Heterocyclylalkynyl substituted amide derivatives, processes for their preparation and use thereof

Abstract

The present application relates to a kind of heterocyclic alkynyl substituted amide derivatives shown in formula (I), its preparation method and application.The present application also relates to pharmaceutical composition comprising the compound as active ingredient and the use of the compound or pharmaceutical composition for treating and / or preventing relevant diseases mediated by PRMT5.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of pharmaceutical chemistry, in particular to a class of heterocyclic alkynyl-substituted amide derivatives, a preparation method thereof and application thereof. BACKGROUND

[0002] Epigenetically regulated gene expression plays an important biological role in protein maturation and cell differentiation, and plays an important role in many human diseases. Protein arginine methylation catalyzed by protein arginine methyltransferase (PRMT) is a common post-translational modification in eukaryotic cells, which affects cell signaling, gene transcription, mRNA translation, DNA recombination and repair, and many other biological processes (Cell Mol. Life Sci. 2015. 72(11): 2041-2059).

[0003] PRMT5 is one of the members of the PRMT family, and the methylation mediated by it plays an important role in maintaining the balance of the normal intracellular environment. However, more and more studies have shown that the abnormal expression of PRMT5 is related to the occurrence of a variety of tumors, and it is overexpressed in a variety of tumors and occurs in different mechanisms in different tumors (Cell Mol. Life Sci. 2015. 72(11): 2041-2059).

[0004] Homozygous loss of tumor suppressor genes is a key driver of tumorigenesis. The loss of tumor suppressor gene CDKN2A located on human chromosome 9p21 is one of the genes with the highest mutation frequency in tumors, with an incidence of 15%. Due to its close proximity to CDKN2A, the methionine adenosine phosphorylase gene (MTAP) is often deleted in tumors, which plays an important role in the methionine and adenine salvage pathway (Cell Reports, 2016, 15: 574-587). The deletion of MTAP can lead to the accumulation of its substrate methylthioadenosine (MTA), and due to the structural similarity between MTA and S-adenosylmethionine (SAM), it can selectively bind to SAM to inhibit the activity of PRMT5, and sensitizes the further inhibition of PRMT5, i.e. synthetic lethality (Science, 2016, VOL 351 ISSUE 6278: 1214-1217).

[0005] However, PRMT5 is a known, essential gene. Knockout or siRNA silencing of PRMT5 in normal tissues can lead to abnormal physiological functions, such as blood cell reduction, infertility, skeletal muscle loss, and cardiac hypertrophy (Journal of Clinical Investigation, 2015, 125(9):3532-44). Currently, no PRMT5 inhibitors in the clinical stage can cause synthetic lethality with MTAP deficiency. Therefore, new strategies are needed to exploit the metabolic vulnerability caused by MTAP deficiency.

[0006] The development of small molecule inhibitors targeting PRMT5·MTA can preferentially act on tumor cells that lack MTAP. Since normal cells do not lack MTAP, the low concentration of MTA has no significant inhibitory effect on normal cells, thereby increasing the therapeutic index (AACR Annual Meeting, 2021, Abstract LB003), providing a new strategy for the treatment of tumors. Summary of the Invention

[0007] Problems to be solved by the invention:

[0008] Although several patent applications for PRMT5 inhibitors have been published, the huge market demand for MTAP- / - cancer patients and the suboptimal clinical efficacy of existing PRMT5 inhibitors necessitate further development of new compounds. Through continuous efforts, the inventors of this application have designed compounds having the structure represented by general formula (I) and discovered that compounds having such structures exhibit excellent PRMT5 inhibitory effects and potential applications.

[0009] Solutions used to solve the problem:

[0010] In order to solve the above problems, the inventors of the present application have conducted intensive studies and found that a class of heterocyclic alkynyl-substituted amide derivatives can achieve the desired purpose, resulting in the completion of the present invention.

[0011] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

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

[0013]

[0014] in,

[0015] is selected from a single bond or a double bond, or does not exist;

[0016] when When it 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 absent, Y is absent, 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] Ring M is selected from 5-10 membered aryl or 5-10 membered heteroaryl, wherein the 5-10 membered 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 Alkoxy is unsubstituted or substituted with one or more R i Substitution; said C 3-6 Cycloalkyl, C 3-6 Cycloalkoxy is unsubstituted or substituted with one or more R j replace;

[0022] R 1 Selected from C 1-6 Alkyl, C 3-10 Cycloalkyl-(C 1-6 alkyl) m -, 4-10 membered heterocyclic-(C 1-6 alkyl) n -, 5-10 membered heteroaryl-(C 1-6 alkyl) p -, 5-10 membered aryl-(C 1-6 alkyl) q-; wherein n = 0 or 1, m = 0 or 1, p = 0 or 1, q = 0 or 1; the heterocyclic group and heteroaryl group each independently contain 1-4 heterocyclic atoms selected from O, N or S; wherein the C 1-6 Alkyl is unsubstituted or substituted with one or more R i Substitution; said C 3-10 Cycloalkyl is unsubstituted or substituted with one or more R j Substituted; the 4-10 membered heterocyclic group is unsubstituted or substituted by one or more R k Substituted; the 5-10 membered aryl, 5-10 membered heteroaryl is unsubstituted or each independently substituted by one or more R l replace;

[0023] R 2 Selected from: 5-10 membered heteroaryl, 4-10 membered heterocyclic group, said 5-10 membered heteroaryl, 4-10 membered heterocyclic group each independently contains 1-4 heteroatoms selected from N, O or S; said 5-10 membered heteroaryl, 4-10 membered heterocyclic group is unsubstituted or replaced by one or more identical or different R b replace;

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

[0025] Each R i 、R j 、R k 、R l Each independently selected from hydroxy, deuterium, halogen, cyano, optionally substituted with one or more C 1-6 Alkyl, optionally substituted by one or more selected from halogen, cyano, hydroxy, -NR'R", C 1-6 Alkoxy-substituted C 1-6 Alkoxy, C 3-10 Cycloalkyl, optionally substituted by one or more C 1-6 Alkyl-substituted 5-10 membered heteroaryl, wherein the 5-10 membered heteroaryl contains 1-4 heteroatoms selected from N, O, and S; wherein R' and R" are each independently selected from H, C 1-6 Alkyl, halogenated C 1-6 Alkyl, or R' and R" together with the nitrogen atom to which they are attached form a 4-8 membered heterocyclic group containing 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, the C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl is unsubstituted or independently substituted by one or more substituents selected from halogen, hydroxy, cyano, C1-C6 alkoxy; wherein -NR'R", R' and R" are independently selected from hydrogen, C 1-6 Alkyl, halogenated C 1-6 Alkyl, or R' and R" together with the nitrogen atom to which they are attached form a 4-8 membered heterocyclic group containing 1-4 heteroatoms selected from N, O, and S.

[0027] In some embodiments, ring M is selected from 5-6 membered aryl or 5-6 membered heteroaryl.

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

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

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

[0031] In some specific embodiments, 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 .

[0032] In some specific embodiments, does not exist, Y does not exist, X is hydrogen, Z is CR 4 R 5 .

[0033] In some specific embodiments, A is selected from O.

[0034] In some embodiments, n1 and n2 are each independently 0 or 1.

[0035] In some embodiments, the group Selected from

[0036] In some embodiments, the group Selected from

[0037] In some embodiments, the group Selected from:

[0038] In some embodiments, the group Selected from:

[0039] In some specific embodiments, R 2 is selected from 5-6 membered heteroaryl and 4-6 membered heterocyclic group; the 5-6 membered heteroaryl and 4-6 membered heterocyclic group each independently contain 1-3 heteroatoms selected from N, O or S; the 5-6 membered heteroaryl and 4-6 membered heterocyclic group are unsubstituted or are each independently replaced by one or more identical or different R b replace.

[0040] In some specific embodiments, R 2 is selected from 5-6 membered heteroaryl, 4-6 membered heterocyclyl, 5-6 membered heteroaryl and 5-6 membered heterocyclyl; the 5-6 membered heteroaryl, 4-6 membered heterocyclyl and 5-6 membered heterocyclyl each independently contain 1, 2 or 3 heteroatoms selected from N, O or S; the 5-6 membered heteroaryl, 4-6 membered heterocyclyl, 5-6 membered heteroaryl and 5-6 membered heterocyclyl are unsubstituted or are each independently replaced by one or more identical or different R b replace.

[0041] In some specific embodiments, R 2 is selected from 5-6 membered heteroaryl, wherein the 5-6 membered heteroaryl contains 1-3 heteroatoms selected from N, O or S; the 5-6 membered heteroaryl is unsubstituted or replaced by one or more identical or different R b replace.

[0042] In some specific embodiments, R 2 is selected from pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, which are unsubstituted or each independently substituted by 1, 2 or 3 R b replace.

[0043] In some specific embodiments, R 2 is selected from 5-6 membered heteroaryl, wherein the 5-6 membered heteroaryl contains 1, 2 or 3 N heteroatoms; the 5-6 membered heteroaryl is unsubstituted or replaced by one or more identical or different R b replace.

[0044] In some specific embodiments, R 2 is selected from pyrazolyl, imidazolyl, pyridinyl, which are unsubstituted or each independently substituted with 1, 2 or 3 R b replace.

[0045] In some embodiments, R 2 is selected from is unsubstituted or independently substituted with 1, 2, or 3 R b substituents.

[0046] In some embodiments, R 2 is selected from 5-membered heteroaryl containing 1-3 heteroatoms selected from N, O, or S, which is unsubstituted or substituted with 1, 2, or 3 R b substituents.

[0047] In some embodiments, R 2 is selected from pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, which is unsubstituted or substituted with 1, 2, or 3 R b substituents.

[0048] In some embodiments, R 2 is selected from 5-membered heteroaryl containing 1-2 heteroatoms selected from N or S, which is unsubstituted or substituted with 1, 2, or 3 R b substituents.

[0049] In some embodiments, R 2 is selected from pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, which is unsubstituted or independently substituted with 1, 2, or 3 R b substituents.

[0050] In some embodiments, R 2 is selected from 5-membered heteroaryl containing 1, 2, or 3 N heteroatoms, which is unsubstituted or substituted with 1, 2, or 3 R b substituents.

[0051] In some embodiments, R 2 is selected from pyrazolyl, imidazolyl, which is unsubstituted or independently substituted with 1, 2, or 3 R b substituents.

[0052] In some embodiments, R 2 is selected from pyrazolyl, which is unsubstituted or independently substituted with 1, 2, or 3 R b substituents.

[0053] In some embodiments, R 2 is selected from which is unsubstituted or independently substituted with 1, 2, or 3 R b substituents.

[0054] In some specific embodiments, R 2 Selected from

[0055]

[0056] In some specific embodiments, R 2 is selected from 6-membered heteroaryl, wherein the 6-membered heteroaryl contains 1, 2 or 3 heteroatoms selected from N, O or S; the 6-membered heteroaryl is unsubstituted or replaced by one or more identical or different R b replace.

[0057] In some specific embodiments, R 2 is selected from 6-membered heteroaryl, said 6-membered heteroaryl containing 1, 2 or 3 N heteroatoms; said 6-membered heteroaryl is unsubstituted or substituted by 1, 2 or 3 identical or different R b replace.

[0058] In some specific embodiments, R 2 is selected from pyridyl, which is unsubstituted or substituted with 1, 2 or 3 identical or different R b replace.

[0059] In some specific embodiments, R 2 Selected from is unsubstituted or substituted with 1, 2 or 3 identical or different R b replace.

[0060] In some specific embodiments, R 2 Selected from

[0061]

[0062] In some specific embodiments, R 2 5-6 membered heteroaryl and 5-6 membered heterocyclic group, wherein the 5-6 membered heteroaryl and 5-6 membered heterocyclic group each independently contain 1, 2 or 3 heteroatoms selected from N, O or S; the 5-6 membered heteroaryl and 5-6 membered heterocyclic group are unsubstituted or replaced by one or more identical or different R b replace.

[0063] In some specific embodiments, R 2 is selected from 5-6 membered heteroaryl and 5-6 membered heterocyclic group, wherein the 5-6 membered heteroaryl and 5-6 membered heterocyclic group each independently contain 1, 2 or 3 N heteroatoms; the 5-6 membered heteroaryl and 5-6 membered heterocyclic group are unsubstituted or replaced by one or more identical or different R b replace.

[0064] In some specific embodiments, R 2selected from 5-membered heteroaryl and 6-membered heterocyclyl, each independently containing 1, 2 or 3 N heteroatoms; said 5-membered heteroaryl and 6-membered heterocyclyl being unsubstituted or substituted by one or more, same or different R b substituted.

[0065] In some specific embodiments, R 2 selected from imidazolyl and piperazinyl; said imidazolyl and piperazinyl being unsubstituted or substituted by 1, 2 or 3, same or different R b substituted.

[0066] In some specific embodiments, R 2 selected from

[0067] In some specific embodiments, R 2 selected from 5-6 membered heteroaryl and 5-6 membered heterocyclyl, 5-6 membered heteroaryl and 5-6 membered heteroaryl, 5-6 membered heteroaryl and phenyl, said 5-6 membered heteroaryl being selected from imidazolyl, pyrazolyl, pyridinyl, said 5-6 membered heterocyclyl being selected from pyrrolidinyl, piperidinyl, piperazinyl; said 5-6 membered heteroaryl and 5-6 membered heterocyclyl, 5-6 membered heteroaryl and 5-6 membered heteroaryl, 5-6 membered heteroaryl and phenyl being unsubstituted or substituted by one or more, same or different R b substituted.

[0068] In some specific embodiments, R 2 selected from 5-6 membered heteroaryl and 5-6 membered heterocyclyl, 5-6 membered heteroaryl and 5-6 membered heteroaryl, 5-6 membered heteroaryl and phenyl, said 5-6 membered heteroaryl being selected from imidazolyl, pyrazolyl, pyridinyl, said 5-6 membered heterocyclyl being selected from pyrrolidinyl, piperidinyl, piperazinyl; said 5-6 membered heteroaryl and 5-6 membered heterocyclyl, 5-6 membered heteroaryl and 5-6 membered heteroaryl, 5-6 membered heteroaryl and phenyl being unsubstituted or substituted by one or more, same or different R b substituted.

[0069] In some specific embodiments, R 2 selected from imidazolyl and piperazinyl; said imidazolyl and piperazinyl being unsubstituted or substituted by 1, 2 or 3, same or different R b substituted.

[0070] In some specific embodiments, R 2 selected from

[0071] In some specific embodiments, R 2is selected from 4-6 membered heterocyclic groups; the 4-6 membered heterocyclic groups contain 1-3 heteroatoms selected from N, O or S; the 4-6 membered heterocyclic groups are unsubstituted or substituted by 1, 2 or 3 identical or different R b replace.

[0072] In some specific embodiments, R 2 is selected from 4-6 membered heterocycloalkyl and 4-6 membered heterocycloalkenyl; the 4-6 membered heterocycloalkyl and 4-6 membered heterocycloalkenyl each independently contain 1-3 heteroatoms selected from N, O or S; the 4-6 membered heterocycloalkyl and 4-6 membered heterocycloalkenyl are unsubstituted or are each independently replaced by 1, 2 or 3 identical or different R b replace.

[0073] In some specific embodiments, R 2 is selected from 4-6 membered heterocycloalkyl and 4-6 membered heterocycloalkenyl; the 4-6 membered heterocycloalkyl and 4-6 membered heterocycloalkenyl each independently contain 1-2 heteroatoms selected from N or O; the 4-6 membered heterocycloalkyl and 4-6 membered heterocycloalkenyl are unsubstituted or are each independently substituted by 1, 2 or 3 identical or different R b replace.

[0074] In some specific embodiments, R 2 is selected from tetrahydro-2H-pyranyl, oxetanyl, 1,2,3,6-tetrahydropyridinyl, 3,6-dihydro-2H-pyranyl; the tetrahydro-2H-pyranyl, oxetanyl, 1,2,3,6-tetrahydropyridinyl, 3,6-dihydro-2H-pyranyl is unsubstituted or each is independently substituted by 1, 2 or 3 identical or different R b replace.

[0075] In some specific embodiments, R 2 Selected from described is unsubstituted or each independently substituted with 1, 2 or 3 identical or different R b replace.

[0076] In some specific embodiments, R 2 Selected from where R 2a With R 2b Together with the carbon atoms to which they are attached, they form a 4-6 membered heterocyclic group, wherein 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 each is independently substituted by 1, 2 or 3 identical or different R b replace.

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

[0078] In some specific embodiments, R 2 Selected from where R 2a With R 2b Together with the carbon atoms to which they are commonly attached, they form a 4-6 membered heterocycloalkyl or a 4-6 membered heterocycloalkenyl group; the 4-6 membered heterocycloalkyl and the 4-6 membered heterocycloalkenyl group each independently contain 1-2 heteroatoms selected from N or O; the 4-6 membered heterocycloalkyl and the 4-6 membered heterocycloalkenyl group are unsubstituted or are each independently substituted by 1, 2 or 3 identical or different R b replace.

[0079] In some specific embodiments, R 2 Selected from where R 2a With R 2b Together with the carbon atoms to which they are commonly attached, they form tetrahydro-2H-pyranyl, oxetanyl, 1,2,3,6-tetrahydropyridinyl, 3,6-dihydro-2H-pyranyl; the tetrahydro-2H-pyranyl, oxetanyl, 1,2,3,6-tetrahydropyridinyl, 3,6-dihydro-2H-pyranyl are unsubstituted or are each independently substituted by 1, 2 or 3 identical or different R b replace.

[0080] In some specific embodiments, R 2 Selected from Among them is unsubstituted or each independently substituted with 1, 2 or 3 identical or different R b replace.

[0081] In some embodiments, each R b Each is independently selected from H, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, and the C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl is unsubstituted or is independently substituted by 1, 2, 3 or 4 halogens.

[0082] In some embodiments, each R bEach is independently selected from H, C1-C3 alkyl, C1-C3 alkoxy, C3-C5 cycloalkyl, and the C1-C3 alkyl, C1-C3 alkoxy, C3-C5 cycloalkyl is unsubstituted or is independently substituted by 1, 2, 3 or 4 substituents selected from F, Cl or Br.

[0083] In some embodiments, each R b Each is independently selected from H, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0084] In some embodiments, each R b Each is independently selected from methyl, methoxy, and cyclopropyl.

[0085] In some embodiments, each R b are each independently selected from cyclopropyl.

[0086] In some embodiments, each R b are each independently selected from methoxy.

[0087] In some embodiments, each R b are each independently selected from methyl.

[0088] In some specific embodiments, R 2 Selected from

[0089]

[0090] In some specific embodiments, R 2 Selected from

[0091] In some embodiments, each R b Each is independently selected from H, deuterium, halogen, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, and the C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl is unsubstituted or is independently substituted by 1, 2, 3 or 4 halogens.

[0092] In some embodiments, each R b Each is independently selected from H, deuterium, halogen, C1-C3 alkyl, C1-C3 alkoxy, C3-C5 cycloalkyl, and the C1-C3 alkyl, C1-C3 alkoxy, C3-C5 cycloalkyl is unsubstituted or is independently substituted by 1, 2, 3 or 4 halogens.

[0093] In some embodiments, each R bEach is independently selected from H, deuterium, F, Cl, Br, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, isopropyloxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, isopropyloxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl is unsubstituted or is each independently substituted with 1, 2, 3 or 4 substituents selected from F, Cl, and Br.

[0094] In some embodiments, 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.

[0095] In some embodiments, each R b Each is independently selected from H, deuterium, F, Cl, methyl, ethyl, isopropyl, methoxy, cyclopropyl, -CHF2, -CF3.

[0096] In some specific embodiments, R b Can be further deuterated.

[0097] In some embodiments, each R b Each is independently selected from H, deuterium, F, Cl, methyl, ethyl, isopropyl, methoxy, cyclopropyl, -CHF2, -CF3, -CD3.

[0098] In some specific embodiments, R 2 Selected from;

[0099]

[0100] In some specific embodiments, R 2 Selected from

[0101]

[0102] In some specific embodiments, R 2 Selected from

[0103]

[0104] In some specific embodiments, R 3 Selected from hydrogen, halogen, C 1-6 Alkyl, C 3-6 Cycloalkyl.

[0105] In some specific embodiments, R 3 Selected from hydrogen, F, Cl, Br, C 1-4 Alkyl, C5-6 Alkyl, C 3-5 Cycloalkyl.

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

[0107] In some specific embodiments, R 3 Selected from hydrogen, F, Cl, Br, methyl, ethyl, propyl, isopropyl, butyl, cyclopropyl, cyclobutyl, cyclopentyl.

[0108] In some specific embodiments, R 3 Selected from hydrogen, F, Cl, methyl, ethyl, cyclopropyl.

[0109] In some specific embodiments, R 3 Selected from hydrogen, F, Cl, Br, C 1-4 alkyl.

[0110] In some specific embodiments, R 3 Selected from hydrogen, F, Cl, methyl, ethyl;

[0111] In some specific embodiments, R 3 Selected from hydrogen, F, methyl.

[0112] In some embodiments, the group Selected from

[0113]

[0114] In some embodiments, the group Selected from

[0115] In some embodiments, the group Selected from:

[0116]

[0117] In some embodiments, the group Selected from:

[0118]

[0119] In some embodiments, the group Selected from:

[0120]

[0121] In some embodiments, the group Selected from

[0122] In some embodiments, the group Selected from

[0123]

[0124] , wherein W is selected from N or CR 7 .

[0125] In some embodiments, W is selected from CR 7 .

[0126] In some specific embodiments, W is selected from N.

[0127] In some specific embodiments, R 4 , R 5 , R 6 , R 7 are each independently selected from hydrogen, halogen, C 1-6 alkyl.

[0128] In some specific embodiments, R 4 , R 5 , R 6 , R 7 are each independently selected from hydrogen, halogen, C 1-4 Alkyl, C 5-6 alkyl.

[0129] In some specific embodiments, R 4 , R 5 , R 6 , R 7 are each independently selected from hydrogen, halogen, C 1-4 alkyl.

[0130] In some specific embodiments, R 4 , R 5 , R 6 , R 7 Each is independently selected from hydrogen, F, Cl, Br, methyl, ethyl, propyl, isopropyl.

[0131] In some specific embodiments, R 4 , R 5 , R 6 , R 7 Each is independently selected from hydrogen, F, Cl, and methyl.

[0132] In some specific embodiments, R 4 , R 5 , R 6 , R 7 Can be further deuterated.

[0133] In some embodiments, R 4 , R 5 , R 6 , R 7 are each independently selected from hydrogen, F, CI, methyl, -CD3.

[0134] In some embodiments, the group is selected from

[0135] wherein W is selected from N or CR 7 .

[0136] In some embodiments, the group is selected from

[0137] wherein W is selected from N or CR 7 , R 7 is selected from H, F, CI, methyl.

[0138] In some embodiments, the group is selected from:

[0139]

[0140] In some embodiments, the group is selected from:

[0141]

[0142] In some embodiments, the group is selected from:

[0143]

[0144] In some embodiments, the group is selected from:

[0145]

[0146] In some embodiments, the group is selected from:

[0147]

[0148] In some embodiments, R 1 is selected from: C 1-6 alkyl, C 3-10 cycloalkyl, C 3-10 cycloalkyl-C 1-6Alkyl-, 5-10 membered heteroaryl-; wherein the C1-C6 alkyl is unsubstituted or substituted by one or more R i Substitution; the C3-C 10 Cycloalkyl is unsubstituted or substituted with one or more R j Substituted; the 5-10 membered heteroaryl is unsubstituted or substituted by one or more R l replace.

[0149] In some specific embodiments, R 1 Selected from C 1-4 Alkyl, C 3-6 Monocyclic alkyl, C 5-8 Bridged cycloalkyl, C 5-8 Spiroalkyl, C 3-6 Monocycloalkyl-C 1-4 Alkyl-, C 5-8 Bridged cycloalkyl-C 1-4 Alkyl-, C 5-8 Spiroalkyl-C 1-4 Alkyl-, 5-6 membered heteroaryl; wherein, the C 1-4 Alkyl is unsubstituted or substituted with one or more R i Substitution; said C 3-6 Monocyclic alkyl, C 5-8 Bridged cycloalkyl, C 5-8 Spiroalkyl is unsubstituted or substituted with one or more R j Substituted; the 5-10 membered heteroaryl is unsubstituted or substituted by one or more R l replace.

[0150] In some specific embodiments, R 1 Selected from methyl, ethyl, propyl, butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, Pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrazolyl, imidazolyl, triazolyl, pyrrolyl, furyl, oxazolyl, isoxazolyl, thiazolyl; wherein the methyl, ethyl, propyl, butyl group is unsubstituted or replaced by one or more R i Substituted; the cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, is unsubstituted or replaced by one or more R j Substituted; the pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrazolyl, imidazolyl, triazolyl, pyrrolyl, furyl, oxazolyl, isoxazolyl, thiazolyl is unsubstituted or substituted by one or more R l replace.

[0151] In some specific embodiments, R 1 Selected from methyl, ethyl, cyclopropyl, cyclobutyl, cyclopentyl, Pyrazolyl, imidazolyl, triazolyl, pyrrolyl; wherein the methyl group and the ethyl group are unsubstituted or replaced by 1, 2 or 3 R i Substituted; the cyclopropyl, cyclobutyl, cyclopentyl, is unsubstituted or replaced by 1, 2 or 3 R j Substituted; the pyrazolyl, imidazolyl, triazolyl, pyrrolyl is unsubstituted or substituted by 1, 2 or 3 R l replace.

[0152] In some specific embodiments, R 1 Selected from methyl, ethyl, cyclopropyl, Pyrazolyl; wherein the methyl group or ethyl group is unsubstituted or replaced by 1, 2 or 3 R i Substituted; the cyclopropyl, is unsubstituted or replaced by 1, 2 or 3 R j Substituted; the pyrazolyl group is unsubstituted or substituted by 1, 2 or 3 R l replace.

[0153] It should be understood that the substituent R 1 In the definition of: R i Always substituted on an alkyl or alkylene group, R j Always substituted on a cycloalkyl group, including monocycloalkyl, bridged cycloalkyl, and spirocycloalkyl; R k Always substituted on a heterocyclic group; R l is always substituted on an aryl or heteroaryl group. For example, If a substituent R j Then, R j Substitution position is cyclopropyl or superior.

[0154] In some specific embodiments, R 1 Selected from methyl, ethyl, cyclopropyl, Pyrazolyl; wherein the methyl, ethyl, cyclopropyl, is unsubstituted; the pyrazolyl group is unsubstituted or substituted by 1, 2 or 3 R l replace.

[0155] In some embodiments, each R i 、R j 、R k 、R l are independently selected from halogen, halogenated C 1-6 Alkyl, C 1-6 Alkyl, C 3-6 Cycloalkyl.

[0156] In some embodiments, each Ri 、R j 、R k 、R l are independently selected from halogen, halogenated C 1-4 Alkyl, C 1-4 Alkyl, C 3-6 Cycloalkyl.

[0157] In some embodiments, each R i 、R j 、R k 、R l are each independently selected from halogen, C 1-4 Alkyl, C 3-6 Cycloalkyl.

[0158] In some embodiments, each R i 、R j 、R k 、R l Each is independently selected from F, Cl, Br, methyl, ethyl, propyl, isopropyl, cyclopropyl, and cyclobutyl.

[0159] In some embodiments, each R i 、R j 、R k 、R l Each is independently selected from F, Cl, methyl, and ethyl.

[0160] In some embodiments, each R i 、R j 、R k 、R l The substituents are each independently selected from methyl.

[0161] In some embodiments, each R l are independently selected from halogen, halogenated C 1-6 Alkyl, C 1-6 Alkyl, C 3-6 Cycloalkyl.

[0162] In some embodiments, each R l are independently selected from halogen, halogenated C 1-4 Alkyl, C 1-4 Alkyl, C 3-6 Cycloalkyl.

[0163] In some embodiments, each R l are each independently selected from halogen, C 1-4 Alkyl, C 3-6 Cycloalkyl.

[0164] In some embodiments, each Rl Each is independently selected from F, Cl, Br, methyl, ethyl, propyl, isopropyl, cyclopropyl, and cyclobutyl.

[0165] In some embodiments, each R l Each is independently selected from F, Cl, methyl, and ethyl.

[0166] In some embodiments, each R l The substituents are each independently selected from methyl.

[0167] In some embodiments, each R l Each is independently selected from D, F, Cl, methyl, and ethyl.

[0168] In some specific embodiments, R 1 Selected from methyl, ethyl, cyclopropyl,

[0169] In some specific embodiments, R 1 Selected from methyl, ethyl, cyclopropyl.

[0170] In some specific embodiments, R 1 Selected from methyl.

[0171] In some specific embodiments, R 1 Selected from -CD3.

[0172] In some embodiments, Formula (I) has the structure depicted in Formula IA:

[0173] Among them, R 1 、R 2 、R 3 ,A,n1,n2,ring M,W,X,Y,Z, Definitions are each as defined above.

[0174] Alternatively, formula (I) has the structure described in formula IB:

[0175] Among them, R 1 、R 2 、R 3 ,A,n1,n2,ring M,W,X,Y,Z, Definitions are each as defined above.

[0176] As an exemplary compound of the present invention, the compound represented by formula (I) is selected from any of the following specific compounds:

[0177]

[0178]

[0179]

[0180]

[0181]

[0182]

[0183]

[0184] Further, the present application also provides the S configuration of the above exemplary compounds:

[0185]

[0186]

[0187]

[0188]

[0189]

[0190]

[0191]

[0192] The term "pharmaceutically acceptable salt" means a salt of a compound of the present application that is found to be suitable for use in pharmaceutical products, which is prepared from a compound of the present application having a particular substituent with a relatively non-toxic acid or base. When the compound of the present application contains a relatively acidic functional group, the base addition salt can be obtained by contacting such a compound in pure form or in a suitable inert solvent with a sufficient amount of the base to produce the salt. When the compound of the present application contains a relatively basic functional group, the acid addition salt can be obtained by contacting such a compound in pure form or in a suitable inert solvent with a sufficient amount of the acid to produce the salt.

[0193] The term "deuterated compound" means that the compound of the present application includes at least one deuterium atom, specifically that one or more hydrogen atoms in the compound of the present application can be replaced or substituted with a deuterium atom. In some embodiments, the compound includes two or more deuterium atoms. In some embodiments, the compound includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 deuterium atoms. Synthetic methods for including isotopes into organic compounds are known in the art.

[0194] Preparation methods:

[0195] The present invention also provides a method for preparing the compound. The preparation of the compound described in formula (I) of the present invention can be achieved by the following illustrative methods and examples, but these methods and examples should not be considered in any way to limit the scope of the present invention. The compounds described in the present invention can also be synthesized by synthetic techniques known to those skilled in the art, or a combination of synthetic methods known in the art and the method described in the present invention. The product obtained in each step of the reaction is obtained by 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 the synthesis can be conventionally synthesized according to the literature (such as provided by Scifinder) or purchased.

[0196] Synthesis process route:

[0197] Route 1:

[0198]

[0199] Step 1: Treating the compound represented by formula I-1 with di-tert-butyl dicarbonate to obtain the compound represented by formula I-2;

[0200] 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 in formula I-3;

[0201] Step 3: Under coupling conditions well known in the art, The compound shown in formula I-3 can be treated with the compound shown in formula I-4;

[0202] Step 4: The compound represented by formula I-4 is subjected to acidic conditions to remove the Boc protecting group to obtain the compound represented by formula I-5;

[0203] Step 5: Under condensation conditions well known in the art, the compound represented by formula I-5 is treated with the compound represented by formula I-6 to obtain the compound represented by formula I;

[0204] Among them, Q 1 , Q 2 Is halogen, X, Y, Z, W, A, R 1 、R 2 、R 3 , n1, n2, and ring M are as defined herein in formula (I).

[0205] Route 2:

[0206]

[0207] Step 1: Under condensation conditions well known in the art, the compound represented by formula II-1 is treated with the compound represented by I-6 to obtain the compound represented by formula II-2;

[0208] Step 2: Under coupling conditions well known in the art, The compound shown in formula II-2 is treated with the compound shown in formula II-3;

[0209] Step 3: The compound represented by formula II-3 is treated with tetrabutylammonium fluoride to obtain the compound represented by formula II-4;

[0210] Step 4: Treating the compound represented by Formula II-4 with the compound represented by R2-Q2 under coupling conditions well known in the art can yield the compound represented by Formula I;

[0211] or,

[0212] Step 2: Under coupling conditions well known in the art, The compound shown in formula II-2 can be treated with the compound shown in formula II-2 to obtain the compound shown in formula I;

[0213] Among them, Q 1 , Q 2 Is halogen, X, Y, Z, W, A, R 1 、R 2 、R 3 , n1, n2, and ring M are as defined herein in formula (I).

[0214] Route 3:

[0215]

[0216] Step 1: Under condensation conditions well known in the art, the compound represented by formula III-1 is treated with the compound represented by I-6 to obtain the compound represented by formula II-4;

[0217] Step 2: Under coupling conditions well known in the art, the compound represented by formula II-4 is treated with the compound represented by R2-Q2 to obtain the compound represented by formula I;

[0218] Among them, Q 2 Is halogen, X, Y, Z, W, A, R 1 、R 2 、R 3 , n1, n2, and ring M are as defined herein in formula (I).

[0219] Pharmaceutical composition:

[0220] The present invention also provides a pharmaceutical composition for treating and / or preventing diseases associated with abnormal PRMT5 expression, comprising a therapeutically and / or preventatively effective amount of the aforementioned compound or its tautomers, stereoisomers, pharmaceutically acceptable salts or deuterated compounds, and optional pharmaceutical excipients.

[0221] In some embodiments, the disease associated with abnormal PRMT5 expression is a tumor or cancer.

[0222] In some embodiments, the disease associated with abnormal PRMT5 expression refers to a disease associated with abnormal PRMT5 expression in which MTAP is deficient.

[0223] In some embodiments, the disease associated with abnormal PRMT5 expression refers to a tumor or cancer with MTAP deficiency.

[0224] Methods for preparing various pharmaceutical compositions containing a certain amount of active ingredient are known or will be apparent to those skilled in the art based on 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.

[0225] The present invention also provides a PRMT5 inhibitor, which comprises a therapeutically and / or preventively effective amount of the aforementioned compound or its tautomer, stereoisomer, pharmaceutically acceptable salt or deuterated compound.

[0226] Medical uses:

[0227] The present invention also provides the use of the aforementioned compound or its tautomers, stereoisomers, pharmaceutically acceptable salts or deuterated compounds, or the aforementioned pharmaceutical composition, in the preparation of PRMT5 inhibitors.

[0228] The present invention also provides the aforementioned compound or its tautomer, stereoisomer, pharmaceutically acceptable salt or deuterated compound, or the aforementioned pharmaceutical composition, for use in the preparation of a drug for treating and / or preventing diseases associated with abnormal PRMT5 expression.

[0229] The present invention also provides the use of the aforementioned compound or its tautomers, stereoisomers, pharmaceutically acceptable salts or deuterated compounds, or the aforementioned pharmaceutical composition, in the treatment and / or prevention of diseases associated with abnormal PRMT5 expression.

[0230] The present invention also provides the aforementioned compound or its tautomer, stereoisomer, pharmaceutically acceptable salt or deuterated compound, or the aforementioned pharmaceutical composition, for use in treating and / or preventing diseases associated with abnormal PRMT5 expression.

[0231] In some embodiments, the disease associated with abnormal PRMT5 expression is a tumor or cancer.

[0232] In some embodiments, the disease associated with abnormal PRMT5 expression refers to a disease associated with abnormal PRMT5 expression in which MTAP is deficient.

[0233] In some embodiments, the disease associated with abnormal PRMT5 expression refers to a tumor or cancer with MTAP deficiency.

[0234] The present invention also provides a method for treating and / or preventing diseases associated with abnormal PRMT5 expression, which comprises administering to an individual in need thereof a therapeutically and / or preventive effective amount of the aforementioned compound or its tautomer, stereoisomer, pharmaceutically acceptable salt or deuterated compound, or the aforementioned pharmaceutical composition.

[0235] In some embodiments, the disease associated with abnormal PRMT5 expression is a tumor or cancer.

[0236] In some embodiments, the disease associated with abnormal PRMT5 expression refers to a disease associated with abnormal PRMT5 expression in which MTAP is deficient.

[0237] In some embodiments, the disease associated with abnormal PRMT5 expression refers to a tumor or cancer with MTAP deficiency.

[0238] As used herein, "treating" generally refers to obtaining a desired pharmacological and / or physiological effect. This effect can be prophylactic, in terms of completely or partially preventing a disease or its symptoms; and / or therapeutic, in terms of partially or completely stabilizing or curing a disease and / or causing side effects due to the disease. As used herein, "treating" encompasses any treatment of a disease in a patient, including: (a) preventing the disease or symptoms from occurring in a patient who has not yet been diagnosed with the disease; (b) suppressing the symptoms of a disease, i.e., arresting their development; or (c) relieving the symptoms of a disease, i.e., causing the disease or symptoms to regress.

[0239] In the present invention, "subject" or "individual in need thereof" refers to a vertebrate. In certain embodiments, the vertebrate refers to a mammal. Mammals include, but are not limited to, livestock (such as cattle), pets (such as cats, dogs, and horses), primates, mice, and rats. In certain embodiments, the mammal refers to a human.

[0240] In the present invention, an "effective amount" refers to an amount that is effective at the necessary dosage and time to achieve the desired therapeutic or preventive effect. The "therapeutically effective amount" of the substance / molecule of the present invention may vary according to factors such as the disease state, age, sex and weight of the individual and the ability of the substance / molecule to elicit the desired response in the individual. A therapeutically effective amount also encompasses an amount in which the therapeutically beneficial effects of the substance / molecule outweigh any toxic or deleterious consequences. A "prophylactically effective amount" refers to an amount that is effective at the necessary dosage and time to achieve the desired preventive effect. Typically, but not necessarily, a prophylactic dose is used in subjects before the onset of disease or in the early stages of the disease, so the prophylactic effective amount will be lower than the therapeutically effective amount. In the case of cancer, a therapeutically effective amount of a drug can reduce the number of cancer cells; reduce tumor size; inhibit (i.e., slow down to a certain extent, preferably stop) cancer cell infiltration into surrounding organs; inhibit (i.e., slow down to a certain extent, preferably stop) tumor metastasis; inhibit tumor growth to a certain extent; and / or alleviate one or more symptoms associated with cancer to a certain extent.

[0241] Definition of terms:

[0242] According to the common practice in this field, The bonds used in the formulae herein to describe the points of attachment of the moiety or substituent to the parent core or structure.

[0243] A dash “-” that does not appear between two letters or symbols is used to indicate the point of attachment of a substituent. For example, C 3-6 Cycloalkyl-(C 1-6 alkyl) r - means through (C 1-6 alkyl) r - connected to the rest of the molecule.

[0244] As used herein, the term "substituted" means that any one or more hydrogens on a designated atom or group are replaced with the selection of a designated group, provided that the normal valence of the designated atom is not exceeded.

[0245] In various parts of this specification, substituents of compounds disclosed herein are disclosed in terms of group types or ranges. It is specifically noted that the present invention includes every independent subcombination of the individual members of these group types and ranges. For example, the term "C 1-6 "Alkyl" specifically refers to methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl and C6 alkyl disclosed independently, or "C 1-4 Alkyl", or independently disclosed "C 1-3 alkyl".

[0246] The term "alkyl" is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having the 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 from one to six carbon atoms. The alkyl group can be unsubstituted or substituted such that one or more hydrogens thereof are replaced by another chemical moiety. 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, t-butyl), pentyl (e.g., n-pentyl, isopentyl, neopentyl), and the like. It will be understood by those skilled in the art that C 1-6 "Alkyl" includes monovalent C 1-6 "Alkyl", divalent C 1-6 "Alkyl", divalent C r "C1-C6 alkyl" in -C1-C6 alkyl- refers to C1-C6 alkylene.

[0247] The term "alkoxy" refers to any of the above alkyl groups (e.g., C 1-6 "Alkyl", C 1-4 "Alkyl", C 1-3 "Alkyl", C

[0248] The term "halo-C 1-6 "Alkyl" or "halo-C 1-6 "Alkoxy" refers to an alkyl group as defined above in which one or more (e.g., 2, 3) hydrogen atoms are replaced by a halogen atom, such as fluorine, chlorine, bromine. In some embodiments, the term "halo-C 1-6 "Alkyl" or "halo-C 1-6 "Alkoxy" refers to an alkyl group as defined above in which one or more (e.g., 2, 3) hydrogen atoms are replaced by a halogen atom, such as fluorine, chlorine, bromine. In some embodiments, the term "halo-C

[0249] The term "hydroxy-substituted C 1-6 "Alkyl" refers to an alkyl group as defined above in which one hydrogen atom is replaced by a hydroxyl group. As an example, the "hydroxy-substituted C 1-6 "Alkyl" can be hydroxymethyl.

[0250] The term "cycloalkyl" refers to a cyclized alkyl group, including monocyclic, bicyclic, or polycyclic ring systems. When the cycloalkyl group is bicyclic or polycyclic, each ring thereof should be a saturated carbocyclic ring or carbocyclic ring residue, and the two or more rings of the bicyclic or polycyclic cycloalkyl group can be connected in a manner including bridged, fused, or spiro. For example, C 3-10 "Cycloalkyl" refers to C3, C4, C5, C6, C7, C8, C9, and C10 Cycloalkyl. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.

[0251] The term "cycloalkenyl" refers to a cycloalkyl group as defined above that has at least one carbon-carbon double bond, for example

[0252] The term "cycloalkoxy" refers to any of the above cycloalkyl groups (e.g., C 3-6 cycloalkyl) which is attached to the rest of the molecule through an oxygen atom (—O—).

[0253] The term "carbocycle" or "carbocycle 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, any one of which may be saturated, partially saturated, unsaturated or aromatic. The bicyclic or polycyclic carbocycles may be connected in a manner such as bridging, fusion or spiro. Examples of these carbocycles include, but are not limited to, cyclopropyl, cyclobutyl, cyclobutenyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptenyl, cycloheptyl, adamantyl, cyclooctyl, phenyl, naphthyl, [2,2,2]bicyclooctane, wait.

[0254] The term "aryl" refers to a monocyclic, bicyclic, or tricyclic aromatic hydrocarbon group having 5 to 14 carbon atoms in the ring portion. When the "aryl" group is bicyclic or tricyclic, each ring is aromatic. The two rings of the bicyclic or tricyclic aryl group can be connected in a manner including bridging, fusion, and spiro connection. Examples include phenyl and naphthyl, each of which can be substituted.

[0255] The terms "heterocycle," "heterocyclic," or "heterocyclyl" 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 1, 2, or 3 heteroatoms selected from O, S, and N. Each heteroatom-containing ring in the group may contain 1 or 2 oxygen or sulfur atoms and / or 1 to 4 nitrogen atoms, provided that the total number of heteroatoms in each ring is 4 or less, and further provided that the ring contains at least one carbon atom. In some preferred embodiments, the heteroatoms are exclusively N or O, and the total number does not exceed 3, preferably only 1-2 heteroatoms. The carbon and sulfur atoms are optionally oxidized, the nitrogen atoms are optionally quaternized, and when valence permits, the ring atoms in the heterocycle are optionally substituted with =O (oxo). (For example: ). The fused rings completing the bicyclic and tricyclic groups may contain only carbon atoms and may be saturated, partially saturated or fully unsaturated, aromatic or non-aromatic. The heterocyclic group may be attached at any available nitrogen or carbon atom. As previously described, heterocyclic groups include "spiroheterocyclyl," "heterobridged ring group," and "heterocycloalkenyl," etc., as described below. Exemplary heterocyclic groups include, but are not limited to, azetidinyl, oxetanyl, pyrrolidinyl, imidazolinyl, oxazolidinyl, isoxazolinyl, thiazolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxoazacycloheptatrienyl, 1-pyridonyl, 4-piperidonyl, tetrahydropyranyl, morpholinyl, 1,3-dioxolane, quinuclidinyl, etc.

[0256] The term "saturated heterocyclic group" refers to the above-mentioned "heterocyclic group" in which the monocyclic group, bicyclic group, or tricyclic group is completely saturated. The "heterocyclic group" is as described above. As an example, the saturated heterocyclic group can be morpholinyl (such as ), piperidinyl (such as ), piperazinyl etc.

[0257] The term "heterocycloalkenyl" refers to a heterocyclic group having at least one carbon-carbon double bond in the heterocycle as defined above, for example

[0258] The term "heteroaryl" refers to substituted and unsubstituted aryl groups described above having at least one heteroatom (O, N, or S) in at least one ring, including aromatic 5-8 membered monocyclic groups, 8-10 membered bicyclic groups, and 10-14 membered tricyclic groups, wherein the heteroatom-containing ring preferably has 1, 2, or 3 heteroatoms selected from O, N, or S. Each heteroatom-containing ring of the heteroaryl group may contain 1 or 2 oxygen or sulfur atoms and / or 1 to 4 nitrogen atoms, provided that the total number of heteroatoms in each ring is 4 or less and each ring has at least one carbon atom. Each ring of a bicyclic or tricyclic heteroaryl group is aromatic.

[0259] The term "spirocyclyl" refers to a bicyclic structure having one common ring atom, wherein each monocyclic ring is a saturated or unsaturated, aromatic or non-aromatic carbocyclic ring having 3-7 carbon atoms. Exemplary spiroalkyl groups include, but are not limited to, spiro[4.5]decane, spiro[3.4]octane, spiro[2.3]hexane, Herein, the spirocyclic group does not include the aryl group defined above.

[0260] The term "saturated spirocyclic group" or "spiroalkyl" refers to a spirocyclic group as defined above in which each monocyclic ring is fully saturated. Exemplary spiroalkyl groups include, but are not limited to, spiro[4.5]decane, spiro[3.4]octane, and spiro[2.3]hexane.

[0261] The term "spiroheterocyclyl" refers to a bicyclic structure having one common ring atom or a tricyclic structure having two independent common ring atoms, wherein each monocyclic ring is a saturated or unsaturated monocyclic group having 3 to 8 ring atoms, wherein at least one ring has 1 or 2 ring atoms selected from N, O or S(O) n wherein n is an integer from 0 to 2 and the remaining ring atoms are C. In addition, one or two ring carbon atoms in the heterocyclyl ring are optionally replaced by a -CO- group. Exemplary spiroheterocyclyls include, but are not limited to, 5-azaspiro[2.3]hexane and 6-oxaspiro[3.4]-7-octanone. Herein, the spiroheterocyclyl does not include heteroaryl as defined above.

[0262] The term "saturated spiroheterocyclyl" or "spiroheteroalkyl" refers to a spiroheterocyclyl group as defined above in which each ring is fully saturated. Exemplary spiroheteroalkyl groups include, but are not limited to:

[0263] The term "bridged ring group" refers to a saturated or unsaturated, aromatic or non-aromatic 3- to 8-membered monocyclic carbocyclic group in which two non-adjacent ring atoms are connected by (CRR) n Group, C 2-6 Alkenyl, C 2-6 alkynyl or bond, wherein n is an integer from 1 to 3, and each R is independently H or methyl (wherein, (CRR) n Group, C 2-6 Alkenyl, C 2-6 Alkynyl or bond is also referred to as bridging group in this application). The bridged ring group is optionally substituted by one or two substituents independently selected from alkyl, halogen, alkoxy, hydroxyl or cyano, wherein alkyl, alkoxy, alkenyl, alkynyl are each defined as above. Examples of bridged ring groups include, but are not limited to, bicyclo [2.2.1] heptane, bicyclo [2.2.2] octane, etc. It should be understood that when two non-adjacent ring atoms of a monocyclic 4 to 7 membered hydrocarbon group are connected by a bond, the "bridged ring group" may be referred to as a "bicyclo" or "fused ring group". Herein, the bridged ring group does not include an aryl group as defined above.

[0264] The term "saturated bridged ring group" or "bridged cycloalkyl" refers to a "bridged ring group" as defined above in which each ring is saturated. Here, saturation should be understood as meaning that each ring carbon atom in the bridged ring group is saturated (including the carbon atoms in the bridging group).

[0265] The term "heterobridged cyclic group" refers to a "bridged cyclic group" as defined above, wherein one, two, three or four ring carbon atoms (including the carbon atoms in the bridging group) are replaced by heteroatoms selected from N, O or S(O)n, wherein n is an integer from 0 to 2. Examples of "heterobridged cyclic groups" include, but are not limited to, 2-azabicyclo[2.2.2]octane, quinuclidine, 7-oxabicyclo[2.2.1]heptane, and the like. Herein, the heterobridged cyclic group does not include heteroaryl groups as defined above.

[0266] The terms "saturated heterobridged ring group" and "heterobridged cycloalkyl" refer to a "heterobridged ring group" as defined above in which each ring is saturated, and saturation here should be understood as meaning that each ring atom in the bridged ring group is saturated (including the carbon / heteroatoms in the bridging group).

[0267] Unless otherwise indicated, when referring to a specifically named aryl (e.g., phenyl), cycloalkyl (e.g., cyclohexyl), heterocycle (e.g., pyrrolidinyl, piperidinyl, morpholinyl) or heteroaryl (e.g., imidazolyl, pyrazolyl, triazolyl) group, the reference is meant to include the ring having 0 to 3, preferably 0 to 2, substituents optionally selected from the substituents described above for aryl, cycloalkyl, heterocycle and / or heteroaryl groups.

[0268] The term "heteroatom" shall include oxygen, sulfur and nitrogen.

[0269] The term "halogen" shall include "F, Cl, Br, I".

[0270] When the term "unsaturated" is used herein to refer to a ring or group, the ring or group may be fully unsaturated or partially unsaturated.

[0271] When the term "saturated" is used herein to refer to a ring or group, unless otherwise specified, the ring or group shall be fully saturated.

[0272] From all the above descriptions, it is obvious to those skilled in the art that any group whose name is a composite name, such as "C 3-10 Cycloalkyl-C(O)-" shall mean a moiety conventionally derived therefrom, e.g., from a C 3-10 The cycloalkyl group is constructed by a carbonyl group substituted by a cycloalkyl group, wherein the cycloalkyl group is as defined above. Other similar compound names can be understood by referring to the above content.

[0273] The term "optionally" means that it can be selected or not. For example, "optionally 1 to 3 R d Substituted C 1-6 Alkyl", which means that the C 1-6 The alkyl group may be substituted with 1 to 3 R d It may be replaced by 1 to 3 R dOther similar definitions can be understood with reference to the above content.

[0274] The term "heterocyclic ring atoms are optionally substituted with oxo" means that the ring atoms of the heterocyclic ring are optionally substituted with =O(oxo) (e.g. The ring atom CH2 undergoes oxygenation to obtain the group ).

[0275] Throughout the specification, groups and substituents thereof may be chosen by one skilled in the art to provide stable moieties and compounds and compounds useful as pharmaceutically acceptable compounds and / or intermediate compounds useful in preparing pharmaceutically acceptable compounds.

[0276] Herein, unless otherwise expressly stated, the description “…are independently selected from” used throughout this document may mean that in different groups, the specific options expressed by the same or different symbols do not affect each other, or that in the same group, the specific options expressed by the same or different symbols do not affect each other.

[0277] The term "XXX is substituted at any substitutable position by one or more substituents selected from YYY" means that XXX can be substituted at any substitutable position by one or more substituents selected from YYY. When XXX is substituted at any substitutable position by multiple substituents selected from YYY, the multiple substituents may be the same or different. The multiple substituents are 2 or more, preferably 2, 3 or 4, more preferably 2 or 3. For example, C 1-6 The alkyl group is substituted at any substitutable position by one or more substituents selected from cyano and hydroxy groups, which means that C 1-6 The alkyl group may be substituted by one or more cyano groups at any substitutable position, may be substituted by one or more hydroxyl groups at any substitutable position, or may be substituted 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 substitutable position.

[0278] In this document, when there is a discrepancy between the structure and the chemical name, the structure prevails.

[0279] Effects of the invention:

[0280] The compounds of the present invention have excellent PRMT5 inhibitory activity and, relative to the MTAP wild-type (WT) HCT-116 cell line, have excellent selective inhibition 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.

[0281] Furthermore, experiments have shown that the compounds of the present invention exhibit varying degrees of advantages in terms of human liver microsomal stability, mouse pK, rat pK, and in vivo anti-tumor effects in a constructed mouse tumor model.

[0282] 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 inhibitory toxicity in terms of hERG and CYP). DETAILED DESCRIPTION

[0283] It should be understood that the terminology used herein is intended to describe specific embodiments and is not intended to be limiting. In addition, although any method, device, and material similar or equivalent to those described herein may be used for implementing or testing the present invention, preferred methods, devices, and materials are now described.

[0284] The structures of the compounds were 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 calculated based on a 10 -6 The units are given in ppm.

[0285] Reaction monitoring and MS measurements were performed using a Thermofisher ESQ (ESI) mass spectrometer.

[0286] HPLC determination was performed using a Thermo Fisher U3000 DAD high pressure liquid chromatograph (GL Sciences ODS-HL HP 3 μm 3.0*100 mm column).

[0287] Qingdao Ocean GF254 silica gel plates were used for thin-layer chromatography (TLC). The silica gel plates used for thin-layer chromatography (TLC) were 0.15-0.2 mm in diameter. High-performance thin-layer chromatography (HPLC) preparative plates were 0.9-1.0 mm in diameter for TLC separation and purification. Column chromatography used Qingdao Ocean 200-300 mesh silica gel as the carrier. The developing solvents used were A: dichloromethane and methanol; B: petroleum ether and ethyl acetate. The solvent volume ratio was adjusted according to the polarity of the compound. A Biotage Isera One preparative liquid phase was used for medium-pressure preparative liquid phase purification.

[0288] In the following examples, unless otherwise specified, all reaction raw materials can be purchased from suppliers in the scifinder database. For example, some reagents in the embodiments of the present invention were purchased from manufacturers such as San Chemical Technology (Shanghai) Co., Ltd., Shanghai Shaoyuan Reagent Co., Ltd., Nanjing Yaoshi Technology Co., Ltd., Jiangsu Aikang Biopharmaceutical R&D Co., Ltd., and Shanghai Bid Pharmaceutical Technology Co., Ltd. Further, unless otherwise specified, the raw materials used in the embodiments of the present invention are all analytically pure. Unless otherwise specified, the ratios of two liquid substances involved in this article are all volume ratios; the percentages of substances involved are all mass percentages.

[0289] 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)

[0290]

[0291] 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 reference WO2022169948A1), N,N-diisopropylethylamine (205.17 μL, 1.24 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium 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 and 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 purified on a normal phase silica gel column (dichloromethane:methanol = 10:1) to give compound 1c (60 mg, 0.13 mmol). ESI-MS (m / z): 452.3 [M+H]. + .

[0292] Preparation of compound 1

[0293] Under nitrogen, to a solution of compound 1c (66 mg, 0.15 mmol) in N-methylpyrrolidone (1 mL) were added 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) in sequence. The reaction was allowed to proceed in a microwave oven at 70°C for 3 hours. After completion of the reaction, water (10 mL) was added and the mixture was extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent removed. Compound 1 (19 mg) was purified by reverse phase column chromatography (acetonitrile: 0.05% aqueous ammonium bicarbonate solution = 0:100%–50%:50%). 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).

[0294] 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)

[0295]

[0296] 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 mixture was allowed to react overnight at 25°C. After completion of the reaction, the mixture was concentrated to afford Compound 2b (1.0 g). ESI-MS (m / z): 240.0 [M+H] + .

[0297] Preparation of Compound 2c: To a solution of trimethylsulfoxide iodide (2.3 g, 10.41 mmol) in tetrahydrofuran (10 mL) was added potassium tert-butoxide (1.16 g, 10.41 mmol). 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. The mixture was allowed to react at room temperature for 1 hour, then heated to 50°C for 3 hours. Potassium tert-butoxide (4.16 mmol, 467 mg) was added and the mixture was allowed to react at 25°C overnight. After completion, the reaction was quenched with water (15 mL) and extracted with ethyl acetate (20 x 3 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 2c (200.0 mg) was obtained by normal phase column chromatography (ethyl acetate / petroleum ether = 0:100%–50%:50%). ESI-MS (m / z): 254.2 [M+H] + .

[0298] Preparation of Compound 2d: To a solution of compound 2c (100 mg, 0.39 mmol) in N,N-dimethylacetamide (5 mL) were added compound 1b (114.4 mg, 0.47 mmol), O-(1H-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). The mixture was allowed to react 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. The filtrate was purified by normal phase column chromatography (methanol / dichloromethane = 0:100%–10%:90%) to afford compound 2d (130.0 mg). ESI-MS (m / z): 478.2 [M+H] + .

[0299] Preparation of compound 2

[0300] Under nitrogen, to a solution of compound 2d (48 mg, 0.1 mmol) in N-methylpyrrolidone (1 mL) were added 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) in sequence. The mixture was microwaved at 70°C for 3 hours. Upon completion, water (10 mL) was added and the mixture was extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent removed. Compound 2 (23 mg) was purified by reverse phase column chromatography (acetonitrile: 0.05% aqueous ammonium bicarbonate solution = 0:100%–40%:60%). 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).

[0301] 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)

[0302]

[0303] Preparation of Compound 3b: To a solution of compound 1a (50 mg, 0.22 mmol) in N,N-dimethylformamide (2 mL) were added compound 3a (50.6 mg, 0.22 mmol, preparation method reference WO2022169948 A1), N,N-diisopropylethylamine (108.93 μL, 0.66 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (208.4 mg, 0.55 mmol), and the mixture was allowed to react overnight at 25°C. Water (50 mL) was added to the reaction solution, which was then extracted with ethyl acetate (3 x 10 mL). The organic phases were washed three times with saturated brine, combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 3b (50 mg) was obtained by normal phase column chromatography (dichloromethane / methanol = 10:1). ESI-MS (m / z): 439.92 [M+H]. + .

[0304] Preparation of compound 3 To a solution of 3b (50 mg, 0.11 mmol) in N-methylpyrrolidone (2 mL) were added cuprous iodide (4.3 mg, 0.02 mmol), tetrakistriphenylphosphine 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). After the addition, the mixture was microwaved at 70 °C under nitrogen protection for 3 h. The reaction mixture was filtered through celite, 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 purified 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 obtain 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).

[0305] 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)

[0306]

[0307] To a solution of 1c (63 mg, 0.14 mmol) in N-methylpyrrolidone (2 mL) were added cuprous iodide (4 mg, 0.02 mmol), tetrakistriphenylphosphine palladium (32 mg, 0.03 mmol), N,N-diisopropylethylamine (90 mg, 0.7 mmol), and 1-cyclopropyl-4-ethynyl-1H-pyrazole (55 mg, 0.42 mmol). The mixture was microwaved at 80°C for 3 hours under nitrogen. The reaction mixture was filtered through celite, water (15 mL) was added, and extraction with ethyl acetate (3 x 10 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 4 (15 mg) was obtained by reverse-phase column chromatography (acetonitrile / 0.05% ammonium bicarbonate solution = 0:100%–50%:50%). 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).

[0308] Compound 4 was separated by SFC (Waters 150Prep-SFC; Chiral AS column; mobile phase A: CO2, B: methanol (containing 0.1% NH3·H2O); B%: 35%) to give compounds 4A and 4B. Compound 4A: SFC analysis method: SHIMADZU LC-20AD; 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%.

[0309] Compound 4B: SFC analysis method: Equipment: SHIMADZU LC-20AD; Chromatographic 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%.

[0310] Example 5: 4-amino-7-fluoro-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 5)

[0311]

[0312] Preparation of compound 5b

[0313] To a solution of compound 1a (50 mg, 0.22 mmol) in N,N-dimethylformamide (2 mL) were added compound 5a (57.2 mg, 0.22 mmol, prepared according to WO2022169948 A1), N,N-diisopropylethylamine (108.93 μL, 0.66 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (208.4 mg, 0.55 mmol). The mixture was allowed to react overnight at 25°C. Water (50 mL) was added to the reaction solution, which was then extracted with ethyl acetate (3 x 10 mL). The organic phases were washed three times with saturated brine, combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 5b (55 mg) was obtained by normal phase column chromatography (dichloromethane / methanol = 10:1). ESI-MS (m / z): 469.95 [M+H]. + .

[0314] Preparation of compound 5 To a solution of compound 5b (55 mg, 0.12 mmol) in N-methylpyrrolidone (2 mL) were added cuprous iodide (4.5 mg, 0.02 mmol), tetrakistriphenylphosphine 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), and the mixture was reacted in a microwave oven at 70 ° C under nitrogen protection for 3 h. The reaction mixture was filtered through celite, 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 purified by normal phase column chromatography (methanol / dichloromethane = 10:1), and then purified by reverse phase column chromatography (acetonitrile / 0.05% ammonium bicarbonate solution = 0:100%–45%:55%) to obtain compound 5 (18 mg). ESI-MS (m / z): 496.0 [M+H] + . 1H NMR (400 MHz, DMSO-d6) δ 8.29 - 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).

[0315] Compound 5 was separated by SFC (Equipment: Waters 150 Prep-SFC; Column: Chiral AS column; Mobile phase A: CO2, B: methanol (containing 0.1% NH3H2O); B%: 35%) to give compound 5A and 5B. Compound 5A: 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: 1.997 min, ee = 99.7%. 1 H NMR (400 MHz, DMSO-d6) δ 8.29 - 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). 1 H NMR (400 MHz, DMSO-d6, 90 °C) δ 8.25 - 8.23 (m, 2H), 7.96 (s, 1H), 7.60 (s, 1H), 7.34 - 7.30 (m, 2H), 7.07 (dd, J = 7.6, 1.4 Hz, 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).

[0316] 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).

[0317] 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-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 5A)

[0318]

[0319] Compound Int-1 was separated by SFC (equipment: Shimadzu Prep-SFC; column: Opti-Chiral A1 column (20*250 mm, 5 μm); mobile phase A: CO2, B: ethanol (containing 0.1% NH3·H2O); B%: 35%) to obtain two components.

[0320] Compound Int-1A: SFC analysis method: Equipment: SHIMADZU LC-20AD; Chromatographic 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: 1.924min, ee = 100%.

[0321] The preparation process of compound Int-3A refers to WO2024131901 A1.

[0322] Confirmation of the absolute configuration of compound Int-2A: The crystal structure of Int-2A was determined by MicroED method. The crystal structure belongs to the triclinic system and the P1 (No.1) space group. The unit cell constant measured by MicroED is α=97.51(12)°,β=89.49(15)°,γ=99.60(18)°,unit cell volume The Z' of the system is 1, and the asymmetric unit of the crystal consists of one API molecule.

[0323] Preparation of compound 1a-A

[0324] 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 spin-dried, slurried with methyl tert-butyl ether (5 mL), and filtered to obtain 1a-A (180 mg). ESI-MS (m / z): 227.96 [M+H] +

[0325] Preparation of compound 5A: The synthetic process for preparing compound 5A from compound 1a-A refers to the synthetic process 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).

[0326] 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)

[0327]

[0328] Preparation of compound 6b

[0329] To a solution of compound 6a (41.9 mg, 0.20 mmol, prepared as described in WO2024131901 A1) in N,N-dimethylacetamide (3 mL) were added compound 1b (48.4 mg, 0.20 mmol), triethylamine (0.07 mL, 0.50 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (156.0 mg, 0.30 mmol). 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-pressure liquid chromatography to obtain compound 6b (5 mg). ESI-MS (m / z): 398.17 [M+H] + .

[0330] Preparation of compound 6

[0331] Under nitrogen protection, to a solution of compound 6b (50 mg, 0.13 mmol) in N,N-dimethylformamide (2 mL) were added 1-methyl-1,2,3,6-tetrahydropyridin-4-yl trifluoromethanesulfonate (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) in sequence, and the mixture was reacted at 25 ° C for 3 hours. The reaction mixture was added 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 removed. Column chromatography (methanol: dichloromethane = 1:20) gave the crude product, which was then purified by reverse phase chromatography (acetonitrile / 0.05% aqueous ammonium bicarbonate 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).

[0332] 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)

[0333]

[0334] Preparation of compound 7b

[0335] 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 on ice for 5 min, followed by the addition of N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (236 mg, 0.62 mol). The reaction was continued for 5 min, and finally, compound 7a (110 mg, 0.45 mol) was added. The reaction was continued for one hour. Water (30 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 ml x 3). The organic phases were washed three times with saturated brine, combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The filtrate was purified by column chromatography (methanol:dichloromethane = 1:20) to afford compound 7b (120 mg). ESI-MS (m / z): 465.91 [M+H]. + .

[0336] Preparation of compound 7

[0337] Under nitrogen, to a solution of compound 7b (120 mg, 0.26 mmol) in N-methylpyrrolidone (3 mL) were added 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) in sequence. The mixture was microwaved at 70°C for three hours. Water (50 mL) was added to the reaction solution, which was then 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 removed. Compound 7 (60 mg) was obtained by column chromatography (methanol:dichloromethane = 1:20). ESI-MS (m / z): 492.22 [M+H]. + . 1H NMR (400 MHz, DMSO) δ 8.28 (s, 2H), 8.08 (s, 1H), 7.69 - 7.59 (m, 3H), 7.48 (d, J = 7.6 Hz, 1H), 7.16 (s, 2H), 7.08 (dd, J = 7.6 1.2 Hz, 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).

[0338] 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)

[0339]

[0340] Preparation of Compound 8b

[0341] To the solution of compound 1a (230 mg, 1.01 mmol) in N,N-dimethylformamide (10 mL) was added compound 8a (279 mg, 1.01 mmol, preparation method refer to WO2022169948 A1), triethylamine (255.1 mg, 2.52 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (576.3 mg, 1.51 mmol), 25 °C reaction overnight. The reaction solution was added with water (50 mL), extracted with ethyl acetate (2*25 ml), the organic phase was washed with saturated brine three times, the combined organic phase was dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated, and purified by normal phase column chromatography (dichloromethane / methanol = 50:1) to obtain compound 8b (280 mg). ESI-MS (m / z): 487.89 [M+H] + .

[0342] Preparation of Compound 8c

[0343] To a solution of compound 8b (280 mg, 0.58 mmol) in N-methylpyrrolidone (3 mL) were added cuprous iodide (21.9 mg, 0.12 mmol), tetrakistriphenylphosphine palladium (66.5 mg, 0.06 mmol), N,N-diisopropylethylamine (371.52 mg, 2.88 mmol), and trimethylsilyl acetylene (113 mg, 1.15 mmol). The mixture was microwaved at 70°C under nitrogen for 3 hours. The reaction mixture was filtered through celite, water (50 mL) was added, and extraction with ethyl acetate (2 x 25 mL) was performed. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 8c (220 mg) was obtained by normal phase column chromatography (dichloromethane / methanol = 20:1). ESI-MS (m / z): 504.3 [M+H]. + .

[0344] Preparation of compound 8d

[0345] To a solution of compound 8c (220 mg, 0.44 mmol) in tetrahydrofuran (5 mL) was added tetrabutylammonium fluoride (870 μL, 0.87 mmol) and the mixture was allowed to react at 25°C for 1 hour. Water (15 mL) was added to the reaction solution, and the mixture was extracted with 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 plate separation (dichloromethane / methanol = 10:1) to afford compound 8d (160 mg). ESI-MS (m / z): 432.03 [M+H]. + .

[0346] Preparation of compound 8

[0347] Under nitrogen, to a solution of compound 8d (43 mg, 0.10 mmol) in N,N-dimethylformamide (2 mL) were added 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) in sequence. The mixture was reacted at 25°C for 3 hours. 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 removed. The crude product was purified by column chromatography (methanol:dichloromethane = 1:20). The crude product was then purified by reverse phase chromatography (acetonitrile / 0.05% aqueous ammonium bicarbonate = 0:100%–40%:60%) to afford 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).

[0348] 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)

[0349]

[0350] The synthetic process for preparing compound 8A from compound 1a-A refers to the synthetic process 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).

[0351] 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)

[0352]

[0353] In a 10 mL microwave tube, reactant 8b (48.6 mg, 0.10 mmol), cuprous iodide (3.8 mg, 0.02 mmol), tetrakistriphenylphosphine palladium (11.5 mg, 0.01 mmol), and N-methylpyrrolidone (2 mL) were added. Nitrogen was bubbled through the tube for 3 minutes, followed by the addition of N,N-diisopropylethylamine (52.94 μL, 0.32 mmol) and N-methyl-4-alkynylpyrazole (21.2 mg, 0.20 mmol). The reaction mixture was stirred in a microwave oven at 70°C for 3 hours. After completion, the reaction was quenched with water and extracted three times with ethyl acetate (30 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography (methanol:dichloromethane = 1:20). Compound 9 (15 mg) was then isolated using a reverse phase preparative column (acetonitrile:0.05% aqueous ammonium bicarbonate = 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).

[0354] Compound 9 was separated by SFC (Waters 150Prep-SFC; Chiral AS column; mobile phase A: CO2, B: methanol (containing 0.1% NH3·H2O); B%: 45%) to yield compounds 9A and 9B. Compound 9A: SFC analysis: SHIMADZU LC-20AD; 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] + .

[0355] Compound 9B: SFC analysis: 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] +.

[0356] 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)

[0357]

[0358] Preparation of compound 10b

[0359] To a solution of compound 1a (210 mg, 0.92 mmol) in N,N-dimethylformamide (10 mL) were added compound 10a (256.3 mg, 0.92 mmol, prepared according to WO2022169948 A1), triethylamine (232.9 mg, 2.30 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (526.2 mg, 1.38 mmol). The mixture was reacted at 25°C overnight. Water (50 mL) was added to the reaction solution, which was then extracted with ethyl acetate (2 x 25 mL). The organic phases were washed three times with saturated brine, combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 10b (360 mg) was obtained by normal phase column chromatography (dichloromethane / methanol = 50:1). ESI-MS (m / z): 468.07 [M+H]. + .

[0360] Preparation of compound 10

[0361] Reactant 10b (46.6 mg, 0.10 mmol), cuprous iodide (3.8 mg, 0.02 mmol), tetrakistriphenylphosphine palladium (11.5 mg, 0.01 mmol), and N-methylpyrrolidone (2 mL) were added to a 10 mL microwave tube. Nitrogen was bubbled through the tube for 3 minutes. N,N-diisopropylethylamine (52.94 μL, 0.32 mmol) and N-methyl-4-alkynylpyrazole (21.2 mg, 0.20 mmol) were then added. The reaction mixture was stirred in a microwave at 70°C for 3 hours. After the reaction, water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate (30 ml). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography (methanol:dichloromethane = 1:20). Compound 10 (15 mg) was then separated using a reverse phase column (acetonitrile:0.05% aqueous ammonium bicarbonate solution = 0:100%–35%:65%). ESI-MS (m / z): 491.96 [M+H] + . 1H NMR (400 MHz, DMSO-d6) δ 8.24 (s, 1H), 8.07-8.04 (m, 2H), 7.69 (s, 1H), 7.52-7.43 (m, 2H), 7.17-6.95 (m, 4H), 6.55-6.48 and 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).

[0362] Example 10A: (S)-4-amino-N, 1,7-trimethyl-N-(6-((1-methyl-1H-pyrazol-4-yl)ethynyl)- 2,3-dihydrobenofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 10A)

[0363]

[0364] The synthetic procedure for preparing Compound 10A from Compound la-A is referenced to the synthetic procedure for preparing Compound 10 from Compound la.

[0365] Compound 10A: ESI-MS (m / z): 492.01 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 8.24 (s, 1H), 8.07-8.04 (m, 2H), 7.69 (s, 1H), 7.52-7.43 (m, 2H), 7.17-6.95 (m, 4H), 6.55-6.48 and 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).

[0366] Example 11: 4-amino-N-(6-((1-cyclopropyl-1H-pyrazol-4-yl)ethynyl)-2,3- dihydrobenofuran-3-yl)-N, 1,7-trimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 11)

[0367]

[0368] Preparation of Compound 11b: To a solution of compound 10a (30.8 mg, 0.12 mmol) in N,N-dimethylacetamide (5 mL) were added 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), and the mixture was allowed to react overnight at 25°C. Water (10 mL) was added to the reaction solution, which was extracted with ethyl acetate (10 x 3 ml). The organic phases were washed three times with saturated brine, combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Reverse-phase column chromatography (acetonitrile / 0.05% aqueous ammonium bicarbonate solution = 0:100% to 50%:50%) afforded compound 11a (7.0 mg). ESI-MS (m / z): 412.15.

[0369] Preparation of compound 11 Under nitrogen protection, to a solution of compound 11a (50 mg, 0.12 mmol) in N,N-dimethylformamide (2 mL) were added 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) in sequence, and the mixture was reacted at 25°C for 3 hours. The reaction mixture was added 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 removed. The crude product was purified by column chromatography (methanol:dichloromethane = 1:20). Compound 11 (20 mg) was prepared by reverse phase chromatography (acetonitrile / 0.05% aqueous ammonium bicarbonate 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).

[0370] 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)

[0371]

[0372] Preparation of Compound 12a

[0373] To a solution of compound 5b (808 mg, 1.72 mmol) in N-methylpyrrolidine (10 mL) was added trimethylsilylethynyl (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) successively under nitrogen protection. After the addition, the mixture was heated at 100 °C for 3 h in a microwave reactor. The reaction mixture was diluted with water (200 mL) and extracted with ethyl acetate (30 mL*3). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (methanol:dichloromethane = 1:20) to give compound 12a (595 mg). ESI-MS (m / z): 488.18 [M+H] + .

[0374] Preparation of Compound 12b

[0375] To a solution of compound 12a (595 mg, 1.22 mmol) in tetrahydrofuran (10 mL) was added tetrabutylammonium fluoride solution (2.44 mL, 1 M) at room temperature. The mixture was stirred for 1 h. The reaction mixture was diluted with water and extracted with ethyl acetate (30 mL*3). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (methanol:dichloromethane = 1:20) to give compound 12b (470 mg). ESI-MS (m / z): 416.12 [M+H] + .

[0376] Preparation of Compound 12

[0377] Under nitrogen, to a solution of compound 12b (62.3 mg, 0.15 mmol) in N-methylpyrrolidone (1.5 mL) were added 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) in sequence. The mixture was reacted in an oil bath at 50°C for three hours. Water (100 mL) was added to the reaction solution, which was then 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 removed. Compound 12 (25 mg) was obtained by column chromatography (methanol:dichloromethane = 1:20). 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).

[0378] 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)

[0379]

[0380] The synthetic process for preparing compound 12A from compound 5b-A refers to the synthetic process for preparing compound 12 from compound 5b.

[0381] 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).

[0382] 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)

[0383]

[0384] Referring to the preparation process of compound 12, 4-iodo-1-isopropyl-1H-pyrazole (70.8 mg, 0.3 mmol) was used to replace 1-cyclopropyl-4-iodo-1H-pyrazole. The reaction was carried out at 50°C under nitrogen protection for 3 hours 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).

[0385] Example 14: 4-amino-7-chloro-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 14)

[0386]

[0387] Referring to the preparation process of compound 8, 1-cyclopropyl-4-iodopyrazole was replaced with 4-iodo-1-(propan-2-yl)pyrazole (21.9 mg, 0.09 mmol). The reaction was carried out at 50°C under nitrogen for 3 hours 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).

[0388] Example 15: 4-amino-7-chloro-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 15)

[0389]

[0390] Referring to the preparation process of compound 8, 1-ethyl-4-iodopyrazole (30.8 mg, 0.14 mmol) was used to replace 1-cyclopropyl-4-iodopyrazole. The reaction was carried out at 50°C under nitrogen protection for 3 hours 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).

[0391] 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)

[0392]

[0393] Preparation of Compound 16b: To a solution of compound 1a (100 mg, 0.44 mmol) in N,N-dimethylformamide (10 mL) were added compound 16a (109 mg, 0.44 mmol, preparation method reference WO2022169948 A1), triethylamine (110.9 mg, 1.10 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (250.5 mg, 0.66 mmol), and the mixture was allowed to react overnight at 25°C. Water (50 mL) was added to the reaction solution, which was then extracted with ethyl acetate (2 x 25 mL). The organic phases were washed three times with saturated brine, combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 16b (118 mg) was obtained by normal phase column chromatography (dichloromethane / methanol = 50:1). ESI-MS (m / z): 459.85 [M+H]. + .

[0394] Preparation of compound 16c

[0395] To a solution of 16b (118 mg, 0.26 mmol) in N-methylpyrrolidone (3 mL) were added cuprous iodide (9.8 mg, 0.05 mmol), tetrakistriphenylphosphine palladium (29.8 mg, 0.03 mmol), triethylamine (104.2 mg, 2.88 mmol), and trimethylsilylacetylene (50.6 mg, 0.51 mmol). The mixture was microwaved at 70°C under nitrogen for 3 hours. The reaction mixture was filtered through celite, water (50 mL) was added, and extraction with ethyl acetate (2 x 25 mL) was performed. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 16c (105 mg) was obtained by normal phase column chromatography (dichloromethane / methanol = 20:1). ESI-MS (m / z): 476.2 [M+H]. + .

[0396] Preparation of Compound 16d: To a solution of compound 16c (105 mg, 0.22 mmol) in tetrahydrofuran (5 mL) was added tetrabutylammonium fluoride (440 μL, 0.44 mmol) and the mixture was allowed to react at 25°C for 1 hour. Water (15 mL) was added to the reaction solution, and the mixture was extracted with 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 afford compound 16d (89.1 mg). ESI-MS (m / z): 404.09 [M+H] + .

[0397] Preparation of compound 16

[0398] To a solution of compound 16d (43 mg, 0.11 mmol) in N,N-dimethylformamide (2 mL) was added 1-cyclopropyl-4-iodopyrazole (49.9 mg, 0.22 mmol), copper iodide (8.10 mg, 0.04 mmol), triethylamine (43.1 mg, 0.43 mmol) and tetrakis(triphenylphosphine)palladium (24.6 mg, 0.02 mmol) sequentially under nitrogen protection. The mixture was stirred at 25 °C for 3 h. The reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (10 mL*3). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (methanol:dichloromethane = 1:20) to give the crude product, which was further purified by reverse phase preparative HPLC (acetonitrile / 0.05% aqueous ammonium bicarbonate solution = 0:100% - 40:60%) to give compound 16 (18 mg). ESI-MS (m / z): 510.18 [M+H] + . 1 HNMR (400 MHz, DMSO-d6) δ 8.19-8.18 (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), 6.45-6.42 and 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), 2.68 and 2.55 (s, 3H), 1.11-0.97 (m, 4H).

[0399] Example 16A: (S)-4-amino-N-(6-((1-cyclopropyl-1H-pyrazol-4-yl)ethynyl)-2,3- dihydrobenofuran-3-yl)-7-fluoro-N-methyl-1,3-dihydrofuro[3,4-c]quinoline-8-carboxamide (Compound 16A)

[0400]

[0401] The synthetic process for preparing compound 16A from compound 1a-A refers to the synthetic process for preparing compound 16 from compound 1a.

[0402] 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).

[0403] 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)

[0404]

[0405] Referring to the preparation process of compound 12, compound 1-ethyl-4-iodo-1H-pyrazole (66.6 mg, 0.3 mmol) was used to replace compound 1-cyclopropyl-4-iodo-1H-pyrazole 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).

[0406] 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)

[0407]

[0408] Referring to the preparation process of compound 8, compound 3-iodo-1-methylpyrazole (41.6 mg, 0.20 mmol) was used to replace compound 1-cyclopropyl-4-iodopyrazole 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).

[0409] 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)

[0410]

[0411] Under nitrogen, to a solution of compound 6b (49.6 mg, 0.125 mmol) in N-methylpyrrolidone (1.5 mL) were added 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). The mixture was reacted in an oil bath at 50°C for three hours. Water (100 mL) was added to the reaction solution, which was then 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 removed. Compound 19 (30 mg) was obtained by column chromatography (methanol:dichloromethane = 1:20). 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).

[0412] 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)

[0413]

[0414] The preparation process of compound 6b-A refers to WO2024131901 A1. The synthesis process for preparing compound 19A from compound 6b-A refers to the synthesis process for preparing compound 19 from compound 6b.

[0415] 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).

[0416] 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)

[0417]

[0418] Referring to the preparation process of compound 19, compound 1-ethyl-4-iodo-1H-pyrazole (55.5 mg, 0.25 mmol) was used to replace compound 4-iodo-1-isopropyl-1H-pyrazole 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).

[0419] 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)

[0420]

[0421] The synthetic process for preparing compound 20A from compound 6b-A refers to the synthetic process for preparing compound 20 from compound 6b.

[0422] 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).

[0423] 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)

[0424]

[0425] Reference the preparation process of compound 12, compound 3-iodo-1-methylpyrazole (21 mg, 0.10 mmol) was used to replace compound 1-cyclopropyl-4-iodo-1H-pyrazole, 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 (400 MHz, DMSO-d6) δ 8.40-8.24 (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), 6.50-5.57 (m, 1H), 4.86-4.65 (m, 2H), 4.44-4.41 (m, 3H), 3.89 (s, 3H), 2.72 and 2.61 (s, 3H).

[0426] Example 21A: (S)-4-amino-7-fluoro-N,1-dimethyl-N-(6-((1-methyl-1H-pyrazol-3-yl)ethynyl)- 2,3-dihydrobenofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 21A)

[0427]

[0428] The synthetic process for preparing compound 21A from compound 12b-A refers to the synthetic process for preparing compound 21 from compound 12b.

[0429] Compound 21A: ESI-MS (m / z): 496.08 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 8.40-8.24 (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), 6.50-5.57 (m, 1H), 4.86-4.65 (m, 2H), 4.44-4.41 (m, 3H), 3.89 (s, 3H), 2.72 and 2.61 (s, 3H).

[0430] Example 22: 4-amino-7-chloro-N-(6-((1,5-dimethyl-1H-pyrazol-4-yl)ethynyl)-2,3- dihydrobenofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 22)

[0431]

[0432] Preparation of Compound 22b: To a solution of compound 22a (1.0 g, 10.41 mmol) in N,N-dimethylformamide (10 mL) was added N-iodosuccinimide (2.5 g, 10.93 mmol), and the mixture was allowed to react 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. The mixture was purified by column chromatography (petroleum ether:ethyl acetate = 6:1) to afford 22b (1.75 g).

[0433] Preparation of compound 22

[0434] Referring to the preparation process of compound 8, compound 22b (41.1 mg, 0.19 mmol) was used to replace compound 1-cyclopropyl-4-iodopyrazole 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).

[0435] 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-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 22A)

[0436]

[0437] The synthesis process of compound 22A prepared from compound 22b and compound 8d-A refers to the synthesis process of compound 22 prepared from compound 22b and compound 8d.

[0438] Compound 22A: ESI-MS (m / z): 525.95 [M+H] + .

[0439] Example 23: 4-amino-N-(6-((1,5-dimethyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 23)

[0440]

[0441] Referring to the preparation process of compound 12, compound 22b (44.4 mg, 0.20 mmol) was used to replace compound 1-cyclopropyl-4-iodo-1H-pyrazole. 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).

[0442] Compound 23 was separated by SFC (Waters 150Prep-SFC; Chiral AS column; mobile phase A: CO2, mobile phase B: methanol (containing 0.1% NH3·H2O); B%: 40%) to give compounds 23A and 23B. Compound 23A: SFC analysis method: SHIMADZU LC-20AD; 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 1mg / mL,DMF).

[0443] 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).

[0444] 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)

[0445]

[0446] Preparation of Compound 24b: To a solution of 3-chloro-1-methylpyrazole (116 mg, 1.0 mmol) in acetonitrile (3 mL) was added N-iodosuccinimide (235.2 mg, 1.05 mmol). The mixture was allowed to react 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 combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent removed. Column chromatography (petroleum ether:ethyl acetate = 6:1) afforded Compound 24b (210 mg). ESI-MS (m / z): 242.89 [M+H] + .

[0447] Preparation of compound 24

[0448] Referring to the preparation process of compound 12, compound 24b (24.2 mg, 0.10 mmol) was used to replace compound 1-cyclopropyl-4-iodo-1H-pyrazole. 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).

[0449] Compound 24 was separated by SFC (Waters 150Prep-SFC; Chiral AS column; mobile phase A: CO2, B: methanol (containing 0.1% NH3·H2O); B%: 40%) to give compounds 24A and 24B. Compound 24A: SFC analysis method: SHIMADZU LC-20AD; 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%.

[0450] Compound 24B: SFC analysis method: Equipment: SHIMADZU LC-20AD; Chromatographic 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%.

[0451] Example 25: 4-amino-N-(6-((1,3-dimethyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 25)

[0452]

[0453] Referring to the preparation process of compound 12, compound 25a (22.2 mg, 0.10 mmol) was used to replace compound 1-cyclopropyl-4-iodo-1H-pyrazole. 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).

[0454] Compound 25 was separated by SFC (Waters 150Prep-SFC; Chiral AS column; mobile phase A: CO2, B: methanol (containing 0.1% NH3·H2O); B%: 40%) to give compounds 25A and 25B. Compound 25A: SFC analysis method: SHIMADZU LC-20AD; 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).

[0455] 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 1 mg / mL,DMF).

[0456] Example 26: 4-amino-7-chloro-N-(6-((1,3-dimethyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 26)

[0457]

[0458] Referring to the preparation process of compound 8, compound 25a (22.2 mg, 0.10 mmol) was used to replace compound 1-cyclopropyl-4-iodopyrazole. 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).

[0459] 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-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 26A)

[0460]

[0461] The synthetic process for preparing compound 26A from compound 8d-A refers to the synthetic process for preparing compound 26 from compound 8d.

[0462] 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).

[0463] 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)

[0464]

[0465] Preparation of compound 27a

[0466] 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 on ice for 5 min, followed by the addition of N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (232.53 mg, 0.61 mol). The reaction was continued for 5 min, and finally, compound 7a (113.54 mg, 0.41 mol) was added. The reaction was continued for one hour. Water (100 mL) was added to the reaction solution, and the solution was extracted with ethyl acetate (20 ml x 3). The organic phases were washed three times with saturated brine, combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The filtrate was purified by column chromatography (methanol:dichloromethane = 1:20) to afford compound 27a (120 mg). ESI-MS (m / z): 484.11 [M+H]. + .

[0467] Preparation of compound 27

[0468] Under nitrogen, to a solution of compound 27a (120 mg, 0.25 mmol) in N-methylpyrrolidone (3 mL) were added 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) in sequence. The reaction 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 organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent removed. Compound 27 (15 mg) was obtained by column chromatography (methanol:dichloromethane = 1:20). ESI-MS (m / z): 510.1 [M+H]. + . 1 H NMR (400MHz, DMSO) δ8.38–8.17(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).

[0469] 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)

[0470]

[0471] Preparation of compound 28a

[0472] Referring to the preparation process of compound 27a, compound 8a (120 mg, 0.43 mol) was used to replace compound 5a to obtain compound 28a (160 mg). ESI-MS (m / z): 499.94 [M+H] + .

[0473] Preparation of compound 28

[0474] Referring to the preparation process of compound 27, compound 28a (100 mg, 0.2 mmol) was used to replace compound 27a to obtain compound 28 (20 mg). ESI-MS (m / z): 526.2 [M+H] + . 1 H 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).

[0475] Example 29: 4-amino-N-(6-((1,5-dimethyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 29)

[0476]

[0477] Referring to the preparation process of compound 19, compound 22b (22.2 mg, 0.10 mmol) was used to replace compound 4-iodo-1-isopropyl-1H-pyrazole. 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] + . 1HNMR(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).

[0478] Example 29A: (S)-4-amino-N-(6-((1,5-dimethyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 29A)

[0479]

[0480] The synthetic process for preparing compound 29A from compound 6b-A refers to the synthetic process for preparing compound 29 from compound 6b.

[0481] 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).

[0482] 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)

[0483]

[0484] Preparation of compound 30b

[0485] To a solution of compound 30a (300 mg, 1.23 mmol) in N-methylpyrrolidone (5 mL) were added cuprous iodide (36 mg, 0.19 mmol), tetrakistriphenylphosphine palladium (288 mg, 0.25 mmol), N,N-diisopropylethylamine (476 mg, 3.69 mmol), and trimethylethynylsilane (242 mg, 2.46 mmol). The mixture was microwaved at 80°C under nitrogen for 3 hours. The reaction mixture was filtered through celite, water (50 mL) was added, and extraction with ethyl acetate (3 x 50 mL) was performed. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 30b (178 mg) was obtained by normal phase column chromatography (methanol / dichloromethane = 1:20). ESI-MS (m / z): 215.1 [M+H]. + .

[0486] Preparation of compound 30c

[0487] To a solution of compound 30b (178 mg, 0.83 mmol) in tetrahydrofuran (5 mL) was added tetrabutylammonium fluoride (434 mg, 1.66 mmol) and the mixture was allowed to react at 25°C for 0.5 h. Water (30 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (3 x 30 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 30c (89 mg) was obtained by normal phase column chromatography (methanol / dichloromethane = 1:15). ESI-MS (m / z): 143.1 [M+H] + .

[0488] Preparation of compound 30

[0489] To a solution of 1c (45 mg, 0.1 mmol) in N-methylpyrrolidone (2 mL) were added cuprous iodide (4 mg, 0.02 mmol), tetrakistriphenylphosphine palladium (23 mg, 0.02 mmol), N,N-diisopropylethylamine (39 mg, 0.3 mmol), and 30c (29 mg, 0.2 mmol). The mixture was microwaved at 80°C under nitrogen for 3 hours. The reaction mixture was filtered through celite, water (20 mL) was added, and extraction with ethyl acetate (3 x 20 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 30 (15 mg) was obtained by reverse-phase column chromatography (acetonitrile / 0.05% ammonium bicarbonate solution = 0:100%–50%:50%). ESI-MS (m / z): 513.9 [M+H] + . 1HNMR(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).

[0490] 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)

[0491]

[0492] The preparation process of compound 1c-A refers to the preparation process of compound 1c. The synthesis process of compound 30A prepared from compound 1c-A refers to the synthesis process of compound 30 prepared from compound 1c.

[0493] 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), 7.6 6–7.59(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–

[0494] 4.61(m,2H),4.42(s,3H),2.68(s,3H).

[0495] Example 31: 4-amino-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 31)

[0496]

[0497] Preparation of compound 31b

[0498] To a solution of compound 31a (500 mg, 2.19 mmol) in N-methylpyrrolidone (10 mL) were added cuprous iodide (63 mg, 0.33 mmol), tetrakistriphenylphosphine palladium (508 mg, 0.44 mmol), N,N-diisopropylethylamine (848 mg, 6.57 mmol), and trimethylethynylsilane (430 mg, 4.38 mmol). The mixture was microwaved at 80°C under nitrogen for 3 hours. The reaction mixture was filtered through celite, water (50 mL) was added, and extraction 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. Compound 31b (350 mg) was obtained by normal phase column chromatography (methanol / dichloromethane = 1:20). ESI-MS (m / z): 247.1 [M+H]. + .

[0499] Preparation of Compound 31c: To a solution of compound 31b (350 mg, 1.42 mmol) in tetrahydrofuran (10 mL) was added tetrabutylammonium fluoride (741 mg, 2.84 mmol) and the mixture was allowed to react at 25°C for 0.5 hours. Water (50 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (3 x 50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 31c (165 mg) was obtained by normal phase column chromatography (methanol / dichloromethane = 1:15). ESI-MS (m / z): 175.1 [M+H] + .

[0500] Preparation of compound 31

[0501] Referring to the preparation process of compound 30, compound 31c (35 mg, 0.2 mmol) was used to replace compound 30c 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).

[0502] 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-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 31A)

[0503]

[0504] The synthetic process for preparing compound 31A from compound 31c and compound 1c-A refers to the synthetic process for preparing compound 31 from compound 1c.

[0505] 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).

[0506] 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)

[0507]

[0508] Referring to the preparation process of compound 5, compound 31c (35 mg, 0.2 mmol) was substituted for compound 4-ethynyl-1-methylpyrazole, and microwave reaction was carried out at 80°C under nitrogen protection for 3 hours 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).

[0509] Example 33: 4-amino-7-fluoro-N,1-dimethyl-N-(6-((1-methyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrofuro[2,3-b]pyridin-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 33)

[0510]

[0511] Preparation of compound 33b

[0512] 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 solution was reacted at 0°C for 30 minutes, and then compound 33a (1.03 g, 5.00 mmol) was added. The reaction solution was then stirred at 25°C for 2 hours. The reaction solution was quenched by adding water (5 mL) and extracted with ethyl acetate (30 mL*3). The organic phases were combined and concentrated to give crude product 33b (1.15 g). MS (m / z): 259.95 [M+H] + .

[0513] Preparation of compound 33c

[0514] 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 mixture was reacted at 25°C for 1 hour, then quenched with water (20 mL) and extracted with ethyl acetate (5 mL x 3). The organic phases were combined and concentrated to obtain the crude product, which was purified by column chromatography (petroleum ether:ethyl acetate = 2:1) to afford 33c (240 mg). ESI-MS (m / z): 228.08 [M+H] + .

[0515] Preparation of compound 33d

[0516] 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 solution was heated to 100°C and stirred for 16 hours. The reaction solution was diluted with ethyl acetate (50 mL) and water (50 mL), and extracted with ethyl acetate (20 mL*3). The organic phases were combined and concentrated to obtain the crude product, which was purified by column chromatography (petroleum ether:ethyl acetate = 5:1) to obtain 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, then diluted with ethyl acetate (20 mL) and water (20 mL). The mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined and concentrated to obtain the crude product, which was purified by column chromatography (petroleum ether:ethyl acetate = 2:1) to afford 33e (98 mg). ESI-MS (m / z): 172.04 [M+H]. +Preparation of Compound 33f: To a solution of compound 33e (700 mg, 4.08 mmol) in tetrahydrofuran (20 mL) were added bis(tert-butyloxycarbonyl)amine (1.06 g, 4.90 mmol), triphenylphosphine (1.60 g, 6.12 mmol), and diisopropyl azodicarboxylate (1.24 g, 6.12 mmol). The mixture was reacted at 25°C under nitrogen for 2 hours. The reaction solution was directly concentrated to obtain the crude product, which was purified by normal phase column chromatography (petroleum ether / ethyl acetate = 10:1) to afford compound 33f (1.10 g). ESI-MS (m / z): 371.15 [M+H] + .

[0517] Preparation of compound 33g

[0518] 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, then quenched by adding water (5 mL) and extracted with ethyl acetate (5 mL*3). The organic phases were combined and concentrated to obtain the crude product, which was purified by column chromatography (petroleum ether:ethyl acetate = 3:1) to give 33 g (610 mg). ESI-MS (m / z): 271.05 [M+H] + .

[0519] Preparation of compound 33h

[0520] Compound 33g (271 mg, 1.00 mmol) was dissolved in tetrahydrofuran (5 mL), cooled to 0°C, and sodium hydroxide (80.0 mg, 2.00 mmol, 60% purity) was added. After the reaction solution was reacted at 0°C for 1 hour, iodomethane (0.10 mg, 1.20 mmol) was added, and the reaction solution was stirred at 25°C for 12 hours. The reaction solution was quenched by adding water (5 mL) and extracted with ethyl acetate (5 mL*3). The organic phases were combined and concentrated to obtain the crude product, which was subjected to thin-layer preparative chromatography (petroleum ether:ethyl acetate = 20:1) to obtain 33h (110 mg).

[0521] Preparation of compound 33i

[0522] 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 the reaction mixture was dried to give compound 33i (40 mg). ESI-MS (m / z): 184.96 [M+H] + .

[0523] Preparation of compound 33j

[0524] To a solution of compound 5a (51.9 mg, 0.20 mmol) in N,N-dimethylformamide (3 mL) were added compound 33i (44.22 mg, 0.20 mmol), triethylamine (80.7 mg, 0.80 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (113.7 mg, 0.30 mmol). The mixture was allowed to react overnight at 25°C. Water (50 mL) was added to the reaction solution, which was then extracted with ethyl acetate (2 x 25 mL). The organic phases were washed three times with saturated brine, combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 33j (40 mg) was obtained by normal phase column chromatography (dichloromethane / methanol = 10:1). ESI-MS (m / z): 426.95 [M+H]. + .

[0525] Preparation of compound 33

[0526] Under nitrogen, to a solution of compound 33j (40 mg, 0.09 mmol) in N,N-dimethylformamide (2 mL) were added 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) in sequence. The mixture was heated to 80°C in a microwave oven for 3 hours. Water (10 mL) was added to the reaction solution, which 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 removed. The crude product was purified by column chromatography (methanol:dichloromethane = 1:20). Compound 33 (5 mg) was then obtained by reverse phase chromatography (acetonitrile / 0.05% aqueous ammonium bicarbonate = 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).

[0527] 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)

[0528]

[0529] Referring to the preparation process of compound 19, 3-chloro-4-iodo-1-methylpyrazole (24b, 24.2 mg, 0.10 mmol) was substituted for 4-iodo-1-isopropyl-1H-pyrazole. 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] + . 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).

[0530] 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)

[0531]

[0532] The synthetic process for preparing compound 34A from compound 6b-A refers to the synthetic process for preparing compound 34 from compound 6b.

[0533] 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).

[0534] 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)

[0535]

[0536] Preparation of compound 35b

[0537] To a solution of compound 35a (321.9 mg, 1.23 mmol) in N,N-dimethylformamide (10 mL) were added compound 1a (325.33 mg, 1.23 mmol), triethylamine (310.6 mg, 3.07 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (701.5 mg, 1.84 mmol). The mixture was allowed to react overnight at 25°C. Water (50 mL) was added to the reaction solution, which was then extracted with ethyl acetate (2 x 25 mL). The organic phases were washed three times with saturated brine, combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 35b (510 mg) was obtained by normal phase column chromatography (dichloromethane / methanol = 10:1). ESI-MS (m / z): 472.02 [M+H]. + .

[0538] Preparation of compound 35

[0539] Under nitrogen, to a solution of compound 35b (47.2 mg, 0.10 mmol) in N,N-dimethylformamide (2 mL) were added 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) in sequence. The reaction mixture was heated to 100°C in a microwave oven for 3 hours. 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 removed. The crude product was purified by column chromatography (methanol:dichloromethane = 1:20). Compound 35 (21 mg) was then obtained by reverse phase chromatography (acetonitrile / 0.05% aqueous ammonium bicarbonate = 0:100%–40%:60%). ESI-MS (m / z): 497.95 [M+H] + . 1H 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).

[0540] Example 36: 4-amino-7-fluoro-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 36)

[0541]

[0542] Referring to the preparation process of compound 12, 5-iodo-1-methylpyrazole (49.9 mg, 0.24 mmol) was used to replace 1-cyclopropyl-4-iodo-1H-pyrazole. The reaction was carried out at 50°C under nitrogen for 3 hours 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).

[0543] Example 37: 4-amino-N-(6-((1,4-dimethyl-1H-pyrazol-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 37)

[0544]

[0545] Referring to the preparation process of compound 12, 3-iodo-1,4-dimethylpyrazole (22.2 mg, 0.10 mmol) was used to replace 1-cyclopropyl-4-iodo-1H-pyrazole. 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).

[0546] 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)

[0547]

[0548] Referring to the preparation process of compound 12, 3-bromo-4-fluoro-1-methylpyrazole (17.9 mg, 0.10 mmol) was used to replace 1-cyclopropyl-4-iodo-1H-pyrazole. The reaction was carried out 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).

[0549] 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)

[0550]

[0551] To a solution of 16b (90 mg, 0.20 mmol) in N-methylpyrrolidone (3 mL) were added cuprous iodide (7.6 mg, 0.04 mmol), tetrakistriphenylphosphine 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). After the addition, the mixture was microwaved at 100 °C under nitrogen protection for 4 h. The reaction mixture was filtered through celite, 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 plate (dichloromethane / methanol = 10:1), and then purified by reverse phase column chromatography (acetonitrile / 0.05% ammonium bicarbonate solution = 0:100%–45%:55%) to obtain 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).

[0552] 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-dihydrofuro[3,4-c]quinoline-8-carboxamide (Compound 39A)

[0553]

[0554] The synthetic process for preparing compound 39A from compound 16b-A refers to the synthetic process for preparing compound 39 from compound 16b.

[0555] 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).

[0556] 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)

[0557]

[0558] Preparation of compound 40b

[0559] To a solution of compound 40a (150 mg, 0.57 mmol, preparation method reference WO2022169948 A1) in N,N-dimethylformamide (5 mL) were added 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'-tetramethyluronium hexafluorophosphate (323.2 mg, 0.85 mmol), and the mixture was reacted at 25°C for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature, water (50 mL) was added, and the mixture was extracted with ethyl acetate (2*25 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. Compound 40b (200 mg) was obtained by normal phase column chromatography (dichloromethane / methanol = 20:1). ESI-MS (m / z): 473.96 [M+H] + .

[0560] Preparation of compound 40

[0561] Referring to the preparation method of compound 39, compound 40b (100 mg, 0.21 mmol) was used to replace compound 16b. The reaction was carried out under nitrogen protection in a microwave oven 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).

[0562] 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-dihydrofurano[3,4-c]quinoline-8-carboxamide (Compound 40A)

[0563]

[0564] The synthetic process for preparing compound 40A from compound 1a-A refers to the synthetic process for preparing compound 40 from compound 1a.

[0565] 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).

[0566] 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)

[0567]

[0568] Referring to the preparation process of compound 12, compound 30a (24.4 mg, 0.10 mmol) was substituted for compound 1-cyclopropyl-4-iodo-1H-pyrazole, and the reaction was carried out under microwave 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).

[0569] Example 42: 4-amino-N-(6-((4-(difluoromethyl)pyridin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 42)

[0570]

[0571] Referring to the preparation process of compound 12, 3-bromo-4-(difluoromethyl)pyridine (50.1 mg, 0.24 mmol) was used to replace 1-cyclopropyl-4-iodo-1H-pyrazole. The reaction was carried out under microwave 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).

[0572] 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)

[0573]

[0574] Preparation of Compound 43a: To a solution of 40b (298 mg, 0.63 mmol) in N-methylpyrrolidone (3 mL) were added cuprous iodide (24.1 mg, 0.13 mmol), tetrakistriphenylphosphine palladium (73.0 mg, 0.06 mmol), N,N-diisopropylethylamine (314.04 μL, 1.90 mmol), and trimethylsilyl acetylene (446.48 μL, 3.16 mmol). The mixture was microwaved at 80°C under nitrogen for 4 hours. The reaction mixture was filtered through celite, water (50 mL) was added, and extraction with ethyl acetate (2 x 25 mL) was performed. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 43a (300 mg) was obtained by normal phase column chromatography (dichloromethane / methanol = 10:1). ESI-MS (m / z): 492.0 [M+H] + .

[0575] Preparation of Compound 43b: To a solution of compound 43a (300 mg, 0.61 mmol) in tetrahydrofuran (3 mL) was added tetrabutylammonium fluoride (1.5 mL, 1.52 mmol) and the mixture was allowed to react at 25°C for 1 hour. Water (15 mL) was added to the reaction solution, and the mixture was extracted with 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) to afford the crude product. The crude product was then purified by reverse-phase column chromatography (acetonitrile / 0.05% ammonium bicarbonate solution = 0:100%–40%:60%) to afford 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) were added cuprous iodide (9.1 mg, 0.05 mmol), tetrakistriphenylphosphine palladium (27.5 mg, 0.02 mmol), triethylamine (99.05 μL, 0.71 mmol), and 1-cyclopropyl-4-iodopyrazole (112 mg, 0.48 mmol). After the addition, the mixture was reacted at 25°C under nitrogen protection for 16 hours. The reaction mixture was filtered through celite, 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 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 afford 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).

[0576] 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-dihydrofuro[3,4-c]quinoline-8-carboxamide (Compound 43A)

[0577]

[0578] The synthetic process for preparing compound 43A from compound 40b-A refers to the synthetic process for preparing compound 43 from compound 40b.

[0579] 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).

[0580] 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)

[0581]

[0582] Referring to the preparation process of compound 12, 3-iodo-4-methylpyridine (22.0 mg, 0.1 mmol) was used to replace 1-cyclopropyl-4-iodo-1H-pyrazole. 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).

[0583] 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)

[0584]

[0585] Referring to the preparation process of compound 12, 3-iodo-2-methylpyridine (43.8 mg, 0.2 mmol) was used to replace 1-cyclopropyl-4-iodo-1H-pyrazole. 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).

[0586] 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)

[0587]

[0588] The synthetic process for preparing compound 45A from compound 12b-A refers to the synthetic process for preparing compound 45 from compound 12b.

[0589] 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),6.54–6.44 and 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).

[0590] 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)

[0591]

[0592] Preparation of Compound 46b: To a solution of 46a (3000 mg, 9.93 mmol) in N,N-dimethylacetamide (5 mL) was added methanesulfonic acid (2-dicyclohexylphosphino-2',4',6'-tri-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 trimethylsilylene (4.21 mL, 29.78 mmol). The reaction mixture was purged with nitrogen three times and microwaved at 70°C for 3 hours. The reaction mixture was filtered through celite, water (100 mL) was added, and the mixture was extracted 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. The filtrate was purified by normal phase column chromatography (dichloromethane / methanol = 20:1) to afford compound 46b (2700 mg). ESI-MS (m / z): 320.11 [M+H] + .

[0593] Preparation of compound 46c

[0594] To a solution of compound 46b (2700 mg, 8.45 mmol) in dichloromethane (20 mL) was added trifluoroacetic acid (6.47 mL, 84.50 mmol) and the mixture was allowed to react at 25°C for 16 hours. After completion of the reaction, the mixture was concentrated to afford crude product 46c (1800 mg), which was used directly in the next step. ESI-MS (m / z): 220.11 [M+H] + .

[0595] Preparation of compound 46d

[0596] To a solution of 46c (1800 mg, 8.20 mmol) in formic acid (5 mL) was added formaldehyde solution (2400.0 mg, 29.60 mmol) and the mixture was allowed to react at 100°C for 1 hour. The reaction mixture was quenched with ammonium bicarbonate, and water (50 mL) was added. 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 afford compound 46d (1000 mg). ESI-MS (m / z): 234.13 [M+H] + .

[0597] Preparation of compound 46e

[0598] To a solution of 46d (1000 mg, 4.28 mmol) in N,N-dimethylformamide (5 mL) was added cesium fluoride (2603.3 mg, 17.14 mmol) and the mixture was reacted at 25°C for 30 minutes. Water (50 mL) was added to the reaction solution, 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 afford compound 46e (450 mg). ESI-MS (m / z): 162.12 [M+H] + .

[0599] Preparation of compound 46

[0600] Referring to the preparation process of compound 5, compound 46e (102.8 mg, 0.64 mmol) was used to replace compound 4-ethynyl-1-methylpyrazole, and the reaction was carried out under microwave at 100°C for 3 hours to obtain compound 46 (7.3 mg). ESI-MS (m / z): 550.8 [M+H] + . 1H NMR (400 MHz, DMSO) δ 8.36 - 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), 2.70 and 2.58 (s, 3H), 2.40 (s, 3H).

[0601] Example 46A: (S)-4-amino-7-fluoro-N, 1 -dimethyl-N-(6-((7-methyl-5, 6,7,8- tetrahydroimidazo[l,2-a]pyrazin-3-yl)ethynyl)-2,3-dihydrobenfuran-3-yl)-lH- pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 46A)

[0602]

[0603] Compound 46A was prepared according to the synthetic procedure described for the preparation of Compound 46 from Compound 46e and Compound 5b-A.

[0604] Compound 46A: ESI-MS (m / z): 551.0 [M+H] + . 1 H NMR (400 MHz, DMSO) δ 8.36 - 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), 2.70 and 2.58 (s, 3H), 2.40 (s, 3H).

[0605] Example 47A: (S)-4-amino-7-fluoro-N, 1 -dimethyl-N-(6-((5-methylpyridin-3- yl)ethynyl)-2,3-dihydrobenfuran-3-yl)-lH-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 47A)

[0606]

[0607] Reference to the procedure for the preparation of Compound 5A, compound 3-ethynyl-5- methylpyridine (23.4 mg, 0.2 mmol) was used in place of compound 4-ethynyl-l- methylpyrazole and the reaction was carried out at 100 °C for 4 h in the microwave to give 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).

[0608] 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)

[0609]

[0610] Referring to the preparation process of compound 5A, 3-ethynylpyridine (52.6 mg, 0.51 mmol) was used to replace 4-ethynyl-1-methylpyrazole. The reaction was carried out under microwave 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).

[0611] Example 49A: (S)-4-amino-N-(6-((1,3-dimethyl-1H-pyrazol-4-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 49A)

[0612]

[0613] Referring to the preparation process of compound 19, compound 4-iodo-1,3-dimethylpyrazole (22.2 mg, 0.1 mmol) was used to replace compound 4-iodo-1-isopropyl-1H-pyrazole, 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 to obtain 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).

[0614] 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-dihydrofuro[3,4-c]quinoline-8-carboxamide (Compound 50A)

[0615]

[0616] Referring to the preparation process of compound 40A, 4-ethynyl-1,3-dimethylpyrazole (75.7 mg, 0.63 mmol) was substituted for 4-ethynyl-1-methylpyrazole. Microwave reaction was performed at 100°C under nitrogen protection 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).

[0617] 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)-1 H- pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 51A)

[0618]

[0619] Reference to the preparation process of Compound 46A, Compound 8b-A (100 mg, 0.21 mmol) was used to replace Compound 5b-A, and microwave reaction was carried out 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 (400 MHz, DMSO) δ 8.43 - 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).

[0620] Example 52A: (S)-4-amino-7-chloro-N-(6-((1,5-dimethyl-1 H-pyrazol-4-yl)ethynyl)- 2,3-dihydrobenzofuran-3-yl)-N-methyl-1,3-dihydrofuro[3,4-c]quinoline-8-carboxamide (Compound 52A)

[0621]

[0622] Reference to the preparation process of Compound 43A, Compound 4-iodo-1,5- dimethylpyrazole (66.2 mg, 0.30 mmol) was used to replace Compound 1 -cyclopropyl- 4-iodopyrazole, and 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).

[0623] 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)

[0624]

[0625] Referring to the preparation process of compound 8A, 1-cyclopropyl-4-iodopyrazole was replaced with 5-iodo-1-methylpyrazole (105.7 mg, 0.51 mmol). The reaction was carried out at room temperature under nitrogen for 16 hours 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).

[0626] 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)

[0627]

[0628] Referring to the preparation process of compound 12A, 2-fluoro-3-iodopyridine (22.3 mg, 0.1 mmol) was substituted for 1-cyclopropyl-4-iodo-1H-pyrazole. 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).

[0629] 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)

[0630]

[0631] Referring to the preparation process of compound 8A, 5-iodo-2-methylpyridine (21.9 mg, 0.10 mmol) was used to replace 1-cyclopropyl-4-iodopyrazole. 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).

[0632] 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)

[0633]

[0634] Referring to the preparation process of compound 8A, 2-fluoro-3-iodopyridine (44.6 mg, 0.20 mmol) was used to replace 1-cyclopropyl-4-iodopyrazole. 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).

[0635] Example 57A: (S)-4-amino-7-chloro-N-methyl-N-(6-((2-methylpyridin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1,3-dihydrofuro[3,4-c]quinoline-8-carboxamide (Compound 57A)

[0636]

[0637] Referring to the preparation process of compound 43A, 3-iodo-2-methylpyridine (31.3 mg, 0.14 mmol) was used to replace 1-cyclopropyl-4-iodopyrazole. The reaction was carried out at room temperature under nitrogen for 3 hours 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).

[0638] Example 58A: (S)-4-amino-7-chloro-N-methyl-N-(6-(6-methylpyridin-3-yl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1,3-dihydrofuro[3,4-c]quinoline-8-carboxamide (Compound 58A)

[0639]

[0640] Referring to the preparation process of compound 43A, 5-iodo-2-methylpyridine (30.7 mg, 0.14 mmol) was used to replace 1-cyclopropyl-4-iodopyrazole. The reaction was carried out at room temperature under nitrogen for 3 hours 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).

[0641] 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)

[0642]

[0643] Referring to the preparation process of compound 12A, 5-iodo-2-methylpyridine (36.9 mg, 0.17 mmol) was used to replace 1-cyclopropyl-4-iodo-1H-pyrazole. 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).

[0644] 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)

[0645]

[0646] Referring to the preparation process of compound 12A, compound 6-iodo-1-methylindazole (41.3 mg, 0.16 mmol) was used to replace compound 1-cyclopropyl-4-iodo-1H-pyrazole. 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 (400 MHz, DMSO-d6) δ 8.39 - 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).

[0647] Example 61A: (S)-4-amino-7-chloro-N-(6-((2-fluoropyridin-3-yl)ethynyl)-2,3- dihydrobenofuran-3-yl)-N-methyl-1,3-dihydrofuro[3,4-c]quinoline-8-carboxamide (Compound 61A)

[0648]

[0649] Reference the procedure for preparing Compound 43A, replace compound 1- cyclopropyl-4-iodopyrazole with compound 2-fluoro-3-iodopyridine (31.2 mg, 0.14 mmol), react for 3 hours at room temperature to obtain Compound 61A (20 mg) ESI-MS (m / z): 515.13 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 8.39 - 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).

[0650] Example 62A: (S)-4-amino-7-chloro-N-methyl-N-(6-((1-methyl-1H-indazol-6-yl)ethynyl)- 2,3-dihydrobenofuran-3-yl)-1,3-dihydrofuro[3,4-c]quinoline-8-carboxamide (Compound 62A)

[0651]

[0652] Referring to the preparation process of compound 43A, 6-iodo-1-methylindazole (36.1 mg, 0.14 mmol) was substituted for 1-cyclopropyl-4-iodopyrazole. The reaction was allowed to proceed 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).

[0653] 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)

[0654]

[0655] Referring to the preparation process of compound 8A, 3-iodo-4-methylpyridine (43.8 mg, 0.2 mmol) was used to replace 1-cyclopropyl-4-iodopyrazole. 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).

[0656] 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)

[0657]

[0658] Referring to the preparation process of compound 8A, 3-iodo-2-methylpyridine (43.8 mg, 0.2 mmol) was used to replace 1-cyclopropyl-4-iodopyrazole. 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).

[0659] 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)

[0660]

[0661] Referring to the preparation process of compound 51A, compound 46e was replaced with 3-ethynyl-5-methylpyridine (23.4 mg, 0.2 mmol). Microwave reaction was carried out at 100°C under nitrogen protection for 4 hours 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).

[0662] 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)

[0663]

[0664] Reference the preparation process of compound 12A, use compound 66a (50.5 mg, 0.20 mmol) to replace compound 1-cyclopropyl-4-iodo-1H-pyrazole, react in microwave at 100 ℃ for three hours to obtain compound 66A (18 mg) ESI-MS (m / z): 536.39 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 8.41-8.22 (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).

[0665] Example 67A: (S)-4-amino-7-chloro-N-(6-((1-(difluoromethyl)-1H-pyrazol-4-yl)ethynyl)-2,3- dihydrobenofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 67A)

[0666]

[0667] Reference the preparation process of compound 8A, use compound 1-(difluoromethyl)-4-iodopyrazole (46.4 mg, 0.19 mmol) to replace compound 1-cyclopropyl-4-iodopyrazole, react at room temperature for 3 hours to obtain compound 67A (8 mg) ESI-MS (m / z): 547.9 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 8.41-8.22 (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).

[0668] 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-dihydrofurano[3,4-c]quinoline-8-carboxamide (Compound 68A)

[0669]

[0670] Referring to the preparation process of compound 43A, compound 24b (38.8 mg, 0.16 mmol) was substituted for 1-cyclopropyl-4-iodopyrazole, 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).

[0671] 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-dihydrofuro[3,4-c]quinoline-8-carboxamide (Compound 69A)

[0672]

[0673] Referring to the preparation process of compound 43A, 1-ethyl-4-iodo-1H-pyrazole (52.9 mg, 0.24 mmol) was substituted for 1-cyclopropyl-4-iodopyrazole. The reaction was allowed to proceed 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).

[0674] Example 70A: (S)-4-amino-7-chloro-N-methyl-N-(6-(pyridin-3-ylethynyl)-2,3-dihydrobenzofuran-3-yl)-1,3-dihydrofuro[3,4-c]quinoline-8-carboxamide (Compound 70A)

[0675]

[0676] Referring to the preparation process of compound 43A, 3-iodopyridine (48.8 mg, 0.24 mmol) was used to replace 1-cyclopropyl-4-iodopyrazole. The reaction was allowed to proceed 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).

[0677] 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-dihydrofuro[3,4-c]quinoline-8-carboxamide (Compound 71A)

[0678]

[0679] Referring to the preparation process of compound 43A, 1-cyclopropyl-4-iodo-5-methyl-1H-pyrazole (59.52 mg, 0.24 mmol, preparation method reference WO2023044171 A1) was substituted for 1-cyclopropyl-4-iodopyrazole. 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).

[0680] 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)

[0681]

[0682] With reference to the preparation method of 9c in WO2024131901 A1, compound 72a was condensed with compound 5c to give compound 72b.

[0683] Under nitrogen, to a solution of compound 72a (48.4 mg, 0.1 mmol) in N,N-dimethylacetamide (2 mL) were added 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) in sequence. The mixture was microwaved at 90°C for 4 hours. Water (50 mL) was added to the reaction solution, which was then 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 removed. Compound 72 (25 mg) was obtained by column chromatography (methanol:dichloromethane = 1:20). 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).

[0684] 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-dihydrofuro[3,4-c]quinoline-8-carboxamide (Compound 73A)

[0685]

[0686] Referring to the preparation process of compound 43A, 1-difluoromethyl-4-iodo-1H-pyrazole (48.8 mg, 0.2 mmol) was substituted for 1-cyclopropyl-4-iodopyrazole. 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).

[0687] 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-dihydrofuro[3,4-c]quinoline-8-carboxamide (Compound 74A)

[0688]

[0689] Referring to the preparation process of compound 43A, compound 66b (40.2 mg, 0.2 mmol) was substituted for 1-cyclopropyl-4-iodopyrazole, 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).

[0690] 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-dihydrofuro[3,4-c]quinoline-8-carboxamide (Compound 75A)

[0691]

[0692] Referring to the preparation process of compound 43A, 3-bromo-6,7-dihydro-5H-pyrrolo[1,2-a]imidazole (37.4 mg, 0.2 mmol) was substituted for 1-cyclopropyl-4-iodopyrazole. 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).

[0693] 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-dihydrofuro[3,4-c]quinoline-8-carboxamide (Compound 76A)

[0694]

[0695] Referring to the preparation process of compound 43A, 6-bromo-1-methylpyrazolo[4,3-b]pyridine (50.5 mg, 0.24 mmol) was used to replace 1-cyclopropyl-4-iodopyrazole. 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).

[0696] 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)

[0697]

[0698] Referring to the preparation process of compound 5A, 2-cyclopropyl-5-ethynylpyridine (70 mg, 0.49 mmol) was used to replace 4-ethynyl-1-methylpyrazole. The reaction was carried out under microwave conditions 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).

[0699] 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-dihydrofuro[3,4-c]quinoline-8-carboxamide (Compound 78A)

[0700]

[0701] Referring to the preparation process of compound 43A, 5-iodo-1-methylpyrazole (59.4 mg, 0.29 mmol) was substituted for 1-cyclopropyl-4-iodopyrazole. The reaction was carried out at room temperature under nitrogen for 3 hours 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).

[0702] 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)

[0703]

[0704] The synthesis process for preparing compound 79A from compound Int-2A was similar to the synthesis 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).

[0705] 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-dihydrofuro[3,4-c]quinoline-8-carboxamide (Compound 80A)

[0706]

[0707] Referring to the preparation process of compound 16A, compound 66a (40.2 mg, 0.2 mmol) was substituted for compound 1-cyclopropyl-4-iodopyrazole, and the reaction was carried out under microwave 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).

[0708] 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-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (Compound 81A)

[0709]

[0710] Referring to the preparation process of compound 8A, 3-iodo-1,4-dimethylpyrazole (35.5 mg, 0.16 mmol) was used to replace 1-cyclopropyl-4-iodopyrazole. The reaction was carried out at room temperature under nitrogen for 16 hours 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).

[0711] 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)

[0712]

[0713] Preparation of compound 82b

[0714] Compound 82a (236.7 mg, 2.57 mmol) was dissolved in N,N-dimethylformamide (10 mL), cooled to 0°C, and sodium hydroxide (205.6 mg, 5.14 mmol, 60% purity) was added. The reaction mixture was reacted at 0°C for 1 hour, and then deuterated iodomethane (745.1 mg, 5.14 mmol) was added. The reaction mixture was stirred at 25°C for 12 hours. The reaction mixture was quenched by adding water (5 mL) and extracted with ethyl acetate (25 mL x 3). The organic phases were washed three times with saturated brine, combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The filtrate was purified by column chromatography (ethyl acetate:petroleum ether = 1:10) to afford compound 82b (250 mg). ESI-MS (m / z): 110.02 [M+H]. + .

[0715] Preparation of Compound 82A

[0716] Referring to the preparation process of compound 5A, compound 82b (21.8 mg, 0.2 mmol) was substituted for compound 4-ethynyl-1-methylpyrazole, and the reaction was carried out under microwave 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).

[0717] 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)

[0718]

[0719] The preparation process of compound 11b-A refers to WO2024131901 A1.

[0720] Preparation of compound 83A

[0721] To a solution of compound 11b-A (60 mg, 0.15 mmol) in N-methylpyrrolidone (2 mL) were added cuprous iodide (5.6 mg, 0.03 mmol), tetrakistriphenylphosphine 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). The mixture was reacted at 25°C under nitrogen for 3 hours. The reaction mixture was filtered through celite, 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 normal phase column chromatography (dichloromethane / methanol = 20:1). The crude compound was then purified by reverse phase column chromatography (acetonitrile / 0.05% ammonium bicarbonate solution = 0:100%–45%:55%) to afford 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).

[0722] 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)

[0723]

[0724] Referring to the preparation process of compound 12A, 3-bromo-6,7-dihydro-5H-pyrrolo[1,2-a]imidazole (45.0 mg, 0.24 mmol) was substituted for 1-cyclopropyl-4-iodo-1H-pyrazole. 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).

[0725] 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)

[0726]

[0727] Preparation of compound 85a

[0728] Under nitrogen, 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). The mixture was microwaved at 90°C for 4 hours. Water (50 mL) was added to the reaction solution, which was then 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 removed. Compound 85a (224 mg) was obtained by column chromatography (methanol:dichloromethane = 1:20). ESI-MS (m / z): 501.7 [M+H]. + .

[0729] Preparation of compound 85b

[0730] To a solution of compound 85a (224 mg, 0.45 mmol) in tetrahydrofuran (5 mL) was added tetrabutylammonium fluoride solution (0.9 mL, 1 M) and 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 combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Column chromatography (methanol:dichloromethane = 1:20) afforded compound 85b (140 mg). ESI-MS (m / z): 430.2 [M+H] + .

[0731] Preparation of compound 85

[0732] Under nitrogen, to a solution of compound 85b (43 mg, 0.1 mmol) in N-methylpyrrolidone (2 mL) were added 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) in sequence. The mixture was reacted in an oil bath at 40°C for 3 hours. Water (100 mL) was added to the reaction solution, which was then 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 removed. The product was purified by column chromatography (methanol:dichloromethane = 1:20) to afford 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).

[0733] 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-dihydrofuro[3,4-c]quinoline-8-carboxamide (Compound 86A)

[0734]

[0735] Referring to the preparation process of compound 40b-A, compound 1a-A was replaced by compound 79b-A to obtain compound 86a-A.

[0736] Referring to the preparation process of compound 43A, compound 86a-A was substituted for compound 40b-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).

[0737] Example 87: 4-amino-7-chloro-N-(7-((1-cyclopropyl-1H-pyrazol-4-yl)ethynyl)isochrome-4-yl)-N-methyl-1,3-dihydrofuro[3,4-c]quinoline-8-carboxamide (Compound 87)

[0738]

[0739] Preparation of compound 87b

[0740] 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 on an ice bath for 5 min, and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (415 mg, 1.09 mmol) was added. Finally, 87a (176.2 mg, 0.73 mmol, preparation method reference WO2024131901 A1) was added. The reaction was continued for 1 hour. Water (100 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 ml*3). 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. Compound 87b (212 mg) was obtained by column chromatography (methanol: dichloromethane = 1:20). ESI-MS (m / z): 488.0 [M+H] + .

[0741] Preparation of Compound 87

[0742] Under nitrogen, to a solution of compound 87b (48.8 mg, 0.1 mmol) in N,N-dimethylacetamide (2 mL) were added 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) in sequence. The reaction mixture was microwaved 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 removed. Compound 87 (30 mg) was obtained by column chromatography (methanol:dichloromethane = 1:20). 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).

[0743] 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-dihydrofuro[3,4-c]quinoline-8-carboxamide (Compound 88A)

[0744]

[0745] Referring to the preparation process of compound 43A, 3-bromo-5,6-dihydro-4H-pyrrolo[2,1-e]pyrazole (44.5 mg, 0.24 mmol) was used to replace 1-cyclopropyl-4-iodopyrazole. The reaction was carried out under microwave conditions 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).

[0746] 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)

[0747]

[0748] Referring to the preparation process of compound 12A, 1-cyclobutyl-4-iodopyrazole (25 mg, 0.1 mmol) was substituted for 1-cyclopropyl-4-iodo-1H-pyrazole. The reaction was carried out at 25°C under nitrogen for 2 hours 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).

[0749] 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-dihydrofurano[3,4-c]quinoline-8-carboxamide (Compound 90A)

[0750]

[0751] Referring to the preparation process of compound 43A, 1-cyclobutyl-4-iodopyrazole (50 mg, 0.20 mmol) was substituted for 1-cyclopropyl-4-iodopyrazole. The reaction was carried out at room temperature under nitrogen for 2 hours 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).

[0752] 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-dihydrofuro[3,4-c]quinoline-8-carboxamide (Compound 91A)

[0753]

[0754] Referring to the preparation process of compound 43A, 3-bromo-4,5,6,7-tetrahydropyrazolo[1,5-a]pyridine (47.8 mg, 0.24 mmol) was substituted for 1-cyclopropyl-4-iodopyrazole. The reaction was carried out under microwave conditions 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).

[0755] Reference Example 1: The preparation method refers to WO2024131901A1.

[0756] Reference Example 2: The preparation method refers to WO2024131901A1.

[0757] Reference Example 3 Preparation method reference WO2022169948 A1

[0758] Reference Example 4 Reference Example 5 Reference Example 6, Preparation method: J. Med. Chem. 2022, 65(3), 1749-1766.

[0759] Reference Example 7 The preparation method is described in WO2022132914 A1.

[0760] Test Example 1 Inhibitory activity of the compounds of the present invention on the proliferation of human colon cancer cells

[0761] 1. Build MTAP - / - HCT-116 cell line

[0762] The colon cancer cell line HCT-116 (purchased from Nanjing Kebai) was co-transfected with the gene editing tool CRISPR / Cas9 and sgRNA, and the MTAP double-allelic inactivation gene 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 synergistic mechanism of MTAP deficiency and PRMT5, including symmetric dimethylarginine (SDMA) level test and cell proliferation test.

[0763] 2. Cell proliferation

[0764] MTAP was cultured in complete medium McCoy's 5A (Gibco, 16600082) / 10% FBS (Gibco, 10099141C) / 1% p / s (Gibco, 15140122). - / - HCT-116 and wild-type HCT-116 cell lines, evaluate the effects of compounds on MTAP - / - Selective inhibition and enhancement of HCT-116 cell proliferation. Day 0, 100 MTAPs 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. Compounds were diluted 3x in a series of dilutions (starting at 20 μM, 3X dilution, a total of 8 concentration points) using DMSO (Sigma, D5879). On Day 1, the compounds were diluted to multiple concentration points and treated with the cells, and the cells were cultured in a 37°C, 5% CO2 incubator for 10 days. On Day 11, 20 μL of MTS (CellTiter A Queous One Solution Cell Proliferation Assay (Promega, G3581) was cultured in a 37°C, 5% CO2 incubator for 2 hours and then read on a Tecan Spark (OD = 490 nM). The data were then analyzed using GraphPad Prism 8 software using the equation "log (inhibitor) vs. normalized response--variable slope" (formula Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC 50 -X)*HillSlope))) perform data analysis to obtain the IC value of the compound 50 Where Y is the inhibition rate, X is the logarithm of the compound concentration, Top refers to the maximum response (inhibition rate when the compound concentration is the maximum), Bottom refers to the baseline response (inhibition rate when the compound concentration is 0), and Hill Slope refers to the IC 50 The slope of the curve, IC 50 The experimental results are shown in Table 1.

[0765] Table 1 IC 50 (μM)

[0766]

[0767]

[0768] The results showed that the specific compounds and their isomers exemplified in the present invention had strong proliferation inhibitory activity against the human MTAP-deficient HCT-116 cell line, and the IC values ​​of most of the exemplified compounds against the human MTAP-deficient HCT-116 cell line were 0. 50 The values ​​were all less than 0.01 μM. In terms of selectivity, the specific compounds and their isomers exemplified in the present invention had good selective inhibition against the human MTAP-deficient HCT-116 cell line relative to the MTAP wild-type (WT) HCT-116 cell line, with most compounds having a selectivity of more than 90 times.

[0769] Test Example 2 Human liver microsome stability test

[0770] Experimental Materials: Human liver microsomes (purchased from BIOIVT) were used in the experiment;

[0771] Reagent preparation:

[0772] PBS: 0.1 M KH2PO4 and K2HPO4 buffer, pH 7.4.

[0773] MgCl2: Weigh a certain amount of MgCl2 and prepare a 16mM MgCl2 solution with PBS.

[0774] NADPH (Chinese name: reduced nicotinamide adenine dinucleotide phosphate, purchased from Sigma, product number: 481973-500mg): Weigh a certain amount of NADPH and prepare NADPH to 4mM with 16mM MgCl2 solution, and the final incubation concentration is 1mM.

[0775] Compounds: Test compounds and positive compounds were prepared to 4 μM in PBS, with a final incubation concentration of 1 μM.

[0776] 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.

[0777] Experimental steps:

[0778] Add the prepared test compound or positive compound to the test tube, then add the prepared NADPH and mix well. Place in a 37°C, 220rpm constant temperature incubator for pre-incubation. After pre-incubation for 5 minutes, add the prepared liver microsomes to start the reaction, and perform duplicate operations. At 0min, 15min, 30min, 45min, and 60min, add a certain volume of ice acetonitrile solution containing internal standard to the corresponding tube to precipitate protein, vortex for 5min, then centrifuge at 4000rpm for 10min, and take the supernatant in a 96-well plate. Put it into LC-MS / MS for analysis. The concentration (peak area ratio) of the example compound was determined by LC-MS / MS, and the rate constant was obtained by plotting "Ln (compound residual amount %)" against "incubation time" in Excel, thereby calculating the half-life and intrinsic clearance of the drug, providing a basis for predicting the clearance rate in vivo. The results of this experiment are shown in Table 2.

[0779] Data Analysis:

[0780] CL int =(0.693 / t 1 / 2, microsomes) × [incubation medium volume (mL) / microsomal protein mass (mg)] × [microsomal protein mass (mg) / liver mass (g)] × [liver mass (g) / body weight (kg)] [1]

[0781] CL H =CL int ×f u ×Q h / (CL int ×f u +Q h )

[0782] Where,

[0783] CL int --Intrinsic clearance (mL / min / kg)

[0784] CL H --Hepatic clearance (mL / min / kg)

[0785] f u --The fraction of plasma protein unbound is 1

[0786] Q h --Hepatic blood flow

[0787] Table 2 Stability of human liver microsomes

[0788] Compound T1 / 2(min) Compound T1 / 2(min) Reference Example 1 16.6 Example 4 169 Reference Example 2 33.6 Compound 5A 66.5 Reference Example 5 25.6 Compound 25A 68.9 Reference Example 7 28.9 Compound 40A 62.9 Compound 50A 49.9 Compound 43A 122 Compound 91A 47.2

[0789] Table 2 shows that the compounds of the present invention have excellent stability in human liver microsomes.

[0790] Experimental Example 3 hERG test

[0791] Experimental reagents:

[0792] Predictor TM hERG Fluorescence Polarization Assay Kit (ThermoFisher Scientific, PV5365).

[0793] Experimental steps:

[0794] Follow the kit instructions: pre-prepare 4×Predictor TM hERG Tracer Red, 4×E-4031, 4×test solution (final concentration is 10 μM). In a 384-well plate, 5 μL Predictor was added to the Assay Blank group. TMhERG FP Assay Buffer and 5μL Predictor TM hERG Membrane. 2.5 μL Predictor TM hERG FP Assay Buffer, 2.5 μL 4×E-4031, 2.5 μL 4×test substance, and then 5 μL Predictor TM hERG Membrane and 2.5 μL 4×Predictor TM hERG Tracer Red. The total experimental volume was 10 μL, with triplicate wells per group. After sample addition, gently tap to mix thoroughly. Incubate at room temperature (25°C) in the dark for 3.5 hours. After incubation, measure fluorescence polarization values ​​(mP) using a microplate reader. The G factor was 2.14.

[0795] Data Analysis:

[0796] The inhibition rate of the test substance was calculated using the following formula:

[0797] Inhibition rate % = (mP negative control - mP test substance) / (mP negative control - mP positive control) * 100%

[0798] Using the above experimental method, the hERG inhibition test results of some compounds of the present invention are shown in Table 3 below.

[0799] Table 3 hERG test

[0800]

[0801] 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).

[0802] Test Example 4 CYP inhibition test

[0803] Experimental Materials: Human liver microsomes (purchased from BIOIVT) were used in the experiment;

[0804] Reagent preparation:

[0805] PBS: 0.1 M KH2PO4 and K2HPO4 buffer, pH 7.4.

[0806] MgCl2: Weigh a certain amount of MgCl2 and prepare a 20mM MgCl2 solution with PBS.

[0807] NADPH (Chinese name: reduced nicotinamide adenine dinucleotide phosphate, purchased from Sigma, product number: 481973-500mg): Weigh a certain amount of NADPH and prepare NADPH to 5mM with 20mM MgCl2 solution, and the final incubation concentration is 1mM.

[0808] Compounds: Test compounds were prepared with DMSO-methanol (v:v = 1:9) solution at concentrations of 3, 1, 0.3, 0.1, 0.03, 0.01, and 0 mM, with final incubation concentrations of 30, 10, 3, 1, 0.3, 0.1, and 0 μM. Positive compound (ketoconazole) was prepared with DMSO-methanol (v:v = 1:9) solution at concentrations of 30, 10, 3, 1, 0.3, and 0 μM, with final incubation concentrations of 0.3, 0.1, 0.03, 0.01, 0.003, and 0 μM.

[0809] Substrate working solution: Prepare substrate working solutions (testosterone and midazolam) to 200 and 15 μM, respectively, in PBS, with final incubation concentrations of 40 and 3 μM.

[0810] Liver microsomes (purchased from BIOIVT, catalog number: X008070): Liver microsomes were diluted to 0.17 mg / mL with PBS, and the final incubation concentration was 0.1 mg / mL.

[0811] Experimental steps:

[0812] Add the prepared test compound or positive compound to the 96-well plate, then add the prepared human liver microsome working solution and substrate working solution and mix well. Place in a 37°C constant temperature incubator for pre-incubation. After pre-incubation for 5 minutes, add the prepared NADPH to start the reaction. Incubate for 5 or 10 minutes. After the incubation, add pre-cooled acetonitrile solution containing internal standard to terminate the reaction. After protein precipitation, vortex for 5 minutes, then centrifuge at 4000rpm for 10 minutes, and take the supernatant to a 96-well plate. The metabolites of the substrate are detected by LC-MS / MS method (peak concentration ratio). The residual activity of the enzyme and the half inhibitory concentration (IC50) are calculated based on the detected content. 50 ).

[0813] Data Analysis:

[0814] Percentage of remaining enzyme activity:

[0815] The activity of each CYP450 subtype enzyme is expressed as the amount of metabolite generated, and the percentage of residual enzyme activity is calculated as follows:

[0816] Residual activity (%) = (activity of the CYP450 enzyme subtype in the presence of the test substance / activity of the solvent control group) × 100%, where the activity of the solvent control group (0 μM) is set as 100%.

[0817] IC50 :

[0818] The concentration of the test substance and the corresponding residual activity percentage were fitted using a nonlinear model using Excel software. The concentration of the test substance that inhibited the formation of metabolites by 50% was the IC 50 The experimental results are shown in Table 4.

[0819] Table 4

[0820]

[0821] Table 4 shows that the compounds of the present invention have lower CYP inhibition toxicity compared to the prior art (such as Reference Example 6).

[0822] Experimental Example 5 Mouse PK Experiment Experimental Materials: The experimental animals were healthy adult BALB / c female mice (provided by Sichuan Weitong Lihua Experimental Animal Technology Co., Ltd.);

[0823] Test process:

[0824] Dosing method and sample collection in mice: BALB / c female mice were orally gavaged with the drug (10 mg / kg, solvent: 0.5% methylcellulose / 0.2% Tween 80). 60 μL of whole blood was collected from the fundus venous plexus of the mice at different time points at 0.25, 0.5, 1, 2, 4, 6, 8, and 24 h after administration, and the plasma was obtained by centrifugation at 4000 rpm for 6 min.

[0825] Sample analysis:

[0826] 10 μL of mouse plasma samples were collected, and 190 μL of acetonitrile solution containing an internal standard was added to precipitate proteins. The samples were vortexed for 10 minutes and then centrifuged at 4000 rpm for 10 minutes. The supernatant was collected and transferred to a 96-well plate for analysis by LC-MS / MS. LC-MS / MS was used to determine plasma drug concentrations at different times after oral administration of the example compounds to mice. The relevant pharmacokinetic parameters were calculated to investigate the pharmacokinetic behavior of the compounds in mice and to evaluate their pharmacokinetic profile. The experimental results are shown in Table 5.

[0827] Table 5 Pharmacokinetic parameters of the compounds of the present invention in mice

[0828]

[0829]

[0830] Note: 1. The solvent is 0.5% MC / 0.2% Tween 80, PO administration

[0831] 2. The PK test method of MRTX1719 in mice is shown in J. Med. Chem. 2022, 65(3), 1749-1766.

[0832] Table 5 shows that the compounds provided by the present invention have excellent pharmacokinetic properties in mice.

[0833] Test Example 6 Rat PK experiment

[0834] Experimental materials: The experimental animals were healthy adult SD male rats (provided by Sichuan Weitonglihua Experimental Animal Technology Co., Ltd.)

[0835] Test process:

[0836] Administration and sample collection in rats: SD male rats were orally gavaged with the drug (10 mg / kg, solvent: 0.5% methylcellulose / 0.2% Tween 80). 200 μL of whole blood was collected from the retinal venous plexus of the rats at different time points at 0.25, 0.5, 1, 2, 4, 6, 8, and 24 h after administration, and the blood was centrifuged at 4000 rpm for 6 min to obtain plasma.

[0837] Sample analysis:

[0838] 10 μL of rat plasma samples were collected, and 190 μL of acetonitrile solution containing an internal standard was added to precipitate proteins. The samples were vortexed for 10 minutes and then centrifuged at 4000 rpm for 10 minutes. The supernatant was collected and transferred to a 96-well plate for analysis using LC-MS / MS. LC-MS / MS was used to determine plasma drug concentrations at different times after oral administration of the example compounds to rats. The relevant pharmacokinetic parameters were calculated to investigate the pharmacokinetic behavior of the compounds in rats and to evaluate their pharmacokinetic profiles. The experimental results are shown in Table 6.

[0839] Table 6 Pharmacokinetic parameters in rats

[0840]

[0841] Note: 1. The solvent is 0.5% MC / 0.2% Tween 80, PO administration

[0842] Table 6 shows that the compounds provided by the present invention have excellent pharmacokinetic properties in rats.

[0843] Experimental Example 7: Evaluation of drug efficacy in a subcutaneous xenograft mouse model of gene-edited MTAP- / - human colon cancer cell line HCT116

[0844] Experimental Materials:

[0845] 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);

[0846] The test substance was administered as a suspension at concentrations of 0.3 and 1 mg / mL using a 0.5% MC + 0.2% Tween 80 aqueous solution. Reference Example 6 (MRTX1719) was administered as a clear solution at a concentration of 5 mg / mL using a 40% PEG400 saline solution containing 2% DMSO. A blank control group received a 40% PEG400 saline solution containing 2% DMSO.

[0847] Experimental methods:

[0848] HCT116 MTAP- / - cells in the logarithmic growth phase were digested and collected, counted, and serum-free medium was used to adjust the cell density to 1×10 8 20-22g Balb / c nude mice (Beijing Huafukang Biotechnology Co., Ltd.) were subcutaneously inoculated with 100 μL of cell suspension per mouse. On the 10th day after inoculation, the tumor volume reached 100-150 mm 3 Animals were randomly divided into groups of 8 according to body weight and tumor volume. The test substance was administered orally at a dose of 10 mg / kg once daily; MRTX1719 was administered orally at a dose of 50 mg / kg once daily. The model control was a vehicle (2% DMSO + 40% PEG400 + 58% saline) administered orally at a dose of 10 mL / kg once daily for a total of 28 days.

[0849] After the animals were inoculated with HCT116 MTAP- / - cells, the body weight and tumor volume of the mice were measured 2-3 times a week. The tumor volume was calculated as length × width. 2 The mice were euthanized on the day of the last administration.

[0850] The tumor inhibition rate TGI of each drug-treated group was calculated according to the formula:

[0851] 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 is the tumor volume of the treatment group at each measurement, V T0 V is the tumor volume measured when the treatment groups were divided; Ct The tumor volume of each measurement in the vehicle group (V Tt Same time), VC0 The resulting tumor volumes were measured when the groups were divided into vehicle groups.

[0852] Data were statistically analyzed using GraphPad Prism 9.0 software and expressed as Mean ± SEM (standard error). Pairwise comparisons between the treatment groups and the control group were analyzed using one-way analysis of variance (ANOVA), with homogeneity of variance tested using the Brown-Forsythe method and unequal variance tested using the Dunnett's T3 method.

[0853] Table 7: Evaluation of the efficacy of HCT116 in subcutaneous xenograft mouse models

[0854]

[0855]

[0856] As shown in Table 7, the in vivo anti-tumor effect of the compound of the present invention in the HCT116 subcutaneous xenograft mouse model was significantly better than that of the clinical molecule MRTX1719.

[0857] Experimental Example 8: Evaluation of drug efficacy in a mouse model transplanted with human lung giant cell carcinoma LU99

[0858] Experimental Materials:

[0859] Human lung giant cell carcinoma cell line LU99 was purchased from Nanjing Kebai Biotechnology Co., Ltd. (CBP61512) and cultured in RPMI-1640 medium (Gibco) containing 10% fetal bovine serum (Gibco);

[0860] The test substance was prepared into suspensions at concentrations of 0.3, 1, and 5 mg / mL using 0.5% MC aqueous solution containing 0.2% Tween 80. Reference Example 6 (MRTX1719) was prepared into a clear solution at a concentration of 5 mg / mL using 40% PEG400 saline containing 2% DMSO.

[0861] Experimental methods:

[0862] LU99 cells in the logarithmic growth phase were digested and collected, counted, and then mixed with serum-free medium and Matrigel (Corning) in a 1:1 ratio to adjust the cell density to 2 × 10 7 18-20 g Balb / c nude mice (Beijing Huafukang Biotechnology Co., Ltd.) were subcutaneously inoculated with 100 μL of cell suspension per mouse. On the 12th day after inoculation, the average tumor volume reached 116 mm 3Animals were randomly divided into groups of seven. The test substance was administered orally at 3, 10, or 50 mg / kg once daily; MRTX1719 was administered orally at 50 mg / kg once daily. A blank group received a 10 mL / kg dose of vehicle (2% DMSO + 40% PEG400 + 58% saline) orally once daily. Dosing lasted a total of 28 days.

[0863] After the animals were inoculated with LU99 cells, the tumor volume was measured 2 to 3 times a week. The tumor volume was calculated as length × width. 2 ×0.5. On the day of the last administration, all mice were euthanized except for the mice with tumor regression, which were then observed after drug withdrawal.

[0864] The tumor inhibition rate TGI of each drug-treated group was calculated according to the formula:

[0865] 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 is the tumor volume of the treatment group at each measurement, V T0 V is the tumor volume measured when the treatment groups were divided; Ct The tumor volume of the model control group at each measurement was V C0 The tumor volume was measured when grouping into the model control group;

[0866] Data were statistically analyzed using GraphPad Prism 9.0 software and expressed as Mean ± SEM (standard error). Pairwise comparisons between each drug-treated group and the model control group were analyzed using one-way analysis of variance (ANOVA). Homogeneity of variance was determined using the Brown-Forsythe method, while inequality of variance was determined using the Dunnett's T3 method.

[0867] Table 8 Evaluation of drug efficacy in LU99 transplantation mouse model

[0868]

[0869] Experimental results and analysis:

[0870] The experimental results are shown in Table 8. The compound of the present invention significantly reduced tumor volume at doses of 3, 10, and 50 mg / kg (P < 0.0001). The anti-tumor effect of the test compound at doses of 10 and 50 mg / kg was significantly superior to that of MRTX1719 at 50 mg / kg, and the test compound at 50 mg / kg caused tumor regression.

[0871] In summary, the experimental results show that: in the human lung giant cell carcinoma LU99 CDX model, the test substance of the present invention has a good anti-tumor effect, and its anti-tumor efficacy at a lower dose is significantly better than that of Yangshen.

[0872] Industrial applicability

[0873] The compounds of the present invention have excellent PRMT5 inhibitory activity and, relative to the MTAP wild-type (WT) HCT-116 cell line, have excellent selective inhibition 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.

[0874] Furthermore, tests such as human liver microsome stability, mouse pK, rat pK, and in vivo antitumor efficacy evaluation in mice revealed that the compounds of the present invention have excellent antitumor application prospects.

[0875] 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 inhibitory toxicity in terms of hERG and CYP).

[0876] In summary, the compounds of the present invention have achieved unexpected technical effects in terms of drug development and safety compared to the existing technologies and have great development potential.

[0877] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

[0878] References

[0879] [1]Davies B, Morris T.Physiological parameters in laboratory animals and humans.Pharm Res.1993;10:1093-5.

Claims

1. A compound represented by formula I or a pharmaceutically acceptable salt thereof: in, Group Selected from Where W is selected from CR 7 , R 7 Selected from hydrogen, halogen, C 1-6 alkyl; Group Selected from R 1 Selected from C 1-4 Alkyl, C 3-6 Monocyclic alkyl; R 2 is selected from 5-10 membered heteroaryl and 4-10 membered heterocyclic group, wherein the 5-10 membered heteroaryl and 4-10 membered heterocyclic group each independently contain 1-4 heteroatoms selected from N, O or S; the 5-10 membered heteroaryl and 4-10 membered heterocyclic group are unsubstituted or replaced by one or more identical or different R b replace; Each R b Each independently selected from deuterium, halogen, C1-C6 alkyl, C 1-6 Alkoxy, C3-C6 cycloalkyl, the C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl is unsubstituted or each independently substituted by one or more substituents selected from halogen; R 3 Selected from hydrogen, halogen, C 1-6 alkyl.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: Group Selected from:

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: Group Selected from 4. The compound according to claim 3 or a pharmaceutically acceptable salt thereof, characterized in that: Group Selected from:

5. The compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof, characterized in that: R 3 Selected from hydrogen, F, Cl, Br, C 1-4 alkyl.

6. The compound according to claim 5 or a pharmaceutically acceptable salt thereof, characterized in that: R 3 Selected from hydrogen, F, Cl, methyl, ethyl.

7. The compound according to claim 6 or a pharmaceutically acceptable salt thereof, characterized in that: R 3 Selected from hydrogen, F, methyl.

8. The compound according to claim 7 or a pharmaceutically acceptable salt thereof, characterized in that: Group Selected from 9. The compound according to claim 8 or a pharmaceutically acceptable salt thereof, characterized in that: Group Selected from 10. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: R 2 is selected from 5-6 membered heteroaryl, 4-6 membered heterocyclyl, 5-6 membered heteroaryl and 5-6 membered heterocyclyl; the 5-6 membered heteroaryl, 4-6 membered heterocyclyl and 5-6 membered heterocyclyl each independently contain 1, 2 or 3 heteroatoms selected from N, O or S; the 5-6 membered heteroaryl, 4-6 membered heterocyclyl, 5-6 membered heteroaryl and 5-6 membered heterocyclyl are unsubstituted or are each independently replaced by one or more identical or different R b replace.

11. The compound according to claim 10 or a pharmaceutically acceptable salt thereof, characterized in that: R 2 is selected from 5-6 membered heteroaryl, wherein the 5-6 membered heteroaryl contains 1, 2 or 3 N heteroatoms; the 5-6 membered heteroaryl is unsubstituted or replaced by one or more identical or different R b replace.

12. The compound according to claim 11 or a pharmaceutically acceptable salt thereof, characterized in that: R 2 is selected from pyrazolyl, imidazolyl, pyridinyl, which are unsubstituted or each independently substituted with 1, 2 or 3 R b replace.

13. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: R 2 is selected from 5-6 membered heteroaryl and 5-6 membered heterocyclyl, 5-6 membered heteroaryl and 5-6 membered heteroaryl, 5-6 membered heteroarylphenyl, said 5-6 membered heteroaryl and 5-6 membered heterocyclyl each independently containing 1, 2 or 3 N heteroatoms; said 5-6 membered heteroaryl and 5-6 membered heterocyclyl, 5-6 membered heteroaryl and 5-6 membered heteroaryl, 5-6 membered heteroarylphenyl is unsubstituted or replaced by one or more identical or different R b replace.

14. The compound according to claim 13 or a pharmaceutically acceptable salt thereof, characterized in that: R 2 is selected from 5-6 membered heteroaryl and 5-6 membered heterocyclic group, 5-6 membered heteroaryl and 5-6 membered heteroaryl, 5-6 membered heteroaryl and phenyl, the 5-6 membered heteroaryl is selected from imidazolyl, pyrazolyl, pyridyl, the 5-6 membered heterocyclic group is selected from pyrrolidinyl, piperidinyl, piperazinyl; the 5-6 membered heteroaryl and 5-6 membered heterocyclic group, 5-6 membered heteroaryl and 5-6 membered heteroaryl, 5-6 membered heteroaryl and phenyl is unsubstituted or replaced by one or more identical or different R b replace.

15. The compound according to claim 14 or a pharmaceutically acceptable salt thereof, characterized in that: R 2 is selected from imidazopyrrolidinyl, imidazopiperidinyl, imidazopiperazinyl, pyrazolophenyl, pyrazolopyridinyl, wherein the imidazopyrrolidinyl, imidazopiperidinyl, imidazopiperazinyl, pyrazolophenyl, pyrazolopyridinyl is unsubstituted or substituted by one or more identical or different R b replace.

16. The compound according to claim 10 or a pharmaceutically acceptable salt thereof, characterized in that: R 2 is selected from 4-6 membered heterocycloalkyl and 4-6 membered heterocycloalkenyl; the 4-6 membered heterocycloalkyl and 4-6 membered heterocycloalkenyl each independently contain 1-3 heteroatoms selected from N, O or S; the 4-6 membered heterocycloalkyl and 4-6 membered heterocycloalkenyl are unsubstituted or are each independently replaced by 1, 2 or 3 identical or different R b replace.

17. The compound according to claim 16 or a pharmaceutically acceptable salt thereof, characterized in that: R 2 is selected from 4-6 membered heterocycloalkyl and 4-6 membered heterocycloalkenyl; the 4-6 membered heterocycloalkyl and 4-6 membered heterocycloalkenyl each independently contain 1-2 heteroatoms selected from N or O; the 4-6 membered heterocycloalkyl and 4-6 membered heterocycloalkenyl are unsubstituted or are each independently substituted by 1, 2 or 3 identical or different R b replace.

18. The compound according to claim 17 or a pharmaceutically acceptable salt thereof, characterized in that: R 2 Selected from described is unsubstituted or each independently substituted with 1, 2 or 3 identical or different R b replace.

19. The compound according to any one of claims 1, 10-18, or a pharmaceutically acceptable salt thereof, characterized in that: Each R b Each is independently selected from H, deuterium, halogen, C1-C3 alkyl, C1-C3 alkoxy, C3-C5 cycloalkyl, and the C1-C3 alkyl, C1-C3 alkoxy, C3-C5 cycloalkyl is unsubstituted or is independently substituted by 1, 2, 3 or 4 halogens.

20. The compound according to claim 19 or a pharmaceutically acceptable salt thereof, characterized in that: 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.

21. The compound according to claim 20 or a pharmaceutically acceptable salt thereof, characterized in that: Each R b Each is independently selected from H, deuterium, F, Cl, methyl, ethyl, isopropyl, methoxy, cyclopropyl, -CHF2, -CF3.

22. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The R b Can be further deuterated.

23. The compound according to claim 22 or a pharmaceutically acceptable salt thereof, characterized in that: Each R b Each is independently selected from H, deuterium, F, Cl, methyl, ethyl, isopropyl, methoxy, cyclopropyl, -CHF2, -CF3, -CD3.

24. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: R 2 Selected from 25. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that Group Selected from:

26. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: R 7 Selected from hydrogen, halogen, C 1-4 alkyl.

27. The compound according to claim 26 or a pharmaceutically acceptable salt thereof, characterized in that: R 7 Each is independently selected from hydrogen, F, Cl, and methyl.

28. The compound according to claim 1, 26 or 27, or a pharmaceutically acceptable salt thereof, characterized in that: R 7 Can be further deuterated.

29. The compound according to claim 28 or a pharmaceutically acceptable salt thereof, characterized in that: R 7 Selected from hydrogen, F, Cl, methyl, -CD3.

30. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: Group Selected from:

31. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: R 1 Selected from methyl, ethyl, cyclopropyl.

32. The compound according to claim 31 or a pharmaceutically acceptable salt thereof, characterized in that: R 1 Selected from methyl.

33. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: R 1 Selected from -CD3.

34. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: Formula (I) has the structure described in Formula IA: Among them, R 1 、R 2 、R 3 ,A,n1,n2,ring M,W,X,Y,Z, Definitions are each as defined in formula I.

35. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound represented by formula (I) is selected from:

36. The compound according to claim 35 or a pharmaceutically acceptable salt thereof, characterized in that: It has the following S configuration:

37. A pharmaceutical composition for treating and / or preventing diseases associated with abnormal PRMT5 expression, characterized in that: The pharmaceutical composition comprises a therapeutically and / or prophylactically effective amount of the compound according to any one of claims 1 to 36 or a pharmaceutically acceptable salt thereof, and optional pharmaceutical excipients.

38. Use of the compound according to any one of claims 1 to 36 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating and / or preventing diseases associated with abnormal PRMT5 expression.

39. The use according to claim 38, wherein the disease associated with abnormal PRMT5 expression is tumor or cancer.

40. The use according to claim 38, wherein the disease associated with abnormal expression of PRMT5 is a disease associated with abnormal expression of PRMT5 caused by MTAP deficiency. The use according to claim 38 , wherein the disease associated with abnormal PRMT5 expression is a tumor or cancer with MTAP deficiency.

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