Alkynyl-substituted heterocyclic amide derivatives, and preparation method and application thereof

Abstract

The invention relates to an alkynyl-substituted heterocyclic amide derivative as shown in a formula (I), and a preparation method and application of the alkynyl-substituted heterocyclic amide derivative. The invention also relates to a pharmaceutical composition containing the compound as an active ingredient and application of the compound or the pharmaceutical composition in treating and / or preventing related diseases mediated by PRMT5. # imgabs0 #

Description

A class of heterocyclic amide derivatives containing alkynyl substitution, preparation method and application thereof Technical Field

[0001] The present invention relates to the field of medicinal chemistry, and in particular to a class of heterocyclic amide derivatives containing alkynyl substitution, a preparation method thereof and an application thereof. Background Art

[0002] Epigenetic regulation of gene expression plays an important biological role in protein maturation and cell differentiation, and plays a crucial role in many human diseases. Arginine guanidinomethylation, catalyzed by protein arginine methyltransferase (PRMT), is a common post-translational modification in eukaryotic cells, affecting a variety of biological processes such as cell signaling, gene transcription, mRNA translation, DNA recombination and repair (Cell Mol. Life Sci. 2015. 72(11): 2041-2059).

[0003] PRMT5 is a member of the PRMT family, and its methylation plays an important role in maintaining normal cellular homeostasis. However, more and more studies have shown that abnormal expression of PRMT5 is associated with the occurrence of various tumors. It is overexpressed in many tumors and has different mechanisms in different tumors (Cell Mol. Life Sci. 2015. 72(11): 2041-2059).

[0004] Homozygous deletion of tumor suppressor genes is a key driver of tumorigenesis. Loss of the tumor suppressor gene CDKN2A, located on human chromosome 9p21, is one of the most frequently mutated genes in tumors, occurring in 15%. Due to its close proximity to CDKN2A, the methylthioadenosine phosphorylase (MTAP) gene is frequently concomitantly deleted in tumors. MTAP plays a crucial role in the methionine and adenine salvage pathway (Cell Reports, 2016, 15:574–587). Loss of MTAP leads to the accumulation of its substrate, methylthioadenosine (MTA). Due to its structural similarity to S-adenosylmethionine (SAM), MTA selectively competes with SAM for binding to PRMT5, partially inhibiting PRMT5 activity and sensitizing further PRMT5 inhibition, resulting in 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 tumor treatment.

[0007] Summary of the Invention

[0008] Problems to be solved by the invention:

[0009] Although several patent applications for PRMT5 inhibitors have been published, new compounds still need to be developed, given the huge market demand for MTAP- / - cancer patients and the suboptimal clinical efficacy and hematotoxicity risks of existing PRMT5 inhibitors. Through continuous efforts, the inventors of this application have designed compounds having the structure represented by general formula (I) and found that compounds having such structures exhibit excellent PRMT5 inhibitory effects and functions, as well as excellent pharmacokinetic properties, and have great prospects for medical applications.

[0010] Solutions used to solve the problem:

[0011] In order to solve the above problems, the inventors of the present application have conducted intensive research and found that a class of heterocyclic amide derivatives containing alkynyl substitution can achieve the desired purpose, resulting in the completion of the present invention.

[0012] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0013] A compound represented by formula I, its tautomers, stereoisomers, pharmaceutically acceptable salts or deuterated compounds thereof:

[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;

[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 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: hydrogen, C 1-6 Alkyl, C 3-6 Cycloalkyl, the C 1-6 Alkyl, C 3-6 The cycloalkyl group is unsubstituted or substituted by one or more identical or different R a 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 a Each independently selected from hydroxy, deuterium, cyano, halogen, -NR'R", optionally substituted by 1, 2, 3 or 4 halogen, hydroxy, cyano, C 1-6 Alkoxy-substituted C 1-6 Alkoxy; wherein R' and R" are each independently selected from hydrogen, C 1-6Alkyl, 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] Herein, 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 embodiments, n1 and n2 are each independently 0 or 1.

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

[0035] In some embodiments, the group Selected from

[0036] In some embodiments, the group Selected from:

[0037] In some embodiments, A is selected from -CH2- or O.

[0038] In some embodiments, the group Selected from

[0039] In some embodiments, the group Selected from:

[0040] Preferably, the group Selected from:

[0041] Preferably, the group Selected from:

[0042] In some specific embodiments, R 2 Selected from hydrogen, C 1-4 Alkyl, C 5-6 Alkyl, C 3-4 Cycloalkyl, C 5- 6 cycloalkyl, the C 1-4 Alkyl, C 5-6 Alkyl, C 3-4 Cycloalkyl, C 5-6 The cycloalkyl group is unsubstituted or substituted by one or more identical or different R a replace.

[0043] In some specific embodiments, R 2 Selected from hydrogen, C 1-4 Alkyl, C 3-4 Cycloalkyl, the C 1-4 Alkyl, C 3-4 The cycloalkyl group is unsubstituted or substituted by one or more identical or different R a replace.

[0044] In some specific embodiments, R 2 is selected from hydrogen, methyl, ethyl, propyl, isopropyl, butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, wherein the methyl, ethyl, propyl, isopropyl, butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl is unsubstituted or replaced by one or more identical or different R a replace.

[0045] In some specific embodiments, R 2 is selected from hydrogen, methyl, ethyl, propyl, cyclopropyl, cyclobutyl, cyclopentyl, said methyl, ethyl, cyclopropyl, cyclobutyl, cyclopentyl being unsubstituted or substituted by one or more identical or different R a replace.

[0046] In some specific embodiments, R 2 is selected from hydrogen, methyl, ethyl or cyclopropyl, said methyl, ethyl or cyclopropyl being unsubstituted or substituted by 1, 2 or 3 identical or different R a replace.

[0047] In some specific embodiments, R 2is selected from methyl or ethyl, the methyl group being unsubstituted or substituted by 1, 2 or 3 identical or different R a replace.

[0048] In some specific embodiments, R 2 isopropyl, which is unsubstituted or substituted by 1, 2 or 3 identical or different R a replace.

[0049] In some specific embodiments, R 2 is selected from cyclopropyl, said cyclopropyl being unsubstituted or substituted by 1, 2 or 3 identical or different R a replace.

[0050] In some specific embodiments, R 2 For hydrogen.

[0051] In some embodiments, each R a Each independently selected from hydroxyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, NR'R", wherein R' and R" are independently selected from hydrogen, C 1-6 Alkyl, halogenated C 1-6 alkyl.

[0052] In some embodiments, each R a Each independently selected from hydroxyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, NR'R", wherein R' and R" are independently selected from hydrogen, C 1-4 alkyl.

[0053] In some embodiments, each R a Each is independently selected from hydroxy, methoxy, ethoxy, propoxy, isopropoxy, butoxy, halogenated methoxy, halogenated ethoxy, halogenated propoxy, halogenated isopropoxy, halogenated butoxy, NR'R", wherein R' and R" are each independently selected from hydrogen, methyl, ethyl, propyl, isopropyl, and butyl.

[0054] In some embodiments, each R a Each is independently selected from hydroxy, methoxy, ethoxy, fluoro- or chloro-substituted methoxy, fluoro- or chloro-substituted ethoxy, NR'R", wherein R' and R" are each independently selected from hydrogen, methyl or ethyl.

[0055] In some embodiments, each R a Each is independently selected from hydroxy, methoxy, -OCF3, -OCHF2, and N,N-dimethylamino.

[0056] Preferably, each R a Each independently selected from hydroxyl, methoxy, -OCF3, -OCHF2, N,N-dimethylamino, N-methylamino

[0057] Preferably, each R a Each independently selected from hydroxy, methoxy, N-methylamino

[0058] In some specific embodiments, R 2 Selected from: hydrogen, methyl, methoxymethyl, trifluoromethoxymethyl, difluoromethoxymethyl, hydroxymethyl, cyclopropyl,

[0059] In some specific embodiments, R 2 Selected from

[0060] In some specific embodiments, R 2 Selected from: hydrogen, methyl, methoxymethyl, trifluoromethoxymethyl, difluoromethoxymethyl, hydroxymethyl, cyclopropyl,

[0061] In some specific embodiments, R 2 Selected from hydrogen, methyl, methoxymethyl, trifluoromethoxymethyl, difluoromethoxymethyl, hydroxymethyl, cyclopropyl,

[0062] In some specific embodiments, R 2 Selected from hydrogen, methoxymethyl,

[0063] In some specific embodiments, R 3 Selected from hydrogen, halogen, C 1-6 Alkyl, C 3-6 Cycloalkyl.

[0064] In some specific embodiments, R 3 Selected from hydrogen, F, Cl, Br, C 1-4 Alkyl, C 5-6 Alkyl, C 3- 5-cycloalkyl.

[0065] In some specific embodiments, R 3 Selected from hydrogen, F, Cl, Br, C 1-4 Alkyl, C 3-5 Cycloalkyl.

[0066] In some specific embodiments, R 3 Selected from hydrogen, F, Cl, Br, methyl, ethyl, propyl, isopropyl, butyl, cyclopropyl, cyclobutyl, cyclopentyl.

[0067] In some specific embodiments, R 3 Selected from hydrogen, F, Cl, methyl, ethyl, cyclopropyl.

[0068] In some specific embodiments, R 3 Selected from hydrogen, F, Cl, Br, C 1-4 alkyl.

[0069] In some specific embodiments, R 3 Selected from hydrogen, F, Cl, methyl, ethyl;

[0070] In some specific embodiments, R 3 Selected from hydrogen, F, methyl.

[0071] In some specific embodiments, R 3 Selected from hydrogen.

[0072] In some embodiments, the group Selected from

[0073] In some embodiments, the group Selected from

[0074] In some embodiments, the group Selected from

[0075] In some embodiments, the group Selected from

[0076] In some embodiments, the group Selected from

[0077] In some embodiments, the group Selected from

[0078] In some embodiments, the group Selected from

[0079] In some embodiments, the group Selected from

[0080] In some embodiments, the group Selected from

[0081] Wherein, W is selected from N or CR 7 .

[0082] In some embodiments, W is selected from CR 7 .

[0083] In some specific embodiments, W is selected from N.

[0084] In some specific embodiments, R 4 , R 5 , R 6 , R 7 are each independently selected from hydrogen, halogen, C 1-6 alkyl.

[0085] 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.

[0086] In some specific embodiments, R 4 , R 5 , R 6 , R 7 are each independently selected from hydrogen, halogen, C 1-4 alkyl.

[0087] 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.

[0088] In some specific embodiments, R 4 , R 5 , R 6 , R 7 Each is independently selected from hydrogen, F, Cl, and methyl.

[0089] In some embodiments, the group Selected from

[0090] Wherein, W is selected from N or CR 7 .

[0091] In some embodiments, the group Selected from

[0092] Wherein, W is selected from N or CR 7 , R7 Selected from H, F, Cl, methyl.

[0093] In some embodiments, the group Selected from:

[0094] In some embodiments, the group Selected from:

[0095] In some embodiments, the group Selected from:

[0096] In some embodiments, the group Selected from:

[0097] In some embodiments, the group Selected from:

[0098] In some specific embodiments, R 1 Selected from: C 1-6 Alkyl, C 3-10 Cycloalkyl, C 3-10 Cycloalkyl-C 1- 6 alkyl-, 5-10 membered heteroaryl-; wherein the C1-C6 alkyl 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.

[0099] 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 5-8Spiroalkyl 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.

[0100] 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 Substitution; 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.

[0101] 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.

[0102] 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.

[0103] It should be understood that the substituent R 1 In the definition of: R i Always substituted on an alkyl or alkylene group, R jAlways 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.

[0104] 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.

[0105] 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.

[0106] In some embodiments, each R i 、R j 、R k 、R l are independently selected from halogen, halogenated C 1-4 Alkyl, C 1-4 Alkyl, C 3-6 Cycloalkyl.

[0107] 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.

[0108] 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.

[0109] In some embodiments, each R i 、R j 、R k 、Rl Each is independently selected from F, Cl, methyl, and ethyl.

[0110] In some embodiments, each R i 、R j 、R k 、R l The substituents are each independently selected from methyl.

[0111] In some embodiments, each R l are independently selected from halogen, halogenated C 1-6 Alkyl, C 1- 6 alkyl, C 3-6 Cycloalkyl.

[0112] In some embodiments, each R l are independently selected from halogen, halogenated C 1-4 Alkyl, C 1- 4 alkyl, C 3-6 Cycloalkyl.

[0113] In some embodiments, each R l are each independently selected from halogen, C 1-4 Alkyl, C 3-6 Cycloalkyl.

[0114] In some embodiments, each R l Each is independently selected from F, Cl, Br, methyl, ethyl, propyl, isopropyl, cyclopropyl, and cyclobutyl.

[0115] In some embodiments, each R l Each is independently selected from F, Cl, methyl, and ethyl.

[0116] In some embodiments, each R l The substituents are each independently selected from methyl.

[0117] In some specific embodiments, R 1 Selected from methyl, ethyl, cyclopropyl,

[0118] In some specific embodiments, R 1 Selected from methyl.

[0119] As an exemplary compound of the present invention, the compound represented by formula (I) is selected from any of the following specific compounds:

[0120] The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention, prepared by reacting the compounds discovered herein with the specified substituents with relatively nontoxic acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the compounds with a sufficient amount of base in neat solution or in a suitable inert solvent. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the compounds with a sufficient amount of acid in neat solution or in a suitable inert solvent.

[0121] The term "deuterated compound" refers to a compound of the present invention that includes at least one deuterium atom, specifically one or more hydrogen atoms in a compound of the present invention that 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 incorporating isotopes into organic compounds are known in the art.

[0122] Preparation method:

[0123] 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.

[0124] Synthesis process route 1:

[0125] Step 1: Treating the compound represented by formula I-1 with di-tert-butyl dicarbonate to obtain the compound represented by formula I-2;

[0126] 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;

[0127] 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;

[0128] 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;

[0129] 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;

[0130] 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).

[0131] Synthesis process route 2:

[0132] Step 1: 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-1 to obtain the compound shown in formula II-1;

[0133] Step 2: The compound represented by formula II-1 is treated with tetrabutylammonium fluoride to obtain the compound represented by formula II-2;

[0134] Step 3: The compound represented by formula II-2 is subjected to acidic conditions to remove the Boc (tert-butyloxycarbonyl) protecting group to obtain the compound represented by formula II-3;

[0135] Step 4: Under condensation conditions well known in the art, the compound represented by formula II-3 is treated with the compound represented by I-6 to obtain the compound represented by formula II;

[0136] Among them, Q 1 Is halogen, X, Y, Z, W, A, R 1 、R 3 , n1, n2, and ring M are as defined herein in formula (I).

[0137] Synthesis process route three:

[0138] 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 III-2;

[0139] Step 2: Under coupling conditions well known in the art, The compound shown in formula III-2 is treated with the compound shown in formula III-3;

[0140] Step 3: The compound represented by formula III-3 is treated with tetrabutylammonium fluoride to obtain the compound represented by formula III (i.e., formula I);

[0141] or,

[0142] Step 4: Under coupling conditions well known in the art, The compound shown in formula III-2 is treated with the compound shown in formula III-4;

[0143] Step 5: The compound represented by formula III-4 is treated with tetrabutylammonium fluoride to obtain the compound represented by formula III (i.e., formula I);

[0144] or,

[0145] Step 6: Under coupling conditions well known in the art, The compound shown in formula III-2 can be treated with the compound shown in formula III-2 to obtain the compound shown in formula III (i.e., formula I);

[0146] Synthesis process route 4:

[0147] Step 1: Under coupling conditions well known in the art, The compound shown in formula III-1 can be treated with the compound shown in formula I-5;

[0148] Step 2: 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 V (i.e., formula I);

[0149] Pharmaceutical composition:

[0150] 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.

[0151] In some embodiments, the disease associated with abnormal PRMT5 expression is a tumor or cancer.

[0152] In some embodiments, the disease associated with abnormal PRMT5 expression refers to a disease associated with abnormal PRMT5 expression in which MTAP is deficient.

[0153] In some embodiments, the disease associated with abnormal PRMT5 expression refers to a tumor or cancer with MTAP deficiency.

[0154] 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.

[0155] 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.

[0156] Medical uses:

[0157] 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.

[0158] 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.

[0159] 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.

[0160] 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.

[0161] In some embodiments, the disease associated with abnormal PRMT5 expression is a tumor or cancer.

[0162] In some embodiments, the disease associated with abnormal PRMT5 expression refers to a disease associated with abnormal PRMT5 expression in which MTAP is deficient.

[0163] In some embodiments, the disease associated with abnormal PRMT5 expression refers to a tumor or cancer with MTAP deficiency.

[0164] 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.

[0165] In some embodiments, the disease associated with abnormal PRMT5 expression is a tumor or cancer.

[0166] In some embodiments, the disease associated with abnormal PRMT5 expression refers to a disease associated with abnormal PRMT5 expression in which MTAP is deficient.

[0167] In some embodiments, the disease associated with abnormal PRMT5 expression refers to a tumor or cancer with MTAP deficiency.

[0168] 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.

[0169] 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.

[0170] 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.

[0171] Definition of terms:

[0172] 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.

[0173] 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.

[0174] 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.

[0175] 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".

[0176] The term "alkyl" refers to both branched and straight chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms. For example, "C1-6 alkyl" refers to C1, C2, C3, C4, C5 and C6. In addition, for example, "C1-6 "Alkyl" refers to an alkyl group having 1 to 6 carbon atoms. The alkyl group may be unsubstituted or substituted so that one or more of its hydrogen atoms are replaced by another chemical group. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl (e.g., n-propyl and isopropyl), butyl (e.g., n-butyl, isobutyl, tert-butyl), pentyl (e.g., n-pentyl, isopentyl, neopentyl), and the like. It will be understood by those skilled in the art that C 1-6 Alkyl groups contain a monovalent C 1-6 Alkyl, divalent C 1-6 Alkylene, such as C3-C6 cycloalkyl-(C1-C6 alkyl) r -The C1-C6 alkyl group refers to a C1-C6 alkylene group.

[0177] The term "alkoxy" refers to any of the above alkyl groups (eg, C1-6 alkyl, C1-4 alkyl, C1-3 alkyl, etc.) that is attached to the rest of the molecule through an oxygen atom (-O-).

[0178] The term "halogenated C 1-6 Alkyl" or "halogenated C 1-6 "Alkoxy" refers to an alkyl or alkoxy group in which one or more (such as 2 or 3) hydrogen atoms are replaced by halogen atoms, such as fluorine, chlorine, or bromine. The alkyl or alkoxy group is defined as above. In some embodiments, the term "halogenated C 1-6 "Alkyl" is preferably fluorinated, for example, -CF3, -CHF2, -CH2F, -CH2CH2F, -CH2CHF2, -CH2CF3, etc. In some embodiments, the term "halogenated C 1-6 The "alkoxy" is preferably fluorinated, for example, it can be -OCF3, -OCHF2, -OCH2F, -OCH2CH2F, -OCH2CHF2, -OCH2CF3, etc.

[0179] The term "hydroxy substituted C 1-6 "C alkyl" means that one of the hydrogen atoms in the alkyl group is replaced by a hydroxyl group, and the definition of the alkyl group is as described above. As an example, the "C alkyl group substituted by a hydroxyl group" 1-6 The "alkyl" may be a hydroxymethyl group.

[0180] 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 should be a saturated carbocyclic ring or a carbocyclic ring residue. The two rings of the bicyclic or polycyclic cycloalkyl group can be connected in a manner including bridging, fusion or spiro connection. For example, C 3-10 Cycloalkyl refers to C3, C4, C5, C6, C7, C8, C9 and C 10 Cycloalkyl. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.

[0181] The term "cycloalkenyl" refers to a cycloalkyl group as defined above that has at least one carbon-carbon double bond, for example

[0182] The term "cycloalkoxy" refers to any of the above cycloalkyl groups (eg, C3-6 cycloalkyl) that is attached to the remainder of the molecule through an oxygen atom (-O-).

[0183] 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.

[0184] 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.

[0185] 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.

[0186] 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.

[0187] The term "heterocycloalkenyl" refers to a heterocyclic group having at least one carbon-carbon double bond in the heterocycle as defined above, for example

[0188] 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.

[0189] 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.

[0190] 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.

[0191] 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.

[0192] 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:

[0193] 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.

[0194] 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).

[0195] 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.

[0196] 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).

[0197] 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.

[0198] The term "heteroatom" shall include oxygen, sulfur and nitrogen.

[0199] The term "halogen" shall include "F, Cl, Br, I".

[0200] 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.

[0201] 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.

[0202] 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.

[0203] 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.

[0204] 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 ).

[0205] 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.

[0206] 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.

[0207] 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.

[0208] In this document, when there is a discrepancy between the structure and the chemical name, the structure prevails.

[0209] Effects of the invention:

[0210] The compound represented by formula (I) of the present invention has excellent PRMT5 inhibitory activity and has strong anti-proliferation activity against the human MTAP-deficient HCT-116 cell line. In addition, compared with the MTAP wild-type (WT) HCT-116 cell line, the compound 1-38 of the present invention has excellent selective inhibition against the human MTAP-deficient HCT-116 cell line, and can be used as a drug for treating or preventing diseases related to this effect. Furthermore, through in vivo pharmacokinetic testing in mice, the compound of the present invention has better in vivo pharmacokinetic advantages compared to the prior art (reference example), and has better drugability and clinical application prospects. DETAILED DESCRIPTION

[0211] 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.

[0212] 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.

[0213] Reaction monitoring and MS measurements were performed using a Thermofisher ESQ (ESI) mass spectrometer.

[0214] 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).

[0215] 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.

[0216] 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.

[0217] Example 1: 4-amino-N-(6-ethynyl-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N,1-dimethyl-

[0218] 1H-Pyrazolo[4,3-c]quinoline-8-carboxamide

[0219] Preparation of tert-butyl (6-bromo-2,3-dihydrobenzofuran-3-yl)carbamate (1b)

[0220] Compound 1a (229 mg, 1.07 mmol), triethylamine (0.30 mL, 2.14 mmol), and di-tert-butyl dicarbonate (0.27 mL, 1.18 mmol) were dissolved in dichloromethane (5 mL) and stirred at 25°C for 2 h. The reaction mixture was quenched with water (5 mL) and extracted with dichloromethane (5 mL x 3). The organic phases were combined and concentrated to obtain the crude product, which was purified by preparative thin-layer chromatography (petroleum ether:ethyl acetate = 10:1) to afford compound 1b (314 mg). E SI-MS (m / z): 258.01 [M+H] + .

[0221] Preparation of tert-butyl (6-bromo-2,3-dihydrobenzofuran-3-yl) (methyl) carbamate (1c)

[0222] Compound 1b (314 mg, 1.00 mmol) was dissolved in tetrahydrofuran (2 mL), cooled to 0°C, and sodium hydride (80.0 mg, 2.00 mmol, 60% purity) was added. The reaction solution was stirred at 0°C for 1 hour, followed by the addition of iodomethane (0.10 mg, 1.20 mmol) and stirring at 25°C for 12 hours. The reaction solution was quenched with water (5 mL) and extracted with ethyl acetate (5 mL*3). The organic phases were combined, concentrated, and purified by thin-layer preparative chromatography (petroleum ether:ethyl acetate = 20:1) to obtain 1c (310 mg). ESI-MS (m / z): 274.26 [M+H] + .

[0223] Preparation of tert-butylmethyl (6-((trimethylsilyl)ethynyl)-2,3-dihydrobenzofuran-3-yl)carbamate (1d)

[0224] Compound 1c (310 mg, 0.94 mmol) was dissolved in N-methylpyrrolidone (2 mL), and ethynyltrimethylsilane (185.5 mg, 1.89 mmol), cuprous iodide (36.0 mg, 0.19 mmol), tetrakistriphenylphosphine palladium (109.2 mg, 0.09 mmol) and N,N-diisopropylethylamine (0.44 mL, 2.64 mmol) were added. The atmosphere was replaced with nitrogen three times, and the mixture was microwaved at 70°C for 3 hours. The reaction solution was diluted with ethyl acetate (5 mL) and water (5 mL) to precipitate a solid, which was filtered and the filtrate was separated. The organic phase was concentrated and purified by thin-layer preparative chromatography (dichloromethane:methanol = 92:8) to give compound 1d (280 mg).

[0225] Preparation of tert-butyl (6-ethynyl-2,3-dihydrobenzofuran-3-yl) (methyl) carbamate (1e)

[0226] Compound 1d (280 mg, 0.81 mmol) was dissolved in tetrahydrofuran (2 mL), and a 1 M solution of tetrabutylammonium fluoride in tetrahydrofuran (1.22 mL, 1.22 mmol) was added. The mixture was stirred at 25°C for 1 h. The reaction mixture was quenched with water (5 mL) and extracted with ethyl acetate (5 mL x 3). The organic phases were combined, concentrated, and purified by preparative thin-layer chromatography (dichloromethane:methanol = 50:1) to afford 1e (208 mg). ESI-MS (m / z): 218.12 [M+H] + .

[0227] Preparation of 6-ethynyl-N-methyl-2,3-dihydrobenzofuran-3-amine (1f)

[0228] Compound 1e (200 mg, 0.73 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 dried to give compound 1f (150 mg).

[0229] Preparation of 4-amino-N-((5-ethynylpyridin-2-yl)methyl)-7-fluoro-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (1)

[0230] To a solution of compound 1f (41.9 mg, 0.20 mmol) in N,N-dimethylacetamide (3 mL) were added compound 1g (52.0 mg, 0.20 mmol, prepared according to WO 2022 / 115377 A1), triethylamine (0.07 mL, 0.50 mmol), and tripyrrolidinylphosphonium bromide hexafluorophosphate (156.0 mg, 0.30 mmol), and the mixture was reacted at 25°C for 18 hours. After completion of the reaction, ethyl acetate (10 mL) and water (10 mL) were added to the reaction solution, the layers were separated, and the organic phase was concentrated and purified by high-pressure liquid chromatography to obtain compound 1 (22 mg). ESI-MS (m / z): 416.23 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ8.28–8.24(m,2H),7.41–7.32(m,4H),7.15–6.98(m,2H),6.47–6.44and 5.59–5.56(m,1H),4.84–4.63(m,2H),4.42–4.23(m,4H),2.69–2.58(s,3H).

[0231] Example 2: 4-amino-N-(6-ethynyl-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide

[0232] To a solution of compound 1f (41.9 mg, 0.20 mmol) in N,N-dimethylacetamide (3 mL) were added compound 2a (48.4 mg, 0.20 mmol, prepared according to WO 2022 / 115377 A1), triethylamine (0.07 mL, 0.50 mmol), and tripyrrolidinylphosphonium bromide hexafluorophosphate (156.0 mg, 0.30 mmol), and the mixture was reacted at 25°C for 18 hours. The reaction solution was diluted with ethyl acetate (10 mL) and water (10 mL), the layers were separated, and the organic phase was concentrated and purified by high-pressure liquid chromatography to obtain compound 2 (5 mg). ESI-MS (m / z): 398.17 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ8.33(s,1H),8.28(s,1H),7.67–7.62(m,2H),7.46–7.44(m,1H),7.22(s,2H),7.1 1–7.09(m,1H),7.00(s,1H),6.52–5.69(m,1H),4.73–4.72(m,2H),4.43(s,3H),4.23(s,1H),2.68(s,3H).

[0233] Example 3: 4-amino-N-(6-ethynyl-2,3-dihydrobenzofuran-3-yl)-N-methyl-1,3-dihydrofuran[4,3-c]quinoline-8-carboxamide

[0234] To a solution of compound 1f (41.9 mg, 0.20 mmol) in N,N-dimethylacetamide (3 mL) were added compound 3a (46.0 mg, 0.20 mmol, prepared according to WO 2022 / 115377 A1), triethylamine (0.07 mL, 0.50 mmol), and tripyrrolidinylphosphonium bromide hexafluorophosphate (156.0 mg, 0.30 mmol), and the reaction mixture was allowed to react at 25°C for 18 hours. The reaction mixture 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 3 (15 mg). ESI-MS (m / z): 386.23 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ7.67–7.61(m,3H),7.42–7.40(m,1H),7.11–7.10(m,1H),7.00(s,1H),6.72(s,2H ),6.48–5.52(m,1H),5.39–5.37(m,2H),5.04–5.03(m,2H),4.73–4.64(m,2H),4.22(s,1H),2.63(s,3H).

[0235] Example 4: 4-amino-N-(6-ethynyl-2,3-dihydro-1-benzofuran-3-yl)-1,7,N-trimethylpyrazolo[4,3-c]quinoline-8-carboxamide

[0236] To a solution of compound 4a (30.8 mg, 0.12 mmol, prepared according to WO 2022 / 115377 A1) in N,N-dimethylacetamide (5 mL) were added compound 1f (20 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. The reaction solution was added with water (10 mL) and extracted with ethyl acetate (10*3 ml). The organic phases were washed three times with saturated brine, combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 4 (7.0 mg) was obtained by reverse phase column chromatography (acetonitrile / 0.05% aqueous ammonium bicarbonate solution = 0:100% to 50%:50%). ESI-MS (m / z): 412.1 [M+H]+ . 1 H NMR(400MHz,DMSO-d6)δ8.22–8.01(m,2H),7.50–7.41(m,2H),7.14–6.98(m,4H),6 .52–5.35(m,1H),4.82–4.58(m,2H),4.38(s,3H),4.21(s,1H),2.68–2.34(m,6H).

[0237] Example 5: 4-amino-N-cyclopropyl-N-(6-ethynyl-2,3-dihydro-1-benzofuran-3-yl)-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide

[0238] Preparation of 5-bromo-2-((cyclopropylimino)methyl)phenol (5b)

[0239] To a solution of 4-bromo-2-hydroxybenzaldehyde (1.0 g, 4.98 mmol) in dichloromethane (10 mL) were added cyclopropylamine (0.38 mL, 5.48 mmol) and acetic acid (0.09 mL, 1.49 mmol) and allowed to react overnight at 25°C. After completion of the reaction, the mixture was concentrated to afford compound 5b (1.0 g). ESI-MS (m / z): 240.0 [M+H] + .

[0240] Preparation of 6-bromo-N-cyclopropyl-2,3-dihydrobenzofuran-3-amine (5c)

[0241] 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 5b (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 allowed to react at 25°C overnight. After completion of the reaction, the mixture 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 5c (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] + .

[0242] Step 3: Preparation of 4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-N-cyclopropyl-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (5d)

[0243] To a solution of 5c (100 mg, 0.39 mmol) in N,N-dimethylacetamide (5 mL) were added compound 2a (114.4 mg, 0.47 mmol), O-(IH-benzotriazol-1-yl)-N,N,N',N'-tetramethylisourea boron tetrafluoride (189.6 mg, 0.59 mmol), and N,N-diisopropylethylamine (0.20 mL, 1.18 mmol). 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. Compound 5d (130.0 mg) was obtained by normal phase column chromatography (methanol / dichloromethane = 0:100%–10%:90%). ESI-MS (m / z): 478.2 [M+H] + .

[0244] Preparation of 4-amino-N-cyclopropyl-1-methyl-N-(6-((trimethylsilyl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (5e)

[0245] To a solution of 5d (50 mg, 0.08 mmol) in N-methylpyrrolidone (4 mL) were added cuprous iodide (6.4 mg, 0.03 mmol), tetrakistriphenylphosphine palladium (19.3 mg, 0.02 mmol), N,N-diisopropylethylamine (0.04 mL, 0.25 mmol), and trimethylsilyl acetylene (9.8 mg, 0.10 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 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. Compound 5e (30.0 mg) was obtained by normal phase column chromatography (methanol / dichloromethane = 20:1). ESI-MS (m / z): 496.2 [M+H]. +

[0246] Preparation of 4-amino-N-cyclopropyl-N-(6-ethynyl-2,3-dihydro-1-benzofuran-3-yl)-1-methylpyrazolo[4,3-c]quinoline-8-carboxamide (5)

[0247] To a solution of compound 5e (30 mg, 0.06 mmol) in tetrahydrofuran (5 mL) was added tetrabutylammonium fluoride (23.7 mg, 0.09 mmol) and the mixture was allowed to react at 25°C for 0.5 h. Water (15 mL) was added to the reaction solution, which was then extracted with ethyl acetate (10 x 3 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Reverse-phase column chromatography (acetonitrile / 0.05% ammonium bicarbonate solution = 0:100%–50%:50%) afforded compound 5 (5.5 mg). ESI-MS (m / z): 424.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.45(d,J=2.0Hz,1H),8.29(s,1H),7.78(dd,J=8.6,2.0Hz,1H),7.60–7.57(m,2H),7.25(s,2H),7.09(dd,J=7.8,1.4Hz,1H ),6.97(s,1H),5.94(dd,J=9.2,4.2Hz,1H),4.83–4.67(m,2H),4.42(s,3 H),4.20(s,1H),2.86–2.82(m,1H),0.42–0.31(m,2H),0.19–0.05(m,2H).

[0248] Example 6: 4-amino-N-[6-(3-methoxyprop-1-ynyl)-2,3-dihydro-1-benzofuran-3-yl]-1,N-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide

[0249] Preparation of 6-(3-methoxyprop-1-yn-1-yl)-N-methyl-2,3-dihydrobenzofuran-3-amine (6b)

[0250] To a solution of compound 6a (100 mg, 0.44 mmol, prepared as described in WO2022169948A1) in N-methylpyrrolidone (5 mL) were added 3-methoxyprop-1-yne (92.5 mg, 1.32 mmol), tetrakistriphenylphosphine palladium (50.8 mg, 0.04 mmol), cuprous iodide (16.8 mg, 0.09 mmol), and N,N-diisopropylethylamine (0.36 mL, 2.20 mmol). The mixture was microwaved at 75°C under nitrogen for 3 hours. After completion of the reaction, water (20 mL) was added, and the mixture was extracted with ethyl acetate (3 x 15 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 6b (70.0 mg) was obtained by normal phase column chromatography (ethyl acetate / petroleum ether = 0:100%–50%:50%). ESI-MS (m / z): 218.0 [MH] + .

[0251] Preparation of 4-amino-N-[6-(3-methoxyprop-1-ynyl)-2,3-dihydro-1-benzofuran-3-yl]-1,N-dimethylpyrazolo[4,3-c]quinoline-8-carboxamide (6)

[0252] To a solution of compound 6b (50 mg, 0.23 mmol) in N,N-dimethylformamide (5 mL) were added compound 2a (66.9 mg, 0.28 mmol), O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethylisourea boron tetrafluoride (110.8 mg, 0.35 mmol), and N,N-diisopropylethylamine (0.11 mL, 0.69 mmol), and the mixture was allowed to react at 25°C for 3 hours. Water (20 mL) was added to the reaction solution, which was then extracted with ethyl acetate (3 x 15 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The mixture was purified by normal phase column chromatography (methanol / dichloromethane = 1:15) to afford compound 6 (10.0 mg). ESI-MS (m / z): 442.2 [M+H]. + . 1 H NMR (400MHz, DMSO-d6) δ8.33(s,1H),8.29–8.27(m,2H),7.65–7.62(m,2H),7.45(d,J=7.8Hz,1H),7.17(s,2H ),7.09(d,J=7.8Hz,1H),6.98(s,1H),4.76–4.72(m,2H),4.44(s,3H),4.34(s,2H),3.35(s,3H),2.69(s,3H).

[0253] Example 7: 4-amino-N-(6-(3-hydroxy-3-methylbut-1-yn-1-yl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide

[0254] Preparation of 4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)N,-1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (7a)

[0255] Compound 6a (86 mg, 0.38 mmol) was dissolved in N,N-dimethylacetamide (3 ml), and compound 2a (94.2 mg, 0.41 mmol), 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 the addition, the mixture was stirred at 25°C for 3 hours. The reaction solution was quenched with water and extracted with ethyl acetate (3*15 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by normal phase silica gel column chromatography (dichloromethane:methanol = 10:1) to give compound 7a (60 mg, 0.13 mmol). ESI-MS (m / z): 452.3 [M+H] + .

[0256] Preparation of 4-amino-N-(6-(3-hydroxy-3-methylbut-1-yn-1-yl)-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (7)

[0257] To a 10 mL microwave tube, reactant 7a (40 mg, 0.09 mmol), cuprous iodide (3.4 mg, 0.02 mmol), tetrakistriphenylphosphine palladium (10.2 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 (43.95 μL, 0.27 mmol) and ((1,1-dimethyl-2-propynyl)oxy)trimethylsilane (785.2 mg, 8.00 mmol). The mixture was stirred in a microwave oven at 70°C for 3 hours. The reaction mixture was quenched with water, extracted with ethyl acetate (3 x 15 mL), washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and isolated by reverse phase column chromatography (acetonitrile:water = 1:4) to afford compound 7 (4.6 mg). ESI-MS (m / z): 455.5 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.37(s,1H),8.26(s,1H),7.80–7.64(m,2H),7.39–7.04(m,3H),6.99(d,J= 7.6Hz,1H),6.86(s,1H),6.28–5.41(m,1H),4.70(s,2H),4.41(s,3H),2.65(s,3H),1.45(s,6H).

[0258] Example 8: 4-amino-7-fluoro-N-[6-(3-hydroxy-3-methylbut-1-ynyl)-2,3-dihydro-1-benzofuran-3-yl]-1,N-dimethylpyrazolo[4,3-c]quinoline-8-carboxamide

[0259] Preparation of 4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (8a)

[0260] To a solution of compound 6a (70 mg, 0.3 mmol) in N,N-dimethylformamide (3 mL) were added compound 1g (80 mg, 0.3 mmol), N,N-diisopropylethylamine (78 mg, 0.6 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (171 mg, 0.45 mmol). The reaction was allowed to proceed at room temperature for 4 hours. Upon completion of the reaction, water (20 mL) was added and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed. The mixture was purified by column chromatography (methanol:dichloromethane = 1:10) to afford compound 8a (81 mg). ESI-MS (m / z): 469.9 [M+H]. + .

[0261] Preparation of 4-amino-7-fluoro-N,1-dimethyl-N-(6-(3-methyl-3-((trimethylsilyl)oxy)but-1-yn-1-yl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (8b) Under nitrogen protection, to a solution of compound 8a (81 mg, 0.17 mmol) in N-methylpyrrolidone (1 mL) were added [(1,1-dimethyl-2-propynyl)oxy]trimethylsilane (53 mg, 0.34 mmol), cuprous iodide (8 mg, 0.04 mmol), N,N-diisopropylethylamine (44 mg, 0.34 mmol) and tetrakis(triphenylphosphine)palladium (23 mg, 0.02 mmol) in sequence. After addition, the mixture was reacted in a microwave oven at 70°C for 3 hours. After the reaction was complete, water (10 mL) was added and the mixture was extracted with ethyl acetate (10 mL*3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed. Column chromatography (methanol:dichloromethane = 1:20) was performed to obtain compound 8b (55 mg). ESI-MS (m / z): 546.1 [M+H] + .

[0262] Preparation of 4-amino-7-fluoro-N-[6-(3-hydroxy-3-methylbut-1-ynyl)-2,3-dihydro-1-benzofuran-3-yl]-1,N-dimethylpyrazolo[4,3-c]quinoline-8-carboxamide (8)

[0263] To a solution of compound 8b (55 mg, 0.1 mmol) in dichloromethane (1 mL) was slowly added a 1,4-dioxane hydrochloride solution (0.5 mL, 4 M) at 0°C. The mixture was allowed to react at room temperature for 30 minutes and then sent for testing. Upon completion of the reaction, a saturated sodium carbonate solution was added to adjust the pH of the system to approximately 8. 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 mixture was purified by reverse phase column chromatography (acetonitrile: 0.05% aqueous ammonium bicarbonate solution = 0:100%–50%:50%) to afford compound 8 (21 mg). ESI-MS (m / z): 474.1 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ 8.40–8.18(m,2H),7.41–7.28(m,4H),7.04–6.99(m,1H),6.93–6.84(m,1H),6.45–5.52(m, 1H),5.46(s,1H),4.81–4.62(m,2H),4.42–4.37(m,3H),2.69and2.57(s,3H),1.47(s,6H).

[0264] Example 9: 4-amino-N-(7-ethynylisochroman-4-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide

[0265] Preparation of 7-bromo-N-methylisochroman-4-amine (9b)

[0266] To a solution of compound 9a (100 mg, 0.44 mmol) in dichloromethane (5 mL) were added methylamine hydrochloride (148.6 mg, 2.20 mmol), N,N-diisopropylethylamine (364.69 μL, 2.20 mmol), and glacial acetic acid (25.20 μL, 0.44 mmol). The mixture was allowed to react overnight at 25°C. Sodium borohydride (25.0 mg, 0.66 mmol) was then added and stirred at room temperature for 2 hours. Saturated sodium bicarbonate (20 mL) was added to the reaction solution, which was extracted with dichloromethane (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 9b (80 mg) was obtained by normal phase column chromatography (petroleum ether / ethyl acetate = 1:1). ESI-MS (m / z): 241.99 [M+H].+ .

[0267] Preparation of 4-amino-N-(7-bromoisochroman-4-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (9c)

[0268] To a solution of compound 9b (50.0 mg, 0.21 mmol) in N,N-dimethylformamide (3 mL) were added compound 2a (50.0 mg, 0.21 mmol), N,N-diisopropylethylamine (102.63 μL, 0.62 mmol), and O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethylisourea boron tetrafluoride (117.8 mg, 0.31 mmol). The mixture was reacted at 25°C overnight. Water (25 mL) was added to the reaction solution, which was then 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. Compound 9c (60 mg) was obtained by normal phase column chromatography (dichloromethane / methanol = 10:1). ESI-MS (m / z): 466.08 [M+H]. + .

[0269] Preparation of 4-amino-N,1-dimethyl-N-(7-((trimethylsilyl)ethynyl)isochroman-4-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (9d)

[0270] To a solution of compound 9c (28 mg, 0.06 mmol) in N-methylpyrrolidone (2 mL) were added cuprous iodide (2.5 mg, 0.01 mmol), tetrakistriphenylphosphine palladium (7.4 mg, 0.01 mmol), N,N-diisopropylethylamine (31.97 μL, 0.19 mmol), and trimethylsilyl acetylene (45.45 μL, 0.32 mmol). The mixture was microwaved at 70°C for 3 hours under nitrogen. The reaction mixture was filtered through celite, water (25 mL) was added, and extraction 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. Compound 9d (15 mg) was obtained by normal phase column chromatography (dichloromethane / methanol = 10:1). ESI-MS (m / z): 484.24 [M+H]. +

[0271] Preparation of 4-amino-N-(6-ethynyl-2,3-dihydrobenzofuran-3-yl)-N-methyl-1,3-dihydrofuran[3,4-c][1,7]naphthyridine-8-carboxamide (9)

[0272] Tetrabutylammonium fluoride (90 μL, 0.09 mmol) was added to a solution of compound 9d (14.5 mg, 0.03 mmol) in tetrahydrofuran (3 mL) and allowed to react at 25°C for 1 hour. Water (10 mL) was added to the reaction solution, which was then extracted with ethyl acetate (2 x 20 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was purified by preparative plate separation (dichloromethane / methanol = 10:1) and then purified by reverse-phase column chromatography (acetonitrile / 0.05% ammonium bicarbonate solution = 0:100%–40%:60%) to afford compound 9 (12 mg). ESI-MS (m / z): 412.05 [M+H] + . 1 H NMR(400MHz,DMSO)δ8.31(d,J=1.5Hz,1H),8.25(s,1H),7.67–7.59(m,2H),7.41(s,2H),7.29(s,1H),7.1 2(s,2H),5.83–4.89(m,1H),4.83–4.54(m,2H),4.39(s,3H),4.19(s,1H),4.19–4.04(m,2H),2.75(s,3H).

[0273] Example 10: Preparation of 4-amino-N-(6-ethynyl-2,3-dihydrobenzofuran-3-yl)-N-methyl-1,3-dihydrofuran[3,4-c][1,7]naphthyridine-8-carboxamide (10)

[0274] Preparation of 4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-N-methyl-1,3-dihydrofuran[3,4-c][1,7]naphthyridine-8-carboxamide (10b)

[0275] To a solution of 6a (100 mg, 0.44 mmol) in N,N-dimethylacetamide (5 mL) were added compound 10a (102 mg, 0.44 mmol, prepared according to WO 2022 / 115377A1), O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethylisourea boron tetrafluoride (251 mg, 0.66 mmol), and N,N-diisopropylethylamine (114 mg, 0.88 mmol). The mixture was reacted 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. Compound 10b (116 mg) was obtained by normal phase column chromatography (methanol / dichloromethane = 1 / 10). ESI-MS (m / z): 441.0 [M+H] + .

[0276] Preparation of 4-amino-N-methyl-N-(6-((trimethylsilyl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1,3-dihydrofuran[3,4-c][1,7]naphthyridine-8-carboxamide (10c)

[0277] To a solution of compound 10b (116 mg, 0.26 mmol) in N-methylpyrrolidone (2 mL) were added cuprous iodide (7.5 mg, 0.04 mmol), tetrakistriphenylphosphine palladium (60 mg, 0.05 mmol), N,N-diisopropylethylamine (107 mg, 0.78 mmol), and trimethylsilyl acetylene (51 mg, 0.52 mmol). The mixture was microwaved at 70°C under nitrogen for 3 hours. The reaction mixture was filtered through celite, water (15 mL) was added, and extraction with ethyl acetate (3 x 10 mL) was performed. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 10c (52 mg) was obtained by normal phase column chromatography (methanol / dichloromethane = 1:20). ESI-MS (m / z): 459.1 [M+H]. + .

[0278] Preparation of 4-amino-N-(6-ethynyl-2,3-dihydrobenzofuran-3-yl)-N-methyl-1,3-dihydrofuran[3,4-c][1,7]naphthyridine-8-carboxamide (10)

[0279] To a solution of compound 10c (52 mg, 0.11 mmol) in tetrahydrofuran (5 mL) was added tetrabutylammonium fluoride (60 mg, 0.22 mmol) and the mixture was allowed to react at 25°C for 0.5 h. Water (15 mL) was added to the reaction solution, which was then extracted with ethyl acetate (10 x 3 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Reverse-phase column chromatography (acetonitrile / 0.05% ammonium bicarbonate solution = 0:100%–50%:50%) afforded compound 10 (23 mg). ESI-MS (m / z): 387.0 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ8.92–8.87(m,1H),8.34(s,1H),7.87–7.82(m,1H),7.51–7.37(m,1H),7.12–6.98 (m,3H),6.43–5.85(m,1H),5.40(s,2H),5.07(s,2H),4.87–4.54(m,2H),4.21(s,1H),2.70–2.67(m,3H).

[0280] Example 11: 4-amino-N-(5-(3-hydroxy-3-methylbut-1-ynyl)-2,3-dihydro-1H-inden-1-yl)-N,1-dimethylpyrazolo[4,3-c]quinoline-8-carboxamide

[0281] Preparation of 5-bromo-N-methyl-2,3-dihydro-1H-inden-1-amine (11b)

[0282] Compound 11a (1.2 g, 5.7 mmol) was dissolved in 10 mL of methanol, followed by the addition of methylamine hydrochloride (425.6 mg, 6.3 mmol) and N,N-diisopropylethylamine (2.99 mL, 17.2 mmol). The reaction mixture was allowed to react at room temperature for 1 hour. The reaction system was then placed in an ice bath, and sodium borohydride (433.5 mg, 11.4 mmol) was added. The reaction mixture was allowed to react for another 1 hour. Water (200 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (50 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 to yield compound 11b (1.25 g). ESI-MS (m / z): 226.1 [M+H]. + .

[0283] Preparation of 4-amino-N-(5-bromo-2,3-dihydro-1H-inden-1-yl)-N,1-dimethylpyrazolo[4,3-c]quinoline-8-carboxamide (11c)

[0284] Compound 2a (112 mg, 0.46 mol) was dissolved in N,N-dimethylformamide (10 mL), and N,N-diisopropylethylamine (0.4 mL, 2.3 mol) was added. The mixture was stirred on ice for five minutes. N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (210 mg, 0.55 mol) was then added. The reaction was continued for five minutes. Finally, 11b (312 mg, 1.38 mol) was added and the reaction was continued for one hour. 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 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 11c (140 mg). ESI-MS (m / z): 450.0 [M+H]. + .

[0285] Preparation of 4-amino-N,1-dimethyl-N-(5-(3-methyl-3-(trimethylsilyloxy)but-1-yn-1-yl)-2,3-dihydro-1H-inden-1-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (11d)

[0286] Under nitrogen, to a solution of compound 11c (70 mg, 0.16 mmol) in N-methylpyrrolidone (2 mL) were added [(1,1-dimethyl-2-propynyl)oxy]trimethylsilane (48.6 mg, 0.31 mmol), cuprous iodide (5.93 mg, 0.03 mmol), N,N-diisopropylethylamine (80.4 mg, 0.62 mmol), and tetrakis(triphenylphosphine)palladium (18.0 mg, 0.016 mmol) in sequence. The reaction mixture was microwaved at 70°C for 3 hours. Water (20 mL) was added to the reaction solution, which was then 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 11d (45 mg) was obtained by column chromatography (methanol:dichloromethane = 1:20). ESI-MS (m / z): 526.3 [M+H]. + .

[0287] Preparation of 4-amino-N-(5-(3-hydroxy-3-methylbut-1-ynyl)-2,3-dihydro-1H-inden-1-yl)-N,1-dimethylpyrazolo[4,3-c]quinoline-8-carboxamide (11)

[0288] To a solution of compound 11d (45 mg, 0.086 mmol) in tetrahydrofuran (2 mL) was added tetrabutylammonium fluoride solution (0.2 mL, 1 M) and allowed to react at room temperature for one hour. Water 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 filtrate was concentrated. Column chromatography (methanol:dichloromethane = 1:20) afforded compound 11 (35 mg). ESI-MS (m / z): 454.1 [M+H]. + . 1 H NMR(400MHz,DMSO)δ8.33(s,1H),8.26(s,1H),7.67–7.60(m,2H),7.34–7.27(m,3H),7.13(s,2H) ,5.55–5.43(m,2H),4.40(s,3H),3.22–3.13(m,2H),2.71(s,3H),1.66–1.52(m,2H),1.47(s,6H).

[0289] Example 12: 4-amino-N,1-dimethyl-N-(6-(3-(methylamino)prop-1-yn-1-yl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide

[0290] Preparation of 4-amino-N,1-dimethyl-N-(6-(3-(methylamino)prop-1-yn-1-yl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (12)

[0291] To a solution of compound 7a (50 mg, 0.11 mmol) in N-methylpyrrolidone (2 mL) were added cuprous iodide (4.2 mg, 0.02 mmol), tetrakistriphenylphosphine palladium (12.8 mg, 0.01 mmol), N,N-diisopropylethylamine (54.93 μL, 0.33 mmol), and N-methylpropargylamine (22.9 mg, 0.33 mmol). After the addition, the atmosphere was replaced with nitrogen and the mixture was microwaved at 70°C 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. The crude product was purified by normal phase column chromatography (dichloromethane / methanol = 10:1), which was then purified by reverse phase column chromatography (acetonitrile / 0.05% ammonium bicarbonate solution = 0:100%–40%:60%) to afford compound 12 (15 mg). ESI-MS (m / z): 441.1 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.30(s,1H),8.26(s,1H),7.62(s,2H),7.40(d,J=7.6Hz,1H),7.13(s,2H),7.02(d,J=7.6Hz, 1H),6.89(s,1H),6.56–5.63(m,1H),4.79–4.63(m,2H),4.41(s,3H),3.5(s,2H),2.66(s,3H),2.43–2.14(m,3H).

[0292] Example 13: 4-amino-N-(6-ethynyl-2,3-dihydrobenzofuran-3-yl)-N,3-dimethyl-3H-pyrazolo[3,4-c]quinoline-8-carboxamide

[0293] Preparation of 4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-N,3-dimethyl-3H-pyrazolo[3,4-c]quinoline-8-carboxamide (13b)

[0294] To a solution of compound 6a (100 mg, 0.44 mmol) in N,N-dimethylformamide (3 mL) were added compound 13a (106.6 mg, 0.44 mmol, prepared according to WO 2022 / 115377A1), N,N-diisopropylethylamine (217.85 μL, 1.32 mmol), and O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethylisourea boron tetrafluoride (250.0 mg, 0.66 mmol), and 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*30 ml). The organic phase was washed three times with saturated brine, combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 13b (110 mg) was obtained by normal phase column chromatography (dichloromethane / methanol = 10:1). ESI-MS (m / z): 452.1 [M+H] + .

[0295] Preparation of 4-amino-N,3-dimethyl-N-(6-((trimethylsilyl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-3H-pyrazolo[3,4-c]quinoline-8-carboxamide (13c)

[0296] To a solution of compound 13b (50 mg, 0.11 mmol) in N-methylpyrrolidone (2 mL) were added cuprous iodide (4.2 mg, 0.02 mmol), tetrakistriphenylphosphine palladium (12.8 mg, 0.01 mmol), N,N-diisopropylethylamine (54.93 μL, 0.33 mmol), and trimethylsilylacetylene (54.3 mg, 0.55 mmol). The mixture was microwave-dried at 70°C for 3 hours under nitrogen. The reaction mixture was filtered through celite, added with water (50 mL), and 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 concentrated. The crude product was purified by normal-phase column chromatography (dichloromethane / methanol = 10:1), which was then purified by reverse-phase column chromatography (acetonitrile / 0.05% ammonium bicarbonate solution = 0:100%–45%:55%) to afford product 13c (25 mg). ESI-MS (m / z): 470.0 [M+H] + .

[0297] Preparation of 4-amino-N-(6-ethynyl-2,3-dihydrobenzofuran-3-yl)-N,3-dimethyl-3H-pyrazolo[3,4-c]quinoline-8-carboxamide (13)

[0298] Tetrabutylammonium fluoride (100 μl, 0.10 mmol) was added to a solution of compound 13c (25 mg, 0.05 mmol) in tetrahydrofuran (2 mL) and allowed to react at 25°C for 1 hour. Water (15 mL) was added to the reaction solution, which was then 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%–35%:65%) to afford compound 13 (12 mg). ESI-MS (m / z): 398.1 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.54(s,1H),8.23(d,J=2.0Hz,1H),7.61(d,J=8.4Hz,1H),7.51(dd,J=8.4,2.0Hz,1H),7.43(d,J=7.6Hz,1H ),7.10(d,J=7.6Hz,1H),6.98(s,1H),6.87(s,2H),6.54–5.57(m,1H),4.77–4.60(m,2H),4.36(s,3H),4.19(s,1H),2.65(s,3H).

[0299] Example 14: 4-amino-N-(2-ethynyl-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide

[0300] Preparation of 2-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-5-one (14b)

[0301] Compound 14a (500 mg, 3.26 mmol) was dissolved in tert-butanol (10 mL), and Mg.SO4.7H2O (974 mg, 6.51 mmol) was added. Potassium permanganate (1.03 g, 6.51 mmol) was dissolved in water (50 mL) and slowly added dropwise to the reaction system under a nitrogen atmosphere. The reaction was allowed to proceed at 25°C for 2 hours. After completion of the reaction, the mixture was filtered through celite, water (50 mL) was added, and the mixture was extracted with ethyl acetate (50 x 3 mL). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 14b (350 mg) was obtained by normal phase column chromatography (ethyl acetate / petroleum ether = 1 / 3). ESI-MS (m / z): 168.0 [M+H] + .

[0302] Preparation of 2-chloro-N-methyl-6,7-dihydro-5H-cyclopenta[b]pyridin-5-amine (14c)

[0303] To a solution of compound 14b (350 mg, 2.10 mmol) in dichloromethane (10 mL) were added methylamine hydrochloride (707 mg, 10.48 mmol), N,N-diisopropylethylamine (2.0 mL, 12.6 mmol), and acetic acid (0.18 mL, 3.2 mmol). The reaction was allowed to proceed at room temperature for 30 minutes, followed by the addition of sodium borohydride (160 mg, 4.2 mmol) and the reaction continued for 2 hours. After completion of the reaction, water (20 mL) was slowly added, and the mixture was extracted with dichloromethane (20 x 3 mL). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 14c (280 mg) was obtained by normal phase column chromatography (ethyl acetate / petroleum ether = 2 / 3). ESI-MS (m / z): 183.1 [M+H] + .

[0304] Preparation of 4-amino-N-(2-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (14d)

[0305] To a solution of compound 14c (91 mg, 0.50 mmol) in N,N-dimethylacetamide (2 mL) were added compound 2a (121 mg, 0.50 mmol), O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethylisourea boron tetrafluoride (285 mg, 0.75 mmol), and N,N-diisopropylethylamine (129 mg, 1.00 mmol). The mixture was reacted at 25°C for 2 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 = 1:10) to afford compound 14d (122.0 mg). ESI-MS (m / z): 407.1 [M+H]. + .

[0306] Preparation of 4-amino-N,1-dimethyl-N-(2-((trimethylsilyl)ethynyl)-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (14e)

[0307] To a solution of compound 14d (122 mg, 0.3 mmol) in N,N-dimethylacetamide (2 mL) were added triethylamine (151 mg, 1.5 mmol), methanesulfonic acid (2-dicyclohexylphosphino-2',4',6'-tri-isopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl) palladium(II) (51 mg, 0.06 mmol), and trimethylsilyl acetylene (89 mg, 0.9 mmol). The mixture was microwave-dried at 70°C for 2 hours under nitrogen. The reaction mixture was filtered through celite, water (20 mL) was added, and extraction with ethyl acetate (20 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 14e (58 mg) was obtained by normal phase column chromatography (methanol / dichloromethane = 1:20). ESI-MS (m / z): 469.1 [M+H]. + .

[0308] Preparation of 4-amino-N-(2-ethynyl-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (14)

[0309] To a solution of compound 14e (58 mg, 0.12 mmol) in tetrahydrofuran (5 mL) was added tetrabutylammonium fluoride (62.6 mg, 0.24 mmol) and the mixture was allowed to react at 25°C for 30 minutes. Water (15 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (10 x 3 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The mixture was purified by reverse-phase column chromatography (acetonitrile / 0.05% ammonium bicarbonate solution = 0:100%–50%:50%) to afford compound 14 (19 mg). ESI-MS (m / z): 397.0 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.35(s,1H),8.27(s,1H),7.84(d,J=7.6Hz,1H),7.71–7.61(m,2H),7.45(d,J=7.6Hz,1H ),7.14(s,2H),5.62(s,1H),4.41(s,3H),4.32–4.30(s,1H),3.10–2.84(m,2H),2.75(s,3H),2.49–2.17(m,2H).

[0310] Example 15: 4-amino-N-(5-ethynyl-2,3-dihydro-1H-inden-1-yl)-N,1-dimethylpyrazolo[4,3-c]quinoline-8-carboxamide

[0311] Preparation of 4-amino-N,1-dimethyl-N-(5-(trimethylsilyl)ethynyl)-2,3-dihydro-1H-inden-1-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (15a)

[0312] Under nitrogen, trimethylsilylacetylene (30.6 mg, 0.31 mmol), cuprous iodide (5.93 mg, 0.03 mmol), N,N-diisopropylethylamine (80.4 mg, 0.62 mmol), and tetrakis(triphenylphosphine)palladium (18.0 mg, 0.016 mmol) were added sequentially to a solution of compound 11c (70 mg, 0.16 mmol) in N-methylpyrrolidone (2 mL). The mixture was microwaved at 70°C for 3 hours. Water (20 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 15a (19 mg) was obtained by column chromatography (methanol:dichloromethane = 1:20). ESI-MS (m / z): 468.1 [M+H]. + .

[0313] Preparation of 4-amino-N-(5-ethynyl-2,3-dihydro-1H-inden-1-yl)-N,1-dimethylpyrazolo[4,3-c]quinoline-8-carboxamide (15)

[0314] To a solution of compound 15a (19 mg, 0.041 mmol) in tetrahydrofuran (1 mL) was added tetrabutylammonium fluoride solution (0.1 mL, 1 M) and the mixture was allowed to react at room temperature for 1 hour. Water 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 filtrate was concentrated. Column chromatography (methanol:dichloromethane = 1:20) afforded compound 15 (3 mg). ESI-MS (m / z): 396.06 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.34(s,1H),8.27(s,1H),7.68–7.61(m,2H),7.38(s,3H),7.15(s,2H),4. 41(s,3H),4.15(s,1H),3.11–2.86(m,2H),2.72(s,3H),2.44–2.30(m,1H),2.24–2.08(m,1H).

[0315] Example 16: 4-amino-7-chloro-N-(6-ethynyl-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide

[0316] Preparation of 4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-7-chloro-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (16b)

[0317] To a solution of compound 6a (400 mg, 1.45 mmol) in N,N-dimethylformamide (5 mL) were added compound 16a (329.7 mg, 1.45 mmol, prepared according to WO 2022 / 115377A1), N,N-diisopropylethylamine (957.81 μL, 5.78 mmol), and O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethylisourea boron tetrafluoride (1099.2 mg, 2.89 mmol), and the mixture was reacted at 25°C overnight. The reaction solution was added with water (100 mL) and extracted with ethyl acetate (2*50 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 16b (600 mg) was obtained by normal phase column chromatography (dichloromethane / methanol = 10:1). ESI-MS (m / z): 485.8 [M+H] + .

[0318] Preparation of 4-amino-7-chloro-N,1-dimethyl-N-(6-((trimethylsilyl)ethynyl)-2,3-dihydrobenzofuran-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (16c)

[0319] To a solution of 16b (600 mg, 1.23 mmol) in N-methylpyrrolidone (5 mL) were added cuprous iodide (47.0 mg, 0.25 mmol), tetrakistriphenylphosphine palladium (142.4 mg, 0.12 mmol), N,N-diisopropylethylamine (612.48 μL, 3.70 mmol), and trimethylsilyl acetylene (605.2 mg, 6.16 mmol). The mixture was microwaved at 70°C under nitrogen for 3 hours. The reaction mixture was filtered through celite, water (100 mL) was added, and extraction with ethyl acetate (2 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. The product 16c (500 mg) was obtained by normal phase column chromatography (dichloromethane / methanol = 10:1). ESI-MS (m / z): 504.1 [M+H]. + .

[0320] Preparation of 4-amino-7-chloro-N-(6-ethynyl-2,3-dihydrobenzofuran-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (16)

[0321] To a solution of compound 16c (504.1 mg, 1.00 mmol) in tetrahydrofuran (10 mL) was added tetrabutylammonium fluoride (2 mL, 2.00 mmol) and the mixture was allowed to react at 25°C for 1 hour. Water (50 mL) was added to the reaction solution, which was then extracted with ethyl acetate (3 x 50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The product was purified by normal phase column chromatography (dichloromethane / methanol = 20:1) to afford 350 mg of the crude product. 20 mg of the crude product was further purified by reverse phase column chromatography (acetonitrile / 0.05% ammonium bicarbonate solution = 0:100%–45%:55%) to afford compound 16 (13.1 mg). ESI-MS (m / z): 432.0 [M+H] + . 1 H NMR (400MHz, DMSO) δ8.38–8.18(m,2H),7.69–7.57(m,1H),7.51–7.22(m,3H),7.14–6.91(m,2H),6.52–5.38(m,1H),4.86–4.51(m,2H),4.44and 4.38(s,3H),4.19–4.17(m,1H),2.69–2.47(m,3H).

[0322] Example 17: 4-amino-N-(6-ethynyl-2,3-dihydrofuro[2,3-b]pyridin-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide

[0323] Preparation of 6-chloro-2-(2-methoxy-2-oxoethoxy)nicotinate (17b)

[0324] Compound methyl glycolate (2.60 g, 5.00 mmol) was dissolved in ethylene glycol dimethyl ether (10 mL), cooled to 0°C, and sodium hydroxide (1.1 g, 5.25 mmol, 60% purity) was added. The reaction solution was reacted at 0°C for 30 minutes, and compound 17a (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 obtain crude product 17b (1.15 g). MS (m / z): 259.95 [M+H] + .

[0325] Preparation of methyl 6-chloro-3-hydroxyfuro[2,3-b]pyridine-2-carboxylate (17c)

[0326] Compound 17b (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 17c (240 mg). ESI-MS (m / z): 228.08 [M+H] + .

[0327] Preparation of 6-chlorofuran[2,3-b]pyridin-3-ol (17d)

[0328] Compound 17c (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 17d (140 mg). ESI-MS (m / z): 170.01 [M+H] + .

[0329] Preparation of 6-chloro-2,3-dihydrofuro[2,3-b]pyridin-3-ol (17e)

[0330] Compound 17d (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 obtain 17e (98 mg). ESI-MS (m / z): 172.04 [M+H] + .

[0331] Preparation of (6-chloro-2,3-dihydrofuro[2,3-b]pyridin-3-yl) 2-{[(2-methylprop-2-yl)oxy]carbonyl}carbamate (17f)

[0332] To a solution of compound 17e (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 17f (1.10 g). ESI-MS (m / z): 371.15 [M+H] + .

[0333] Preparation of tert-butyl (6-chloro-2,3-dihydrofuro[2,3-b]pyridin-3-yl)carbamate (17 g)

[0334] Compound 17f (1.10 g, 2.97 mmol) was dissolved in acetonitrile (20 mL), and lithium bromide (0.80 g, 8.90 mmol, 60% purity) was added. The reaction solution was reacted at 65°C for 16 hours, and then the reaction solution was quenched by adding water (5 mL). The mixture was extracted with ethyl acetate (5 mL*3). The organic phases were combined and concentrated to obtain the crude product. Column chromatography (petroleum ether:ethyl acetate = 3:1) gave 17 g (610 mg). ESI-MS (m / z): 271.05 [M+H] + .

[0335] Preparation of tert-butyl (6-chloro-2,3-dihydrofuran[2,3-b]pyridin-3-yl)(methyl)carbamate (17h): Compound 17g (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), extracted with ethyl acetate (5 mL*3), and the organic phases were combined and concentrated to obtain a crude product, which was subjected to thin layer preparative chromatography (petroleum ether: ethyl acetate = 20:1) to obtain 17h (110 mg).

[0336] Preparation of 6-chloro-N-methyl-2,3-dihydrofuro[2,3-b]pyridin-3-amine hydrochloride (17i)

[0337] Compound 17h (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 17i (40 mg). ESI-MS (m / z): 184.96 [M+H] + .

[0338] Preparation of 4-amino-N-(6-chloro-2,3-dihydrofluoro[2,3-b]pyridin-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (17j)

[0339] To a solution of 17i (40 mg, 0.22 mmol) in N,N-dimethylformamide (5 mL) were added compound 2a (52.5 mg, 0.22 mmol), O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethylisourea boron tetrafluoride (82.4 mg, 0.22 mmol), and triethylamine (6.06 mg, 0.66 mmol). The mixture was allowed to react at 25°C for 3 hours. Water (10 mL) was added, and the mixture was extracted with ethyl acetate (20 x 5 mL). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 17j (30.0 mg) was obtained by normal phase column chromatography (methanol / dichloromethane = 0:100%–10%:90%). ESI-MS (m / z): 408.99 [M+H] + .

[0340] Preparation of 4-amino-N,1-dimethyl-N-(6-((trimethylsilyl)ethynyl)-2,3-dihydrofluoro[2,3-b]pyridin-3-yl)-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (17k)

[0341] To a solution of 17j (20 mg, 0.05 mmol) in N,N-dimethylformamide (2 mL) was added methanesulfonic acid (2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl) palladium(II) (8.5 mg, 0.01 mmol), triethylamine (0.03 mL, 0.20 mmol), and trimethylsilyl acetylene (9.8 mg, 0.10 mmol). The mixture was microwave-dried 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) was performed. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The mixture was purified by normal phase column chromatography (dichloromethane / methanol = 20:1) to afford compound 17k (15.0 mg). ESI-MS (m / z): 471.3 [M+H] +

[0342] Preparation of 4-amino-N-(6-ethynyl-2,3-dihydrofuro[2,3-b]pyridin-3-yl)-N,1-dimethyl-1H-pyrazolo[4,3-c]quinoline-8-carboxamide (17)

[0343] Tetrabutylammonium fluoride (20.1 mg, 0.06 mmol) was added to a solution of compound 17k (15 mg, 0.03 mmol) in tetrahydrofuran (2 mL), and the mixture was allowed to react at 25°C for 0.5 h. Water (5 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (5 x 3 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 17 (5.0 mg) was obtained by reverse-phase column chromatography (acetonitrile / 0.05% ammonium bicarbonate solution = 0:100%–50%:50%). ESI-MS (m / z): 399.2 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 8.34–8.30 (m, 1H), 8.28 (s, 1H), 7.93 (dd, J = 7.6, 1.2 Hz, 1H), 7.68–7.60 (m, 2H), 7.22 (d, J = 7.6 Hz, 1H), 7.17 (s, 2H), 6.38–6.01 (m, 1H), 4.88–4.68 (m, 2H), , 4.43 (s, 3H), 4.34 (s, 1H), 2.74 (s, 3H). Reference Example

[0344] The preparation method refers to WO2022169948A1.

[0345] Test Example 1 Inhibitory activity of the compounds of the present invention on the proliferation of human colon cancer cells

[0346] 1. Build MTAP - / - HCT-116 cell line

[0347] The colon cancer cell line HCT-116 (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 testing and cell proliferation testing.

[0348] 2. Cell proliferation

[0349] 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 is the concentration of compound that results in half inhibition.

[0350] Table 1 IC 50 (μM)

[0351] The results showed that the compounds of the present invention had strong proliferation inhibitory activity against the human MTAP-deficient HCT-116 cell line. The IC values ​​of compounds 1 to 38 against the human MTAP-deficient HCT-116 cell line were 50 The values ​​were all less than 0.04 μM. The IC values ​​of compounds 1 to 9, 16, and 17 were 50 The values ​​were all less than 0.05 μM; the IC 50The value can be less than 10 nM. Compared with the MTAP wild-type (WT) HCT-116 cell line, the compounds of the present invention have good selective inhibition against the human MTAP-deficient HCT-116 cell line. The selective inhibition of compounds 1 to 38 against the human MTAP-deficient HCT-116 cell line is greater than 50 times, among which compound 3 has a selective inhibition of up to 301 times against the human MTAP-deficient HCT-116 cell line.

[0352] Test Example 2 Pharmacokinetics of the compounds of the present invention in mice

[0353] Materials and methods

[0354] The experimental animals were healthy adult BALB / c female mice (provided by Beijing Huafukang Biotechnology Co., Ltd. and Sichuan Weitonglihua Laboratory Animal Technology Co., Ltd.);

[0355] BALB / c female mice were given a single oral gavage (10 mg / kg). 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 blood was centrifuged at 4000 rpm for 6 min to obtain plasma.

[0356] Sample analysis:

[0357] 10 μL of mouse plasma samples were collected and protein was precipitated with 190 μL of acetonitrile solution containing an internal standard. The samples were vortexed for 10 minutes and then centrifuged at 4000 rpm for 10 minutes. The supernatant was then transferred to a 96-well plate and analyzed by LC-MS / MS (Shimadzu LC-30A, AB API 4500). 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 profiles.

[0358] Experimental results:

[0359] Table 2 Pharmacokinetic parameters of the compounds of the present invention in mice

[0360] Note: 1. The solvent used is 0.5% MC / 0.2% Tween 80

[0361] The results showed that the compound 1-38 of the present invention had better AUC and higher Cmax in mouse pharmacokinetics than the reference example.

[0362] Industrial applicability

[0363] The compounds of the present invention have excellent PRMT5 inhibitory activity and strong anti-proliferation activity against the human MTAP-deficient HCT-116 cell line. Furthermore, the compounds of the present invention have excellent selective inhibition against the human MTAP-deficient HCT-116 cell line relative to the MTAP wild-type (WT) HCT-116 cell line, and may be used as drugs for treating or preventing diseases associated with this effect. Furthermore, in vivo pharmacokinetic testing in mice demonstrated that the compounds of the present invention have superior in vivo pharmacokinetic advantages compared to the prior art (reference examples), resulting in improved drugability and clinical application prospects.

[0364] 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.

Claims

1. The compound represented by Formula I, its tautomers, stereoisomers, pharmaceutically acceptable salts or deuterated compounds thereof: in, is selected from a single bond or a double bond, or does not exist; 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 ; when When absent, Y is absent, X is hydrogen, and Z is CR 4 R 5 ; W is selected from CR 7 or N; A is selected from CR 8 R 9 ,NR 8 or O; Ring M is selected from 5-10 membered aryl or 5-10 membered heteroaryl; 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; 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 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; R 2 Selected from: hydrogen, C 1-6 Alkyl, C 3-6 Cycloalkyl, the C 1-6 Alkyl, C 3-6 The cycloalkyl group is unsubstituted or substituted by one or more identical or different R a replace; n1, n2 are each independently 0, 1, 2; 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; Each R a Each independently selected from hydroxy, deuterium, cyano, halogen, -NR'R", optionally substituted by 1, 2, 3 or 4 halogen, hydroxy, cyano, C 1-6 Alkoxy-substituted C 1-6 Alkoxy; wherein R' and R" are each 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.

2. The compound according to claim 1, its tautomers, stereoisomers, pharmaceutically acceptable salts or deuterated compounds thereof, characterized in that: Ring M is selected from 5-6 membered aryl or 5-6 membered heteroaryl; Preferably, ring M is selected from phenyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrazolyl, imidazolyl, triazolyl, pyrrolyl, furanyl, oxazolyl, isoxazolyl, thiazolyl; Further preferably, ring M is selected from phenyl or pyridyl; Preferably, A is selected from O; Preferably, n1 and n2 are each independently 0 or 1; Preferably, the group Selected from Preferably, the group Selected from: Or preferably, A is selected from -CH2- or O; Preferably, the group Selected from Preferably, the group Selected from: Preferably, the group Selected from: Preferably, the group Selected from:

3. The compound according to claim 1, its tautomers, stereoisomers, pharmaceutically acceptable salts or deuterated compounds thereof, characterized in that: R 2 Selected from hydrogen, C 1-4 Alkyl, C 5-6 Alkyl, C 3-4 Cycloalkyl, C 5-6 Cycloalkyl, the C 1-4 Alkyl, C 5-6 Alkyl, C 3-4 Cycloalkyl, C 5-6 The cycloalkyl group is unsubstituted or substituted by one or more identical or different R a replace; Preferably, R 2 Selected from hydrogen, C 1-4 Alkyl, C 3-4 Cycloalkyl, the C 1-4 Alkyl, C 3-4 The cycloalkyl group is unsubstituted or substituted by one or more identical or different R a replace; Preferably, R 2 is selected from hydrogen, methyl, ethyl, propyl, isopropyl, butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, wherein the methyl, ethyl, propyl, isopropyl, butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl is unsubstituted or replaced by one or more identical or different R a replace; Preferably, R 2 is selected from hydrogen, methyl, ethyl, propyl, cyclopropyl, cyclobutyl, cyclopentyl, said methyl, ethyl, cyclopropyl, cyclobutyl, cyclopentyl being unsubstituted or substituted by one or more identical or different R a replace; Preferably, R 2 is selected from hydrogen, methyl, ethyl or cyclopropyl, said methyl, ethyl or cyclopropyl being unsubstituted or substituted by 1, 2 or 3 identical or different R a replace; Preferably, R 2 is selected from methyl or ethyl, the methyl group being unsubstituted or substituted by 1, 2 or 3 identical or different R a replace; Preferably, R 2 isopropyl, which is unsubstituted or substituted by 1, 2 or 3 identical or different R a replace; Preferably, R 2 is selected from cyclopropyl, said cyclopropyl being unsubstituted or substituted by 1, 2 or 3 identical or different R a replace; Preferably, R 2 is hydrogen; Preferably, each R a Each independently selected from hydroxyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, NR'R", wherein R' and R" are independently selected from hydrogen, C 1-6 Alkyl, halogenated C 1-6 alkyl; Preferably, each R a Each independently selected from hydroxyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, NR'R", wherein R' and R" are independently selected from hydrogen, C 1-4 alkyl; Preferably, each R a Each is independently selected from hydroxy, methoxy, ethoxy, propoxy, isopropoxy, butoxy, halogenated methoxy, halogenated ethoxy, halogenated propoxy, halogenated isopropoxy, halogenated butoxy, NR'R", wherein R' and R" are each independently selected from hydrogen, methyl, ethyl, propyl, isopropyl, butyl; Preferably, each R a are each independently selected from hydroxy, methoxy, ethoxy, fluoro- or chloro-substituted methoxy, fluoro- or chloro-substituted ethoxy, NR'R", wherein R' and R" are each independently selected from hydrogen, methyl or ethyl; Preferably, each R a Each independently selected from hydroxy, methoxy, -OCF3, -OCHF2, N,N-dimethylamino; Preferably, each R a Each independently selected from hydroxy, methoxy, -OCF3, -OCHF2, N,N-dimethylamino, N-methylamino; Preferably, each R a Each independently selected from hydroxy, methoxy, N-methylamino; Preferably, R 2 Selected from: hydrogen, methyl, methoxymethyl, trifluoromethoxymethyl, difluoromethoxymethyl, hydroxymethyl, cyclopropyl, Preferably, R 2 Selected from Preferably, R 2 Selected from: hydrogen, methyl, methoxymethyl, trifluoromethoxymethyl, difluoromethoxymethyl, hydroxymethyl, cyclopropyl, Preferably, R 2 Selected from hydrogen, methyl, methoxymethyl, trifluoromethoxymethyl, difluoromethoxymethyl, hydroxymethyl, cyclopropyl, Preferably, R 2 Selected from hydrogen, methoxymethyl, 4. The compound according to any one of claims 1 to 3, its tautomers, stereoisomers, pharmaceutically acceptable salts or deuterated compounds thereof, characterized in that: R 3 Selected from hydrogen, halogen, C 1-6 Alkyl, C 3-6 Cycloalkyl; Preferably, R 3 Selected from hydrogen, F, Cl, Br, C 1-4 Alkyl, C 5-6 Alkyl, C 3-5 Cycloalkyl; Preferably, R 3 Selected from hydrogen, F, Cl, Br, C 1-4 Alkyl, C 3-5 Cycloalkyl; Preferably, R 3 is selected from hydrogen, F, Cl, Br, methyl, ethyl, propyl, isopropyl, butyl, cyclopropyl, cyclobutyl, cyclopentyl; Preferably, R 3 is selected from hydrogen, F, Cl, methyl, ethyl, cyclopropyl; Or preferably, R 3 Selected from hydrogen, F, Cl, Br, C 1-4 alkyl; Preferably, R 3 Selected from hydrogen, F, Cl, methyl, ethyl; Preferably, R 3 is selected from hydrogen, F, methyl; Preferably, R 3 selected from hydrogen; Preferably, the group Selected from Preferably, the group Selected from Preferably, the group Selected from Preferably, the group Selected from Or preferably, the group Selected from Preferably, the group Selected from Preferably, the group Selected from Preferably, the group Selected from 5. The compound according to claim 1, its tautomers, stereoisomers, pharmaceutically acceptable salts or deuterated compounds thereof, characterized in that: Group Selected from Wherein, W is selected from N or CR 7 ; Preferably, R 4 , R 5 , R 6 , R 7 are each independently selected from hydrogen, halogen, C 1-6 alkyl; Preferably, R 4 , R 5 , R 6 , R 7 are each independently selected from hydrogen, halogen, C 1-4 Alkyl, C 5-6 alkyl base; Preferably, R 4 , R 5 , R 6 , R 7 are each independently selected from hydrogen, halogen, C 1-4 alkyl; Preferably, R 4 , R 5 , R 6 , R 7 Each is independently selected from hydrogen, F, Cl, Br, methyl, ethyl, propyl, isopropyl; Preferably, R 4 , R 5 , R 6 , R 7 Each is independently selected from hydrogen, F, Cl, methyl; Preferably, the group Selected from Wherein, W is selected from N or CR 7 , R 7 Selected from H, F, Cl, methyl; Preferably, the group Selected from: Preferably, the group Selected from: Or preferably, the group Selected from: Preferably, the group Selected from: Preferably, the group Selected from:

6. The compound according to claim 1, its tautomers, stereoisomers, pharmaceutically acceptable salts or deuterated compounds thereof, characterized in that: R 1 Selected from: C 1-6 Alkyl, C 3-10 Cycloalkyl, C 3-10 Cycloalkyl-C 1-6 Alkyl-, 5-10 membered heteroaryl-; wherein the C1-C6 alkyl 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; Preferably, 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; Preferably, 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 Substitution; 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; Preferably, 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; Preferably, 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; Preferably, 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; Preferably, each R l are independently selected from halogen, halogenated C 1-6 Alkyl, C 1-6 Alkyl, C 3-6 Cycloalkyl; Preferably, each R l are independently selected from halogen, halogenated C 1-4 Alkyl, C 1-4 Alkyl, C 3-6 Cycloalkyl; Preferably, each R l are each independently selected from halogen, C 1-4 Alkyl, C 3-6 Cycloalkyl; Preferably, each R l Each is independently selected from F, Cl, Br, methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl; Preferably, each R l Each independently selected from F, Cl, methyl, ethyl; Preferably, each R l are each independently selected from methyl; Preferably, R 1 Selected from methyl, ethyl, cyclopropyl, Preferably, R 1 Selected from methyl.

7. The compound according to any one of claims 1 to 6, its tautomers, stereoisomers, pharmaceutically acceptable salts or deuterated compounds thereof, wherein the compound represented by formula (I) is selected from:

8. 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 7 or its tautomer, stereoisomer, pharmaceutically acceptable salt or deuterated compound, and optional pharmaceutical excipients; Preferably, the disease associated with abnormal PRMT5 expression is tumor or cancer; Preferably, the disease associated with abnormal PRMT5 expression refers to a disease associated with abnormal PRMT5 expression caused by MTAP deficiency; Preferably, the disease associated with abnormal PRMT5 expression refers to a tumor or cancer with MTAP deficiency.

9. Use of the compound or tautomer, stereoisomer, pharmaceutically acceptable salt or deuterated compound according to any one of claims 1 to 7, or the pharmaceutical composition according to claim 8, in the preparation of a medicament for treating and / or preventing diseases associated with abnormal PRMT5 expression, and / or in treating and / or preventing diseases associated with abnormal PRMT5 expression; Preferably, the disease associated with abnormal PRMT5 expression is tumor or cancer; Preferably, the disease associated with abnormal PRMT5 expression refers to a disease associated with abnormal PRMT5 expression caused by MTAP deficiency; Preferably, the disease associated with abnormal PRMT5 expression refers to a tumor or cancer with MTAP deficiency.

10. The compound or tautomer, stereoisomer, pharmaceutically acceptable salt or deuterated compound according to any one of claims 1 to 7, or the pharmaceutical composition according to claim 8, for treating and / or preventing diseases associated with abnormal PRMT5 expression. Preferably, the disease associated with abnormal PRMT5 expression is tumor or cancer; Preferably, the disease associated with abnormal PRMT5 expression refers to a disease associated with abnormal PRMT5 expression caused by MTAP deficiency; Preferably, the disease associated with abnormal PRMT5 expression refers to a tumor or cancer with MTAP deficiency.

Images