WO2020259478A1

WO2020259478A1 - Tricyclic compound as prmt5 inhibitor and application thereof

Info

Publication number: WO2020259478A1
Application number: PCT/CN2020/097652
Authority: WO
Inventors: 王勇; 赵立文; 全旭; 郑国闯; 孙韡; 詹康宁; 杨婷婷
Original assignee: 南京圣和药物研发有限公司
Priority date: 2019-06-24
Filing date: 2020-06-23
Publication date: 2020-12-30
Also published as: CN112125886A; CN112125886B

Abstract

A tricyclic compound as represented by formula (I) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, preparation methods therefor, and a pharmaceutical composition containing the compounds, and use of the compounds or composition for treating PRMT5-mediated diseases. The compound shows remarkable inhibitory activity on PRMT5.

Description

Tricyclic compounds as PRMT5 inhibitors and their applications

Technical field

The present invention belongs to the field of medicinal chemistry, and specifically relates to tricyclic compounds or isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs thereof as PRMT5 inhibitors, their preparation methods and pharmaceutical combinations containing these compounds And the use of these compounds or compositions for the treatment of PRMT5-mediated diseases.

Background technique

DNA modification plays a central role in triggering gene expression programs at different stages of cell growth and development. Arginine methylation plays an important role in cell processes, including signal transduction, transcription, RNA processing, DNA recombination and repair . Protein arginine methyltransferases (PRMTs) catalyze the methylation of specific arginine residues by transferring methyl groups from S-adenosylmethionine (SAM) to the guanidine nitrogen of arginine. The different arginine methylation methods can divide PRMTs into three categories: Type I (PRMT 1, 2, 3, 4, 6 and 8) catalyzes monomethylation and asymmetric dimethylation, and type II (PRMT5). And PRMT9) catalyze monomethylation and symmetric dimethylation, while type III (PRMT7) only performs monomethylation.

Among them, PRMT5 specifically binds to methyltransferase complex protein 50 (MEP50), which can symmetrically methylate histones H3 and H4, and regulate the transcription of specific target genomes. PRMT5 catalyzed histone H3 arginine 8 (R8) and H4R3 symmetric dimethylation has been shown to inhibit the expression of several tumor suppressor genes, such as tumor suppressor gene 7 (ST7), retinoblastoma (RB) tumor suppressor genes Family and receptor type O protein tyrosine phosphatase (PTPROt).

In addition to the ability to methylate histones, PRMT5 can also methylate several important transcription factors, making it play an important role in cell regulation. PRMT5 can methylate p53 and change its DNA binding activity, thereby triggering changes in the gene expression program controlled by p53. PRMT5 has also been shown to methylate N-MYC and change its protein stability and enhance its oncogenic activity in neuroblastoma. PRMT5 can also directly methylate transcription factors, including E2F-1 and NF-κB/p65, and induce the expression of its target genes. PRMT5 can not only modify nuclear transcription factors, but also methylate cytoplasmic proteins such as golgin and ribosomal protein S10 (RPS10). Therefore, in addition to its ability to directly regulate its own target genes, PRMT5 can also indirectly affect global gene expression through symmetric methylation of key transcription factors, thereby affecting cell growth, proliferation and differentiation.

Numerous studies have confirmed that PRMT5 is overexpressed in different types and aggressive cancers, including B-cell and T-cell lymphoma, metastatic melanoma, neuroblastoma and glioblastoma, germ cell tumors, ovarian cancer, Nasopharyngeal cancer, breast cancer, colorectal cancer and gastric cancer. Current research shows that PRMT5 plays an important role in controlling cell growth and proliferation, and its overexpression promotes cell transformation.

The enhanced expression of PRMT5 in cancer cells is related to the transcriptional silencing of its target tumor suppressor genes. PRMT5 can promote the growth of cancer cells by methylation of the promoter histones H3R8 and H4R3 and by modifying specific arginine residues of key transcription factors including E2F1 and NF-kB/p65 to cause global chromatin changes. PRMT5 also interacts with programmed cell death 4 (PDCD4), causing it to become methylated at R110 and lose its tumor suppressor activity in MCF-7 cells. In general, the overexpression of PRMT5 may make it interact with growth promoting proteins and tumor suppressor proteins to facilitate the growth, survival and metastasis of cancer cells.

In summary, PRMT5 inhibitors have a clear mechanism in the treatment of tumors and other related diseases, and have great potential to become a new treatment method in the field of tumor treatment. Therefore, it is necessary to develop safer and more effective PRMT5 inhibitors to meet clinical needs. demand.

Summary of the invention

An object of the present invention is to provide a class of compounds with PRMT5 inhibitory activity represented by general formula (I) or isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs thereof,

among them,

Ring A is selected from aryl, heteroaryl, cycloalkyl and heterocyclyl, and the aryl, heteroaryl, cycloalkyl and heterocyclic group may be one or more selected from halogen, hydroxy, alkyl, Haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxy, cyano, amino, monoalkylamino, alkylacylamino, alkylacyl, alkylsulfonyl, amino Substitution of acyl, alkylaminoacyl, dialkylamino, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and oxo groups;

Cy is selected from the group consisting of aryl, heteroaryl, cycloalkyl, and heterocyclyl. The aryl, heteroaryl, cycloalkyl, and heterocyclic group may be one or more selected from halogen, hydroxy, alkyl, and halogenated alkyl. Group, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxy, cyano, amino, monoalkylamino, alkylacylamino, alkylacyl, alkylsulfonyl, aminoacyl , Alkylamino acyl, dialkylamino, alkenyl, alkynyl, haloalkyl acyl, hydroxyalkyl acyl, cycloalkyl acyl, heterocyclyl acyl, cycloalkyl, heterocyclic, aryl, heteroaryl And oxo group substitution;

R ¹ , R ² , R ³ , and R ⁴ are each independently selected from hydrogen, halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxy, Cyano, amino, monoalkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl and dialkylamino;

R ^{5 is} selected from hydrogen, alkyl, haloalkyl, hydroxyalkyl, alkylacyl, aminoacyl and alkylaminoacyl;

X ¹ , X ² , and X ³ are each independently selected from N and C (R ⁶ ), wherein R ^{6 is} selected from hydrogen, halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, Hydroxyalkoxy, nitro, carboxy, cyano, amino, monoalkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl, dialkylamino, and cycloalkyl; and

L is selected from -NH-, -S-, -O-, -C(O)NH-, -NHC(O)-, -OC(O)NH-, -NHC(O)O-, -S(O ) ₂ NH-, -NHS(O) ₂ -, -C(O)- and -NHC(O)NH-.

Another object of the present invention is to provide a method for preparing the compound of general formula (I) of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug.

Another object of the present invention is to provide a composition comprising the compound of the general formula (I) of the present invention or its isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs and a pharmaceutically acceptable carrier, and The compound of the general formula (I) of the present invention or its isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs and a combination of another or more drugs.

Another object of the present invention is to provide a method for treating PRMT5-mediated diseases with the compound of general formula (I) or isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs thereof of the present invention, and the present invention Application of the compound of general formula (I) or its isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs in the preparation of medicines for treating PRMT5-mediated diseases.

For the above-mentioned purpose of the invention, the present invention provides the following technical solutions:

In the first aspect, the present invention provides a compound represented by general formula (I) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof,

among them,

Cy is selected from the group consisting of aryl, heteroaryl, cycloalkyl, and heterocyclyl. The aryl, heteroaryl, cycloalkyl, and heterocyclic group may be one or more selected from halogen, hydroxy, alkyl, and halogenated alkyl. Group, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxy, cyano, amino, monoalkylamino, alkylacylamino, alkylacyl, alkylsulfonyl, aminoacyl , Alkylamino acyl, dialkylamino, alkenyl, alkynyl, haloalkyl acyl, hydroxyalkyl acyl, cycloalkyl acyl, heterocyclyl acyl, cycloalkyl, heterocyclyl, aryl, heteroaryl And oxo group substitution;

In some preferred embodiments, the compound of the present invention is a compound of general formula (I) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein:

Ring A is selected from C _6-12 aryl, 5-12 membered heteroaryl, C _3-12 cycloalkyl and 3-12 membered heterocyclic group, the C _6-12 aryl, 5-12 membered heteroaryl Group, C _3-12 cycloalkyl and 3-12 membered heterocyclic group can be selected by one or more selected from halogen, hydroxy, C _1-6 alkyl, halo C _1-6 alkyl, hydroxy C _1-6 Alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy, hydroxy C _1-6 alkoxy, nitro, carboxy, cyano, amino, mono C _1-6 alkylamino, C _1-6 alkyl acylamino, C _1-6 alkyl acyl, C _1-6 alkylsulfonyl, amino acyl, C _1-6 alkylamino acyl, bis C _1-6 alkylamino, C _2-10 Alkenyl, C _2-10 alkynyl, C _3-12 cycloalkyl, 3-12 membered heterocyclic group, 6-12 membered aryl, 5-12 membered heteroaryl and oxo group substitution;

Further preferably, ring A is selected from C _6-10 aryl, 5-10 membered heteroaryl, C _3-10 cycloalkyl and 3-10 membered heterocyclic group, the C _6-10 aryl, 5- The 10-membered heteroaryl group, C _3-10 cycloalkyl group and 3-10 membered heterocyclic group may be one or more selected from halogen, hydroxy, C _1-3 alkyl, halogenated C _1-3 alkyl, hydroxy C _1-3 alkyl, C _1-3 alkoxy, halogenated C _1-3 alkoxy, hydroxy C _1-3 alkoxy, nitro, carboxy, cyano, amino, mono C _1-3 alkane amino, acylamino C _1-3 alkyl, C _1-3 alkyl group, C _1-3 alkyl sulfonyl, amino group, C _1-3 alkylamino group, a bis C _1-3 alkyl group, C _2-6 alkenyl, C _2-6 alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic group, C _6-8 aryl, 5-8 membered heteroaryl and oxo groups Group substitution

More preferably, ring A is selected from C _6-8 aryl, 5-6 membered heteroaryl, C _3-6 cycloalkyl, 3-6 membered azaheterocyclyl, 3-6 membered oxoheterocyclyl, 3-6 membered sulfur heterocyclic group, the C _6-8 aryl group, 5-6 membered heteroaryl group, C _3-6 cycloalkyl group, 3-6 membered azacyclic group, 3-6 membered oxygen heterocyclic group Group, 3-6 membered sulfur heterocyclic group can be selected by one or more selected from halogen, hydroxy, C _1-3 alkyl, halo C _1-3 alkyl, hydroxy C _1-3 alkyl, C _1-3 Alkoxy, halogenated C _1-3 alkoxy, hydroxy C _1-3 alkoxy, nitro, carboxyl, cyano, amino, mono C _1-3 alkylamino, C _1-3 alkyl acylamino , C _1-3 alkyl acyl, C _1-3 alkyl sulfonyl, amino acyl, C _1-3 alkyl amino acyl, double C _1-3 alkyl amino, C _2-6 alkenyl, C _2-6 Group substitution of alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic group, C _6-8 aryl, 5-8 membered heteroaryl and oxo group;

Still further preferably, ring A is selected from phenyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, aziridine Group, azetidinyl, tetrahydropyrrolyl, dihydropyrrolyl, pyrrolyl, piperidinyl, tetrahydropyridyl, dihydropyridyl, pyridyl, propylene oxide, oxetanyl , Tetrahydrofuranyl, dihydrofuranyl, furanyl, tetrahydropyranyl, dihydropyranyl, pyranyl, sulfiethane, sulfide cyclobutanyl, tetrahydrothienyl, dihydrothienyl, Thienyl, cyclopentanyl sulfide, pyrazolyl, dihydropyrazolyl, pyrazolyl, imidazolyl, dihydroimidazolyl, imidazolyl, oxazolyl, dihydrooxazolyl, oxazolyl , Thiazolyl, dihydrothiazolyl, thiazolyl, isoxazolidinyl, dihydroisoxazolyl, isoxazolyl, isothiazolidinyl, dihydroisothiazolyl, isothiazolyl, hexahydropyrimidinyl , Tetrahydropyrimidinyl, dihydropyrimidinyl, pyrimidinyl, hexahydropyridazinyl, tetrahydropyridazinyl, dihydropyridazinyl, pyridazinyl, piperazinyl, tetrahydropyrazinyl, dihydropyrazine Group, pyrazinyl, morpholinyl, thiomorpholinyl, dioxane, dithiohexacyclic, tauryl, bicyclo[2.2.1]heptyl, azaspiro[2.3]hexane Group, oxobicyclo[3.1.0] hexane group, the group may be one or more selected from halogen, hydroxyl, C _1-3 alkyl, halogenated C _1-3 alkyl, hydroxy C _{1- 3} alkyl, C _1-3 alkoxy, halogenated C _1-3 alkoxy, hydroxy C _1-3 alkoxy, nitro, carboxyl, cyano, amino, mono C _1-3 alkylamino, C _1-3 alkyl acyl amino, C _1-3 alkyl acyl, C _1-3 alkyl sulfonyl, amino acyl, C _1-3 alkyl amino acyl, bis C _1-3 alkyl amino, C _2- Group substitution of ₆ alkenyl, C _2-6 alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic group, C _6-8 aryl, 5-8 membered heteroaryl and oxo group .

Cy is selected from C _6-12 aryl, 5-12 membered heteroaryl, C _3-12 cycloalkyl and 3-12 membered heterocyclic group, the C _6-12 aryl, 5-12 membered heteroaryl , C _3-12 cycloalkyl and 3-12 membered heterocyclic group can be selected by one or more selected from halogen, hydroxy, C _1-6 alkyl, halogenated C _1-6 alkyl, hydroxy C _1-6 alkane Group, C _1-6 alkoxy, halogenated C _1-6 alkoxy, hydroxy C _1-6 alkoxy, nitro, carboxy, cyano, amino, mono C _1-6 alkylamino, C _{1 -6} alkyl acylamino, C _1-6 alkyl acyl, C _1-6 alkyl sulfonyl, amino acyl, C _1-6 alkyl amino acyl, double C _1-6 alkyl amino, C _2-10 alkene Group, C _2-10 alkynyl, halogenated C _1-6 alkyl acyl, hydroxy C _1-6 alkyl acyl, C _3-12 cycloalkyl acyl, 3-12 membered heterocyclyl acyl, C _3-12 Cycloalkyl, 3-12 membered heterocyclic group, 6-12 membered aryl, 5-12 membered heteroaryl and oxo group substitution;

Further preferably, Cy is selected from C _6-10 aryl, 5-10 membered heteroaryl, C _3-10 cycloalkyl and 3-10 membered heterocyclic group, the C _6-10 aryl, 5-10 One or more membered heteroaryl groups, C _3-10 cycloalkyl groups and 3-10 membered heterocyclic groups may be selected from halogen, hydroxy, C _1-3 alkyl, halogenated C _1-3 alkyl, hydroxy C _1-3 alkyl, C _1-3 alkoxy, halogenated C _1-3 alkoxy, hydroxy C _1-3 alkoxy, nitro, carboxy, cyano, amino, mono C _1-3 alkyl Amino, C _1-3 alkyl acylamino, C _1-3 alkyl acyl, C _1-3 alkyl sulfonyl, amino acyl, C _1-3 alkyl amino acyl, bis C _1-3 alkyl amino, C _2-6 alkenyl, C _2-6 alkynyl, halogenated C _1-3 alkyl acyl, hydroxy C _1-3 alkyl acyl, C _3-8 cycloalkyl acyl, 3-8 membered heterocyclic acyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic group, 6-8 membered aryl, 5-8 membered heteroaryl and oxo group substitution;

More preferably, Cy is selected from phenyl, 5-6 membered heteroaryl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and 3-6 membered heterocyclyl, the phenyl, 5-6 Member heteroaryl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and 3-6 membered heterocyclic group can be selected by one or more selected from halogen, hydroxy, C _1-3 alkyl, halogenated C _{1 -3} alkyl, hydroxy C _1-3 alkyl, C _1-3 alkoxy, halogenated C _1-3 alkoxy, hydroxy C _1-3 alkoxy, nitro, carboxy, cyano, amino, Mono-C _1-3 alkylamino, C _1-3 alkyl acylamino, C _1-3 alkyl acyl, C _1-3 alkylsulfonyl, amino acyl, C _1-3 alkylamino acyl, double C _{1 -3} alkylamino, C _2-6 alkenyl, C _2-6 alkynyl, halogenated C _1-3 alkyl acyl, hydroxy C _1-3 alkyl acyl, C _3-8 cycloalkyl acyl, 3- 8-membered heterocyclyl acyl, C _3-8 cycloalkyl, 3-8 membered heterocyclyl, 6-8 membered aryl, 5-8 membered heteroaryl and oxo group are substituted.

R ¹ , R ² , R ³ , and R ⁴ are each independently selected from hydrogen, halogen, hydroxy, C _1-6 alkyl, halo C _1-6 alkyl, hydroxy C _1-6 alkyl, C _1-6 Alkoxy, halogenated C _1-6 alkoxy, hydroxy C _1-6 alkoxy, nitro, carboxyl, cyano, amino, mono C _1-6 alkylamino, C _1-6 alkyl acylamino , C _1-6 alkyl acyl, amino acyl, C _1-6 alkyl amino acyl and double C _1-6 alkyl amino;

Further preferably, R ¹ , R ² , R ³ , and R ⁴ are each independently selected from hydrogen, halogen, hydroxyl, C _1-3 alkyl, halogenated C _1-3 alkyl, hydroxy C _1-3 alkyl, C _1-3 alkoxy, halogenated C _1-3 alkoxy, hydroxyl C _1-3 alkoxy, nitro, carboxyl, cyano, amino, mono C _1-3 alkylamino, C _1-3 Alkyl acylamino, C _1-3 alkyl acyl, amino acyl, C _1-3 alkylamino acyl and bis C _1-3 alkylamino;

More preferably, R ¹ , R ² , R ³ , and R ⁴ are each independently selected from hydrogen, halogen, hydroxyl, methyl, ethyl, propyl, isopropyl, halogenated C _1-3 alkyl, hydroxyl C _1-3 alkyl, C _1-3 alkoxy, halogenated C _1-3 alkoxy, hydroxy C _1-3 alkoxy, nitro, carboxy, cyano, amino, mono C _1-3 alkane Alkylamino, C _1-3 alkyl acylamino, C _1-3 alkyl acyl, amino acyl, C _1-3 alkylamino acyl and bis C _1-3 alkylamino.

R ^{5 is} selected from hydrogen, C _1-6 alkyl, halogenated C _1-6 alkyl, hydroxy C _1-6 alkyl, C _1-6 alkyl acyl, amino acyl, C _1-6 alkylamino acyl;

More preferably, R ^{5 is} selected from hydrogen, C _1-3 alkyl, halogenated C _1-3 alkyl, hydroxy C _1-3 alkyl, C _1-3 alkyl acyl, amino acyl, C _1-3 alkane Aminoacyl;

More preferably, R ^{5 is} selected from hydrogen, methyl, ethyl, propyl, trifluoromethyl, hydroxymethyl, hydroxyethyl, methylacyl, ethylacyl, propylacyl, aminoacyl, methyl Aminoacyl, ethylaminoacyl and propylaminoacyl.

R ^{6 is} selected from hydrogen, halogen, hydroxy, C _1-6 alkyl, halogenated C _1-6 alkyl, hydroxy C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy Group, hydroxy C _1-6 alkoxy, nitro, carboxy, cyano, amino, mono C _1-6 alkylamino, C _1-6 alkyl acylamino, C _1-6 alkyl acyl, amino acyl, C _1-6 alkylaminoacyl, bis-C _1-6 alkylamino and C _3-12 cycloalkyl;

More preferably, R ^{6 is} selected from hydrogen, halogen, hydroxy, C _1-3 alkyl, halogenated C _1-3 alkyl, hydroxy C _1-3 alkyl, C _1-3 alkoxy, halogenated C _{1 -3} alkoxy, hydroxy C _1-3 alkoxy, nitro, carboxy, cyano, amino, mono C _1-3 alkylamino, C _1-3 alkyl acylamino, C _1-3 alkyl acyl , Aminoacyl, C _1-3 alkylamino acyl, double C _1-3 alkylamino and C _3-8 cycloalkyl;

More preferably, R ^{6 is} selected from hydrogen, halogen, hydroxy, methyl, ethyl, propyl, isopropyl, halogenated C _1-3 alkyl, hydroxy C _1-3 alkyl, C _1-3 alkane Oxy, halogenated C _1-3 alkoxy, hydroxy C _1-3 alkoxy, nitro, carboxy, cyano, amino, mono C _1-3 alkylamino, C _1-3 alkyl acylamino, C _1-3 alkyl acyl, amino acyl, C _1-3 alkyl amino acyl, bis C _1-3 alkylamino and C _3-8 cycloalkyl.

In some embodiments, the present invention provides a compound of general formula (Ia) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof,

Among them, Cy, L and ring A have the definitions described in the general formula (I) above.

In some embodiments, the present invention provides a compound of general formula (Ib) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof,

In some embodiments, the present invention provides a compound of general formula (Ic) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof,

In some embodiments, the present invention provides a compound of general formula (Id) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof,

In some preferred embodiments, the compound of the present invention is a compound of general formula (I), (Ia), (Ib), (Ic) or (Id) or its isomers, pharmaceutically acceptable salts, solvents Compounds, crystals or prodrugs, of which:

Cy is selected from

The present invention provides the following specific compounds or isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs thereof:

In another aspect, the present invention provides a method for preparing the compound of general formula (I) of the present invention, which includes the step of reacting the compound of formula 1 and the compound of formula 2:

Wherein, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , Cy, L, X ¹ , X ² , X ³ , and ring A have the definitions described in the general formula (I), and the compound of formula 1 and formula 2 The compound of is a commercially available compound or can be synthesized by other technical means commonly used by those skilled in the art.

In a third aspect, the present invention provides a pharmaceutical composition comprising the compound of the present invention or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof.

In some embodiments, the present invention provides the compound of the present invention or its isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs and the compounds of the present invention or its isomers, pharmaceutically acceptable Pharmaceutical compositions of salts, solvates, crystals or prodrugs, said compounds or pharmaceutical compositions are used to treat PRMT5-mediated diseases.

In some embodiments, the present invention provides a pharmaceutical composition comprising a compound of the present invention or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, and a pharmaceutically acceptable carrier.

The compound of the present invention or its isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs can be mixed with pharmaceutically acceptable carriers, diluents or excipients to prepare pharmaceutical preparations suitable for oral administration or Parenteral administration. Administration methods include, but are not limited to intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal and oral routes. The preparation can be administered by any route, for example, by infusion or bolus injection, by the route of absorption through the epithelium or skin mucosa (such as oral mucosa or rectum, etc.). Administration can be systemic or local. Examples of preparations for oral administration include solid or liquid dosage forms, specifically, tablets, pills, granules, powders, capsules, syrups, emulsions, suspensions, and the like. The preparation can be prepared by a method known in the art, and contains a carrier, diluent or excipient conventionally used in the field of pharmaceutical preparations.

In the fourth aspect, the present invention provides a compound represented by formula (I), (Ia), (Ib), (Ic) or (Id) or its isomers, pharmaceutically acceptable salts, solvates, crystals Or a prodrug, or a pharmaceutical composition containing the same for the method of treating PRMT5-mediated diseases and the use in the preparation of a medicine for treating PRMT5-mediated diseases.

In some preferred embodiments, the present invention provides a compound represented by formula (I), (Ia), (Ib), (Ic) or (Id) of the present invention or an isomer, pharmaceutically acceptable salt thereof, Solvates, crystals or prodrugs, or pharmaceutical compositions containing them are used to treat PRMT5-mediated diseases and their use in the preparation of drugs for the treatment of PRMT5-mediated diseases, wherein the PRMT5-mediated diseases include But not limited to: proliferative diseases, metabolic diseases or blood diseases. In some embodiments, the PRMT5-mediated disease of the present invention is cancer.

In some embodiments, PRMT5-mediated diseases of the present invention include but are not limited to: acoustic neuroma, adenocarcinoma, adrenal carcinoma, anal cancer, angiosarcoma (eg, lymphangiosarcoma, lymphatic endothelial sarcoma, angiosarcoma) , Accessory cancer, benign monoclonal gammopathy, bile cancer (for example, cholangiocarcinoma), bladder cancer, breast cancer (for example, breast adenocarcinoma, breast papillary cancer, breast cancer, breast medullary cancer, triple negative breast cancer ), brain cancer (for example, meningioma; glioma, such as astrocytoma, oligodendroglioma; medulloblastoma), bronchial carcinoma, carcinoid tumor, cervical cancer (for example, cervical adenocarcinoma) , Choriocarcinoma, chordoma, craniopharyngioma, colorectal cancer (for example, colon cancer, rectal cancer, colorectal adenocarcinoma), epithelial cancer, ependymoma, endothelial sarcoma (for example, Kaposi’s sarcoma ( Kaposi's sarcoma), multiple idiopathic hemorrhagic sarcoma), endometrial cancer (e.g., uterine cancer, uterine sarcoma), esophageal cancer (e.g., esophageal adenocarcinoma, Barrett's adenocarinoma) , Ewing sarcoma, eye cancer (for example, intraocular melanoma, retinoblastoma), familial eosinophilia, gallbladder cancer, gastric cancer (for example, gastric adenocarcinoma), gastrointestinal GIST, head and neck cancer (e.g., head and neck squamous cell carcinoma, oral cancer (e.g., oral squamous cell carcinoma (OSCC)), throat cancer (e.g., laryngeal cancer, pharyngeal cancer, nasopharyngeal cancer, Oropharyngeal cancer)), hematopoietic cancer (e.g., leukemia such as acute lymphoblastic leukemia (ALL) (e.g., B-cell ALL, T-cell ALL), acute myeloid leukemia (AML) (e.g., B-cell AML, T-cell AML), chronic myelogenous leukemia (CML) (e.g., B-cell CML, T-cell CML), and chronic lymphocytic leukemia (CLL) (e.g., B-cell CLL, T-cell CLL) ); Lymphomas such as Hodgkin’s lymphoma (HL) (for example, B-cell HL, T-cell HL) and non-Hodgkin’s lymphoma (NHL) (for example, B-cell NHL such as diffuse large cell lymphoma (DLCL) (for example, diffuse large B-cell lymphoma (DLBCL)), follicular lymphoma, chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL), mantle cell lymphoma (MCL) , Marginal zone B-cell lymphoma (e.g., mucosal associated lymphoid tissue (MALT) lymphoma, nodular marginal zone B-cell lymphoma, splenic marginal zone B-cell lymphoma), primary mediastinal B-cell lymphoma Tumor, Burkitt lymphoma (Burkittlymphoma), lymphoplasmacytic lymphoma (ie, "Waldanstrom macroglobulinemia (

macroglobulinemia)”), hairy cell leukemia (HCL), immunoblastic large cell lymphoma, precursor B-lymphoblastic lymphoma and primary central nervous system (CNS) lymphoma; and T-cell NHL such as Precursor T-lymphoblastic lymphoma/leukemia, peripheral T-cell lymphoma (PTCL) (e.g., cutaneous T-cell lymphoma (CTCL) (e.g., mycosis fungiodes, Sizari syndrome) Syndrome (Sezary syndrome), hemangioimmunoblastic T-cell lymphoma, extranodal natural killer T-cell lymphoma, enteropathy type T-cell lymphoma, subcutaneous panniculitis-like T-cell lymphoma, interstitial Degenerative large cell lymphoma); one or more leukemia/lymphoma mixtures as described above; and multiple myeloma (MM)), heavy chain diseases (for example, alpha chain disease, gamma chain disease, mu chain disease ), hemangioblastoma, inflammatory myofibroblastoma, immune cell amyloidosis, kidney cancer (for example, Wilms’ tumor, also known as Wilms’ tumor, renal cell carcinoma), liver cancer ( For example, hepatocellular carcinoma (HCC), malignant hepatoma), lung cancer (e.g., bronchial carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), lung adenocarcinoma), leiomyosarcoma (LMS), mast cell Hyperplasia (e.g., systemic mastocytosis), myelodysplastic syndrome (MDS), mesothelioma, myeloproliferative disease (MPD) (e.g., polycythemia vera (PV), idiopathic thrombocytosis) (ET), idiopathic myelofibrosis (AMM), also known as myelofibrosis (MF), chronic idiopathic myelofibrosis, chronic myelogenous leukemia (CML), chronic neutrophil leukemia (CNL), addiction Acidic leukocytosis (HES)), neuroblastoma, neurofibromas (e.g., type 1 or type 2 multiple neurofibroma (NF), Schwannomatosis (schwannomatosis)), neuroendocrine carcinoma (e.g. , Gastrointestinal pancreatic neuroendocrine tumors (GEP-NET), carcinoid tumors), osteosarcoma, ovarian cancer (e.g., cystadenocarcinoma, ovarian embryonic carcinoma, ovarian adenocarcinoma, ovarian clear cell carcinoma, ovarian serous cystadenocarcinoma ,), papillary adenocarcinoma, pancreatic cancer (e.g., pancreatic adenocarcinoma, intraductal papillary myxoma (IPMN), islet cell tumor), penile cancer (e.g., Paget's disease of the penis and scrotum), Pineal tumor, primary neuroectodermal tumor (PNT), prostate cancer (for example, prostate adenocarcinoma), rectal cancer, rhabdomyosarcoma, salivary duct cancer, skin cancer (for example, squamous cell carcinoma (SCC), angle Acanthoma (KA), melanoma, basal cell carcinoma (BCC)), small bowel cancer (e.g., adnexal carcinoma), soft tissue sarcoma (e.g., malignant fibrous histiocytoma (MFH), liposarcoma, malignant peripheral nerve sheath tumor (MPNST ), chondrosarcoma, fibrosarcoma, myxosarcoma), sebaceous gland cancer, sweat gland cancer, synovial tumor, testicular cancer (e.g., seminoma, testicular embryonic carcinoma), thyroid cancer (e.g., papillary thyroid carcinoma, papillary Thyroid cancer (PTC), medullary thyroid cancer), urethral cancer, vaginal cancer and vulvar cancer (for example, Paget's disease of the vulva), medulloblastoma, adenoid cystic carcinoma, melanoma, glioblastoma .

In some embodiments, the PRMT5-mediated diseases of the present invention include metabolic disorders such as diabetes or obesity.

In some embodiments, the PRMT5-mediated diseases of the present invention include hemoglobinopathies such as sickle cell disease or β-thalassemia.

In some embodiments, the PRMT5-mediated diseases of the present invention include inflammatory and autoimmune diseases.

In some preferred embodiments, the present invention provides a compound represented by general formula I of the present invention or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, or a pharmaceutical composition containing it for use Method for treating PRMT5-mediated diseases and use in the preparation of drugs for treating PRMT5-mediated diseases, wherein the PRMT5-mediated diseases include but not limited to: breast cancer, esophageal cancer, bladder cancer, lung cancer, hematopoietic system Cancer, lymphoma, medulloblastoma, medulloblastoma, rectal adenocarcinoma, colon cancer, stomach cancer, pancreatic cancer, liver cancer, adenoid cystic carcinoma, prostate cancer, lung cancer, head and neck squamous cell carcinoma, brain Carcinoma, hepatocellular carcinoma, melanoma, oligodendroglioma, glioblastoma, testicular cancer, ovarian clear cell carcinoma, ovarian serous cystadenocarcinoma, thyroid cancer, multiple myeloma (AML), kidney cells Cancer, mantle cell lymphoma, triple negative breast cancer, non-small cell lung cancer, hemoglobinopathies, diabetes, and obesity.

Definition of Terms

Unless stated to the contrary, the terms used in the specification and claims have the following meanings.

The "hydrogen", "carbon" and "oxygen" in the compounds of the present invention include all their isotopes. Isotopes should be understood to include those atoms having the same atomic number but different mass numbers. For example, hydrogen isotopes include protium, tritium, and deuterium, carbon isotopes include ¹² C, ¹³ C, and ¹⁴ C, and oxygen isotopes include ¹⁶ O and ¹⁸ O.

"Isomers" in the present invention refer to molecules with the same atomic composition and connection mode but different three-dimensional arrangements, including but not limited to diastereomers, enantiomers, cis-trans isomers, and their Mixtures, such as racemic mixtures. Many organic compounds exist in optically active forms, that is, they have the ability to rotate the plane of plane-polarized light. When describing optically active compounds, the prefixes D, L or R, S are used to indicate the absolute configuration of the molecular chiral center. The prefixes D, L or (+), (-) are used to name the symbols of the plane-polarized light rotation of the compound. (-) or L means the compound is levorotatory, and the prefix (+) or D means the compound is dextrorotatory. The chemical structures of these stereoisomers are the same, but their stereostructures are different. A specific stereoisomer may be an enantiomer, and a mixture of isomers is usually called an enantiomeric mixture. A 50:50 mixture of enantiomers is called a racemic mixture or a racemate, which may result in no stereoselectivity or stereospecificity during chemical reactions. The terms "racemic mixture" and "racemate" refer to an equimolar mixture of two enantiomers, lacking optical activity.

Depending on the choice of starting materials and methods, the compound of the present invention can be one of the possible isomers or a mixture of them, such as racemates and diastereoisomer mixtures (depending on the number of asymmetric carbon atoms) The form exists. Optically active (R)- or (S)-isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques.

Any resulting mixture of stereoisomers can be separated into pure or substantially pure geometric isomers, enantiomers, and diastereomers based on differences in the physical and chemical properties of the components, for example, by chromatography Method and/or fractional crystallization method.

The "halogen" in the present invention means fluorine, chlorine, bromine, and iodine. The "halo" in the present invention refers to substitution by fluorine, chlorine, bromine or iodine.

The "alkyl" in the present invention refers to a linear or branched saturated aliphatic hydrocarbon group, preferably a linear or branched group containing 1 to 6 carbon atoms, and more preferably a linear or branched group containing 1 to 3 carbon atoms Chain groups, non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethyl Propyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, etc. The alkyl group may be substituted or unsubstituted, and when substituted, the substituent may be at any available point of attachment.

Both "carbonyl" and "acyl" in the present invention refer to -C(O)-.

The "sulfonyl group" in the present invention means -S(O) ₂ -.

The "sulfonamide group" in the present invention means -S(O) ₂ NH-.

The "haloalkyl" in the present invention refers to an alkyl group substituted with at least one halogen.

The "hydroxyalkyl" in the present invention refers to an alkyl group substituted with at least one hydroxy group.

"Alkoxy" in the present invention refers to -O-alkyl. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, propoxy, n-propoxy, isopropoxy, isobutoxy, sec-butoxy and the like. The alkoxy group may be optionally substituted or unsubstituted, and when substituted, the substituent may be at any available point of attachment.

The "cycloalkyl" in the present invention refers to a cyclic saturated hydrocarbon group. Suitable cycloalkyl groups may be substituted or unsubstituted monocyclic, bicyclic or tricyclic saturated hydrocarbon groups with 3-12 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

The "heterocyclic group" of the present invention refers to a 3- to 12-membered non-aromatic group having 1 to 4 ring heteroatoms (wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus and silicon) Groups of ring systems ("3-12 membered heterocyclyl"). In heterocyclyl groups containing one or more nitrogen atoms, the point of attachment may be a carbon or nitrogen atom, as long as the valence permits. Heterocyclyl groups can either be monocyclic ("monocyclic heterocyclyl") or fused, bridged or spiro ring systems (e.g. bicyclic systems (also known as "bicyclic heterocyclyl")) And it can be saturated or partially unsaturated. Suitable heterocyclic groups include but are not limited to piperidinyl, azetidinyl, aziridinyl, tetrahydropyrrolyl, piperazinyl, dihydroquinazolinyl, oxepanyl, oxa Cyclobutyl, tetrahydrofuranyl, tetrahydropyranyl,

Wait. Each instance of the heterocyclic group may be optionally substituted or unsubstituted, and when substituted, the substituent may be at any available point of attachment.

The "aryl" of the present invention refers to an aromatic system that can contain a single ring or a fused polycyclic ring, preferably a single ring or a fused bicyclic aromatic system, which contains 6 to 12 carbon atoms, preferably about 6 to about 10 carbon atoms. Suitable aryl groups include, but are not limited to, phenyl, naphthyl, anthryl, fluorenyl, indanyl. The aryl group may be optionally substituted or unsubstituted, and when substituted, the substituent may be at any available point of attachment.

The "heteroaryl group" in the present invention refers to an aryl group in which at least one carbon atom is replaced by a heteroatom, preferably composed of 5-12 atoms (5-12 membered heteroaryl), and more preferably composed of 5-10 atoms (5-10 membered heteroaryl), the heteroatoms are O, S, N. The heteroaryl groups include, but are not limited to, imidazolyl, pyrrolyl, furyl, thienyl, pyrazolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, Tetrazolyl, indolyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, triazinyl, isoindolyl, benzopyrazolyl, benzimidazolyl, benzofuranyl, benzopyridine Pyryl, benzothienyl, benzoxazolyl, benzothiazolyl, benzisoxazolyl, benzisothiazolyl, quinolinyl, isoquinolinyl, quinazolinyl, cinnolinyl, Quinoxalinyl, benzoxazinyl, benzothiazinyl, imidazopyridyl, pyrimidopyrazolyl, pyrimidimidazolyl, etc. Heteroaryl groups can be optionally substituted or unsubstituted, and when substituted, the substituents can be at any available point of attachment.

The "pharmaceutically acceptable salt" of the present invention refers to the salt of the compound of the present invention. Such salt is safe and effective when used in mammals, and has due biological activity.

The "solvate" in the present invention refers to a complex formed by the combination of a solute (such as an active compound, a salt of an active compound) and a solvent (such as water) in a conventional sense. The solvent refers to a solvent known or easily determined by those skilled in the art. If it is water, the solvate is usually referred to as a hydrate, such as hemihydrate, monohydrate, dihydrate, trihydrate, or alternative amounts thereof.

The in vivo effects of the compound of formula (I) can be partly exerted by one or more metabolites formed in the body of the human or animal after the compound of formula (I) is administered. As described above, the in vivo effects of the compound of formula (I) can also be exerted through the metabolism of precursor compounds ("prodrugs"). The "prodrug" of the present invention refers to a compound that is converted into a compound of the invention by reaction with enzymes, gastric acid, etc. under physiological conditions in an organism, that is, a compound that is converted into a compound of the invention by enzyme oxidation, reduction, hydrolysis, etc. /Or a compound or the like that is converted into a compound of the invention by a hydrolysis reaction of gastric acid or the like.

The "crystalline" in the present invention refers to a solid whose internal structure is formed by repeating constituent atoms (or groups thereof) regularly in three dimensions, which is different from an amorphous solid that does not have such a regular internal structure.

The "pharmaceutical composition" of the present invention refers to containing any of the compounds of the present invention, including corresponding isomers, prodrugs, solvates, pharmaceutically acceptable salts or chemically protected forms thereof, and a Or a mixture of multiple pharmaceutically acceptable carriers and/or another one or more drugs. The purpose of the pharmaceutical composition is to facilitate the administration of the compound to the organism. The composition is generally used to prepare drugs for the treatment and/or prevention of diseases mediated by one or more kinases.

The "pharmaceutically acceptable carrier" in the present invention refers to a carrier that does not cause significant irritation to organisms and does not interfere with the biological activity and properties of the administered compound, and includes all solvents, diluents or other excipients, dispersants, and surface activities. Isotonic agent, thickener or emulsifier, preservative, solid binder, lubricant, etc. Unless any conventional carrier medium is incompatible with the compound of the present invention. Some examples of pharmaceutically acceptable carriers include, but are not limited to, sugars, such as lactose, glucose, and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, And cellulose and cellulose acetate; malt, gelatin, etc.

The "excipient" of the present invention refers to an inert substance added to a pharmaceutical composition to further facilitate the administration of a compound. Excipients may include calcium carbonate, calcium phosphate, various sugars and various types of starch, cellulose derivatives, gelatin, vegetable oil, and polyethylene glycol.

The "PRMT5" of the present invention may be any mutant or variant of wild-type PRMT5 or PRMT5, and the mutant or variant of PRMT5 contains one or more mutations (for example, conservative substitutions).

Detailed ways

The present invention will be further described in detail below in conjunction with examples, but the present invention is not limited to these examples. The materials used in the following examples are commercially available unless otherwise specified.

Example 1: (R)-7'-((1-Acetylpiperidin-4-yl)amino)-2'-(3-(3,4-dihydroisoquinolin-2(1H)-yl )-2-hydroxypropyl)-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-1'-one

Step 1: Preparation of methyl 2-(1-cyanocyclopropyl)benzoate

Place sodium hydride (4.46g, 111mmol) in a three-necked flask, add 20mL of anhydrous N,N-dimethylformamide at 0°C, stir for 5min, and slowly add methyl 2-(cyanomethyl)benzoate (7.80g, 44.6mmol) of N,N-dimethylformamide solution (80mL), stirred at 0°C for 30min, and then slowly added 1,2-dibromoethane (10.0g, 53.5mmol) dropwise. Then move to room temperature and react for 2h. After the reaction is complete, add 20mL saturated ammonium chloride solution to quench, extract with ethyl acetate (30mL×3), combine the organic phases, wash with water (10mL×2), saturated brine, dry over anhydrous sodium sulfate, and evaporate under reduced pressure Solvent and column chromatography to obtain the title compound. LC-MS m/z: [M+H] ⁺ =202.

Step 2: Preparation of 2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-1'-one

Dissolve methyl 2-(1-cyanocyclopropyl)benzoate (13.3g, 66.1mmol) in 150mL of absolute ethanol, add cobalt chloride hexahydrate (31.5g, 132mmol), add in portions at 0°C Sodium borohydride (7.54g, 198mmol), moved to room temperature and reacted for 1h, and then reacted at 80°C for 2h. After the reaction is completed, suction filtration, the filtrate is evaporated to remove the solvent under reduced pressure, and the title compound is obtained by column chromatography. LC-MS m/z: [M+H] ⁺ =174.

Step 3: Preparation of 7'-nitro-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-1'-one

Dissolve 2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-1'-one (6.33g, 36.6mmol) in concentrated sulfuric acid cooled in an ice bath ( 30mL), add potassium nitrate (3.69g, 36.6mmol) in batches at -10°C, and move to room temperature to react for 1h. After the reaction is complete, pour it into ice water, a solid precipitates out, suction filtration, and the filter cake is dried to obtain the title compound. LC-MS m/z: [M+H] ⁺ =219.

Step 4: Preparation of 7'-amino-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-1'-one

Dissolve 7'-nitro-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-1'-one (4.57g, 210mmol) in 40mL ethanol and In a mixed solvent of 10 mL of water, iron powder (2.93 g, 52.4 mmol) and ammonium chloride (3.33 g, 62.9 mmol) were added and reacted at 80° C. for 2 hours. After the reaction is complete, suction filtration, the filtrate is concentrated under reduced pressure, extracted with dichloromethane (30 mL×3), the organic phases are combined, dried over anhydrous sodium sulfate, the solvent is evaporated under reduced pressure, and the title compound is separated by column chromatography. LC-MS m/z: [M+H] ⁺ =189.

Step 5: 7'-((1-Acetylpiperidin-4-yl)amino)-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]- Preparation of 1′-ketone

The 7'-amino-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-1'-one (1.55g, 8.24mmol), 1-acetyl -4-piperidone (1.16 g, 8.24 mmol) was dissolved in 20 mL of methanol, glacial acetic acid (0.472 mL, 8.24 mmol) was added, and the reaction was carried out at room temperature for 2 hours. Then borane pyridine complex (1.24 mL, 12.4 mmol) was slowly added dropwise at 0°C, and the mixture was moved to room temperature for 2 h. After the reaction is complete, adjust the pH to alkaline with saturated sodium bicarbonate solution, extract with dichloromethane (20mL×3), combine the organic phases, dry over anhydrous sodium sulfate, evaporate the solvent under reduced pressure, and separate by column chromatography to obtain the title compound . LC-MS m/z: [M+H] ⁺ =314.

Step 6: (R)-7′-((1-Acetylpiperidin-4-yl)amino)-2′-(3-(3,4-dihydroisoquinolin-2(1H)-yl) -2-hydroxypropyl)-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-1'-one

The 7′-((1-acetylpiperidin-4-yl)amino)-2′,3′-dihydro-1′H-spiro[cyclopropane-1,4′-isoquinoline]-1′ -Ketone (1.56g, 4.98mmol) was dissolved in 10mL of anhydrous N,N-dimethylformamide solution, sodium hydride (0.299g, 7.48mmol) was added in portions at 0°C, stirred for 30min, and (R) was added. 2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline (0.942g, 4.98mmol) in N,N-dimethylformamide solution (5mL), Move to room temperature and react overnight. After the reaction was completed, 8 mL of saturated ammonium chloride solution was added for quenching, the solvent was evaporated under reduced pressure, the column chromatography was separated, and the title compound was prepared and separated by prep-HPLC (preparative liquid phase). ¹ H NMR (400MHz, DMSO-d ₆ ) δ7.16-7.23 (m, 1H), 7.05-7.16 (m, 3H), 6.99-7.05 (m, 1H), 6.65-6.77 (m, 2H), 5.57 (d, 1H), 4.75 (s, 1H), 4.19 (d, 1H), 3.94-4.10 (m, 1H), 3.71-3.87 (m, 2H), 3.62 (s, 2H), 3.42-3.54 (m , 2H), 3.34-3.41(m, 1H), 3.11-3.25(m, 2H), 2.75-2.94(m, 3H), 2.62-2.74(m, 2H), 2.36-2.48(m, 2H), 2.00 (s, 3H), 1.79-1.96 (m, 2H), 1.12-1.36 (m, 2H), 0.75-0.97 (m, 4H). LC-MS m/z: [M+H] ⁺ =503.

Example 2: (R)-7'-((1-Acetylpiperidin-4-yl)amino)-2'-(3-(3,4-dihydroisoquinolin-2(1H)-yl )-2-hydroxypropyl)-2′,3′-dihydro-1′H-spiro[cyclobutane-1,4′-isoquinoline]-1′-one

Step 1: Preparation of methyl 2-(1-cyanocyclobutyl)benzoate

Put sodium hydride (4.46g, 111mmol) in a 500mL three-necked flask, add 200mL anhydrous N,N-dimethylformamide at 0°C, stir for 5 minutes, slowly add 2-(cyanomethyl)benzoic acid A solution of methyl ester (10.0g, 57.1mmol) in N,N-dimethylformamide (40mL), stirred at 0°C for 30 minutes, and then slowly added 1,3-dibromopropane (13.8g, 68.5mmol) dropwise, After dripping, it was moved to room temperature and reacted for 2h. After monitoring the completion of the reaction, the reaction solution was poured into 200 mL saturated ammonium chloride solution for quenching, extracted with ethyl acetate (100 mL×3), the organic phases were combined, dried, the solvent was evaporated under reduced pressure, and the title compound was separated by column chromatography. LC-MS m/z: [M+H] ⁺ =216.

Step 2: Preparation of 2',3'-dihydro-1'H-spiro[cyclobutane-1,4'-isoquinoline]-1'-one

Dissolve methyl 2-(1-cyanocyclobutyl)benzoate (5.60g, 25.9mmol) in 150mL of absolute ethanol, add cobalt chloride hexahydrate (12.3g, 51.8mmol), and batch at 0°C Add sodium borohydride (2.95g, 77.7mmol), move to room temperature and react for 30 minutes, then react at 80°C overnight (16 hours). After monitoring the completion of the reaction, it was filtered, the filtrate was evaporated under reduced pressure to remove the solvent, and the title compound was obtained by column chromatography.

LC-MS m/z: [M+H] ⁺ =188.

Step 3: Preparation of 7'-nitro-2',3'-dihydro-1'H-spiro[cyclobutane-1,4'-isoquinoline]-1'-one

Dissolve 2',3'-dihydro-1'H-spiro[cyclobutane-1,4'-isoquinoline]-1'-one (4.00g, 21.3mmol) in concentrated sulfuric acid cooled in an ice bath (40mL), add potassium nitrate (2.14g, 21.3mmol) in batches at -10°C, then move to room temperature and react for 1 hour. After the reaction is complete, pour into ice water to precipitate a solid, filter with suction, and dry the filter cake to obtain the title compound. LC-MS m/z: [M+H] ⁺ =233.

Step 4: Preparation of 7'-amino-2',3'-dihydro-1'H-spiro[cyclobutane-1,4'-isoquinoline]-1'-one

Dissolve 7'-nitro-2',3'-dihydro-1'H-spiro[cyclobutane-1,4'-isoquinoline]-1'-one (4.26g, 18.2mmol) in 40mL In a mixed solvent of ethanol and 10 mL of water, iron powder (5.10 g, 91.4 mmol) and ammonium chloride (4.84 g, 91.4 mmol) were added to react at 80°C for 1.5 hours. After the reaction is complete, filter, dilute the filtrate with water (200 mL), extract with dichloromethane (50 mL×3), combine the organic phases, dry over anhydrous sodium sulfate, evaporate the solvent under reduced pressure, and separate by column chromatography to obtain the title compound. LC-MS m/z: [M+H] ⁺ =203.

Step 5: 7'-((1-Acetylpiperidin-4-yl)amino)-2',3'-dihydro-1'H-spiro[cyclobutane-1,4'-isoquinoline] Preparation of -1′-ketone

The 7'-amino-2',3'-dihydro-1'H-spiro[cyclobutane-1,4'-isoquinoline]-1'-one (250mg, 1.23mmol), 1-acetyl -4-piperidone (173 mg, 1.23 mmol) was dissolved in 10 mL of methanol, 3 drops of glacial acetic acid were added, and the reaction was carried out at room temperature for 30 minutes. Then borane pyridine complex (0.180 mL, 1.84 mmol) was slowly added dropwise at 0°C, and the mixture was moved to room temperature for 2 h. After the reaction was completed, the pH was adjusted to alkaline with saturated sodium bicarbonate solution, extracted with dichloromethane (20 mL×3), the organic phases were combined, dried, the solvent was evaporated under reduced pressure, and the title compound was separated by column chromatography.

LC-MS m/z: [M+H] ⁺ =328.

Step 6: (R)-7′-((1-Acetylpiperidin-4-yl)amino)-2′-(3-(3,4-dihydroisoquinolin-2(1H)-yl) -2-hydroxypropyl)-2',3'-dihydro-1'H-spiro[cyclobutane-1,4'-isoquinoline]-1'-one

7′-((1-Acetylpiperidin-4-yl)amino)-2′,3′-dihydro-1′H-spiro[cyclobutane-1,4′-isoquinoline]-1 '-Ketone (200mg, 0.610mmol) was dissolved in 5mL of anhydrous N,N-dimethylformamide solution, sodium hydride (36.6mg, 0.910mmol) was added in batches at 0℃, stirred for 30 minutes and then added (R) -2-(oxirane-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline (115mg, 0.610mmol) in N,N-dimethylformamide (5mL), Move to room temperature and react overnight. After monitoring the completion of the reaction, the reaction solution was poured into 30mL saturated ammonium chloride solution for quenching, extracted with dichloromethane (20mL×3), combined the organic phases, evaporated the solvent under reduced pressure to obtain the crude product, prepared and separated by prep-HPLC to obtain the title compound . ¹ H NMR (400MHz, DMSO-d ₆ ) δ 7.27-7.29 (m, 1H), 7.04-7.14 (m, 5H), 6.79-6.81 (m, 1H), 5.62-5.64 (m, 1H), 4.75 -4.76 (m, 1H), 4.17-4.20 (m, 1H), 3.98-4.05 (m, 1H), 3.48-3.85 (m, 8H), 3.16-3.26 (m, 2H), 2.65-2.86 (m, 5H), 1.85-2.15 (m, 12H), 1.18-1.31 (m, 2H). LC-MS m/z: [M+H] ⁺ =517.

Example 3: (R)-7′-((1-Acetylpiperidin-4-yl)amino)-2′-(3-(3,4-dihydroisoquinolin-2(1H)-yl )-2-hydroxypropyl)-2',3'-dihydro-1'H-spiro[cyclopentane-1,4'-isoquinoline]-1'-one

The preparation method is similar to the preparation method of Example 1, except that the raw material 1,2-dibromoethane in step 1 is replaced with 1,4-dibromobutane to obtain the title compound. ¹ H NMR (300MHz, DMSO-d ₆ ) δ7.00-7.40 (m, 6H), 6.75 (d, 1H), 5.59 (d, 1H), 4.72 (brs, 1H), 4.18 (d, 1H), 4.02 (s, 1H), 3.70-3.80 (m, 2H), 3.62 (s, 2H), 3.45-3.51 (m, 2H), 3.30-3.41 (m, 2H), 3.10-3.20 (m, 2H), 2.60-2.80 (m, 5H), 2.30-2.50 (m, 2H), 2.00 (s, 3H), 1.80-1.95 (m, 2H), 1.50-1.80 (m, 7H), 1.10-1.30 (m, 2H) ). LC-MS m/z: [M+H] ⁺ =531.

Example 4: (R)-N-(2'-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-1'-oxo-2 ′,3′-Dihydro-1′H-spiro[cyclopropane-1,4′-isoquinoline]-7′-yl)benzamide

Step 1: Preparation of N-(1'-oxo-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-7'-yl)benzamide

Dissolve 7′-amino-2′,3′-dihydro-1′H-spiro[cyclopropane-1,4′-isoquinoline]-1′-one (0.210g, 1.12mmol) in 10mL dichloromethane To the methane, add N,N-diisopropylethylamine (289mg, 2.24mmol), slowly add benzoyl chloride (172mg, 1.23mmol) dropwise at 0°C, and move to room temperature to react for 1h after the dripping. After the reaction is complete, add 5 mL of water, extract with dichloromethane (10 mL×3), combine the organic phases, dry over anhydrous sodium sulfate, evaporate the solvent under reduced pressure, and separate by column chromatography to obtain the title compound. LC-MS m/z: [M+H] ⁺ =293.

Step 2: (R)-N-(2'-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-1'-oxo-2' , 3'-Dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-7'-yl)benzamide

The N-(1′-oxo-2′,3′-dihydro-1′H-spiro[cyclopropane-1,4′-isoquinoline]-7′-yl)benzamide (115mg, 0.394 mmol) was dissolved in 2mL of anhydrous N,N-dimethylformamide solution, sodium hydride (23.6mg, 0.591mmol) was added at 0℃, stirred for 30min, and (R)-2-(ethylene oxide-2 -Methyl)-1,2,3,4-tetrahydroisoquinoline (74.4mg, 0.394mmol) in N,N-dimethylformamide solution (2mL), moved to room temperature and reacted overnight. After the reaction was completed, 3 mL of saturated ammonium chloride solution was added for quenching, the solvent was evaporated under reduced pressure, and the column chromatography was separated to obtain the title compound. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 10.35 (s, 1H), 8.32 (s, 1H), 7.98 (d, 2H), 7.92 (d, 1H), 7.57-7.64 (m, 1H), 7.47-7.56 (m, 2H), 7.06-7.14 (m, 3H), 6.98-7.06 (m, 2H), 4.81 (d, 1H), 3.98-4.12 (m, 1H), 3.77-3.87 (m, 1H) ), 3.63 (s, 2H), 3.54-3.60 (m, 1H), 3.43-3.50 (m, 1H), 3.25 (dd, 1H), 2.92-2.61 (m, 6H), 1.02 (d, 4H). LC-MS m/z: [M+H] ⁺ =482.

Example 5: (R)-N-(2'-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-1'-oxo-2 ′,3′-Dihydro-1′H-spiro[cyclopropane-1,4′-isoquinoline]-7′-yl)benzenesulfonamide

Step 1: Preparation of N-(1'-oxo-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-7'-yl)benzenesulfonamide

Dissolve 7'-amino-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-1'-one (0.300g, 1.59mmol) in 10mL dichloro To the methane, add N,N-diisopropylethylamine (0.526mL, 3.19mmol), slowly add benzenesulfonyl chloride (0.310g, 1.75mmol) dropwise at 0°C, and move to room temperature to react for 2h after the drop. After the reaction is complete, add 5 mL of water, extract with dichloromethane (10 mL×3), combine the organic phases, dry over anhydrous sodium sulfate, evaporate the solvent under reduced pressure, and separate by column chromatography to obtain the title compound. LC-MS m/z: [M+H] ⁺ =329.

Step 2: (R)-N-(2'-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-1'-oxo-2' Preparation of 3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-7'-yl)benzenesulfonamide

The N-(1′-oxo-2′,3′-dihydro-1′H-spiro[cyclopropane-1,4′-isoquinoline]-7′-yl)benzenesulfonamide (80.0mg, 0.244mmol) was dissolved in 2mL of anhydrous N,N-dimethylformamide solution, sodium hydride (14.6mg, 0.366mmol) was added at 0°C, stirred for 30min, and (R)-2-(ethylene oxide- A solution (1 mL) of 2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline (46.1 mg, 0.244 mmol) in N,N-dimethylformamide was moved to room temperature and reacted overnight. After the reaction was completed, 3 mL of saturated ammonium chloride solution was added for quenching, the solvent was evaporated under reduced pressure, and the column chromatography was separated to obtain the title compound. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 10.35 (s, 1H), 7.75 (d, 2H), 7.62-7.69 (m, 1H), 7.45-7.61 (m, 3H), 7.14-7.21 (m , 1H), 7.05-7.14 (m, 3H), 6.99-7.05 (m, 1H), 6.83-6.92 (m, 1H), 4.74 (d, 1H), 3.95-4.03 (m, 1H), 3.71-3.79 (m, 1H), 3.60(s, 2H), 3.45-3.52(m, 1H), 3.36-3.41(m, 1H), 3.13-3.20(m, 1H), 2.75-2.86(m, 3H), 2.65 -2.74 (m, 3H), 0.84-1.06 (m, 4H). LC-MS m/z: [M+H] ⁺ =518.

Example 6: (R)-2'-(3-(3,4-Dihydroisoquinoline-2(1H)-yl)-2-hydroxypropyl)-7'-(piperidin-4-yl Amino)-2′,3′-dihydro-1′H-spiro[cyclopropane-1,4′-isoquinoline]-1′-one

Step 1: 4-((1'-oxo-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-7'-yl)amino)piperidine Preparation of 1-tert-butyl carboxylate

The 7'-amino-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-1'-one (200mg, 1.06mmol), 4-oxopiper Tert-butyl pyridine-1-carboxylate (233 mg, 1.17 mmol) was dissolved in 8 mL of methanol, a drop of glacial acetic acid was added, and the reaction was carried out at room temperature for 0.5 h. Then, borane pyridine complex (0.160 mL, 1.60 mmol) was slowly added dropwise at 0° C. and moved to room temperature for 2 h. After the reaction was completed, the solvent was evaporated under reduced pressure, and the title compound was obtained by column chromatography. LC-MS m/z: [M+H] ⁺ =372.

Step 2: (R)-4-((2'-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-1'-oxo-2 Preparation of',3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-7'-yl)amino)piperidine-1-carboxylic acid tert-butyl ester

4-((1′-oxo-2′,3′-dihydro-1′H-spiro[cyclopropane-1,4′-isoquinoline]-7′-yl)amino)piperidine-1 -Tert-butyl carboxylate (340mg, 0.916mmol) was dissolved in 3mL of anhydrous N,N-dimethylformamide solution, sodium hydride (55.0mg, 1.37mmol) was added at 0°C, stirred for 30min, then (R) was added -2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline (173mg, 0.916mmol) in N,N-dimethylformamide (2mL), Move to room temperature and react overnight. After the reaction was completed, 4 mL of saturated ammonium chloride solution was added for quenching, the solvent was evaporated under reduced pressure, and the column chromatography was separated to obtain the title compound. LC-MS m/z: [M+H] ⁺ =561.

Step 3: (R)-2'-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-7'-(piperidin-4-ylamino )-2',3'-Dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-1'-one

Put (R)-4-((2'-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-1'-oxo-2', 3′-Dihydro-1′H-spiro[cyclopropane-1,4′-isoquinoline]-7′-yl)amino)piperidine-1-carboxylic acid tert-butyl ester (300mg, 0.535mmol) placed in the single Add 5 mL of 4.0M hydrogen chloride dioxane solution to the neck flask, and react at room temperature overnight. After the completion of the reaction, the solvent was evaporated under reduced pressure, and the title compound was obtained through preparation and separation. ¹ H NMR (400MHz, DMSO-d ₆ ) δ7.06-7.22 (m, 4H), 6.99-7.05 (m, 1H), 6.62-6.76 (m, 2H), 5.51 (d, 1H), 4.71-4.82 (m, 1H), 3.97-4.07(m, 1H), 3.74-3.84(m, 1H), 3.61(s, 2H), 3.44-3.51(m, 1H), 3.36-3.42(m, 1H), 3.10 -3.25(m, 2H), 2.88-2.99(m, 2H), 2.76-2.85(m, 2H), 2.64-2.75(m, 2H), 2.52-2.59(m, 2H), 2.38-2.48(m, 3H), 1.76-1.88 (m, 2H), 1.25-1.14 (m, 2H), 0.77-0.94 (s, 4H). LC-MS m/z: [M+H] ⁺ =461.

Example 7: (R)-7′-((1-(cyclopropanecarbonyl)piperidin-4-yl)amino)-2′-(3-(3,4-dihydroisoquinoline-2(1H )-Yl)-2-hydroxypropyl)-2′,3′-dihydro-1′H-spiro[cyclopentane-1,4′-isoquinoline]-1′-one

Step 1: Preparation of 1-(cyclopropanecarbonyl)piperidin-4-one

Dissolve piperidin-4-one (1.00g, 10.1mmol) in 20mL of anhydrous dichloromethane, add cyclopropylformyl chloride (1.05g, 10.1mmol) and triethylamine (1.53g, 15.2mmol) at 0°C , Reaction at room temperature for 1h. After the reaction was completed, the solvent was evaporated under reduced pressure to obtain the title compound. ¹ H NMR (400MHz, CDCl ₃ -d ₆ ) δ 3.94 (t, 4H), 2.52 (t, 4H), 1.79-1.85 (m, 1H), 0.92-0.94 (m, 4H). LC-MS m /z: [M+1] ⁺ ＝168.

Step 2: 7'-((1-(cyclopropanecarbonyl)piperidin-4-yl)amino)-2',3'-dihydro-1'H-spiro[cyclopentane-1,4'-iso Preparation of quinoline]-1′-one

Combine 1-(cyclopropanecarbonyl)piperidin-4-one (200mg, 1.20mmol) and 7′-amino-2′,3′-dihydro-1′H-spiro[cyclopentane-1,4′- Isoquinoline]-1′-one (250mg, 1.20mmol) was dissolved in 10mL methanol, glacial acetic acid (70mg, 1.20mmol) was added dropwise, reacted at room temperature for 1h, and pyridineborane (160mg, 1.72mmol) was added dropwise at 0°C, React at room temperature for 1h. After the reaction is complete, add sodium bicarbonate solution to adjust the pH to 8, add dichloromethane (10 mL×2) for extraction, dry with anhydrous sodium sulfate, evaporate the solvent under reduced pressure, and separate the title compound by column chromatography. LC-MS m/z: [M+H] ⁺ =368.

Step 3: (R)-7′-((1-(cyclopropanecarbonyl)piperidin-4-yl)amino)-2′-(3-(3,4-dihydroisoquinoline-2(1H) -Yl)-2-hydroxypropyl)-2',3'-dihydro-1'H-spiro[cyclopentane-1,4'-isoquinoline]-1'-one

Dissolve sodium hydride (30.0mg, 0.735mmol) in 3mL of anhydrous N,N-dimethylformamide, and slowly add 2mL of 7′-((1-(cyclopropanecarbonyl)piperidin-4-yl )Amino)-2',3'-dihydro-1'H-spiro[cyclopentane-1,4'-isoquinoline]-1'-one (180mg, 0.49mmol) of N,N-dimethyl Base formamide solution, react at 0°C for 0.5h. Add (R)-2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline (102mg, 0.49mmol) N,N-dimethyl under ice bath The formamide (2mL) solution was added dropwise and reacted at room temperature for 16h. After the reaction is complete, slowly add ammonium chloride solution dropwise to quench the sodium hydride, evaporate the solvent under reduced pressure, add 10 mL of dichloromethane to dissolve, filter under reduced pressure, collect the filtrate, and evaporate the solvent under reduced pressure to obtain the crude product. The crude product is prepared and separated to obtain the title Compound. ¹ H NMR (400MHz, CDCl ₃ ) δ7.00-7.40 (m, 6H), 6.75 (d, 1H), 5.59 (d, 1H), 4.72 (brs, 1H), 4.18 (d, 1H), 4.02 ( s, 1H), 3.70-3.80 (m, 2H), 3.62 (s, 2H), 3.45-3.51 (m, 2H), 3.30-3.41 (m, 2H), 3.10-3.20 (m, 2H), 2.60- 2.80 (m, 5H), 2.30-2.50 (m, 4H), 2.00 (s, 3H), 1.80-1.95 (m, 2H), 1.50-1.80 (m, 7H), 1.10-1.30 (m, 2H). LC-MS m/z: [M+H] ⁺ =557.

Example 8: (R)-N-(2'-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-1'-oxo-2 ′,3′-Dihydro-1′H-spiro[cyclobutane-1,4′-isoquinoline]-7′-yl)-1-methyl-1H-pyrazole-4-carboxamide

Step 1: 1-Methyl-N-(1′-oxo-2′,3′-dihydro-1′H-spiro[cyclobutane-1,4′-isoquinoline]-7′-yl ) Preparation of -1H-pyrazole-4-carboxamide

The 7'-amino-2',3'-dihydro-1'H-spiro[cyclobutane-1,4'-isoquinoline]-1'-one (200mg, 0.99mmol), 1-methyl -1H-pyrazole-4-carboxylic acid (125mg, 0.99mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate ( 566 mg, 1.49 mmol) was dissolved in 10 mL of dichloromethane, N,N-diisopropylethylamine (255 mg, 1.98 mmol) was added at room temperature, and the reaction was carried out at room temperature for 1 h. After the reaction is complete, wash with water (10 mL×3), dry with anhydrous sodium sulfate, evaporate the solvent under reduced pressure, and separate the title compound by column chromatography. LC-MS m/z: [M+H] ⁺ =311.

Step 2: (R)-N-(2'-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-1'-oxo-2' , 3′-Dihydro-1′H-spiro[cyclobutane-1,4′-isoquinoline]-7′-yl)-1-methyl-1H-pyrazole-4-carboxamide

Sodium hydride (78.0mg, 1.94mmol) was dissolved in 3mL of anhydrous N,N-dimethylformamide, and 2mL of 1-methyl-N-(1'-oxo-2' was slowly added dropwise at 0°C, 3'-Dihydro-1'H-spiro[cyclobutane-1,4'-isoquinoline]-7'-yl)-1H-pyrazole-4-carboxamide (300mg, 0.97mmol) of N, The N-dimethylformamide solution was reacted at 0°C for 0.5h. Add (R)-2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline (200mg, 0.97mmol) N,N-dimethyl under ice bath The formamide (2mL) solution was added dropwise and reacted at room temperature for 16h. After the reaction was completed, ammonium chloride solution was slowly added dropwise to quench the sodium hydride, the solvent was evaporated under reduced pressure, 10 mL of dichloromethane was added for dissolution, filtered under reduced pressure, the filtrate was collected, the solvent was evaporated under reduced pressure, and the title compound was obtained after preparation and separation. 1H NMR (400MHz, CDCl ₃ ) δ 8.28-8.30 (m, 1H), 7.96-7.98 (m, 2H), 7.91 (s, 1H), 7.82-7.85 (m, 1H), 7.49 (d, 1H) , 7.11-7.13 (m, 3H), 7.00 (d, 1H), 4.07-4.16 (m, 1H), 3.95 (s, 3H), 3.74-3.90 (m, 5H), 3.58-3.62 (m, 1H) , 3.46-3.50(m, 1H), 2.86-2.93(m, 3H), 2.69-2.72(m, 1H), 2.59-2.62(m, 1H), 2.49-2.54(m, 1H), 2.26-2.35( m, 2H), 2.13-2.23 (m, 2H), 2.02-2.08 (m, 2H). LC-MS m/z: [M+H] ⁺ =500.

Example 9: (R)-N-(2'-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-1'-oxo-2 ′,3′-Dihydro-1′H-spiro[cyclopropane-1,4′-isoquinoline]-7′-yl)-1-methyl-1H-pyrazole-4-carboxamide

Step 1: 1-Methyl-N-(1′-oxo-2′,3′-dihydro-1′H-spiro[cyclopropane-1,4′-isoquinoline]-7′-yl) Preparation of -1H-pyrazole-4-carboxamide

The 7'-amino-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-1'-one (180mg, 0.96mmol), 1-methyl- 1H-pyrazole-4-carboxylic acid (132mg, 1.05mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (547mg , 1.44 mmol) was dissolved in 10 mL of dichloromethane, N,N-diisopropylethylamine (246 mg, 1.92 mmol) was added at room temperature, and the reaction was carried out at room temperature for 1 h. After the reaction is complete, wash with water (10 mL×3), dry with anhydrous sodium sulfate, evaporate the solvent under reduced pressure, and separate the title compound by column chromatography. LC-MS m/z: [M+H] ⁺ =297.

Step 2: (R)-N-(2'-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-1'-oxo-2' , 3′-Dihydro-1′H-spiro[cyclopropane-1,4′-isoquinoline]-7′-yl)-1-methyl-1H-pyrazole-4-carboxamide

Sodium hydride (67.0mg, 1.68mmol) was dissolved in 3mL of anhydrous N,N-dimethylformamide, and 2mL of 1-methyl-N-(1'-oxo-2' was slowly added dropwise at 0°C, 3′-Dihydro-1′H-spiro[cyclopropane-1,4′-isoquinoline]-7′-yl)-1H-pyrazole-4-carboxamide (250mg, 0.84mmol) N, N -Dimethylformamide solution, react at 0°C for 0.5h. Add (R)-2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline (174mg, 0.84mmol) N,N-dimethyl under ice bath The formamide (2mL) solution was added dropwise and reacted at room temperature for 16h. After the reaction was completed, ammonium chloride solution was slowly added dropwise to quench the sodium hydride, the solvent was evaporated under reduced pressure, 10 mL of dichloromethane was added for dissolution, filtered under reduced pressure, the filtrate was collected, the solvent was evaporated under reduced pressure, and the title compound was obtained after preparation and separation. ¹ H NMR (400MHz, CDCl ₃ ) δ 8.11-8.20 (m, 2H), 7.90-7.97 (m, 3H), 7.10-7.13 (m, 3H), 6.98-7.00 (m, 1H), 7.84 (d , 1H), 4.04-4.09 (m, 1H), 3.93 (s, 3H), 3.71-3.86 (m, 3H), 3.50-3.62 (m, 3H), 3.39-3.44 (m, 1H), 2.89-2.92 (m, 3H), 2.65-2.73 (m, 1H), 2.47-2.59 (m, 2H), 0.97-1.07 (m, 4H). LC-MS m/z: [M+H] ⁺ =486.

Example 10: (R)-1-Acetyl-N-(2'-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-1' -Oxo-2',3'-dihydro-1'H-spiro[cyclobutane-1,4'-isoquinoline]-7'-yl)piperidine-4-carboxamide

Step 1: 1-Acetyl-N-(1′-oxo-2′,3′-dihydro-1′H-spiro[cyclobutane-1,4′-isoquinoline]-7′-yl ) Preparation of piperidine-4-carboxamide

The 7'-amino-2',3'-dihydro-1'H-spiro[cyclobutane-1,4'-isoquinoline]-1'-one (300mg, 1.47mmol), 1-acetyl Piperidine-4-carboxylic acid (287mg, 1.62mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (837mg, 2.20 mmol) was dissolved in dry dichloromethane (10 mL), and triethylamine (296 mg, 2.94 mmol) was added and reacted at room temperature for 3 hours. After the completion of the reaction monitored by LCMS, the reaction solution was poured into 10 mL of water, extracted with dichloromethane (15 mL×3), the organic phases were combined, the crude product was dried and spin-dried to be slurried with ethyl acetate (5 mL), and filtered to obtain the title compound. LC-MS m/z: [M+H] ⁺ =356.

Step 2: (R)-1-Acetyl-N-(2′-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-1′- Preparation of oxo-2',3'-dihydro-1'H-spiro[cyclobutane-1,4'-isoquinoline]-7'-yl)piperidine-4-carboxamide

The 1-acetyl-N-(1′-oxo-2′,3′-dihydro-1′H-spiro[cyclobutane-1,4′-isoquinoline]-7′-yl)piper Pyridine-4-carboxamide (100mg, 0.280mmol) was dissolved in 8mL of anhydrous N,N-dimethylformamide solution. Sodium hydride (20.0mg, 0.420mmol) was added in batches at 0°C, stirred for 20min and then added ( R)-2-(Ethylene oxide-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline (64.2mg, 0.330mmol) in N,N-dimethylformamide solution ( 5mL), move to room temperature and react overnight. The reaction solution was poured into 10 mL saturated ammonium chloride solution and quenched, extracted with dichloromethane (15 mL×3), combined the organic phases, and evaporated the solvent under reduced pressure to obtain the crude product, which was prepared and separated by prep-HPLC to obtain the title compound. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 10.03 (s, 1H), 8.06 (s, 1H), 7.83-7.85 (m, 1H), 7.51-7.53 (m, 1H), 7.07-7.12 (m , 2H), 7.01-7.04 (m, 1H), 4.77-4.82 (m, 1H), 4.38-4.41 (m, 1H), 4.04-4.09 (m, 1H), 3.83-3.89 (m, 2H), 3.63 -3.76(m, 4H), 3.25-3.28(m, 2H), 3.03-3.10(m, 1H), 2.78-2.85(m, 2H), 2.67-2.76(m, 2H), 2.56-2.62(m, 2H), 2.04-2.24(m, 5H), 2.01(s, 3H), 1.90-1.96(m, 1H), 1.71-1.86(m, 2H), 1.54-1.64(m, 2H), 1.38-1.48( m, 2H).LC-MS m/z: [M+H] ⁺ ＝545

Example 11: (R)-1-Acetyl-N-(2'-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-1' -Oxo-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-7'-yl)piperidine-4-carboxamide

Step 1: 1-Acetyl-N-(1′-oxo-2′,3′-dihydro-1′H-spiro[cyclopropane-1,4′-isoquinoline]-7′-yl) Preparation of piperidine-4-carboxamide

The 7'-amino-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-1'-one (200mg, 1.06mmol), (1-acetyl Piperidine-4-carboxylic acid (206mg, 1.16mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (604mg, 1.59 mmol) was dissolved in dry dichloromethane (10mL), triethylamine (214mg, 2.12mmol) was added and reacted at room temperature for 2 hours. After the completion of the reaction was monitored by LCMS, the reaction solution was poured into 10mL of water and dichloromethane (15mL× 3) Extract, combine the organic phases, dry and spin-dry the crude product to be separated and purified by column chromatography to obtain the title compound. LC-MS m/z: [M+H] ⁺ =342.

Step 2: (R)-1-Acetyl-N-(2′-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-1′- Preparation of oxo-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-7'-yl)piperidine-4-carboxamide

1-Acetyl-N-(1′-oxo-2′,3′-dihydro-1′H-spiro[cyclopropane-1,4′-isoquinoline]-7′-yl)piperidine -4-formamide (120mg, 0.350mmol) was dissolved in 5mL of anhydrous N,N-dimethylformamide solution, sodium hydride (21.0mg, 0.530mmol) was added in batches at 0°C, stirred for 20min, and then added (R )-2-(Ethylene oxide-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline (80.0mg, 0.420mmol) in N,N-dimethylformamide solution (5mL ), move to room temperature and react overnight. The reaction solution was poured into 10 mL saturated ammonium chloride solution and quenched, extracted with dichloromethane (15 mL×3), combined the organic phases, evaporated the solvent under reduced pressure to obtain the crude product and separated by prep-HPLC to obtain the title compound. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 10.00 (s, 1H), 8.09 (s, 1H), 7.72-7.74 (m, 1H), 7.04-7.09 (m, 3H), 6.92-6.94 (m , 1H), 4.77-4.78 (m, 1H), 4.38-4.41 (m, 1H), 3.99-4.06 (m, 1H), 3.77-3.88 (m, 2H), 3.60-3.64 (m, 2H), 3.52 -3.55 (m, 2H), 3.41-3.45 (m, 2H), 3.20-3.25 (m, 2H), 3.03-3.09 (m, 2H), 2.80-2.90 (m, 2H), 2.67-2.72 (m, 2H), 2.52-2.62(m, 3H), 2.01(s, 3H), 1.76-1.83(m, 2H), 1.54-1.65(m, 2H), 1.40-1.48(m, 2H).LC-MS m /z: [M+H] ⁺ =531.

Example 12: (R)-N-(2'-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-1'-oxo-2 ′,3′-Dihydro-1′H-spiro[cyclopropane-1,4′-isoquinoline]-7′-yl)tetrahydro-2H-pyran-4-carboxamide

Step 1: N-(1′-oxo-2′,3′-dihydro-1′H-spiro[cyclopropane-1,4′-isoquinoline]-7′-yl)tetrahydro-2H- Preparation of pyran-4-carboxamide

The 7'-amino-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-1'-one (190mg, 1.01mmol), tetrahydropyran- 4-formic acid (156mg, 1.20mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (570mg, 1.50mmol) was dissolved in In 10 mL of dichloromethane, add N,N-diisopropylethylamine (258 mg, 2.02 mmol) at room temperature, and react at room temperature for 2 h. After the reaction is complete, wash with water (10 mL×3), dry with anhydrous sodium sulfate, evaporate the solvent under reduced pressure, and separate and purify by column to obtain 250 mg of yellow solid. LC-MS m/z: [M+H] ⁺ =301.

Step 2: (R)-N-(2'-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-1'-oxo-2' , 3'-Dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-7'-yl)tetrahydro-2H-pyran-4-carboxamide

N-(1′-oxo-2′,3′-dihydro-1′H-spiro[cyclopropane-1,4′-isoquinoline]-7′-yl)tetrahydro-2H-pyran -4-formamide (250mg, 0.830mmol) was dissolved in 5mL of anhydrous N,N-dimethylformamide, sodium hydride (67mg, 1.67mmol) was added at 0℃, and the reaction was continued for 1 hour at 0℃; then added (R)-2-(Ethylene oxide-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline (159mg, 0.83mmol) in N,N-dimethylformamide solution ( 2mL). The reaction was stirred overnight at room temperature, the reaction solution was poured into 10 mL of saturated ammonium chloride solution for quenching, extracted with dichloromethane, the organic phases were combined, and the solvent was evaporated under reduced pressure to obtain the crude product, which was prepared and separated by prep-HPLC to obtain the title compound. ¹ H NMR (400MHz, CDCl ₃ ) δ 8.04 (d, 1H), 7.69 (s, 1H), 7.27 (s, 1H), 7.06 (d, 3H), 6.93 (d, 1H), 6.76 (d, 1H), 3.95-4.09(m, 3H), 3.72-3.82(m, 2H), 3.51-3.62(m, 2H), 3.36-3.50(m, 4H), 2.85(s, 3H), 2.39-2.71( m, 5H), 1.77-1.88 (m, 4H), 0.95 (d, 4H). LC-MS m/z: [M+H] ⁺ =490.

According to the synthesis method of Example 1 of the present invention, the compounds of Examples 13-34 were synthesized using different commercially available raw materials. The characterization parameters of these compounds are shown in Table 1:

Table 1

Experimental example 1 Compound in vitro kinase activity evaluation

1. Experimental materials

Compounds: The compounds of the present invention prepared in the above examples. Each compound was formulated into 10mM mother liquor with DMSO, and finally diluted to 10 concentrations for detection. The final concentrations were 10000.00nM, 3333.33nM, 1111.11nM, 370.37nM, 123.46nM, 41.15 nM, 13.72nM, 4.57nM, 1.52nM, 0.51nM.

Reagents and consumables: PRMT5, purchased from Active Motif, item number 31921; peptide substrate H4(1-21)S1ac, purchased from Gil Biochemical (Shanghai) Co., Ltd., item number 342095; [ ³ H]-SAM, Purchased from PerkinElmer, the article number is NET155V001MC; SAM, purchased from Sigma, the article number is A7007-100MG; SAH, purchased from Sigma Company, the article number is A9384-25MG; DTT, purchased from Shenggong Bioengineering (Shanghai) Co., Ltd. Limited company, the article number is A620058-0005. Corning-3657, purchased from Corning Company, the article number is 3657; Echo Qualified 384-Well, purchased from Labcyte Company, the article number is P-05525; FlashPlate, purchased from Perkin Elmer, the article number is SMP410J001PK.

Instruments: Scintillation counter, purchased from PerkinElmer, model MicroBeta2; ultrasonic nanoliter liquid processing system, purchased from Labcyte, model Echo 550.

2. Experimental method

2.1. Preparation of reaction buffer and reaction termination solution: 1x reaction buffer liquid components are 10mM Tris-HCl, pH 8.0; 0.01% Tween-20; 1mM DTT. The composition of the reaction stop solution was 125 μM ³ H-SAM solution.

2.2 Compound preparation

2.2.1 Compound dilution

The compound was dissolved in 100% DMSO into a 10 mM stock solution, and then the compound was diluted to the required concentration on an Echo384-well plate.

2.2.2 Transfer compounds to 384 reaction plate

Use the Echo550 instrument to transfer 250nL of compound from the diluted Echo384-well plate to the 384-well reaction plate.

2.3 Enzymatic reaction

2.3.1 Preparation of 1.67 times enzyme solution

Add PRMT5 to 1x reaction buffer to form a 1.67x enzyme solution.

2.3.2 Preparation of 2.5 times the substrate solution

The peptide substrate and [ ³ H]-SAM were added to the 1-fold reaction buffer to form a 2.5-fold substrate solution (final concentrations of 100 nM and 250 nM, respectively).

2.3.3 Add enzyme solution to 384-well plate

Add 15 μL of 1.67 times enzyme solution to the wells of the 384-well reaction plate. For the control wells without enzyme activity, replace the enzyme solution with 15 μL of 1X reaction buffer. Centrifuge at 1000 rpm for 1 min and incubate at room temperature for 15 minutes.

2.3.4 Add the substrate solution to the 384-well plate to start the enzymatic reaction

Add 10 μL of 2.5-fold substrate solution to each well of a 384-well reaction plate. Centrifuge at 1000 rpm for 1 min. React at 25°C for 60 minutes.

2.3.5 Termination of enzymatic reaction

Add 5μL of reaction stop solution to each well of the 384-well reaction plate to stop the reaction. Take 25uL from each well of the test plate and transfer it to Flashplate, and place it at room temperature for 1h. Then wash the Flashpate plate 3 times with 0.1% Tween-20 solution.

2.4 MicroBeta 2 Read data

2.4 Calculation of inhibition rate

Copy data from Microbeta 2. Convert the data into inhibition rate data. The maximum value refers to the conversion rate of the DMSO control, and the minimum value refers to the conversion rate of the control without enzyme activity. Inhibition rate (%)=(maximum-sample value)/(maximum-minimum)×100%.

GraphPad data import, and use the "log (inhibitor) vs.response - Variable slope" curve fitting to obtain IC _50. The IC ₅₀ results of some compounds are shown in Table 2.

Table 2

"-" means untested

From the above experimental results, it can be seen that the IC50 value of the compound of the present invention against PRMT5 is at the nM level, and exhibits significant inhibitory activity against PRMT5.

Although the present invention has been described in detail above, those skilled in the art understand that various modifications and changes can be made to the present invention without departing from the spirit and scope of the present invention. The scope of rights of the present invention is not limited to the detailed description made above, but should belong to the claims.

Claims

A compound represented by general formula (I) or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug,

among them,

Ring A is selected from aryl, heteroaryl, cycloalkyl and heterocyclyl, and the aryl, heteroaryl, cycloalkyl and heterocyclic group may be one or more selected from halogen, hydroxy, alkyl, Haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxy, cyano, amino, monoalkylamino, alkylacylamino, alkylacyl, alkylsulfonyl, amino Substitution of acyl, alkylaminoacyl, dialkylamino, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and oxo groups;

Cy is selected from the group consisting of aryl, heteroaryl, cycloalkyl, and heterocyclyl. The aryl, heteroaryl, cycloalkyl, and heterocyclic group may be one or more selected from halogen, hydroxy, alkyl, and halogenated alkyl. Group, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxy, cyano, amino, monoalkylamino, alkylacylamino, alkylacyl, alkylsulfonyl, aminoacyl , Alkylamino acyl, dialkylamino, alkenyl, alkynyl, haloalkyl acyl, hydroxyalkyl acyl, cycloalkyl acyl, heterocyclyl acyl, cycloalkyl, heterocyclic, aryl, heteroaryl And oxo group substitution;

R 1 , R 2 , R 3 , and R 4 are each independently selected from hydrogen, halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxy, Cyano, amino, monoalkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl and dialkylamino;

R 5 is selected from hydrogen, alkyl, haloalkyl, hydroxyalkyl, alkylacyl, aminoacyl and alkylaminoacyl;

X 1 , X 2 , and X 3 are each independently selected from N and C (R 6 ), wherein R 6 is selected from hydrogen, halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, Hydroxyalkoxy, nitro, carboxy, cyano, amino, monoalkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl, dialkylamino, and cycloalkyl; and

L is selected from -NH-, -S-, -O-, -C(O)NH-, -NHC(O)-, -OC(O)NH-, -NHC(O)O-, -S(O ) 2 NH-, -NHS(O) 2 -, -C(O)- and -NHC(O)NH-.
The compound according to claim 1, or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof,

Wherein ring A is selected from C 6-12 aryl, 5-12 membered heteroaryl, C 3-12 cycloalkyl and 3-12 membered heterocyclic group, the C 6-12 aryl, 5-12 membered heterocyclic group Aryl, C 3-12 cycloalkyl and 3-12 membered heterocyclic group can be selected by one or more selected from halogen, hydroxyl, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1- 6 alkyl, C 1-6 alkoxy, halogenated C 1-6 alkoxy, hydroxy C 1-6 alkoxy, nitro, carboxy, cyano, amino, mono C 1-6 alkylamino, C 1-6 alkyl acylamino, C 1-6 alkyl acyl, C 1-6 alkylsulfonyl, amino acyl, C 1-6 alkylamino acyl, bis C 1-6 alkylamino, C 2- Group substitution of 10 alkenyl, C 2-10 alkynyl, C 3-12 cycloalkyl, 3-12 membered heterocyclic group, 6-12 membered aryl, 5-12 membered heteroaryl and oxo group .
The compound or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug according to claim 1 or 2, wherein Cy is selected from C 6-12 aryl, 5-12 membered heteroaryl, C 3-12 cycloalkyl group and 3-12 membered heterocyclic group, the C 6-12 aryl group, 5-12 membered heteroaryl group, C 3-12 cycloalkyl group and 3-12 membered heterocyclic group can be One or more selected from halogen, hydroxy, C 1-6 alkyl, halogenated C 1-6 alkyl, hydroxy C 1-6 alkyl, C 1-6 alkoxy, halogenated C 1-6 alkoxy Group, hydroxy C 1-6 alkoxy, nitro, carboxy, cyano, amino, mono C 1-6 alkylamino, C 1-6 alkyl acylamino, C 1-6 alkyl acyl, C 1- 6 alkylsulfonyl group, an amino group, a C 1-6 alkylamino group, a di-C 1-6 alkylamino, C 2-10 alkenyl, C 2-10 alkynyl group, halo C 1-6 alkyl group , Hydroxy C 1-6 alkyl acyl, C 3-12 cycloalkyl acyl, 3-12 membered heterocyclyl acyl, C 3-12 cycloalkyl, 3-12 membered heterocyclic group, 6-12 membered aryl group , 5-12 membered heteroaryl and oxo group substitutions.
The compound or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof according to any one of claims 1-3, wherein R 1 , R 2 , R 3 , and R 4 are each independently Selected from hydrogen, halogen, hydroxy, C 1-6 alkyl, halogenated C 1-6 alkyl, hydroxy C 1-6 alkyl, C 1-6 alkoxy, halogenated C 1-6 alkoxy, Hydroxy C 1-6 alkoxy, nitro, carboxy, cyano, amino, mono C 1-6 alkylamino, C 1-6 alkyl acylamino, C 1-6 alkyl acyl, amino acyl, C 1 -6 alkylaminoacyl and bis-C 1-6 alkylamino.
The compound according to any one of claims 1 to 3, or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein R 5 is selected from hydrogen, C 1-6 alkyl, halogen Substituted C 1-6 alkyl, hydroxy C 1-6 alkyl, C 1-6 alkyl acyl, amino acyl, C 1-6 alkyl amino acyl.
The compound or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof according to any one of claims 1 to 3, wherein the general formula (I) has the structure of the following general formula (Ia) ,

Among them, Cy, L and ring A have the definitions described in claims 1-3.
The compound according to claim 6 or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, wherein Cy is selected from
The compound according to claim 1, or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein the compound is a compound selected from:
A pharmaceutical composition comprising the compound according to any one of claims 1 to 8 or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, and a pharmaceutically acceptable carrier.
The compound of any one of claims 1-8 or its isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs or the pharmaceutical composition of claim 9 is prepared for the treatment of PRMT5 mediation The application of drugs for leading diseases.