7.30(s,2H),7.16–6. 93(m,2H),6.22–5.31(m,1H),4.91–4.57(m,2H),4.51–4.37(m,3H),3.87(s,3H),3.32–2.91(m,2H),1.14–0.75(m,3H).

[0461] Example 29: 4-Amino-N-(6-((1,5-dimethyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1Hpyrazolo[4,3-c]quinoline-8-carboxamide (Compound 29)

[0462]

[0463] Following the preparation procedure of compound 19, compound 4-iodo-1-isopropyl-1H-pyrazole was replaced with compound 22b (22.2 mg, 0.10 mmol), and the reaction was carried out at 25 °C for 3 hours to obtain compound 29 (15 mg). ESI-MS (m / z): 492.14 [M+H] + . 1 HNMR(400MHz,DMSO-d6)δ8.33(s,1H),8.28(s,1H),7.68–7.61(m,2H),7.58(s,1H),7.44(d,J=8.0Hz,1H),7.17(s,2H),7.1 1(d,J=8.0Hz,1H),7.00(s,1H),6.64–5.80(m,1H),4.82–4.66(m,2H),4.44(s,3H),3.77(s,3H),2.70(s,3H),2.37(s,3H).

[0464] Example 29A: (S)-4-amino-N-(6-((1,5-dimethyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1Hpyrazolo[4,3-c]quinoline-8-carboxamide (Compound 29A)

[0465]

[0466] The synthetic process for preparing compound 29A from compound 6b-A is the same as the synthetic process for preparing compound 29 from compound 6b.

[0467] Compound 29A: ESI-MS (m / z): 492.16 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.33(s,1H),8.28(s,1H),7.68–7.61(m,2H),7.58(s,1H),7.44(d,J=8.0Hz,1H),7.18(s,2H),7.1 1(d,J=8.0Hz,1H),7.00(s,1H),6.64–5.80(m,1H),4.80–4.69(m,2H),4.44(s,3H),3.77(s,3H),2.70(s,3H),2.37(s,3H).

[0468] Example 30: 4-amino-N-(6-((1-(difluoromethyl)-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazino[4,3-c]quinoline-8-carboxamide (Compound 30)

[0469]

[0470] Preparation of compound 30b

[0471] To a solution of compound 30a (300 mg, 1.23 mmol) in N-methylpyrrolidone (5 mL), cuprous iodide (36 mg, 0.19 mmol), tetrakis(triphenylphosphine)palladium (288 mg, 0.25 mmol), N,N-diisopropylethylamine (476 mg, 3.69 mmol), and trimethylethynylsilane (242 mg, 2.46 mmol) were added. The mixture was then microwaved at 80 °C for 3 hours under nitrogen protection. The reaction solution was filtered through diatomaceous earth, and water (50 mL) was added. Extraction was performed with ethyl acetate (3 x 50 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Normal-phase column chromatography (methanol / dichloromethane = 1:20) was performed to give compound 30b (178 mg). ESI-MS (m / z): 215.1 [M+H] + .

[0472] Preparation of compound 30c

[0473] To a tetrahydrofuran (5 mL) solution of compound 30b (178 mg, 0.83 mmol), tetrabutylammonium fluoride (434 mg, 1.66 mmol) was added, and the reaction was carried out at 25 °C for 0.5 h. The reaction mixture was then extracted with water (30 mL) and ethyl acetate (3 x 30 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and subjected to normal-phase column chromatography (methanol / dichloromethane = 1:15) to give compound 30c (89 mg). ESI-MS (m / z): 143.1 [M+H] + .

[0474] Preparation of compound 30

[0475] To a solution of N-methylpyrrolidone (45 mg, 0.1 mmol) at 1°C (2 mL), cuprous iodide (4 mg, 0.02 mmol), tetrakis(triphenylphosphine)palladium (23 mg, 0.02 mmol), N,N-diisopropylethylamine (39 mg, 0.3 mmol), and 3°C (29 mg, 0.2 mmol) were added. The mixture was then microwaved at 80°C for 3 hours under nitrogen protection. The reaction solution was filtered through diatomaceous earth, and water (20 mL) was added. Extraction was performed with ethyl acetate (3 x 20 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Reversed-phase column chromatography (acetonitrile / 0.05% ammonium bicarbonate solution = 0:100%–50%:50%) yielded compound 30 (15 mg). ESI-MS (m / z): 513.9 [M+H] + . 1 HNMR(400MHz,DMSO-d6)δ8.67(s,1H),8.34(s,1H),8.28(s,1H),8.08(s,1H),7.86(t,J=58.8Hz,1H),7.65(s,2H ),7.53–7.42(m,1H),7.17(s,3H),7.04(s,1H),6.57–5.50(m,1H),4.83–4.69(m,2H),4.44(s,3H),2.71(s,3H).

[0476] Example 30A: (S)-4-amino-N-(6-((1-(difluoromethyl)-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazino[4,3-c]quinoline-8-carboxamide (Compound 30A)

[0477]

[0478] The preparation process of compound 1c-A is the same as that of compound 1c. The synthesis process for compound 30A from compound 1c-A is the same as that for compound 30 from compound 1c.

[0479] Compound 30A: ESI-MS (m / z): 513.98 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.65 (s, 1H), 8.34–8.24 (m, 2H), 8.05 (s, 1H), 7.83 (t, J = 59Hz, 1H), 7.66–7.59 (m, 2H), 7.51 –7.41(m,1H),7.14–7.12(m,3H),7.02(s,1H),6.63–5.60(m,1H),4.84–4.61(m,2H),4.42(s,3H),2.68(s,3H).

[0480] Example 31: 4-Amino-N,1-Dimethyl-N-(6-((1-methyl-3-(trifluoromethyl)-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazol[4,3-c]quinoline-8-carboxamide (Compound 31)

[0481]

[0482] Preparation of compound 31b

[0483] To a solution of compound 31a (500 mg, 2.19 mmol) in N-methylpyrrolidone (10 mL), cuprous iodide (63 mg, 0.33 mmol), tetratetraphenylphosphine palladium (508 mg, 0.44 mmol), N,N-diisopropylethylamine (848 mg, 6.57 mmol), and trimethylethynylsilane (430 mg, 4.38 mmol) were added. The mixture was then microwaved at 80 °C for 3 hours under nitrogen protection. The reaction solution was filtered through diatomaceous earth, and water (50 mL) was added. Extraction was performed with ethyl acetate (3 x 50 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Normal-phase column chromatography (methanol / dichloromethane = 1:20) was then performed to give compound 31b (350 mg). ESI-MS (m / z): 247.1 [M+H] + .

[0484] Preparation of compound 31c: Tetrabutylammonium fluoride (741 mg, 2.84 mmol) was added to a tetrahydrofuran (10 mL) solution of compound 31b (350 mg, 1.42 mmol), and the reaction was carried out at 25 °C for 0.5 h. The reaction solution was then extracted with water (50 mL) and ethyl acetate (3 x 50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and subjected to normal-phase column chromatography (methanol / dichloromethane = 1:15) to give compound 31c (165 mg). ESI-MS (m / z): 175.1 [M+H]+ .

[0485] Preparation of compound 31

[0486] Following the preparation procedure of compound 30, compound 30c was replaced with compound 31c (35 mg, 0.2 mmol) to obtain compound 31 (15 mg). ESI-MS (m / z): 546.2 [M+H] + . 1 HNMR (400MHz, DMSO-d6) δ8.33(s,2H),8.28(s,1H),7.64(s,2H),7.48(d,J=7.6Hz,1H),7.16(s,2H),7.11( d,J=7.6Hz,1H),6.98(s,1H),6.47–5.70(m,1H),4.82–4.68(m,2H),4.43(s,3H),3.96(s,3H),2.70(s,3H).

[0487] Example 31A: (S)-4-amino-N,1-dimethyl-N-(6-((1-methyl-3-(trifluoromethyl)-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazol[4,3-c]quinoline-8-carboxamide (Compound 31A)

[0488]

[0489] The synthetic process for preparing compound 31A from compounds 31c and 1c-A is the same as the synthetic process for preparing compound 31 from compound 1c.

[0490] Compound 31A: ESI-MS (m / z): 546.17 [M+H] + . 1 H NMR(400MHz,DMSO)δ8.31(s,2H),8.26(s,1H),7.67–7.60(m,2H),7.50–7.43(m,1H),7.14(s,2H),7.11– 7.06(m,1H),6.97(s,1H),6.44–5.74(m,1H),4.82–4.70(m,2H),4.42(s,3H),3.94(s,3H),2.68(s,3H).

[0491] Example 32: 4-Amino-7-fluoro-N,1-dimethyl-N-(6-((1-methyl-3-(trifluoromethyl)-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 32)

[0492]

[0493] Following the preparation procedure of compound 5, compound 4-ethynyl-1-methylpyrazole was replaced with compound 31c (35 mg, 0.2 mmol), and the mixture was microwaved at 80 °C for 3 hours under nitrogen protection to obtain compound 32 (15 mg). ESI-MS (m / z): 563.6 [M+H] + . 1 HNMR(400MHz,DMSO-d6)δ8.48–8.16(m,3H),7.55–7.19(m,4H),7.18–6.96(m,2H),6.4 8–5.60(m,1H),4.87–4.61(m,2H),4.43–4.40(m,3H),3.96(s,3H),2.72–2.60(m,3H).

[0494] Example 33: 4-Amino-7-fluoro-N,1-dimethyl-N-(6-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrofuran[2,3-b]pyridin-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 33)

[0495]

[0496] Preparation of compound 33b

[0497] Methyl glycolate (450 mg, 5.00 mmol) was dissolved in ethylene glycol dimethyl ether (10 mL), cooled to 0 °C, and sodium hydroxide (210 mg, 5.25 mmol, 60% purity) was added. The reaction mixture was reacted at 0 °C for 30 minutes, followed by the addition of compound 33a (1.03 g, 5.00 mmol). The reaction mixture was then stirred at 25 °C for 2 hours. The reaction mixture was quenched with water (5 mL), extracted with ethyl acetate (30 mL x 3), and the organic phases were combined and concentrated to give crude product 33b (1.15 g). MS (m / z): 259.95 [M+H] + .

[0498] Preparation of compound 33c

[0499] Compound 33b (416 mg, 1.60 mmol) was dissolved in tetrahydrofuran (20 mL), and potassium tert-butoxide (215 mg, 1.92 mmol) was added. The reaction solution was reacted at 25 °C for 1 hour, then quenched with water (20 mL). The mixture was extracted with ethyl acetate (5 mL x 3), and the organic phases were combined. The organic phase was concentrated to obtain a crude product, which was then subjected to column chromatography (petroleum ether: ethyl acetate = 2:1) to give 33c (240 mg). ESI-MS (m / z): 228.08 [M+H]+ .

[0500] Preparation of compound 33d

[0501] Compound 33c (228 mg, 1.00 mmol) was dissolved in methanol (20 mL), and 6 M hydrochloric acid solution (5 mL) was added. The reaction mixture was heated to 100 °C and stirred for 16 hours. The mixture was then diluted with ethyl acetate (50 mL) and water (50 mL), and extracted with ethyl acetate (20 mL x 3). The organic phases were combined and concentrated to obtain the crude product, which was then subjected to column chromatography (petroleum ether:ethyl acetate = 5:1) to give 33d (140 mg). ESI-MS (m / z): 170.01 [M+H] + Preparation of Compound 33e: Compound 33d (138 mg, 0.81 mmol) was dissolved in methanol (5 mL), and sodium borohydride (46 mg, 1.22 mmol) was added. The reaction solution was stirred at 25 °C for 1 hour. The reaction solution was then diluted with ethyl acetate (20 mL) and water (20 mL), and extracted with ethyl acetate (10 mL * 3). The organic phases were combined and concentrated to obtain the crude product, which was then subjected to column chromatography (petroleum ether: ethyl acetate = 2:1) to give 33e (98 mg). ESI-MS (m / z): 172.04 [M+H] + Preparation of Compound 33f: Bis(tert-butyloxycarbonyl)amine (1.06 g, 4.90 mmol), triphenylphosphine (1.60 g, 6.12 mmol), and diisopropyl azodicarbonate (1.24 g, 6.12 mmol) were added to a tetrahydrofuran (20 mL) solution of compound 33e (700 mg, 4.08 mmol). The reaction was carried out at 25 °C for 2 hours under nitrogen protection. The reaction solution was directly concentrated to obtain the crude product, which was then subjected to normal-phase column chromatography (petroleum ether / ethyl acetate = 10:1) to give compound 33f (1.10 g). ESI-MS (m / z): 371.15 [M+H] + .

[0502] Preparation of compound 33g

[0503] Compound 33f (1.10 g, 2.97 mmol) was dissolved in acetonitrile (20 mL), and lithium bromide (0.80 g, 8.90 mmol, 60% purity) was added. The reaction solution was reacted at 65 °C for 16 hours, and then quenched with water (5 mL). The mixture was extracted with ethyl acetate (5 mL x 3), and the organic phases were combined. The organic phase was concentrated to obtain a crude product, which was then subjected to column chromatography (petroleum ether: ethyl acetate = 3:1) to give 33 g (610 mg). ESI-MS (m / z): 271.05 [M + H] + .

[0504] Preparation of compound 33h

[0505] 33 g (271 mg, 1.00 mmol) of compound was dissolved in tetrahydrofuran (5 mL), cooled to 0 °C, and sodium hydrogen (80.0 mg, 2.00 mmol, 60% purity) was added. After reacting at 0 °C for 1 hour, iodomethane (0.10 mg, 1.20 mmol) was added, and the reaction mixture was stirred at 25 °C for 12 hours. The reaction mixture was quenched with water (5 mL), extracted with ethyl acetate (5 mL * 3), and the organic phases were combined. The organic phases were concentrated to obtain the crude product, which was then subjected to preparative thin-layer chromatography (petroleum ether: ethyl acetate = 20:1) to obtain 33 h (110 mg).

[0506] Preparation of compound 33i

[0507] Compound 33h (70 mg, 0.25 mmol) was dissolved in tetrahydrofuran (2 mL), and 4 M dioxane hydrochloride (0.5 mL) was added. The reaction mixture was stirred at 25 °C for 1 hour, and then evaporated to dryness to give compound 33i (40 mg). ESI-MS (m / z): 184.96 [M+H] + .

[0508] Preparation of compound 33j

[0509] Compound 33i (44.22 mg, 0.20 mmol), triethylamine (80.7 mg, 0.80 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (113.7 mg, 0.30 mmol) were added to a solution of compound 5a (51.9 mg, 0.20 mmol) in N,N-dimethylformamide (3 mL). The reaction mixture was reacted overnight at 25 °C. The reaction solution was then diluted with water (50 mL), extracted with ethyl acetate (2 x 25 mL), and the organic phase was washed three times with saturated brine. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and subjected to normal-phase column chromatography (dichloromethane / methanol = 10:1) to give compound 33j (40 mg). ESI-MS (m / z): 426.95 [M+H] + .

[0510] Preparation of compound 33

[0511] Under nitrogen protection, 4-ethynyl-1-methylpyrazole (9.9 mg, 0.09 mmol), XPhos Pd G3 95% (15.9 mg, 0.02 mmol), and triethylamine (37.9 mg, 0.37 mmol) were added sequentially to a solution of compound 33j (40 mg, 0.09 mmol) in N,N-dimethylformamide (2 mL). After the addition was complete, the mixture was heated to 80 °C in a microwave oven and reacted for 3 hours. The reaction solution was then diluted with water (10 mL) and extracted with ethyl acetate (10 mL * 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by evaporation. The crude product was obtained by column chromatography (methanol:dichloromethane = 1:20), and compound 33 (5 mg) was obtained by reverse-phase chromatography (acetonitrile / 0.05% ammonium bicarbonate aqueous solution = 0:100%–40%:60%). ESI-MS (m / z): 496.96 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ8.39–8.23(m,2H),8.16(s,1H),7.93–7.81(m,1H),7.76(s,1H),7.40–7.1 7(m,4H),6.46–5.60(m,1H),4.97–4.59(m,2H),4.46–4.40(m,3H),3.89(s,3H),2.77–2.65(m,3H).

[0512] Example 34: 4-amino-N-(6-((3-chloro-1-methyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 34)

[0513]

[0514] Following the preparation procedure of compound 19, compound 4-iodo-1-isopropyl-1H-pyrazole was replaced with compound 3-chloro-4-iodo-1-methylpyrazole (24b, 24.2 mg, 0.10 mmol), and the reaction was carried out at 25 °C for 3 hours to obtain compound 34 (22 mg). ESI-MS (m / z): 511.92 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ8.34(s,1H),8.28(s,1H),8.18(s,1H),7.68–7.62(m,2H),7.48(d,J=8.0Hz,1H),7.18(s,2H ),7.13(d,J=8.0Hz,1H),7.01(s,1H),6.57–5.70(m,1H),4.83–4.70(m,2H),4.44(s,3H),3.85(s,3H),2.70(s,3H).

[0515] Example 34A: (S)-4-amino-N-(6-((3-chloro-1-methyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 34A)

[0516]

[0517] The synthetic process for preparing compound 34A from compound 6b-A is the same as the synthetic process for preparing compound 34 from compound 6b.

[0518] Compound 34A: ESI-MS (m / z): 512.02 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.34(s,1H),8.28(s,1H),8.18(s,1H),7.68–7.62(m,2H),7.48(d,J=8.0Hz,1H),7.18(s,2H ),7.13(d,J=8.0Hz,1H),7.01(s,1H),6.57–5.70(m,1H),4.83–4.70(m,2H),4.44(s,3H),3.85(s,3H),2.70(s,3H).

[0519] Example 35: 4-Amino-7-fluoro-N,3-dimethyl-N-(6-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1,3-dihydrofuran[3,4-c]quinoline-8-carboxamide (Compound 35)

[0520]

[0521] Preparation of compound 35b

[0522] Compound 1a (325.33 mg, 1.23 mmol), triethylamine (310.6 mg, 3.07 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (701.5 mg, 1.84 mmol) were added to a solution of compound 35a (321.9 mg, 1.23 mmol) in N,N-dimethylformamide (10 mL), and the mixture was reacted overnight at 25 °C. The reaction mixture was then diluted with water (50 mL), extracted with ethyl acetate (2 x 25 mL), and washed three times with saturated brine. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and subjected to normal-phase column chromatography (dichloromethane / methanol = 10:1) to give compound 35b (510 mg). ESI-MS (m / z): 472.02 [M+H] + .

[0523] Preparation of compound 35

[0524] Under nitrogen protection, 4-ethynyl-1-methylpyrazole (21.2 mg, 0.20 mmol), cuprous iodide (7.7 mg, 0.04 mmol), triethylamine (40.9 mg, 0.40 mmol), and tetrakis(triphenylphosphine)palladium (23.4 mg, 0.02 mmol) were added sequentially to a solution of compound 35b (47.2 mg, 0.10 mmol) in N,N-dimethylformamide (2 mL). After the addition was complete, the mixture was heated to 100 °C in a microwave oven and reacted for 3 hours. The reaction solution was then diluted with water (10 mL) and extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by evaporation. The crude product was obtained by column chromatography (methanol:dichloromethane = 1:20), followed by reverse-phase chromatography (acetonitrile / 0.05% ammonium bicarbonate aqueous solution = 0:100%–40%:60%) to obtain compound 35 (21 mg). ESI-MS (m / z): 497.95 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.08–8.06(m,1H),7.90–7.60(m,2H),7.40–7.27(m,2H),7.15–7.06(m,1H),6.99(d,J=8.0Hz,1H),6.76( s,2H),6.45–6.41and5.54–5.48(m,1H),5.48–5.23(m,3H),4.83–4.53(m,2H),3.87(s,3H),2.68–2.55(m,3H),1.43–1.41(m,3H).

[0525] Example 36: 4-Amino-7-fluoro-N,1-dimethyl-N-(6-((1-methyl-1H-pyrazol-5-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazol[4,3-c]quinoline-8-carboxamide (Compound 36)

[0526]

[0527] Following the preparation procedure of compound 12, compound 1-cyclopropyl-4-iodo-1H-pyrazole was replaced with compound 5-iodo-1-methylpyrazole (49.9 mg, 0.24 mmol), and the reaction was carried out at 50 °C for 3 hours under nitrogen protection to obtain compound 36 (18 mg). ESI-MS (m / z): 496.0 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.44–8.18(m,2H),7.51(s,1H),7.48–7.08(m,6H),6.61(s,1H),6.51–6.40and 5.69–5.54(m,1H),4.87–4.61(m,2H),4.41and 4.39(s,3H),3.93(s,3H),2.70–2.59(m,3H).

[0528] Example 37: 4-Amino-N-(6-((1,4-dimethyl-1H-pyrazol-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N,1-dimethyl-1Hpyrazolo[4,3-c]quinoline-8-carboxamide (Compound 37)

[0529]

[0530] Following the preparation procedure of compound 12, compound 1-cyclopropyl-4-iodo-1H-pyrazole was replaced with compound 3-iodo-1,4-dimethylpyrazole (22.2 mg, 0.10 mmol), and the reaction was carried out at 25 °C for 3 hours to obtain compound 37 (5 mg). ESI-MS (m / z): 510.14 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ8.39–8.23(m,2H),7.58(s,1H),7.49–7.30(m,4H),7.22–7.11(m,1H),7.09–7.00(m,1H),6.50–6.44and 5.62–5.57(m,1H),4.88–4.60(m,2H),4.43–4.41(m,3H),3.81(s,3H),2.72–2.61(m,3H),2.09(s,3H).

[0531] Example 38: 4-Amino-7-fluoro-N-(6-((4-fluoro-1-methyl-1H-pyrazol-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 38)

[0532]

[0533] Following the preparation procedure of compound 12, compound 1-cyclopropyl-4-iodo-1H-pyrazole was replaced with compound 3-bromo-4-fluoro-1-methylpyrazole (17.9 mg, 0.10 mmol), and the mixture was reacted in a microwave oven at 80 °C for 3 hours to obtain compound 38 (5 mg). ESI-MS (m / z): 514.21 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ8.39–8.22(m,2H),8.00(d,J=4.8Hz,1H),7.54–7.27(m,4H),7.23–7.14(m,1H),7.12–7.03(m,1H),6.51–6.44and 5.63–5.58(m,1H),4.90–4.61(m,2H),4.43–4.41(m,3H),3.83(s,3H),2.72–2.61(m,3H).

[0534] Example 39: 4-Amino-7-fluoro-N-methyl-N-(6-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1,3-dihydrofuran[3,4-c]quinoline-8-carboxamide (Compound 39)

[0535]

[0536] Add cuprous iodide (7.6 mg, 0.04 mmol), tetrakis(triphenylphosphine)palladium (23.1 mg, 0.02 mmol), N,N-diisopropylethylamine (99.38 μL, 0.60 mmol), and 4-ethynyl-1-methylpyrazole (63.7 mg, 0.60 mmol) to a 3 mL solution of N-methylpyrrolidone (90 mg, 0.20 mmol) of 16b. After addition, microwave reaction at 100 °C for 4 hours under nitrogen protection. The reaction solution was filtered through diatomaceous earth, water (50 mL) was added, and the mixture was extracted with ethyl acetate (2 x 25 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude compound was purified by preparative chromatography (dichloromethane / methanol = 10:1), followed by reversed-phase column chromatography (acetonitrile / 0.05% ammonium bicarbonate solution = 0:100%–45%:55%) to give compound 39 (22 mg). ESI-MS (m / z): 483.9 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.08–8.05(m,1H),7.75–7.60(m,2H),7.39–7.27(m,2H),7.14–7.04(m,1H),6.96(d,J=17.2Hz,1H),6.82(s,2H),6.45–6.36and 5.54–5.44(m,1H),5.37–5.31(m,2H),5.00(s,2H),4.83–4.51(m,2H),3.85(s,3H),2.66–2.53(m,3H).

[0537] Example 39A: (S)-4-amino-7-fluoro-N-methyl-N-(6-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1,3-dihydrofuran[3,4-c]quinoline-8-carboxamide (Compound 39A)

[0538]

[0539] The synthetic process for preparing compound 39A from compound 16b-A is the same as the synthetic process for preparing compound 39 from compound 16b.

[0540] Compound 39A: ESI–MS (m / z): 484.00 [M+H] + . 1H NMR (400MHz, DMSO) δ8.10–8.07(m,1H),7.74–7.62(m,2H),7.42–7.28(m,2H),7.15–7.06(m,1H),7.05–6.95(m,1H),6.85(s,2H),6.48–6.39and 5.55–5.46(m,1H),5.42–5.33(m,2H),5.03(s,2H),4.85–4.57(m,2H),3.88(s,3H),2.69–2.56(m,3H).

[0541] Example 40: 4-Amino-7-chloro-N-methyl-N-(6-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1,3-dihydrofuran[3,4-c]quinoline-8-carboxamide (Compound 40)

[0542]

[0543] Preparation of compound 40b

[0544] Compound 1a (149.9 mg, 0.57 mmol), N,N-diisopropylethylamine (366.1 mg, 2.83 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (323.2 mg, 0.85 mmol) were added to a 5 mL solution of N,N-dimethylformamide containing compound 40a (150 mg, 0.57 mmol, preparation method referred to WO2022169948 A1). The reaction was carried out at 25 °C for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature, and water (50 mL) was added. The solution was extracted with ethyl acetate (2 x 25 mL), and the organic phase was washed three times with saturated brine. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and subjected to normal phase column chromatography (dichloromethane / methanol = 20:1) to obtain compound 40b (200 mg). ESI-MS (m / z): 473.96 [M+H] + .

[0545] Preparation of compound 40

[0546] Following the preparation method of compound 39, compound 16b was replaced with compound 40b (100 mg, 0.21 mmol), and the mixture was reacted under nitrogen protection at 100 °C for 4 hours to obtain compound 40 (40 mg). ESI-MS (m / z): 500.1 [M+H] + . 1H NMR (400MHz, DMSO) δ8.07–8.03(m,1H),7.86–7.56(m,3H),7.44–7.28(m,1H),7.13–7.02(m,1H),7.00–6.9 1(m,1H),6.83(s,2H),6.50–5.27(m,3H),5.07–4.94(m,2H),4.83–4.44(m,2H),3.88–3.81(m,3H),2.67and 2.45(s,3H).

[0547] Example 40A: (S)-4-amino-7-chloro-N-methyl-N-(6-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1,3-dihydrofuran[3,4-c]quinoline-8-carboxamide (Compound 40A)

[0548]

[0549] The synthetic process for preparing compound 40A from compound 1a-A is the same as the synthetic process for preparing compound 40 from compound 1a.

[0550] Compound 40A: ESI-MS (m / z): 499.95 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.08–8.03(m,1H),7.85–7.56(m,3H),7.44–7.26(m,1H),7.12–7.02(m,1H),7.00–6.91(m,1H ),6.83(s,2H),6.52–5.28(m,3H),5.07–4.94(m,2H),4.84–4.45(m,2H),3.85–3.84(s,3H),2.67and2.45(s,3H).

[0551] Example 41: 4-Amino-N-(6-((1-(difluoromethyl)-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N,1-dimethyl-1H-pyrazino[4,3-c]quinoline-8-carboxamide (Compound 41)

[0552]

[0553] Following the preparation procedure of compound 12, compound 1-cyclopropyl-4-iodo-1H-pyrazole was replaced with compound 30a (24.4 mg, 0.10 mmol), and the mixture was reacted in a microwave oven at 80 °C for 3 hours to obtain compound 41 (35 mg). ESI-MS (m / z): 531.98 [M+H]+ . 1 H NMR(400MHz,DMSO-d6)δ8.66(s,1H),8.40–8.22(m,2H),8.07(s,1H),7.86(t,J=58 .8Hz,1H),7.56–7.27(m,4H),7.21–7.10(m,1H),7.08–7.00(m,1H),6.50–6.42and 5.63–5.56(m,1H),4.91–4.58(m,2H),4.43–4.41(m,3H),2.71–2.60(m,3H).

[0554] Example 42: 4-amino-N-(6-((4-(difluoromethyl)pyridin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N,1-dimethyl-1H-pyrazole[4,3-c]quinoline-8-carboxamide (Compound 42)

[0555]

[0556] Following the preparation procedure of compound 12, compound 1-cyclopropyl-4-iodo-1H-pyrazole was replaced with compound 3-bromo-4-(difluoromethyl)pyridine (50.1 mg, 0.24 mmol), and the mixture was reacted in a microwave oven at 100 °C for 4 hours to obtain compound 42 (8 mg). ESI-MS (m / z): 543.2 [M+H] + . 1 H NMR(400MHz, DMSO)δ8.90(s,1H),8.76(d,J=5.2Hz,1H),8.41–8.21(m,2H),7.69(d,J=5.2Hz,1H),7.48–7.12(m,7H),6.53–6.41and 5.68–5.55(m,1H),4.90–4.60(m,2H),4.42–4.39(m,3H),2.71–2.59m,3H).

[0557] Example 43: 4-Amino-7-chloro-N-(6-(((1-cyclopropyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N-methyl-1,3-dihydrofuran[3,4-c]quinoline-8-carboxamide (Compound 43)

[0558] Preparation of Compound 43a: Cuprous iodide (24.1 mg, 0.13 mmol), tetrakis(triphenylphosphine)palladium (73.0 mg, 0.06 mmol), N,N-diisopropylethylamine (314.04 μL, 1.90 mmol), and trimethylsilylacetylene (446.48 μL, 3.16 mmol) were added to a solution of 40b (298 mg, 0.63 mmol) in 3 mL of N-methylpyrrolidone. The mixture was then microwaved at 80 °C for 4 hours under nitrogen protection. The reaction solution was filtered through diatomaceous earth, and water (50 mL) was added. Extraction was performed with ethyl acetate (2 x 25 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Normal-phase column chromatography (dichloromethane / methanol = 10:1) yielded compound 43a (300 mg). ESI-MS (m / z): 492.0 [M+H] + .

[0559] Preparation of Compound 43b: Tetrabutylammonium fluoride (1.5 mL, 1.52 mmol) was added to a tetrahydrofuran (3 mL) solution of compound 43a (300 mg, 0.61 mmol), and the reaction was carried out at 25 °C for 1 hour. The reaction solution was then extracted with water (15 mL) and ethyl acetate (2 x 25 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was purified by preparative chromatography (dichloromethane / methanol = 10:1), and then subjected to reversed-phase column chromatography (acetonitrile / 0.05% ammonium bicarbonate solution = 0:100%–40%:60%) to obtain compound 43b (210 mg). ESI-MS (m / z): 419.94 [M+H] + Preparation of Compound 43: To a solution of compound 43b (100 mg, 0.24 mmol) in N-methylpyrrolidone (2 mL), cuprous iodide (9.1 mg, 0.05 mmol), tetrakis(triphenylphosphine)palladium (27.5 mg, 0.02 mmol), triethylamine (99.05 μL, 0.71 mmol), and 1-cyclopropyl-4-iodopyrazole (112 mg, 0.48 mmol) were added. The reaction was carried out under nitrogen protection at 25 °C for 16 hours. The reaction solution was filtered through diatomaceous earth, water (50 mL) was added, and the mixture was extracted with ethyl acetate (2 x 25 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude compound was obtained by normal-phase column chromatography (dichloromethane / methanol = 20:1), followed by reverse-phase column chromatography (acetonitrile / 0.05% ammonium bicarbonate solution = 0:100%–40%:60%) to give compound 43 (36 mg). ESI-MS (m / z): 526.1 [M+H] + . 1H NMR (400MHz, DMSO) δ8.18–8.13(m,1H),7.86–7.56(m,3H),7.44–7.27(m,1H),7.12–7.01(m,1H),6.99–6.9 1(m,1H),6.83(s,2H),6.51–5.33(m,3H),5.03–4.96(m,2H),4.84–4.45(m,2H),3.79–3.71(m,1H),2.67and 2.45(s,3H),1.10–0.93(m,4H).

[0560] Example 43A: (S)-4-amino-7-chloro-N-(6-((1-cyclopropyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N-methyl-1,3-dihydrofuran[3,4-c]quinoline-8-carboxamide (Compound 43A)

[0561] The synthetic process for preparing compound 43A from compound 40b-A is the same as the synthetic process for preparing compound 43 from compound 40b.

[0562] Compound 43A: ESI-MS (m / z): 526.1 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.16–8.13(m,1H),7.84–7.58(m,3H),7.42–7.29(m,1H),7.10–7.01(m,1H),6.99–6.9 1(m,1H),6.83(s,2H),6.49–5.32(m,3H),5.01–4.98(m,2H),4.84–4.45(m,2H),3.79–3.71(m,1H),2.67and 2.45(s,3H),1.09–0.96(m,4H). 1 H NMR (400MHz, DMSO-d6, 90℃) δ8.05(s,1H),7.62–7.53(m,3H),7.34(d,J=2.8Hz,1H),7.05(d,J=2.8Hz,1H),6.90(s,1H),6. 53(s,2H),6.44–5.32(m,3H),5.01(s,2H),4.77–4.58(m,2H),3.75–3.70(m,1H),2.68–2.49(m,3H),1.06–0.94(m,4H).[α] D 25 179.6 (c1 mg / mL, DMF).

[0563] Example 44: 4-Amino-7-fluoro-N,1-dimethyl-N-(6-((4-methylpyridin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 44)

[0564]

[0565] Following the preparation procedure of compound 12, compound 1-cyclopropyl-4-iodo-1H-pyrazole was replaced with compound 3-iodo-4-methylpyridine (22.0 mg, 0.1 mmol), and the reaction was carried out in an oil bath at 40 °C for three hours to obtain compound 44 (15 mg). ESI-MS (m / z): 506.95 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.66(s,1H),8.49–8.41(m,1H),8.36–8.24(m,2H),7.56–7.32(m,5H),7.29–7. 09(m,2H),6.52–5.56(m,1H),4.92–4.65(m,2H),4.48–4.33(m,3H),2.77–2.56(m,3H),2.48(s,3H).

[0566] Example 45: 4-Amino-7-fluoro-N,1-dimethyl-N-(6-((2-methylpyridin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 45)

[0567]

[0568] Following the preparation procedure of compound 12, compound 1-cyclopropyl-4-iodo-1H-pyrazole was replaced with compound 3-iodo-2-methylpyridine (43.8 mg, 0.2 mmol), and the reaction was carried out at 40 °C for three hours to obtain compound 45 (20 mg). ESI-MS (m / z): 507.02 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.51–8.42(m,1H),8.35–8.22(m,2H),7.96–7.85(m,1H),7.54–7. 05(m,7H),6.53–5.55(m,1H),4.91–4.62(m,2H),4.48–4.34(m,3H),2.74–2.62(m,6H).

[0569] Example 45A: (S)-4-amino-7-fluoro-N,1-dimethyl-N-(6-((2-methylpyridin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 45A)

[0570]

[0571] The synthetic process for preparing compound 45A from compound 12b-A is the same as the synthetic process for preparing compound 45 from compound 12b.

[0572] Compound 45A: ESI-MS (m / z): 506.99 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.51–8.45(m,1H),8.39–8.20(m,2H),7.94–7.86(m,1H),7.54–7.07(m,7H),6.54–6.44and 5.67–5.57(m,1H),4.92–4.79and 4.74–4.61(m,2H),4.43(s,3H),2.75–2.60(m,6H).

[0573] Example 46: 4-Amino-7-fluoro-N,1-dimethyl-N-(6-((7-methyl-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 46)

[0574]

[0575] Preparation of Compound 46b: Methanesulfonic acid (2-dicyclohexylphosphino-2',4',6'-tris-isopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (565.2 mg, 0.66 mmol), triethylamine (4.13 mL, 29.78 mmol), and trimethylsilyne (4.21 mL, 29.78 mmol) were added to a solution of 46a (3000 mg, 9.93 mmol) in N,N-dimethylacetamide (5 mL). The reaction mixture was purged with nitrogen three times and reacted in a microwave oven at 70 °C for 3 hours. The reaction mixture was filtered through diatomaceous earth, and water (100 mL) was added. Extraction was performed with ethyl acetate (2 x 100 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by normal-phase column chromatography (dichloromethane / methanol = 20:1) to obtain compound 46b (2700 mg). ESI-MS (m / z): 320.11 [M+H] + .

[0576] Preparation of compound 46c

[0577] A solution of compound 46b (2700 mg, 8.45 mmol) in dichloromethane (20 mL) was added with trifluoroacetic acid (6.47 mL, 84.50 mmol), and the reaction was carried out at 25 °C for 16 hours. After the reaction was complete, the product was concentrated to obtain crude product 46c (1800 mg), which was used directly in the next step. ESI-MS (m / z): 220.11 [M+H] + .

[0578] Preparation of compound 46d

[0579] Formaldehyde solution (2400.0 mg, 29.60 mmol) was added to a formic acid solution (5 mL) containing 1800 mg (8.20 mmol). The reaction mixture was then incubated at 100 °C for 1 hour. The reaction solution was quenched with ammonium bicarbonate, water (50 mL) was added, and the mixture was extracted with ethyl acetate (2 x 50 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by normal-phase column chromatography (dichloromethane / methanol = 20:1) to give compound 46d (1000 mg). ESI-MS (m / z): 234.13 [M+H] + .

[0580] Preparation of compound 46e

[0581] Cesium fluoride (2603.3 mg, 17.14 mmol) was added to a solution of 46d (1000 mg, 4.28 mmol) in N,N-dimethylformamide (5 mL). After addition, the reaction mixture was reacted at 25 °C for 30 min. The reaction solution was then diluted with water (50 mL), extracted with ethyl acetate (2 x 50 mL), and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by normal-phase column chromatography (dichloromethane / methanol = 20:1) to give compound 46e (450 mg). ESI-MS (m / z): 162.12 [M+H] + .

[0582] Preparation of compound 46

[0583] Following the preparation procedure for compound 5, compound 4-ethynyl-1-methylpyrazole was replaced with compound 46e (102.8 mg, 0.64 mmol), and the mixture was reacted in a microwave oven at 100 °C for 3 hours to obtain compound 46 (7.3 mg). ESI-MS (m / z): 550.8 [M+H] + . 1H NMR (400MHz, DMSO) δ8.37–8.20(m,2H),7.47–7.04(m,7H),6.51–5.53(m,1H),4.87–4.56(m,2H) ,4.45–4.33(m,3H),3.99(s,2H),3.56(s,2H),2.86–2.79(m,2H),2.69–2.58m,3H),2.39(s,3H).

[0584] Example 46A: (S)-4-amino-7-fluoro-N,1-dimethyl-N-(6-((7-methyl-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 46A)

[0585]

[0586] The synthetic process for preparing compound 46A from compounds 46e and 5b-A is the same as the synthetic process for preparing compound 46 from compounds 46e and 5b.

[0587] Compound 46A: ESI-MS (m / z): 551.0 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.36–8.19(m,2H),7.46–7.28(m,5H),7.19–6.99(m,2H),6.49–5.54(m,1H), 4.85–4.61(m,2H),4.45–4.37(m,3H),4.02–3.97(m,2H),3.54(s,2H),2.86–2.81(m,2H),2.70and 2.58(s,3H),2.40(s,3H).

[0588] Example 47A: (S)-4-amino-7-fluoro-N,1-dimethyl-N-(6-((5-methylpyridin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 47A)

[0589]

[0590] Following the preparation procedure of compound 5A, compound 4-ethynyl-1-methylpyrazole was replaced with compound 3-ethynyl-5-methylpyridine (23.4 mg, 0.2 mmol), and the mixture was reacted in a microwave oven at 100 °C for 4 hours to obtain compound 47A (25 mg). ESI-MS (m / z): 507.27 [M+H]+ . 1 H NMR (400MHz, DMSO) δ8.73–8.23(m,4H),7.83(s,1H),7.60–7.03(m,6H),6.54–5. 57(m,1H),4.91–4.60(m,2H),4.51–4.35(m,3H),2.77–2.58(m,3H),2.34(s,3H).

[0591] Example 48A: (S)-4-amino-7-fluoro-N,1-dimethyl-N-(6-(pyridin-3-ylethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 48A)

[0592]

[0593] Following the preparation procedure of compound 5A, compound 4-ethynyl-1-methylpyrazole was replaced with compound 3-ethynylpyridine (52.6 mg, 0.51 mmol), and the mixture was reacted in a microwave oven at 100 °C for 4 hours to obtain compound 48A (40 mg). ESI-MS (m / z): 493.1 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.75(s,1H),8.60–8.59(m,1H),8.39–8.22(m,2H),8.00–7.94(m,1H),7.52–7.16( m,6H),7.15–7.05(m,1H),6.54–5.53(m,1H),4.88–4.61(m,2H),4.41–4.39(m,3H),2.70–2.59(m,3H).

[0594] Example 49A: (S)-4-amino-N-(6-((1,3-dimethyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1Hpyrazolo[4,3-c]quinoline-8-carboxamide (Compound 49A)

[0595]

[0596] Following the preparation procedure of compound 19, compound 4-iodo-1-isopropyl-1H-pyrazole was replaced with compound 4-iodo-1,3-dimethylpyrazole (22.2 mg, 0.1 mmol), and compound 6b-A (39.7 mg, 0.12 mmol) was used to replace compound 6b. The reaction was carried out at 25 °C for three hours, and the result was compound 49A (23 mg). ESI-MS (m / z): 492.19 [M+H]+. 1H NMR(400MHz,DMSO-d6)δ8.34–8.33(m,1H),8.28(s,1H),7.95(s,1H),7.67–7.62(m,2H),7.45(d,J=7.6Hz,1H),7.19(s,2H),7 .10(d,J=7.6Hz,1H),6.99(s,1H),6.40–5.56(m,1H),4.88–4.60(m,2H),4.44(s,3H),3.78(s,3H),2.70(s,3H),2.24(s,3H).

[0597] Example 50A: (S)-4-amino-7-chloro-N-(6-((1,3-dimethyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N-methyl-1,3-dihydrofurano[3,4-c]quinoline-8-carboxamide (Compound 50A)

[0598]

[0599] Following the preparation procedure of compound 40A, compound 4-ethynyl-1-methylpyrazole was replaced with compound 4-ethynyl-1,3-dimethylpyrazole (75.7 mg, 0.63 mmol). The mixture was reacted under nitrogen protection at 100 °C with microwave for 3 hours to obtain compound 50A (50 mg). ESI-MS (m / z): 513.96 [M+H] + . 1 H NMR (400MHz, DMSO) δ7.95–7.88(m,1H),7.86–7.55(m,2H),7.44–7.26(m,1H),7.13–7.02(m,1H),7.01–6.9 1(m,1H),6.83(s,2H),6.52–5.28(m,3H),5.06–4.92(m,2H),4.84–4.44(m,2H),3.76–3.75(m,3H),2.67and 2.45(s,3H),2.22–2.21(s,3H).

[0600] Example 51A: (S)-4-amino-7-chloro-N,1-dimethyl-N-(6-((7-methyl-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 51A)

[0601]

[0602] Following the preparation procedure of compound 46A, compound 5b-A was replaced with compound 8b-A (100 mg, 0.21 mmol), and the mixture was microwaved at 100 °C for 3 hours under nitrogen protection to obtain compound 51A (41 mg). ESI-MS (m / z): 566.97 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.43–8.19(m,2H),7.72–7.62(m,1H),7.58–7.29(m,4H),7.22–7.01(m,2H),6.60–5.40(m,1H),4.90–4. 56(m,2H),4.46–4.41(m,3H),4.05–3.97(m,2H),3.58–3.57(m,2H),2.88–2.81(m,2H),2.73–2.51(m,3H),2.42–2.41(m,3H).

[0603] Example 52A: (S)-4-amino-7-chloro-N-(6-((1,5-dimethyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N-methyl-1,3-dihydrofurano[3,4-c]quinoline-8-carboxamide (Compound 52A)

[0604]

[0605] Following the preparation procedure of compound 43A, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 4-iodo-1,5-dimethylpyrazole (66.2 mg, 0.30 mmol), and the reaction was carried out at room temperature for 16 hours under nitrogen protection to obtain compound 52A (51 mg). ESI-MS (m / z): 513.96 [M+H] + . 1H NMR (400MHz, DMSO) δ7.85–7.55(m,3H),7.45–7.25(m,1H),7.14–7.03(m,1H),7.02–6.93(m,1H),6.83(s,2H), 6.50–5.27(m,3H),5.08–4.95(m,2H),4.88–4.40(m,2H),3.75(s,3H),2.67and2.45(s,2H),2.35–2.34(s,3H).

[0606] Example 53A: (S)-4-amino-7-chloro-N,1-dimethyl-N-(6-((1-methyl-1H-pyrazol-5-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 53A)

[0607]

[0608] Following the preparation procedure of compound 8A, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 5-iodo-1-methylpyrazole (105.7 mg, 0.51 mmol), and the reaction was carried out at room temperature for 16 hours under nitrogen protection to obtain compound 53A (55 mg). ESI-MS (m / z): 511.99 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.44–8.17(m,2H),7.70–7.59(m,1H),7.58–7.33(m,2H),7.27–7.08(m,4H),6.65–6. 58(m,1H),6.55–5.41(m,1H),4.88–4.55(m,2H),4.45–4.39(s,3H),3.94–3.92(m,3H),2.72–2.47(m,3H).

[0609] Example 54A: (S)-4-amino-7-fluoro-N-(6-((2-fluoropyridin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 54A)

[0610]

[0611] Following the preparation procedure of compound 12A, compound 1-cyclopropyl-4-iodo-1H-pyrazole was replaced with compound 2-fluoro-3-iodopyridine (22.3 mg, 0.1 mmol), and the reaction was carried out at 40 °C for three hours to obtain compound 54A (15 mg). ESI-MS (m / z): 511.07 [M+H]+ . 1 H NMR (400MHz, DMSO) δ8.44–8.17(m,4H),7.57–7.43(m,2H),7.43–7.19(m,4H),7.18–7. 07(m,1H),6.56–5.57(m,1H),4.92–4.63(m,2H),4.49–4.33(m,3H),2.78–2.57(m,3H).

[0612] Example 55A: (S)-4-amino-7-chloro-N,1-dimethyl-N-(6-(6-methylpyridin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 55A)

[0613]

[0614] Following the preparation procedure of compound 8A, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 5-iodo-2-methylpyridine (21.9 mg, 0.10 mmol), and the reaction was carried out at 25 °C for 3 hours to obtain compound 55A (15 mg). ESI-MS (m / z): 522.91 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.69–8.59(m,1H),8.44–8.20(m,2H),7.90–7.84(m,1H),7.72–7.63(m,1H),7.5 9–7.05(m,6H),6.57–5.44(m,1H),4.91–4.57(m,2H),4.48–4.42(s,3H),3.34(s,3H),2.76–2.54(m,3H).

[0615] Example 56A: (S)-4-amino-7-chloro-N-(6-((2-fluoropyridin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 56A)

[0616]

[0617] Following the preparation procedure of compound 8A, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 2-fluoro-3-iodopyridine (44.6 mg, 0.20 mmol), and the reaction was carried out at 25 °C for 3 hours to obtain compound 56A (10 mg). ESI-MS (m / z): 526.73 [M+H] + . 1H NMR(400MHz,DMSO-d6)δ8.45–8.17(m,4H),7.72–7.64(m,1H),7.62–7.35(m,2H),7.34–7 .07(m,1H),6.57–5.45(m,1H),4.94–4.57(m,2H),4.47–4.41(s,3H),2.76–2.53(m,3H).

[0618] Example 57A: (S)-4-amino-7-chloro-N-methyl-N-(6-((2-methylpyridin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1,3-dihydrofurano[3,4-c]quinoline-8-carboxamide (Compound 57A)

[0619]

[0620] Following the preparation procedure of compound 43A, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 3-iodo-2-methylpyridine (31.3 mg, 0.14 mmol). The reaction was carried out at room temperature for 3 hours under nitrogen protection to obtain compound 57A (13 mg). ESI-MS (m / z): 510.92 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.50–8.46(m,1H),7.93–7.87(m,1H),7.70–7.57(m,2H),7.52–7.17(m,3H),7.17–7.07(m ,1H),6.86(s,2H),6.55–5.32(m,3H),5.05–4.98(m,2H),4.89–4.49(m,2H),2.71–2.68(m,3H),2.66–2.67(m,3H).

[0621] Example 58A: (S)-4-amino-7-chloro-N-methyl-N-(6-(6-methylpyridin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1,3-dihydrofurano[3,4-c]quinoline-8-carboxamide (Compound 58A)

[0622]

[0623] Following the preparation procedure of compound 43A, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 5-iodo-2-methylpyridine (30.7 mg, 0.14 mmol), and the reaction was carried out at room temperature for 3 hours under nitrogen protection to obtain compound 58A (15 mg). ESI-MS (m / z): 510.93 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ8.66–8.61(m,1H),7.89–7.58(m,3H),7.52–7.32(m,2H),7.25–7.15(m,1H),7.1 3–7.06(m,1H),6.86(s,2H),6.53–5.30(m,3H),5.05–4.98(m,2H),4.89–4.48(m,2H),2.72–2.47(m,6H).

[0624] Example 59A: (S)-4-amino-7-fluoro-N,1-dimethyl-N-(6-(6-methylpyridin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 59A)

[0625]

[0626] Following the preparation procedure of compound 12A, compound 1-cyclopropyl-4-iodo-1H-pyrazole was replaced with compound 5-iodo-2-methylpyridine (36.9 mg, 0.17 mmol), and the reaction was carried out at 25 °C for three hours to obtain compound 59A (15 mg). ESI-MS (m / z): 507.21 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ8.64(s,1H),8.29–8.24(m,2H),7.89–7.84(m,1H),7.69–7.07(m,6H), 6.52–5.57(m,1H),4.92–4.64(m,2H),4.43–4.41(s,3H),3.33–3.24(m,3H),2.72–2.61(m,3H).

[0627] Example 60A: (S)-4-amino-7-fluoro-N,1-dimethyl-N-(6-((1-methyl-1H-indazol-6-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 60A)

[0628]

[0629] Following the preparation procedure of compound 12A, compound 1-cyclopropyl-4-iodo-1H-pyrazole was replaced with compound 6-iodo-1-methylindazole (41.3 mg, 0.16 mmol), and the reaction was carried out at 25 °C for three hours to obtain compound 60A (8 mg). ESI-MS (m / z): 546.08 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ8.39–8.24(m,2H),8.12(s,1H),8.02(s,1H),7.75–7.70(m,1H),7.58–7.52(m,1H),7.47–7.31(m,4H), 7.26–7.14(m,1H),7.13–7.05(m,1H),6.52–5.56(m,1H),4.92–4.60(m,2H),4.44–4.41(m,3H),4.08(s,3H),2.73–2.61(m,3H).

[0630] Example 61A: (S)-4-amino-7-chloro-N-(6-((2-fluoropyridin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N-methyl-1,3-dihydrofurano[3,4-c]quinoline-8-carboxamide (Compound 61A)

[0631]

[0632] Following the preparation procedure of compound 43A, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 2-fluoro-3-iodopyridine (31.2 mg, 0.14 mmol), and the reaction was carried out at room temperature for 3 hours to obtain compound 61A (20 mg). ESI-MS (m / z): 515.13 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.32–8.28(m,1H),8.26–8.18(m,1H),7.70–7.56(m,2H),7.52–7.36(m,2H),7.28–7. 17(m,1H),7.15–7.07(m,1H),6.86(s,2H),6.56–5.28(m,3H),5.07–4.97(m,2H),4.92–4.44(m,2H),2.70and 2.48(s,3H).

[0633] Example 62A: (S)-4-amino-7-chloro-N-methyl-N-(6-((1-methyl-1H-indazol-6-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1,3-dihydrofuran[3,4-c]quinoline-8-carboxamide (Compound 62A)

[0634]

[0635] Following the preparation procedure of compound 43A, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 6-iodo-1-methylindazole (36.1 mg, 0.14 mmol), and the reaction was carried out at room temperature for 3 hours to obtain compound 62A (13 mg). ESI-MS (m / z): 549.92 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ8.12–8.08(m,1H),7.96–7.92(m,1H),7.88–7.59(m,3H),7 .52–7.34(m,1H),7.30–7.17(m,2H),7.14–7.07(m,1H),6.86(s,2H),6.54–6.46and 5.43–5.40(m,1H),5.39–5.29(m,2H),5.06–4.98(m,2H),4.89–4.48(m,2H),4.09–4.08(m,3H),2.71and 2.49(s,3H).

[0636] Example 63A: (S)-4-amino-7-chloro-N,1-dimethyl-N-(6-((4-methylpyridin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 63A)

[0637]

[0638] Following the preparation procedure of compound 8A, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 3-iodo-4-methylpyridine (43.8 mg, 0.2 mmol), and the reaction was carried out at 40 °C for three hours to obtain compound 63A (15 mg). ESI-MS (m / z): 522.89 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.69–8.64(m,1H),8.48–8.21(m,3H),7.71–7.63(m,1H),7.41–7. 24(m,6H),6.59–5.42(m,1H),4.89–4.62(m,2H),4.47–4.40(m,3H),2.77–2.47(m,6H).

[0639] Example 64A: (S)-4-amino-7-chloro-N,1-dimethyl-N-(6-((2-methylpyridin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 64A)

[0640]

[0641] Following the preparation procedure of compound 8A, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 3-iodo-2-methylpyridine (43.8 mg, 0.2 mmol), and the reaction was carried out at 40 °C for three hours to obtain compound 64A (15 mg). ESI-MS (m / z): 522.92 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.51–8.45(m,1H),8.42–8.19(m,2H),7.93–7.86(m,1H),7.70–7.61(m,1H) ,7.58–7.07(m,6H),6.57–5.43(m,1H),4.88–4.64(m,2H),4.47–4.40(m,3H),2.74–2.66(m,6H).

[0642] Example 65A: (S)-4-amino-7-chloro-N,1-dimethyl-N-(6-((5-methylpyridin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 65A)

[0643]

[0644] Following the preparation procedure of compound 51A, compound 46e was replaced with compound 3-ethynyl-5-methylpyridine (23.4 mg, 0.2 mmol), and the mixture was microwaved at 100 °C for 4 hours under nitrogen protection to obtain compound 65A (12 mg). ESI-MS (m / z): 522.96 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.59–8.55(m,1H),8.48–8.21(m,3H),7.84–7.81(m,1H),7.71–7.64(m,1H) ,7.60–7.08(m,5H),6.58–5.42(m,1H),4.88–4.54(m,2H),4.47–4.41(m,3H),2.75–2.33(m,6H).

[0645] Example 66A: (S)-4-amino-7-fluoro-N,1-dimethyl-N-(6-((5,6,7,8-tetrahydroimidazo[1,2-a]pyridin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 66A)

[0646]

[0647] Following the preparation procedure of compound 12A, compound 1-cyclopropyl-4-iodo-1H-pyrazole was replaced with compound 66a (50.5 mg, 0.20 mmol), and the mixture was reacted in a microwave oven at 100 °C for three hours to obtain compound 66A (18 mg). ESI-MS (m / z): 536.39 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.41–8.22(m,2H),7.47–7.26(m,5H),7.22–7.13(m,1H),7.10–7.03(m,1H),6.51–5.56(m,1H),4.90–4 .67(m,2H),4.43–4.41(s,3H),4.01–3.95(m,2H),2.82–2.75(m,2H),2.72–2.60(m,3H),2.01–1.94(m,2H),1.91–1.83(m,2H).

[0648] Example 67A: (S)-4-amino-7-chloro-N-(6-((1-(difluoromethyl)-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 67A)

[0649]

[0650] Following the preparation procedure of compound 8A, compound 1-cyclopropyl-4-iopyrazole was replaced with compound 1-(difluoromethyl)-4-iopyrazole (46.4 mg, 0.19 mmol), and the reaction was carried out at room temperature (25 °C) for 3 hours to obtain compound 67A (8 mg). ESI-MS (m / z): 547.9 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ8.69–8.63(m,1H),8.42–8.18(m,2H),8.10–8.04(m,1H),7.99–7.62(m,2H),7.58–7 .29(m,3H),7.20–6.96(m,2H),6.55–5.41(m,1H),4.86–4.55(m,2H),4.46–4.40(m,3H),2.74–2.47(m,3H).

[0651] Example 68A: (S)-4-amino-7-chloro-N-(6-((3-chloro-1-methyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N-methyl-1,3-dihydrofuran[3,4-c]quinoline-8-carboxamide (Compound 68A)

[0652]

[0653] Following the preparation procedure of compound 43A, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 24b (38.8 mg, 0.16 mmol), and the reaction was carried out at room temperature for 3 hours to obtain compound 68A (10 mg). ESI-MS (m / z): 533.89 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.20–8.17(m,1H),7.89–7.61(m,2H),7.54–7.32(m,1H),7.18–7.06(m,1H),7.04–6.97(m,1H) ,6.86(s,2H),6.53–5.31(m,3H),5.05–4.98(m,2H),4.86–4.48(m,2H),4.19–4.11(m,2H),3.85–3.84(s,3H),2.69and 2.47(s,3H).

[0654] Example 69A: (S)-4-amino-7-chloro-N-(6-((1-ethyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N-methyl-1,3-dihydrofurano[3,4-c]quinoline-8-carboxamide (Compound 69A)

[0655]

[0656] Following the preparation procedure of compound 43A, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 1-ethyl-4-iodo-1H-pyrazole (52.9 mg, 0.24 mmol), and the reaction was carried out at room temperature for 3 hours to obtain compound 69A (10 mg). (13 mg) ESI-MS (m / z): 514.0 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ8.15–8.11(m,1H),7.89–7.61(m,3H),7.47–7.26(m,1H),7.14–7.03(m,1H),7.02–6. 92(m,1H),6.85(s,2H),6.51–5.30(m,3H),5.05–4.97(m,2H),4.84–4.44(m,2H),4.19–4.11(m,2H),2.69and 2.46(s,3H),1.42–1.34(m,3H).

[0657] Example 70A: (S)-4-amino-7-chloro-N-methyl-N-(6-(pyridin-3-ylethynyl)-2,3-dihydrobenzofuran-3-yl)-1,3-dihydrofurano[3,4-c]quinoline-8-carboxamide (Compound 70A)

[0658]

[0659] Following the preparation procedure of compound 43A, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 3-iodopyridine (48.8 mg, 0.24 mmol), and the reaction was carried out at room temperature for 3 hours to obtain compound 70A (24 mg). ESI-MS (m / z): 497.0 [M+H] + . 1 HNMR(400MHz,DMSO-d6)δ8.79–8.74(m,1H),8.63–8.58(m,1H),8.02–7.96(m,1H),7.90–7.57(m,2H),7.54–7 .34(m,2H),7.28–7.07(m,2H),6.86(s,2H),6.54–5.33(m,3H),5.08–4.97(m,2H),4.89–4.48(m,2H),2.70and 2.48(s,3H).

[0660] Example 71A: (S)-4-amino-7-chloro-N-(6-((1-cyclopropyl-5-methyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N-methyl-1,3-dihydrofuran[3,4-c]quinoline-8-carboxamide (Compound 71A)

[0661]

[0662] Following the preparation procedure of compound 43A, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 1-cyclopropyl-4-iodopyrazole (59.52 mg, 0.24 mmol, preparation method according to WO2023044171 A1). The reaction was carried out at room temperature for 3 hours to obtain compound 71A (8 mg). ESI-MS (m / z): 540.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ7.90–7.60(m,2H),7.57–7.52(m,1H),7.48–7.28(m,1H),7.16–7.05(m,1H),7.04–6. 95(m,1H),6.86(s,2H),6.52–5.28(m,3H),5.07–4.97(m,2H),4.86–4.46(m,2H),3.60–3.53(m,1H),2.69and 2.47(s,3H),2.45–2.44(m,3H),1.08–1.01(m,4H).

[0663] Example 72: 4-Amino-7-fluoro-N,1-dimethyl-N-(7-((1-methyl-1H-pyrazol-4-yl)ethynyl)isochroman-4-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 72)

[0664]

[0665] Referring to the preparation method of 9c in WO2024131901 A1, compound 72a and compound 5c were condensed to obtain compound 72b.

[0666] Under nitrogen protection, 4-ethynyl-1-methyl-1H-pyrazole (21.2 mg, 0.2 mmol), Xphos Pd G3 (8.5 mg, 0.01 mmol), and triethylamine (40.5 mg, 0.4 mmol) were added sequentially to a solution of compound 72a (48.4 mg, 0.1 mmol) in N,N-dimethylacetamide (2 mL). After addition, the mixture was reacted in a microwave oven at 90 °C for 4 hours. Water (50 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated. The solution was then subjected to column chromatography (methanol:dichloromethane = 1:20) to give compound 72 (25 mg). ESI-MS (m / z): 510.01 [M+H] + . 1H NMR (400MHz, DMSO) δ8.40–8.26(m,2H),8.10–8.05(m,1H),7.72–7.67(m,1H),7.49–7.23(m,6H) ,5.84–4.52(m,3H),4.44–4.39(m,3H),4.23–3.97(m,2H),3.89–3.86(m,3H),2.85–2.65(m,3H).

[0667] Example 73A: (S)-4-amino-7-chloro-N-(6-((1-(difluoromethyl)-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N-methyl-1,3-dihydrofurano[3,4-c]quinoline-8-carboxamide (Compound 73A)

[0668]

[0669] Following the preparation procedure of compound 43A, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 1-difluoromethyl-4-iodo-1H-pyrazole (48.8 mg, 0.2 mmol), and the reaction was carried out at 40 °C for 3 hours to obtain compound 73A (20 mg). ESI-MS (m / z): 536.12 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.71–8.64(m,1H),8.12–8.04(m,1H),8.02–7.82(m,1H),7.73–7.58(m,2H),7.51–7.3 1(m,1H),7.20–6.99(m,2H),6.86(s,2H),6.55–5.29(m,3H),5.08–4.99(m,2H),4.88–4.45(m,2H),2.71and 2.46(s,1H).

[0670] Example 74A: (S)-4-amino-7-chloro-N-methyl-N-(6-((5,6,7,8-tetrahydroimidazo[1,2-a]pyridin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1,3-dihydrofurano[3,4-c]quinoline-8-carboxamide (Compound 74A)

[0671]

[0672] Following the preparation procedure of compound 43A, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 66b (40.2 mg, 0.2 mmol), and the reaction was carried out at 100 °C for 3 hours to obtain compound 74A (25 mg). ESI-MS (m / z): 540.25 [M+H]+ . 1 H NMR (400MHz, DMSO) δ7.90–7.58(m,2H),7.47–7.32(m,1H),7.29–7.23(m,1H),7.21–7.00(m,2H),6.86(s,2H),6.54– 6.42(m,1H),6.51–5.28(m,3H),5.07–4.97(m,2H),4.86–4.48(m,2H),4.03–3.93(m,2H),2.82–2.73(m,2H),2.71and 2.46(s,3H),2.01–1.93(m,2H),1.90–1.82(m,2H).

[0673] Example 75A: (S)-4-amino-7-chloro-N-(6-((6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N-methyl-1,3-dihydrofurano[3,4-c]quinoline-8-carboxamide (Compound 75A)

[0674]

[0675] Following the preparation procedure of compound 43A, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 3-bromo-6,7-dihydro-5H-pyrrolo[1,2-a]imidazole (37.4 mg, 0.2 mmol), and the reaction was carried out at 100 °C for 4 hours to obtain compound 75A (25 mg). ESI-MS (m / z): 526.08 [M+H] + . 1 H NMR (400MHz, DMSO) δ7.87–7.58(m,2H),7.49–7.26(m,2H),7.18–6.97(m,H),6.85(s,2H),6.53–5.29( m,3H),5.06–4.97(m,2H),4.86–4.46(m,2H),4.09–3.96(m,2H),2.86–2.75(m,2H),2.72–2.45(m,5H).

[0676] Example 76A: (S)-4-amino-7-chloro-N-methyl-N-(6-((1-methyl-1H-pyrazolo[4,3-b]pyridin-6-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1,3-dihydrofurano[3,4-c]quinoline-8-carboxamide (Compound 76A)

[0677]

[0678] Following the preparation procedure of compound 43A, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 6-bromo-1-methylpyrazolo[4,3-b]pyridine (50.5 mg, 0.24 mmol), and the reaction was carried out at 100 °C for 4 hours to obtain compound 76A (18 mg). ESI-MS (m / z): 551.0 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.69–8.64(m,1H),8.51–8.43(m,1H),8.36–8.31(m,1H),7.90–7.36(m,3H),7.30–7. 09(m,2H),6.85(s,2H),6.56–5.30(m,3H),5.07–4.97(m,2H),4.89–4.50(m,2H),4.12–4.11(m,3H),2.71and 2.50(s,3H).

[0679] Example 77A: (S)-4-amino-N-{6-[(6-cyclopropylpyridin-3-yl)ethynyl]-2,3-dihydro-1-benzofuran-3-yl}-7-fluoro-1,N-dimethylpyrazolo[4,3-c]quinoline-8-carboxamide (Compound 77A)

[0680]

[0681] Following the preparation procedure of compound 5A, compound 4-ethynyl-1-methylpyrazole was replaced with compound 2-cyclopropyl-5-ethynylpyridine (70 mg, 0.49 mmol), and the mixture was reacted in a microwave oven at 80 °C for 4 hours to obtain compound 77A (33 mg). ESI-MS (m / z): 533.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.58 (s, 1H), 8.31–8.23 (m, 2H), 7.82 (d, J = 8.0Hz, 1H), 7. 49–7.05(m,7H),6.53–5.55(m,1H),4.89–4.61(m,2H),4.50–4.30(m,3H),2.72and 2.60(s,3H),2.21–2.12(m,1H),1.07–0.93(m,4H).

[0682] Example 78A: (S)-4-amino-7-chloro-N-methyl-N-(6-((1-methyl-1H-pyrazol-5-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1,3-dihydrofurano[3,4-c]quinoline-8-carboxamide (Compound 78A)

[0683]

[0684] Following the preparation procedure of compound 43A, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 5-iodo-1-methylpyrazole (59.4 mg, 0.29 mmol), and the reaction was carried out at room temperature for 3 hours under nitrogen protection to obtain compound 78A (23 mg). ESI-MS (m / z): 499.91 [M+H] + . 1 H NMR (400MHz, DMSO) δ7.88–7.62(m,1H),7.62–7.59(m,1H),7.55–7.50(m,1H),7.49–7.33(m,1H),7.27–7.0 7(m,2H),6.84(s,2H),6.65–6.59(m,1H),6.57–5.27(m,3H),5.05–4.96(m,2H),4.85–4.47(m,2H),3.94and 3.93(s,3H),2.68and 2.46(s,3H).

[0685] Example 79A: (S)-4-amino-7-fluoro-1-methyl-N-(methyl-D3)-N-(6-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 79A)

[0686]

[0687] The synthetic process for preparing compound 79A from compound Int-2A is the same as the synthetic process for preparing compound 5A from compound Int-2A. Compound 79A (25 mg). ESI-MS (m / z): 499.00 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.38–8.20(m,2H),8.08(s,1H),7.69(s,1H),7.45–7.25(m,4H),7. 17–6.93(m,2H),6.50–5.54(m,1H),4.87–4.61(m,2H),4.46–4.37(m,3H),3.87(s,3H).

[0688] Example 80A: (S)-4-amino-7-fluoro-N-methyl-N-(6-((5,6,7,8-tetrahydroimidazo[1,2-a]pyridin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1,3-dihydrofurano[3,4-c]quinoline-8-carboxamide (Compound 80A)

[0689]

[0690] Following the preparation procedure of compound 16A, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 66a (40.2 mg, 0.2 mmol), and the mixture was reacted in a microwave oven at 100 °C for 4 hours to obtain compound 80A (15 mg). ESI-MS (m / z): 524.32 [M+H] + . 1 HNMR(400MHz,DMSO)δ7.68–7.62(m,1H),7.42–7.29(m,2H),7.28–7.24(m,1H ),7.21–7.11(m,1H),7.09–7.02(m,1H),6.85(s,2H),6.49–5.50(m,1H),5.4 3–5.30(m,2H),5.06–4.98(m,2H),4.85–4.57(m,2H),4.02–3.93(m,2H),2.8 1–2.74(m,2H),2.70and–2.55(s,3H),2.01–1.94(m,2H),1.90–1.82(m,2H).

[0691] Example 81A: (S)-4-amino-7-chloro-N-(6-((1,4-dimethyl-1H-pyrazol-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1Hpyrazolo[4,3-c]quinoline-8-carboxamide (Compound 81A)

[0692]

[0693] Following the preparation procedure of compound 8A, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 3-iodo-1,4-dimethylpyrazole (35.5 mg, 0.16 mmol), and the reaction was carried out at room temperature for 16 hours under nitrogen protection to obtain compound 81A (15 mg). ESI-MS (m / z): 526.2 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ8.42–8.21(m,2H),7.72–7.26(m,5H),7.21–7.10(m,1H),7.10–6.99(m,1H),6.55–5.43(m,1H),4.89–4.57(m,2H),4.47and 4.41(s,3H),3.82–3.81(m,3H),2.75–2.51(m,3H),2.10–2.08(m,3H).

[0694] Example 82A: (S)-4-amino-7-fluoro-N,1-dimethyl-N-(6-((1-(methyl-D3)-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 82A)

[0695]

[0696] Preparation of compound 82b

[0697] Compound 82a (236.7 mg, 2.57 mmol) was dissolved in N,N-dimethylformamide (10 mL). The solution was cooled to 0 °C and sodium hydride (205.6 mg, 5.14 mmol, 60% purity) was added. After reacting at 0 °C for 1 hour, deuterated iodomethane (745.1 mg, 5.14 mmol) was added. The reaction mixture was then stirred at 25 °C for 12 hours. The reaction mixture was quenched with water (5 mL), extracted with ethyl acetate (25 mL x 3), and the organic phase was washed three times with saturated brine. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The filtrate was then subjected to column chromatography (ethyl acetate: petroleum ether = 1:10) to give compound 82b (250 mg). ESI-MS (m / z): 110.02 [M+H] + .

[0698] Preparation of compound 82A

[0699] Following the preparation procedure of compound 5A, compound 4-ethynyl-1-methylpyrazole was replaced with compound 82b (21.8 mg, 0.2 mmol), and the mixture was reacted in a microwave oven at 100 °C for 4 hours to obtain compound 82A (25 mg). ESI-MS (m / z): 498.98 [M+H] + . 1 HNMR(400MHz,DMSO)δ8.40–8.22(m,2H),8.07(s,1H),7.69(s,1H),7.46–7.24(m,4H),7.17–7.06(m, 1H),7.05–6.93(m,1H),6.50–5.52(m,1H),4.88–4.64(m,2H),4.48–4.35(m,3H),2.74–2.57(m,3H).

[0700] Example 83A: (S)-4-amino-N-(6-((1-(difluoromethyl)-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 83A)

[0701]

[0702] The preparation process of compound 11b-A is referenced in WO2024131901 A1.

[0703] Preparation of compound 83A

[0704] To a solution of compound 11b-A (60 mg, 0.15 mmol) in N-methylpyrrolidone (2 mL), cuprous iodide (5.6 mg, 0.03 mmol), tetrakis(triphenylphosphine)palladium (16.8 mg, 0.01 mmol), triethylamine (60.64 μL, 0.44 mmol), and 1-(difluoromethyl)-4-iodopyrazole (106.7 mg, 0.44 mmol) were added, and the mixture was reacted at 25 °C for 3 hours under nitrogen protection. The reaction solution was filtered through diatomaceous earth, water (50 mL) was added, and the mixture was extracted with ethyl acetate (2 x 25 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude compound was obtained by normal-phase column chromatography (dichloromethane / methanol = 20:1), followed by reverse-phase column chromatography (acetonitrile / 0.05% ammonium bicarbonate solution = 0:100%–45%:55%) to give compound 83A (18 mg). ESI-MS (m / z): 528.17 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.67(s,1H),8.24(s,1H),8.11–8.04(m,2H),7.85(t,J=58Hz,1H),7.59–7.41(m, 2H),7.25–7.13(m,1H),7.09–6.95(m,3H),6.58–5.48(m,1H),4.87–4.55(m,2H),4.39(s,3H),2.72and 2.37(s,3H).

[0705] Example 84A: (S)-4-amino-N-(6-((6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 84A)

[0706]

[0707] Following the preparation procedure of compound 12A, compound 1-cyclopropyl-4-iodo-1H-pyrazole was replaced with compound 3-bromo-6,7-dihydro-5H-pyrrolo[1,2-a]imidazole (45.0 mg, 0.24 mmol), and the reaction was carried out at 100 °C for three hours to obtain compound 84A (19 mg). ESI-MS (m / z): 522.0 [M+H] +. 1 H NMR (400MHz, DMSO) δ8.38–8.19(m,2H),7.43–7.27(s,5H),7.19–7.07(m,1H),7.06–6.95(m,1H),6.50–5.52(m,1 H),4.86–4.59(m,2H),4.47–4.32(m,3H),4.07–3.93(m,2H),2.84–2.75(m,2H),2.69(s,2H),2.60–2.53(m,3H).

[0708] Example 85: 4-Amino-N-(7-(((1-cyclopropyl-1H-pyrazol-4-yl)ethynyl)isochroman-4-yl)-7-fluoro-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 85)

[0709] Preparation of compound 85a

[0710] Under nitrogen protection, trimethylsilylacetylene (86.9 mg, 0.88 mmol), Xphos Pd G3 (37.4 mg, 0.044 mmol), and triethylamine (179 mg, 1.77 mmol) were added sequentially to a solution of compound 72a (214 mg, 0.44 mmol) in N,N-dimethylacetamide (5 mL). After addition, the mixture was reacted in a microwave at 90 °C for 4 hours. Water (50 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation. The solution was then subjected to column chromatography (methanol:dichloromethane = 1:20) to give compound 85a (224 mg). ESI-MS (m / z): 501.7 [M+H] + .

[0711] Preparation of compound 85b

[0712] A solution of tetrabutylammonium fluoride (0.9 mL, 1 M) in tetrahydrofuran (5 mL) of compound 85a (224 mg, 0.45 mmol) was added, and the mixture was allowed to react at room temperature for 1 hour. Water was added, and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and subjected to column chromatography (methanol:dichloromethane = 1:20) to give compound 85b (140 mg). ESI–MS (m / z): 430.2 [M+H] + .

[0713] Preparation of compound 85

[0714] Under nitrogen protection, 1-cyclopropyl-4-iodo-1H-pyrazole (46.8 mg, 0.2 mmol), cuprous iodide (3.8 mg, 0.02 mmol), N,N-diisopropylethylamine (51.7 mg, 0.4 mmol), and tetrakis(triphenylphosphine)palladium (11.6 mg, 0.01 mmol) were added sequentially to a 2 mL solution of N-methylpyrrolidone containing compound 85b (43 mg, 0.1 mmol). The mixture was reacted in an oil bath at 40 °C for 3 hours after the additions. The reaction mixture was then diluted with water (100 mL), extracted with ethyl acetate (20 mL x 3), and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated. The solution was then subjected to column chromatography (methanol:dichloromethane = 1:20) to give compound 85 (20 mg). ESI-MS (m / z): 535.84 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.40–8.25(m,2H),8.23–8.16(m,1H),7.71–7.67(m,1H),7.47–7.2 6(m,6H),5.82–4.51(m,3H),4.42(s,3H),4.23–3.96(m,2H),3.83–3.72(m,1H),2.82and 2.69(s,3H),1.13–0.97(m,4H).

[0715] Example 86A: (S)-4-amino-7-chloro-N-(6-((1-cyclopropyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N-(methyl-D3)-1,3-dihydrofurano[3,4-c]quinoline-8-carboxamide (Compound 86A)

[0716]

[0717] Following the preparation process of compound 40b-A, compound 1a-A was replaced with compound 79b-A to obtain compound 86a-A.

[0718] Following the preparation procedure of compound 43A, compound 40b-A was replaced with compound 86a-A to obtain compound 86A (30 mg). ESI-MS (m / z): 528.96 [M+H] + . 1H NMR (400MHz, DMSO) δ8.20–8.15(m,1H),7.87–7.57(m,3H),7.47–7.29(m,1H),7.15–7.03(m,1H),7.02–6.92(m,1 H),6.85(s,2H),6.54–5.28(m,3H),5.08–4.96(m,2H),4.85–4.43(m,2H),3.81–3.71(m,1H),1.10–0.97(m,4H).

[0719] Example 87: 4-Amino-7-chloro-N-(7-((1-cyclopropyl-1H-pyrazol-4-yl)ethynyl)isochromochromo-4-yl)-N-methyl-1,3-dihydrofurano[3,4-c]quinoline-8-carboxamide (Compound 87)

[0720]

[0721] Preparation of compound 87b

[0722] Compound 40a (193 mg, 0.73 mmol) was dissolved in N,N-dimethylformamide (10 mL), and N,N-diisopropylethylamine (0.63 mL, 3.64 mmol) was added. The mixture was stirred in an ice bath for 5 min, and then N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (415 mg, 1.09 mmol) was added. Finally, 87a (176.2 mg, 0.73 mmol, preparation method referred to WO2024131901 A1) was added, and the reaction was continued for 1 hour. The reaction solution was added with water (100 mL), extracted with ethyl acetate (20 mL * 3), and the organic phase was washed three times with saturated brine. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and subjected to column chromatography (methanol:dichloromethane = 1:20) to obtain compound 87b (212 mg). ESI-MS (m / z): 488.0 [M+H] + .

[0723] Preparation of compound 87

[0724] Under nitrogen protection, 1-cyclopropyl-4-ethynyl-1H-pyrazole (26.4 mg, 0.2 mmol), cuprous iodide (3.8 mg, 0.02 mmol), N,N-diisopropylethylamine (0.07 mL, 0.4 mmol), and tetrakis(triphenylphosphine)palladium (11.6 mg, 0.01 mmol) were added sequentially to a solution of compound 87b (48.8 mg, 0.1 mmol) in N,N-dimethylacetamide (2 mL, 0.4 mmol). The mixture was then reacted in a microwave oven at 100 °C for 4 hours. Water (50 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation. The solution was then subjected to column chromatography (methanol:dichloromethane = 1:20) to give compound 87 (30 mg). ESI-MS (m / z): 539.96 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.21–8.17(m,1H),7.76–7.63(m,3H),7.51–7.38(m,2H),7.34–7.24(m,1H),6.85(s,2H),5.84–5.76(m,1H),5. 40–5.31(m,2H),5.07–4.98(m,2H),4.89–4.51(m,2H),4.22–3.91(m,2H),3.84–3.73(m,1H),2.79–2.57(m,3H),1.12–0.98(m,4H).

[0725] Example 88A: (S)-4-amino-7-chloro-N-(6-((5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N-methyl-1,3-dihydrofurano[3,4-c]quinoline-8-carboxamide (Compound 88A)

[0726]

[0727] Following the preparation procedure of compound 43A, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 3-bromo-5,6-dihydro-4H-pyrrolo[2,1-e]pyrazole (44.5 mg, 0.24 mmol), and the mixture was reacted in a microwave oven at 100 °C for 4 hours to obtain compound 88A (10 mg). ESI-MS (m / z): 525.9 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ7.90–7.58(m,3H),7.45–7.27(m,1H),7.15–7.02(m,1H),6.99–6.93(m,1H),6.85(s,2H), 6.50–5.28(m,3H),5.06–4.99(m,2H),4.87–4.45(m,2H),4.16–4.08(m,2H),2.98–2.90(m,2H),2.69–2.46(m,5H).

[0728] Example 89A: (S)-4-amino-N-(6-((1-cyclobutyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N,1-dimethyl-1H-pyrazino[4,3-c]quinoline-8-carboxamide (Compound 89A)

[0729]

[0730] Following the preparation procedure of compound 12A, compound 1-cyclopropyl-4-iodo-1H-pyrazole was replaced with compound 1-cyclobutyl-4-iodo-1H-pyrazole (25 mg, 0.1 mmol), and the reaction was carried out at 25 °C for 2 hours under nitrogen protection to obtain compound 89A (32 mg). ESI-MS (m / z): 536.02 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.42–8.20(m,3H),7.73(s,1H),7.47–7.31(m,4H),7.15–7.05(m,1H),7.03–6.94(m,1H), 6.50–5.54(m,1H),4.92–4.56(m,3H),4.53–4.29(m,3H),2.75–2.56(m,3H),2.50–2.29(m,4H),1.86–1.74(m,2H).

[0731] Example 90A: (S)-4-amino-7-chloro-N-(6-((1-cyclobutyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N-methyl-1,3-dihydrofuran[3,4-c]quinoline-8-carboxamide (Compound 90A)

[0732]

[0733] Following the preparation procedure of compound 43A, compound 1-cyclopropyl-4-iopyrazole was replaced with compound 1-cyclobutyl-4-iopyrazole (50 mg, 0.20 mmol), and the reaction was carried out at room temperature for 2 hours under nitrogen protection to obtain compound 90A (24 mg). ESI-MS (m / z): 540.00 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.24–8.20(m,1H),7.88–7.58(m,3H),7.47–7.29(m,1H),7.15–7.03(m,1H),7.02–6.93(m ,1H),6.86(s,2H),6.52–5.29(m,3H),5.07–4.98(m,2H),4.93–4.46(m,3H),2.69–2.35(m,7H),1.87–1.74(m,2H).

[0734] Example 91A: (S)-4-amino-7-chloro-N-methyl-N-(6-((4,5,6,7-tetrahydropyrazolo[1,5-a]pyridin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1,3-dihydrofuran[3,4-c]quinoline-8-carboxamide (Compound 91A)

[0735]

[0736] Following the preparation procedure of compound 43A, compound 1-cyclopropyl-4-iodopyrazole was replaced with compound 3-bromo-4,5,6,7-tetrahydropyrazolo[1,5-a]pyridine (47.8 mg, 0.24 mmol), and the mixture was microwaved at 100 °C for 4 hours to obtain compound 91A (8 mg). ESI-MS (m / z): 539.87 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ7.93–7.54(m,3H),7.47–7.31(m,1H),7.14–6.88(m,4H),6.52–5.29 (m,3H),5.06–4.98(m,2H),4.85–4.45(m,2H),4.12–4.04(m,2H),2.85–2.81(m,2H),2.69and 2.47(s,3H),2.04–1.94(m,2H),1.88–1.80(m,2H).

[0737] Example 1 for reference: The preparation method is based on WO2024131901A1.

[0738] Reference Example 2: The preparation method is based on WO2024131901A1.

[0739] See Example 3 The preparation method refers to WO2022169948 A1

[0740] See Example 4 See Example 5

[0741] See Example 6. The preparation method is referenced in J.Med.Chem.2022,65(3),1749-1766.

[0742] See Example 7 The preparation method is based on WO2022132914 A1.

[0743] Experimental Example 1: Inhibitory activity of the compound of the present invention against the proliferation of human colon cancer cells.

[0744] 1. Build MTAP - / - HCT-116 cell line

[0745] HCT-116 colon cancer cell line (purchased from Nanjing Kebai) was co-transfected with gene editing tools CRISPR / Cas9 and sgRNA. MTAP biallelic inactivated MTAP was obtained by Western blotting, Sanger sequencing, and other detection methods. - / - HCT-116 cell line. MTAP - / - The HCT-116 cell line was used for selective screening and evaluation of compounds based on the MTAP deficiency and PRMT5 synergistic mechanism, including symmetric dimethylarginine (SDMA) level assay and cell proliferation assay.

[0746] 2. Cell proliferation

[0747] MTAP was cultured in complete culture medium McCoy's 5A (Gibco, 16600082) / 10% FBS (Gibco, 10099141C) / 1% p / s (Gibco, 15140122). - / - HCT-116 and wild-type HCT-116 cell lines were used to evaluate the effects of compounds on MTAP.- / - The selective inhibition and enhancement of HCT-116 cell proliferation. Day 0, 100 MTAP molecules were added to each well of a 96-well cell culture plate (Corning, 3599). - / - HCT-116 cells or wild-type HCT-116 cells were cultured in a 37°C, 5% CO2 incubator. The compound was serially diluted 3x with DMSO (Sigma, D5879) (starting concentration 20 μM, 3x dilution, for a total of 8 concentration points). On Day 1, the compound was diluted to multiple concentration points and used to treat cells separately, and the cells were cultured for another 10 days at 37°C, 5% CO2. On Day 11, 20 μL of MTS (CellTiter) was added to each well. A Queous One Solution Cell Proliferation Assay (Promega, G3581), after incubation at 37°C and 5% CO2 for 2 hours, readings were taken using Tecan Spark (OD = 490 nM). Data analysis was then performed using GraphPad Prism 8 software, employing the equation "log(inhibitor) vs. normalized response-variable slope" (formula Y = Bottom + (Top - Bottom) / (1 + 10^(LogIC)). 50 Data analysis was performed using -X)*HillSlope))) to obtain the IC50 of the compound. 50 Value. Where Y is the inhibition rate, X is the logarithm of the compound concentration, Top refers to the maximum response (inhibition rate at the highest compound concentration), Bottom refers to the baseline response (inhibition rate at zero compound concentration), and Hill Slope refers to the IC50 value. 50 The slope of the curve, IC 50 The concentration of the compound at half-maximal inhibition is given. The experimental results are shown in Table 1.

[0748] Table 1IC 50 (μM)

[0749]

[0750]

[0751] The results showed that the specific compounds and their isomers of the present invention exhibited strong inhibitory activity against the human MTAP-deficient HCT-116 cell line, and most of the example compounds showed an IC50 inhibitory effect on the human MTAP-deficient HCT-116 cell line. 50The values ​​were all less than 0.01 μM. In terms of selectivity, compared with the MTAP wild-type (WT) HCT-116 cell line, the specific compounds and their isomers of the present invention exhibited better selective inhibition against the human MTAP-deficient HCT-116 cell line, with most compounds showing a selective inhibition of more than 90-fold.

[0752] Experimental Example 2: Human Liver Microsomal Stability Test

[0753] Experimental materials: Human liver microsomes used in the experiment (purchased from BIOIVT);

[0754] Reagent preparation:

[0755] PBS: 0.1M KH2PO4 and K2HPO4 buffer, pH 7.4.

[0756] MgCl2: Weigh a certain amount of MgCl2 and prepare a 16mM MgCl2 solution using PBS.

[0757] NADPH (Chinese name: reduced nicotinamide adenine dinucleotide phosphate, purchased from Sigma, catalog number: 481973-500mg): Weigh a certain amount of NADPH and prepare it to 4mM with 16mM MgCl2 solution. The final incubation concentration is 1mM.

[0758] Compounds: The test compound and positive compound were prepared to 4 μM with PBS, and the final incubation concentration was 1 μM.

[0759] Liver microsomes (purchased from BIOIVT, catalog number: X008070): Liver microsomes were diluted to 1 mg / mL with PBS, and the final incubation concentration was 0.5 mg / mL.

[0760] Experimental steps:

[0761] The prepared test compound or positive compound was added to a test tube, followed by the prepared NADPH solution, and mixed thoroughly. The tube was pre-incubated at 37°C and 220 rpm for 5 minutes. After pre-incubation, the prepared liver microsomes were added to initiate the reaction, and the reaction was repeated in replicates. At 0, 15, 30, 45, and 60 minutes, a certain volume of ice-cold acetonitrile solution containing an internal standard was added to the corresponding tube to precipitate the protein. The tube was vortexed for 5 minutes, followed by centrifugation at 4000 rpm for 10 minutes. The supernatant was collected and transferred to a 96-well plate for analysis by LC-MS / MS. The concentration (peak area ratio) of the compounds in the examples was determined by LC-MS / MS. The rate constant was obtained by plotting "Ln (residual amount of compound %)" against "incubation time" in Excel, thereby calculating the drug's half-life and intrinsic clearance rate, providing a basis for predicting in vivo clearance. The results of this experiment are shown in Table 2.

[0762] Data Analysis:

[0763] CL int =(0.693 / t) 1 / 2 Microsomes) × [Incubation liquid volume (mL) / Microsome protein content (mg)] × [Microsome protein content (mg) / Liver weight (g)] × [Liver weight (g) / Body weight (kg)] [1]

[0764] CL H =CL int ×f u ×Q h / (CL int ×f u +Q h )

[0765] In the formula,

[0766] CL int --Inherent clearance rate (mL / min / kg)

[0767] CL H --Liver clearance rate (mL / min / kg)

[0768] f u --Plasma protein unbound fraction is 1

[0769] Q h --Hepatic blood flow

[0770] Table 2. Stability of human liver microsomes

[0771] compound T1 / 2(min) compound T1 / 2(min) Reference Example 1 16.6 Example 4 169 See Example 2 33.6 Compound 5A 66.5 See Example 5 25.6 Compound 25A 68.9 See Example 7 28.9 Compound 40A 62.9 Compound 50A 49.9 Compound 43A 122 Compound 91A 47.2

[0772] Table 2 shows that the compounds of the present invention have excellent stability in human liver microsomes.

[0773] Experimental Example 3: hERG Test

[0774] Experimental reagents:

[0775] Predictor TM hERG Fluorescence Polarization Assay Kit (ThermoFisher Scientific, PV5365).

[0776] Experimental steps:

[0777] Follow the kit instructions: Prepare 4×Predictor kits in advance. TMhERG Tracer Red, 4×E-4031, and 4× test solution (final detection concentration 10 μM). In a 384-well plate, 5 μL of lecithin was added sequentially to the Assay Blank group. TM hERG FP Assay Buffer and 5μL Predictor TM hERG Membrane. Add 2.5 μL of Predictor to each of the negative control, positive control, and test group. TM Add hERG FP Assay Buffer, 2.5 μL 4×E-4031, 2.5 μL 4× test substance, and then add 5 μL Predictor sequentially. TM hERG Membrane and 2.5μL 4×Predictor TM hERG Tracer Red. The total volume of the experimental system was 10 μL, with three replicates per group. After adding the sample, gently tap to mix, and incubate at room temperature (25℃) in the dark for 3.5 h. After incubation, the fluorescence polarization value (mP) was measured using a microplate reader, and the G Factor was 2.14.

[0778] Data Analysis:

[0779] The inhibition rate of the test substance is calculated using the following formula:

[0780] Inhibition rate % = (mP negative control - mP test substance) / (mP negative control - mP positive control) * 100% Using the above experimental method, the hERG inhibition experimental results of some compounds of the present invention are shown in Table 3 below.

[0781] Table 3 hERG Test

[0782] Table 3 shows that the compounds of the present invention have lower hERG inhibitory toxicity compared to the prior art (such as Reference Example 6).

[0783] Experimental Example 4: CYP Inhibition Test

[0784] Experimental materials: Human liver microsomes used in the experiment (purchased from BIOIVT);

[0785] Reagent preparation:

[0786] PBS: 0.1M KH2PO4 and K2HPO4 buffer, pH 7.4.

[0787] MgCl2: Weigh a certain amount of MgCl2 and prepare a 20mM MgCl2 solution using PBS.

[0788] NADPH (Chinese name: reduced nicotinamide adenine dinucleotide phosphate, purchased from Sigma, catalog number: 481973-500mg): Weigh a certain amount of NADPH, and use 20mM MgCl2 solution to prepare NADPH to 5mM. The final incubation concentration is 1mM.

[0789] Compounds: The test compounds were prepared to concentrations of 3, 1, 0.3, 0.1, 0.03, 0.01, and 0 mM using DMSO-methanol (v:v = 1:9), with final incubation concentrations of 30, 10, 3, 1, 0.3, 0.1, and 0 μM. The positive compound (ketoconazole) was prepared to concentrations of 30, 10, 3, 1, 0.3, and 0 μM using DMSO-methanol (v:v = 1:9), with final incubation concentrations of 0.3, 0.1, 0.03, 0.01, 0.003, and 0 μM.

[0790] Substrate working solutions: Prepare substrate working solutions (testosterone and midazolam) to 200 and 15 μM respectively using PBS, with final incubation concentrations of 40 and 3 μM.

[0791] Liver microsomes (purchased from BIOIVT, catalog number: X008070): Liver microsomes were diluted with PBS to 0.17 mg / mL, and the final incubation concentration was 0.1 mg / mL.

[0792] Experimental steps:

[0793] Add the prepared test compound or positive compound to a 96-well plate, followed by the prepared human liver microsomal working solution and substrate working solution, and mix thoroughly. Incubate at 37°C for 5 minutes, then add the prepared NADPH initiation reaction solution. Incubate for 5 or 10 minutes. After incubation, add pre-cooled acetonitrile solution containing internal standard to terminate the reaction. After protein precipitation, vortex for 5 minutes, then centrifuge at 4000 rpm for 10 minutes. Collect the supernatant in a 96-well plate. Detect the content (peak concentration ratio) of the substrate metabolites using LC-MS / MS. Calculate the residual enzyme activity and half-maximal inhibitory concentration (IC50) based on the detected content. 50 ).

[0794] Data Analysis:

[0795] Percentage of remaining enzyme activity:

[0796] The activity of each CYP450 subtype enzyme is expressed as the amount of metabolites produced. The formula for calculating the percentage of remaining enzyme activity is as follows:

[0797] Remaining activity (%) = (Activity of this CYP450 enzyme subtype in the presence of the test substance ÷ Activity of the solvent control group) × 100%, with the activity of the solvent control group (0 μM) set at 100%.

[0798] IC50 :

[0799] The concentration of the test substance and the corresponding percentage of residual activity were fitted using a nonlinear model in Excel software. The concentration at which the test substance achieved 50% inhibition of metabolite formation was defined as the IC50. 50 Values. The results of this experiment are shown in Table 4.

[0800] Table 4

[0801] Table 4 shows that the compounds of the present invention have lower CYP inhibitory toxicity compared to the prior art (such as Reference Example 6).

[0802] Experimental Example 5: Mouse PK Experiment. Experimental materials: The experimental animals were healthy adult BALB / c female mice (provided by Sichuan Vital River Laboratory Animal Technology Co., Ltd.);

[0803] Experimental procedure:

[0804] Administration and sample collection in mice: BALB / c female mice were administered the drug orally by gavage (10 mg / kg, solvent: 0.5% methylcellulose / 0.2% Tween 80). 60 μL of whole blood from the fundus venous plexus of the mice was collected at different time points (0.25, 0.5, 1, 2, 4, 6, 8 and 24 h) after administration. Plasma was collected by centrifugation at 4000 rpm for 6 min.

[0805] Sample analysis:

[0806] 10 μL of mouse plasma was collected, and 190 μL of acetonitrile solution containing internal standard was added to precipitate proteins. The mixture was vortexed for 10 min, followed by centrifugation at 4000 rpm for 10 min. The supernatant was collected into a 96-well plate and analyzed by LC-MS / MS. The drug concentration in the plasma of mice after gavage administration of the compound was determined at different time points using LC-MS / MS, and relevant pharmacokinetic parameters were calculated to study the pharmacokinetic behavior of the compound in mice and evaluate its pharmacokinetic characteristics. The experimental results are shown in Table 5.

[0807] Table 5. Pharmacokinetic parameters of the compounds of this invention in mice.

[0808]

[0809] Note: 1. The solvent used is 0.5% MC / 0.2% Tween 80, PO for administration.

[0810] 2. The test method for PK in MRTX1719 mice is based on J.Med.Chem.2022,65(3),1749-1766.

[0811] Table 5 shows that the compounds provided by this invention have excellent pharmacokinetic properties in mice.

[0812] Experiment 6: Rat PK Experiment Experimental Materials: Healthy adult male SD rats (provided by Sichuan Vitonlihua Laboratory Animal Technology Co., Ltd.)

[0813] Experimental procedure:

[0814] Administration and sample collection in rats: Male SD rats were administered the drug orally by gavage (10 mg / kg, solvent: 0.5% methylcellulose / 0.2% Tween 80). Whole blood (200 μL) from the fundus venous plexus was collected at different time points (0.25, 0.5, 1, 2, 4, 6, 8, and 24 h) after administration. Plasma was collected by centrifugation at 4000 rpm for 6 min. Sample analysis:

[0815] 10 μL of rat plasma sample was taken, and 190 μL of acetonitrile solution containing internal standard was added to precipitate proteins. The mixture was vortexed for 10 min, followed by centrifugation at 4000 rpm for 10 min. The supernatant was collected into a 96-well plate and analyzed by LC-MS / MS. The drug concentration in the plasma of rats after gavage administration of the compound was determined at different time points using LC-MS / MS, and relevant pharmacokinetic parameters were calculated to study the pharmacokinetic behavior of the compound in rats and evaluate its pharmacokinetic characteristics. The experimental results are shown in Table 6.

[0816] Table 6. Pharmacokinetic parameters of rats

[0817] Note: 1. The solvent used is 0.5% MC / 0.2% Tween 80, PO for administration.

[0818] Table 6 shows that the compounds provided by this invention have excellent pharmacokinetic properties in rats.

[0819] Example 7: Efficacy evaluation in a mouse model of subcutaneous xenograft of gene-edited MTAP- / - human colon cancer cell line HCT116.

[0820] Experimental materials:

[0821] Gene-edited MTAP- / - human colon cancer cell line HCT116 (purchased from Nanjing Kebai) was cultured in McCoys 5A medium (Invitrogen) containing 10% fetal bovine serum (Gibco);

[0822] The test substance was administered after being prepared into a suspension with a concentration of 0.3 mg / mL using an aqueous solution containing 0.5% MC and 0.2% Tween 80. Reference Example 6 (MRTX1719) was administered after being prepared into a clear solution with a concentration of 5 mg / mL using 40% PEG400 saline containing 2% DMSO. The control group used 40% PEG400 saline containing 2% DMSO.

[0823] Experimental methods:

[0824] HCT116 MTAP- / - cells in logarithmic growth phase were digested and collected, counted, and then adjusted to a cell density of 1×10⁻⁶ cells using serum-free medium. 8 Cells / mL. 20–22 g Balb / c nude mice (Beijing Huafukang Biotechnology Co., Ltd.) were subcutaneously inoculated with 100 μL of cell suspension per mouse. On day 10 after inoculation, the tumor volume reached 100–150 mmHg. 3 Animals were randomly assigned to groups of eight based on body weight and tumor volume. The test substance was administered by gavage at a dose of 10 mg / kg once daily; MRTX1719 was administered by gavage at a dose of 50 mg / kg once daily. The control group was administered by gavage at a dose of 10 mL / kg using a solvent (2% DMSO + 40% PEG400 + 58% saline) once daily. The administration period was 28 days.

[0825] After animals were inoculated with HCT116 MTAP- / - cells, mouse body weight and tumor volume were measured 2-3 times per week. Tumor volume was calculated as length × width. 2 Calculated by multiplying by 0.5. Mice were euthanized on the day following the last administration.

[0826] The tumor inhibition rate (TGI) for each treatment group was calculated using the following formula:

[0827] The formula for calculating TGI (tumor volume) is as follows: TGI TV =1-((V) Tt -V T0 ) / (V Ct -V C0 ))×100%. Where V Tt V represents the tumor volume at each measurement in the treatment group. T0 Tumor volume was measured when grouping the treatment group; V Ct The group represents the tumor volume (as opposed to V) at each measurement in the solvent group. Tt (Same time), V C0 The tumor volume was measured when the solvent group was grouped.

[0828] Data were statistically analyzed using GraphPad Prism 9.0 software, and are expressed as Mean ± SEM (standard error). One-way ANOVA was used for pairwise comparisons between each treatment group and the control group. Homogeneity of variance was determined using the Brown-Forsythe method, and unequal variances were determined using Dunnett's T3 method.

[0829] Table 7. Efficacy evaluation in the HCT116 subcutaneous xenograft mouse model.

[0830]

[0831]

[0832] As shown in Table 7, the compounds of this invention exhibit significantly better in vivo antitumor effects than the clinical molecule MRTX1719 in the HCT116 subcutaneous xenograft mouse model.

[0833] Efficacy evaluation in a mouse model of human giant cell lung cancer LU99 transplantation, Case 8

[0834] Experimental materials:

[0835] The human lung giant cell carcinoma line LU99 was purchased from Nanjing Kebai Biotechnology Co., Ltd. (CBP61512) and cultured in RPMI-1640 medium (Gibco) containing 10% fetal bovine serum (Gibco).

[0836] The test substance was prepared into suspensions with concentrations of 0.3, 1, and 5 mg / mL using a 0.5% MC aqueous solution containing 0.2% Tween 80. Reference Example 6 (MRTX1719) was prepared into a clear solution with a concentration of 5 mg / mL using 40% PEG400 physiological saline containing 2% DMSO.

[0837] Experimental methods:

[0838] LU99 cells in the logarithmic growth phase were digested and collected, counted, and then adjusted to a cell density of 2 × 10⁶ cells using 1:1 serum-free medium and Corning gel. 7 Cells / mL. 18–20 g Balb / c nude mice (Beijing Huafukang Biotechnology Co., Ltd.) were subcutaneously inoculated with 100 μL of cell suspension per mouse. On day 12 after inoculation, the average tumor volume reached 116 mm². 3 Animals were randomly divided into groups of 7. The test substance was administered by gavage at doses of 3, 10, and 50 mg / kg once daily; MRTX1719 was administered by gavage at a dose of 50 mg / kg once daily. The control group was administered by gavage at a dose of 10 mL / kg of the solvent (2% DMSO + 40% PEG400 + 58% saline) once daily. The total administration period was 28 days.

[0839] After animals were inoculated with LU99 cells, tumor volume was measured 2-3 times per week, with the tumor volume calculated as length × width. 2 Calculated by multiplying by 0.5. On the last day of administration, except for mice whose tumors had regressed and were observed after drug withdrawal, the remaining mice were euthanized.

[0840] The tumor inhibition rate (TGI) for each treatment group was calculated using the following formula:

[0841] The formula for calculating TGI (tumor volume) is as follows: TGI TV =1-((V) Tt -V T0 ) / (V Ct -V C0 ))×100%. Where V Tt V represents the tumor volume at each measurement in the treatment group. T0 Tumor volume was measured when grouping the treatment group; V Ct The group represents the tumor volume at each measurement, V (Vmodel). C0 Tumor volumes were measured when grouping the model control group;

[0842] Data were statistically analyzed using GraphPad Prism 9.0 software, and are expressed as Mean ± SEM (standard error). Pairwise comparisons between each treatment group and the model control group were performed using one-way ANOVA. Homogeneity of variance was determined using the Brown-Forsythe method, and heterogeneity of variance was determined using Dunnett's T3 method.

[0843] Table 8. Efficacy evaluation of LU99 in mouse transplantation models.

[0844]

[0845] Experimental Results and Analysis:

[0846] The experimental results are shown in Table 8. The compounds of the present invention significantly reduced tumor volume at concentrations of 3, 10, and 50 mg / kg (P<0.0001). The antitumor effects of the test substance at 10 and 50 mg / kg were significantly better than those of MRTX1719 at 50 mg / kg, and the test substance at 50 mg / kg caused tumor regression.

[0847] In summary, the experimental results show that the test substance of this invention has a good anti-tumor effect in the human lung giant cell carcinoma LU99 CDX model, and its anti-tumor efficacy is significantly better than that of Yangshen at a lower dose.

[0848] Industrial applicability

[0849] The compounds of this invention have excellent PRMT5 inhibitory activity, and compared with the MTAP wild-type (WT) HCT-116 cell line, the compounds of this invention have excellent selective inhibitory activity against the human MTAP-deficient HCT-116 cell line. They can be used to develop small molecule drugs targeting PRMT5·MTA and preferentially act on MTAP-deficient tumor cells.

[0850] Furthermore, through experiments such as human liver microsomal stability, mouse pK, rat pK, and in vivo antitumor efficacy evaluation in mice, it can be found that the compounds of the present invention have excellent antitumor application prospects.

[0851] Furthermore, the compounds of the present invention have a higher safety window than existing compounds and are significantly superior to existing technologies in terms of safety (such as inhibition of toxicity in hERG, CYP, etc.).

[0852] In summary, the compounds of this invention have achieved unexpected technical effects in terms of drug development and safety compared to existing technologies, and have great development potential.

[0853] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

[0854] References

[0855] [1] Davies B, Morris T. Physiological parameters in laboratory animals and humans. Pharm Res. 1993; 10: 1093-5.

Claims

1. The compound represented by Formula I, its tautomers, stereoisomers, pharmaceutically acceptable salts, or deuterated compounds thereof: in, Selected from single or double bonds, or It does not exist; when When X is a single bond or a double bond, X, Y, and Z are independently selected from CR. 4 R 5 NR 6 O, N, S, CR 4 ; when When Y does not exist, X is hydrogen and Z is CR. 4 R 5 ; W is selected from CR 7 Or N; A is selected from CR 8 R 9 NR 8 Or O; The ring M is selected from 5-10 aryl or 5-10 heteroaryl, wherein the 5-10 heteroaryl contains 1-4 heteroatoms selected from N, O or S; R 3 R 4 R 5 R 6 R 7 R 8 R 9 Independently selected from hydrogen, deuterium, halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 cycloalkyl, C 3-6 Cycloalkoxy; wherein, the C 1-6 Alkyl, C 1-6 The alkoxy group is unsubstituted or surrounded by one or more R groups. i Replace; the C 3-6 cycloalkyl, C 3-6 The cycloalkoxy group is unsubstituted or surrounded by one or more R groups. j replace; R 1 Selected from C 1-6 Alkyl, C 3-10 cycloalkyl-(C 1-6 alkyl) m -, 4-10 membered heterocyclic group-(C 1-6 alkyl) n -, 5-10 quinone heteroaryl-(C 1-6 alkyl) p -, 5-10 aryl-(C 1-6 alkyl) q -; where n = 0 or 1, m = 0 or 1, p = 0 or 1, q = 0 or 1; the heterocyclic group and the heteroaryl group each independently contain 1-4 heterocyclic atoms selected from O, N or S; wherein, the C 1-6 The alkyl group is unsubstituted or contains one or more R groups. i Replace; the C 3-10 The cycloalkyl group is unsubstituted or contains one or more R groups. j Substitution; the 4-10 membered heterocyclic group is unsubstituted or replaced by one or more R groups. k Substitution; the 5-10 aryl and 5-10 heteroaryl groups are unsubstituted or independently substituted by one or more R groups. l replace; R 2 Selected from: 5-10-membered heteroaryl groups and 4-10-membered heterocyclic groups, wherein each of the 5-10-membered heteroaryl group and the 4-10-membered heterocyclic group independently contains 1-4 heteroatoms selected from N, O, or S; wherein the 5-10-membered heteroaryl group and the 4-10-membered heterocyclic group are unsubstituted or converted by one or more identical or different R atoms. b replace; n1 and n2 are each independently 0, 1, and 2; Each R i R j R k R l Each C is independently selected from hydroxyl, deuterium, halogen, cyano, and optionally substituted by one or more C groups selected from halogen, cyano, or hydroxyl. 1-6 Alkyl group, optionally surrounded by one or more groups selected from halogen, cyano, hydroxyl, -NR'R", C 1-6 alkoxy-substituted C 1-6 Alkoxy, C 3-10 cycloalkyl, optionally with one or more C 1-6 Alkyl-substituted 5-10-membered heteroaryl groups, wherein the 5-10-membered heteroaryl group contains 1-4 heteroatoms selected from N, O, and S; wherein R' and R" are each independently selected from H and C. 1-6 Alkyl, halogenated C 1-6 Alkyl groups, or R' and R" together with the nitrogen atom they are attached to, form 4-8 membered heterocyclic groups, wherein the 4-8 membered heterocyclic groups contain 1-4 heteroatoms selected from N, O, and S; Each R b Each is independently selected from hydroxyl, deuterium, cyano, halogen, -NR'R", C1-C6 alkyl, C 1-6 Alkoxy, C3-C6 cycloalkyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, and C3-C6 cycloalkyl are unsubstituted or each is independently substituted by one or more substituents selected from halogen, hydroxyl, cyano, and C1-C6 alkoxy; wherein in -NR'R”, R' and R” are each independently selected from hydrogen, C 1-6 Alkyl, Halogenated C 1-6 Alkyl groups, or R' and R" together with the nitrogen atom to which they are attached, form 4-8 membered heterocyclic groups, which contain 1-4 heteroatoms selected from N, O, and S.

2. The compound according to claim 1, wherein its tautomers, stereoisomers, pharmaceutically acceptable salts, or deuterated compounds thereof, are characterized in that: Ring M is selected from 5-6 aryl or 5-6 heteroaryl; Preferably, ring M is selected from phenyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrazolyl, imidazolyl, triazolyl, pyrroleyl, furanyl, oxazolyl, isoxazolyl, and thiazolyl; More preferably, ring M is selected from phenyl or pyridinyl; Preferably, A is selected from O; Preferably, n1 and n2 are each independently 0 or 1; Preferred, group Selected from Preferred, group Selected from Preferred, group Selected from: Preferred, group Selected from:

3. The compound according to claim 1, its tautomers, stereoisomers, pharmaceutically acceptable salts, or deuterated compounds thereof, characterized in that: R 2 The compounds are selected from 5-6-membered heteroaryl groups and 4-6-membered heterocyclic groups; each of the 5-6-membered heteroaryl group and the 4-6-membered heterocyclic group independently contains 1-3 heteroatoms selected from N, O, or S; the 5-6-membered heteroaryl group and the 4-6-membered heterocyclic group are unsubstituted or independently converted by one or more identical or different R atoms. b replace; Or, R 2 The group is selected from 5-6-membered heteroaryl, 4-6-membered heterocyclic, and 5-6-membered heteroaryl-5-6-membered heterocyclic groups; each of the 5-6-membered heteroaryl, 4-6-membered heterocyclic, and 5-6-membered heterocyclic groups independently contains 1, 2, or 3 heteroatoms selected from N, O, or S; the 5-6-membered heteroaryl, 4-6-membered heterocyclic, and 5-6-membered heteroaryl-5-6-membered heterocyclic groups are unsubstituted or independently converted by one or more identical or different R atoms. b replace; Preferred, R 2 The aryl group is selected from 5-6-membered heteroaryl groups, wherein the 5-6-membered heteroaryl group contains 1-3 heteroatoms selected from N, O, or S; the 5-6-membered heteroaryl group is unsubstituted or converted by one or more identical or different R atoms. b Replacement; preferred, R 2 The group is selected from pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyridyl, pyrazinyl, pyrimidinyl, and pyridazinyl, and is either unsubstituted or independently bound by one, two, or three R groups. b replace; Or preferably, R 2 Selected from 5-6-membered heteroaryl groups, wherein the 5-6-membered heteroaryl group contains one, two, or three N heteroatoms; the 5-6-membered heteroaryl group is unsubstituted or converted by one or more identical or different R atoms. b replace; Preferred, R 2 Selected from pyrazolyl, imidazole, and pyridyl groups, which are unsubstituted or independently bound by one, two, or three R groups. b replace; Preferred, R 2 Selected from It is either unreplaced or independently controlled by one, two, or three Rs. b replace; Or preferably, R 2 The 5-membered heteroaryl group is selected from 5-membered heteroaryl groups, wherein the 5-membered heteroaryl group contains 1-3 heteroatoms selected from N, O, or S, and the 5-membered heteroaryl group is unsubstituted or surrounded by 1, 2, or 3 R atoms. b replace; Preferred, R 2 Selected from pyrazolyl, imidazole, thiazolyl, isothiazolyl, oxazolyl, and isoxazolyl, which are unsubstituted or surrounded by one, two, or three R groups. b replace; Or preferably, R 2 Selected from 5-membered heteroaryl groups, wherein the 5-membered heteroaryl group contains 1-2 heteroatoms selected from N or S, and wherein the 5-membered heteroaryl group is unsubstituted or converted by 1, 2 or 3 R atoms. b replace; Preferred, R 2 Selected from pyrazolyl, imidazole, thiazolyl, and isothiazolyl groups, which are unsubstituted or independently bound by one, two, or three R groups. b replace; Or preferably, R 2 Selected from 5-membered heteroaryl groups, wherein the 5-membered heteroaryl group contains 1, 2, or 3 N heteroatoms, and wherein the 5-membered heteroaryl group is unsubstituted or converted by 1, 2, or 3 identical or different R atoms. b replace; Preferred, R 2 Selected from pyrazolyl or imidazole groups, which are unsubstituted or independently bound by one, two, or three R groups. b replace; Preferred, R 2 Selected from pyrazolyl groups, which are unsubstituted or independently composed of one, two, or three R groups. b replace; Preferred, R 2 Selected from It is either unreplaced or independently controlled by one, two, or three Rs. b replace; Preferred, R 2 Selected from Or preferably, R 2 The group is selected from 6-membered heteroaryl groups, wherein the 6-membered heteroaryl group contains one, two, or three heteroatoms selected from N, O, or S; the 6-membered heteroaryl group is unsubstituted or substituted with one or more identical or different R atoms. b replace; Preferred, R 2 Selected from 6-membered heteroaryl groups, wherein the 6-membered heteroaryl group contains 1, 2, or 3 N heteroatoms; wherein the 6-membered heteroaryl group is unsubstituted or converted by 1, 2, or 3 identical or different R atoms. b replace; Preferred, R 2 Selected from pyridinyl, wherein the pyridinyl group is unsubstituted or converted by one, two, or three identical or different R groups. b replace; Preferred, R 2 Selected from It is unsubstituted or replaced by one, two, or three identical or different Rs. b replace; Preferred, R 2 Selected from Or preferably, R 2 The compounds are selected from 5-6-membered heteroaryl and 5-6-membered heterocyclic groups, wherein each of the 5-6-membered heteroaryl and 5-6-membered heterocyclic groups independently contains 1, 2, or 3 heteroatoms selected from N, O, or S; the 5-6-membered heteroaryl and 5-6-membered heterocyclic groups are unsubstituted or modified by one or more identical or different R atoms. b replace; Preferred, R 2 Selected from 5-6-membered heteroaryl and 5-6-membered heterocyclic groups, wherein each of the 5-6-membered heteroaryl and 5-6-membered heterocyclic group independently contains 1, 2, or 3 N heteroatoms; wherein the 5-6-membered heteroaryl and 5-6-membered heterocyclic group is unsubstituted or converted by one or more identical or different R atoms. b replace; Preferred, R 2 Selected from 5-membered heteroaryl and 6-membered heterocyclic groups, wherein each of the 5-membered heteroaryl and 6-membered heterocyclic group independently contains 1, 2, or 3 N heteroatoms; wherein the 5-membered heteroaryl and 6-membered heterocyclic group is unsubstituted or converted by one or more identical or different R atoms. b replace; Preferred, R 2 Selected from imidazolidenepiperazinyl; said imidazolidenepiperazinyl group is unsubstituted or converted by one, two, or three identical or different R groups. b replace; Preferred, R 2 Selected from Or preferably, R 2 The compounds are selected from 5-6-membered heteroaryl and 5-6-membered heterocyclic groups, 5-6-membered heteroaryl and 5-6-membered heteroaryl groups, and 5-6-membered heteroaryl phenyl groups, wherein each of the 5-6-membered heteroaryl and 5-6-membered heterocyclic groups independently contains 1, 2, or 3 heteroatoms selected from N, O, or S; and wherein the 5-6-membered heteroaryl and 5-6-membered heterocyclic groups, 5-6-membered heteroaryl and 5-6-membered heteroaryl phenyl groups are unsubstituted or converted by one or more identical or different R atoms. b replace; Preferred, R 2 The molecule is selected from 5-6-membered heteroaryl and 5-6-membered heterocyclic groups, 5-6-membered heteroaryl and 5-6-membered heteroaryl groups, and 5-6-membered heteroaryl phenyl groups, wherein the 5-6-membered heteroaryl group is selected from imidazolyl, pyrazolyl, and pyridinyl, and the 5-6-membered heterocyclic group is selected from pyrrolidinyl, piperidinyl, and piperazinyl; the 5-6-membered heteroaryl and 5-6-membered heterocyclic group, 5-6-membered heteroaryl and 5-6-membered heteroaryl phenyl groups are unsubstituted or substituted with one or more identical or different R groups. b replace; Preferred, R 2 The compound is selected from imidazopyrrolidinyl, imidazopiperidinyl, imidazopiperazinyl, pyrazolophenyl, and pyrazolopyridinyl, wherein the imidazopyrrolidinyl, imidazopiperidinyl, imidazopiperazinyl, pyrazolophenyl, and pyrazolopyridinyl are unsubstituted or substituted with one or more identical or different R groups. b replace; Preferred, R 2 Selected from Or preferably, R 2 The group is selected from 4-6 membered heterocyclic groups; the 4-6 membered heterocyclic group contains 1-3 heteroatoms selected from N, O or S; the 4-6 membered heterocyclic group is unsubstituted or converted by 1, 2 or 3 identical or different R atoms. b replace; Preferred, R 2 Selected from 4-6-membered heterocyclic alkyl groups and 4-6-membered heterocyclic alkenyl groups; each of the 4-6-membered heterocyclic alkyl groups and 4-6-membered heterocyclic alkenyl groups independently contains 1-3 heteroatoms selected from N, O, or S; the 4-6-membered heterocyclic alkyl groups and 4-6-membered heterocyclic alkenyl groups are unsubstituted or independently separated by 1, 2, or 3 identical or different R atoms. b replace; Preferred, R 2 Selected from 4-6-membered heterocyclic alkyl groups and 4-6-membered heterocyclic alkenyl groups; each of the 4-6-membered heterocyclic alkyl groups and 4-6-membered heterocyclic alkenyl groups independently contains 1-2 heteroatoms selected from N or O; the 4-6-membered heterocyclic alkyl groups and 4-6-membered heterocyclic alkenyl groups are unsubstituted or independently separated by 1, 2 or 3 identical or different R atoms. b replace; Preferred, R 2 Selected from tetrahydro-2H-pyranyl, oxetyl, 1,2,3,6-tetrahydropyridyl, and 3,6-dihydro-2H-pyranyl; wherein the tetrahydro-2H-pyranyl, oxetyl, 1,2,3,6-tetrahydropyridyl, and 3,6-dihydro-2H-pyranyl are unsubstituted or independently converted by one, two, or three identical or different R groups. b replace; Preferred, R 2 Selected from The It is either unsubstituted or independently replaced by one, two, or three identical or different Rs. b replace; Or preferably, R 2 Selected from Where R 2a With R 2b Together with the carbon atoms they are attached to, they form a 4-6 membered heterocyclic group, which contains 1-3 heteroatoms selected from N, O, or S; the 4-6 membered heterocyclic group is unsubstituted or independently bound by 1, 2, or 3 identical or different R atoms. b replace; Preferred, R 2 Selected from Where R 2a With R 2b Together with the carbon atoms they are connected to, they form 4-6 membered heterocyclic alkyl groups or 4-6 membered heterocyclic alkenyl groups; each of the 4-6 membered heterocyclic alkyl groups and 4-6 membered heterocyclic alkenyl groups independently contains 1-3 heteroatoms selected from N, O, or S; the 4-6 membered heterocyclic alkyl groups and 4-6 membered heterocyclic alkenyl groups are unsubstituted or independently bound by 1, 2, or 3 identical or different R atoms. b replace; Preferred, R 2 Selected from Where R 2a With R 2b Together with the carbon atoms they are connected to, they form 4-6 membered heterocyclic alkyl groups or 4-6 membered heterocyclic alkenyl groups; each of the 4-6 membered heterocyclic alkyl groups and 4-6 membered heterocyclic alkenyl groups independently contains 1-2 heteroatoms selected from N or O; the 4-6 membered heterocyclic alkyl groups and 4-6 membered heterocyclic alkenyl groups are unsubstituted or independently bound by 1, 2 or 3 identical or different R atoms. b replace; Preferred, R 2 Selected from Where R 2a With R 2b Together with the carbon atoms they are linked to, they form tetrahydro-2H-pyranyl, oxetyl, 1,2,3,6-tetrahydropyridyl, and 3,6-dihydro-2H-pyranyl; said tetrahydro-2H-pyranyl, oxetyl, 1,2,3,6-tetrahydropyridyl, and 3,6-dihydro-2H-pyranyl are unsubstituted or independently bound by one, two, or three identical or different R atoms. b replace; Preferred, R 2 Selected from Among them, groups It is either unsubstituted or independently replaced by one, two, or three identical or different Rs. b replace; Preferably, each R b Each of the following is independently selected from H, C1-C6 alkyl, C1-C6 alkoxy, and C3-C6 cycloalkyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, and C3-C6 cycloalkyl are unsubstituted or are independently substituted by one, two, three, or four halogens; Preferably, each R b Each of the following is independently selected from H, C1-C3 alkyl, C1-C3 alkoxy, and C3-C5 cycloalkyl, wherein the C1-C3 alkyl, C1-C3 alkoxy, and C3-C5 cycloalkyl are unsubstituted or are independently substituted by one, two, three, or four substituents selected from F, Cl, or Br; Preferably, each R b Each is independently selected from H, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl; Preferably, each R b Each is independently selected from methyl, methoxy, and cyclopropyl; Preferably, each R b Each is independently selected from cyclopropyl; Preferably, each R b Each is independently selected from methoxy groups; Preferably, each R b Each is independently selected from methyl groups; Preferred, R 2 Selected from Preferred, R 2 Selected from Or preferably, each R b Each of the following is independently selected from H, deuterium, halogen, C1-C6 alkyl, C1-C6 alkoxy, and C3-C6 cycloalkyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, and C3-C6 cycloalkyl are unsubstituted or are independently substituted by one, two, three, or four halogens. Further preferred, each R b Each of the following is independently selected from H, deuterium, halogen, C1-C3 alkyl, C1-C3 alkoxy, and C3-C5 cycloalkyl, wherein the C1-C3 alkyl, C1-C3 alkoxy, and C3-C5 cycloalkyl are unsubstituted or are independently substituted by one, two, three, or four halogens. Further preferred, each R b Each of the following groups is independently selected from H, deuterium, F, Cl, Br, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl, wherein the methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl groups are unsubstituted or independently substituted by one, two, three, or four substituents selected from F, Cl, and Br; Further preferred, each R b Each is independently selected from H, deuterium, F, Cl, Br, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -CHF2, -CF3; Further preferred, each R b Each is independently selected from H, deuterium, F, Cl, methyl, ethyl, isopropyl, methoxy, cyclopropyl, -CHF2, -CF3; More preferably, the R b It can be further replaced by deuterium; Preferably, each R b Each is independently selected from H, deuterium, F, Cl, methyl, ethyl, isopropyl, methoxy, cyclopropyl, -CHF2, -CF3, -CD3; Further preferred, R 2 Selected from; Or preferably, R 2 Selected from Or preferably, R 2 Selected from 4. The compound according to any one of claims 1-3, wherein its tautomer, stereoisomer, pharmaceutically acceptable salt, or deuterated compound is characterized in that: R 3 Selected from hydrogen, halogens, C 1-6 Alkyl, C 3-6 cycloalkyl; Preferred, R 3 Selected from hydrogen, F, Cl, Br, C 1-4 Alkyl, C 5-6 Alkyl, C 3-5 cycloalkyl; Preferred, R 3 Selected from hydrogen, F, Cl, Br, C 1-4 Alkyl, C 3-5 cycloalkyl; Preferred, R 3 Selected from hydrogen, F, Cl, Br, methyl, ethyl, propyl, isopropyl, butyl, cyclopropyl, cyclobutyl, and cyclopentyl; Preferred, R 3 Selected from hydrogen, F, Cl, methyl, ethyl, and cyclopropyl; Or preferably, R 3 Selected from hydrogen, F, Cl, Br, C 1-4 alkyl; Preferred, R 3 Selected from hydrogen, F, Cl, methyl, ethyl; Preferred, R 3 Selected from hydrogen, F, and methyl; Preferred, group Selected from Preferred, group Selected from Preferred, group Selected from: Preferred, group Selected from: Or preferably, the group Selected from: Preferred, group Selected from 5. The compound according to claim 1, wherein its tautomers, stereoisomers, pharmaceutically acceptable salts, or deuterated compounds thereof, are characterized in that: Group Selected from Wherein, W is selected from N or CR 7 ; Preferred, R 4 R 5 R 6 R 7 Each is independently selected from hydrogen, halogen, and C. 1-6 alkyl; Preferred, R 4 R 5 R 6 R 7 Each is independently selected from hydrogen, halogen, and C. 1-4 Alkyl, C 5-6 alkyl; Preferred, R 4 R 5 R 6 R 7 Each is independently selected from hydrogen, halogen, and C. 1-4 alkyl; Preferred, R 4 R 5 R 6 R 7 Each is independently selected from hydrogen, F, Cl, Br, methyl, ethyl, propyl, and isopropyl; Preferred, R 4 R 5 R 6 R 7 Each is independently selected from hydrogen, F, Cl, and methyl; Further preferred, the above R 4 R 5 R 6 R 7 It can be further replaced by deuterium; Preferred, R 4 R 5 R 6 R 7 Each is independently selected from hydrogen, F, Cl, methyl, and -CD3; Preferred, group Selected from Wherein, W is selected from N or CR 7 R 7 Selected from H, F, Cl, and methyl groups; Preferred, group Selected from: Preferred, group Selected from: Or preferably, the group Selected from: Preferred, group Selected from: Preferred, group Selected from:

6. The compound according to claim 1, its tautomers, stereoisomers, pharmaceutically acceptable salts, or deuterated compounds thereof, characterized in that: R 1 Selected from: C 1-6 Alkyl, C 3-10 cycloalkyl, C 3-10 cycloalkyl-C 1-6 Alkyl-, 5-10-membered heteroaryl-; wherein the C1-C6 alkyl group is unsubstituted or converted by one or more R i Replacement; the C3-C 10 The cycloalkyl group is unsubstituted or contains one or more R groups. j Substitution; the 5-10 membered heteroaryl group is unsubstituted or replaced by one or more R groups. l replace; Preferred, R 1 Selected from C 1-4 Alkyl, C 3-6 Monocycloalkyl, C 5-8 Bridged cycloalkyl, C 5-8 Spirocycloalkyl, C 3-6 Monocycloalkyl-C 1-4 Alkyl-, C 5-8 Bridged cycloalkyl-C 1-4 Alkyl-, C 5-8 Spirocycloalkyl-C 1-4 Alkyl-, 5-6-membered heteroaryl; wherein, the C 1-4 The alkyl group is unsubstituted or contains one or more R groups. i Replace; the C 3-6 Monocycloalkyl, C 5-8 Bridged cycloalkyl, C 5-8 Spirocycloalkyl is unsubstituted or converted by one or more R j Substitution; the 5-10 membered heteroaryl group is unsubstituted or replaced by one or more R groups. l replace; Preferred, R 1 Selected from methyl, ethyl, propyl, butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, Pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrazolyl, imidazoleyl, triazolyl, pyrroleyl, furanyl, oxazolyl, isoxazolyl, thiazolyl; wherein the methyl, ethyl, propyl, and butyl groups are unsubstituted or substituted with one or more R groups. i Substitution; the cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, It is unreplaced or replaced by one or more R j Substitution; the pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrazolyl, imidazoleyl, triazolyl, pyrroleyl, furanyl, oxazolyl, isoxazolyl, and thiazolyl groups are unsubstituted or substituted by one or more R groups. l replace; Preferred, R 1 Selected from methyl, ethyl, cyclopropyl, cyclobutyl, cyclopentyl, Pyrazolyl, imidazole, triazolyl, pyrroleyl; wherein the methyl or ethyl group is unsubstituted or is surrounded by one, two, or three R groups. i Substitution; the cyclopropyl, cyclobutyl, cyclopentyl, Is it unreplaced or replaced by 1, 2 or 3 Rs? j Substitution; the pyrazolyl, imidazole, triazolyl, and pyrroleyl groups are unsubstituted or replaced by one, two, or three R groups. l replace; Preferred, R 1 Selected from methyl, ethyl, cyclopropyl, Pyrazolyl; wherein the methyl or ethyl group is unsubstituted or is occupied by one, two, or three R groups. i Substitution; the cyclopropyl, Is it unreplaced or replaced by 1, 2 or 3 Rs? j Substitution; the pyrazol group is unsubstituted or replaced by one, two, or three R groups. l replace; Preferred, R 1 Selected from methyl, ethyl, cyclopropyl, Pyrazolyl; wherein the methyl, ethyl, cyclopropyl, It is unsubstituted; the pyrazol group is unsubstituted or surrounded by one, two, or three R groups. l replace; Preferably, each R l Each is independently selected from halogens and halogenated C. 1-6 Alkyl, C 1-6 Alkyl, C 3-6 cycloalkyl; Preferably, each R l Each is independently selected from halogens and halogenated C. 1-4 Alkyl, C 1-4 Alkyl, C 3-6 cycloalkyl; Preferably, each R l Each is independently selected from halogens, C 1-4 Alkyl, C 3-6 cycloalkyl; Preferably, each R l Each is independently selected from F, Cl, Br, methyl, ethyl, propyl, isopropyl, cyclopropyl, and cyclobutyl; Preferably, each R l Each is independently selected from F, Cl, methyl, and ethyl; Preferably, each R l Each is independently selected from methyl groups; Preferably, each R l Each is independently selected from D, F, Cl, methyl, and ethyl; Preferred, R 1 Selected from methyl, ethyl, cyclopropyl, Preferred, R 1 Selected from methyl, ethyl, and cyclopropyl; Preferred, R 1 Selected from methyl; Preferred, R 1 Selected from -CD3.

7. The compound according to any one of claims 1-6, wherein its tautomer, stereoisomer, pharmaceutically acceptable salt, or deuterated compound is characterized in that: Equation (I) has the structure described in Equation IA: Among them, R 1 R 2 R 3 ,A,n1,n2,ring M,W,X,Y,Z, Each is defined as described in Equation I; Alternatively, formula (I) has the structure described in formula IB: Among them, R 1 R 2 R 3 ,A,n1,n2,ring M,W,X,Y,Z, Each is defined as described in Equation I.

8. The compound according to any one of claims 1-7, wherein its tautomers, stereoisomers, pharmaceutically acceptable salts or deuterated compounds thereof, and the compound represented by formula (I) is selected from: Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12, Compound 13, Compound 14, Compound 15, Compound 16, Compound 17, Compound 18, Compound 19, Compound 20, Compound 21, Compound 22, Compound 23, Compound 24, Compound 25, Compound 26, Compound 27, Compound 28, Compound 29, Compound 30, Compound 31, Compound 32, Compound 33, Compound 34, Compound 35, Compound 36, Compound 37, Compound 38, Compound 39, Compound 40, Compound 41 Compounds 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, and 79 are listed. Compounds 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 93, 94, 95, 98, 100, 101, 103, 104, 105, 106, 107, 108, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, and 125. Compounds 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, or the S configuration of any of the above compounds.

9. A pharmaceutical composition for treating and / or preventing diseases associated with abnormal PRMT5 expression, characterized in that: The pharmaceutical composition comprises a therapeutic and / or preventative amount of the compound as described in any one of claims 1-8 or its tautomers, stereoisomers, pharmaceutically acceptable salts or their deuterated compounds, and optionally pharmaceutical excipients. Preferably, the disease associated with abnormal PRMT5 expression is a tumor or cancer; Preferably, the diseases associated with abnormal PRMT5 expression refer to diseases with MTAP deficiency that are associated with abnormal PRMT5 expression. Preferably, the disease associated with abnormal PRMT5 expression refers to tumors or cancers with MTAP deficiency.

10. The compound or tautomer, stereoisomer, pharmaceutically acceptable salt or deuterated compound of any one of claims 1-8, or the pharmaceutical composition of claim 9, in the preparation of a medicament for treating and / or preventing diseases associated with abnormal PRMT5 expression, and / or in the treatment and / or prevention of diseases associated with abnormal PRMT5 expression; Preferably, the disease associated with abnormal PRMT5 expression is a tumor or cancer; Preferably, the disease associated with abnormal PRMT5 expression refers to a disease with MTAP deficiency that is associated with abnormal PRMT5 expression. Preferably, the disease associated with abnormal PRMT5 expression refers to tumors or cancers with MTAP deficiency.

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