WO2023143210A1

WO2023143210A1 - Phthalazinone compounds, preparation method therefor, pharmaceutical composition containing phthalazinone compounds and uses thereof

Info

Publication number: WO2023143210A1
Application number: PCT/CN2023/072449
Authority: WO
Inventors: 陈旭星; 李京; 陈艳红
Original assignee: 上海优理惠生医药有限公司
Priority date: 2022-01-26
Filing date: 2023-01-16
Publication date: 2023-08-03
Also published as: CN116903611A; CN118574829A

Abstract

The present invention provides phthalazinone compounds, a preparation method therefor, a pharmaceutical composition containing the phthalazinone compounds and the uses thereof. Specifically, the compounds have a structure as shown in formula I, may synergistically inhibit PRMT5 with MTA, and may be used for preparing drugs for treating MTAP-/- related cancers.

Description

A phthalazinone compound, its preparation method, its pharmaceutical composition and its application

technical field

The present invention relates to the field of medicinal chemistry, and more specifically, the present invention relates to a phthalazinone compound, which can be used as an MTA synergistic PRMT5 inhibitor to prepare a drug for treating MTAP ^-/- related cancers.

Background technique

Protein arginine N-methyltransferase (PRMT) is classified into type I, type II and type III according to the kind of methyl arginine produced. Type I mainly catalyzes the transfer of the methyl group from S-adenosyl-L-methionine (SAM) to the omega ammonia nitrogen of the guanidine group of protein L-arginine to generate monomethylarginine (MMA) and the second The transfer of two methyl groups to the same ω ammonia nitrogen of the guanidino group produces asymmetric dimethylarginine (aDMA); type II mainly catalyzes the formation of MMA and the transfer of the second methyl group to another ω ammonia nitrogen to produce asymmetric dimethylarginine arginine (sDMA); type III only catalyzes the formation of MMA. Among them, PRMT5 is a type II protein arginine methyltransferase that plays an important role in regulating cellular processes, including DNA repair, cell cycle progression, transcriptional regulation, and RNA splicing. Upregulation of PRMT5 can lead to a variety of cancers, including liver cancer, breast cancer, skin cancer, pancreatic cancer, head and neck cancer, bowel cancer, lung cancer, gastric cancer, esophageal cancer, kidney cancer, bladder cancer, urethral cancer, prostate cancer, testicular cancer, uterine cancer Cancer, Ovarian Cancer, Vaginal Cancer, Fallopian Tube Cancer, Cholangiocarcinoma, Multiple Myeloma, Spinal Neurofibroma, Astrocytoma, Glioma, Acute Lymphocytic Leukemia, Chronic Lymphocytic Leukemia, Hodgkin Lymphoma, Non-Hodgkin's lymphoma, sarcoma.

Methylthioadenosine phosphorylase (MTAP) is located near the p16/CDKN2a gene and is co-deleted with p16/CDKN2a in approximately 15% of human cancers. This co-deletion leads to poor prognosis in malignancies and lacks effective molecularly targeted therapies. MTAP is a metabolizing enzyme of methylthio adenosine (MTA), which catalyzes the conversion of MTA to adenosine and 5-methylthioribose-1-phosphate. In tumor cells, the loss of MTAP leads to the accumulation of MTA, which can competitively inhibit PRMT5 and form a PRMT5-MTA complex due to its structural similarity to SAM. The small molecule inhibitor designed based on this complex can selectively kill tumor cells with little effect on normal cells and improve drug safety.

We believe that in MTAP-deficient cancers, cooperative inhibition of PRMT5 activity by MTAs will provide therapeutic benefit for multiple cancers. Currently, there is a lack of effective molecularly targeted therapies, so it is necessary to develop new MTA-synergistic PRMT5 inhibitors, which can inhibit PRMT5 activity in the presence of elevated MTA concentrations, for the treatment of MTAP-deficient related cancers.

Contents of the invention

The purpose of the present invention is to provide a more efficient and better druggable MTA synergistic PRMT5 inhibitor.

The first aspect of the present invention provides a compound represented by formula I, or a pharmaceutically acceptable salt thereof, enantiomer isomers, diastereomers, tautomers, cis-trans isomers, solvates, polymorphs, deuteriums, or combinations thereof,

in,

R ^1a and R ^1b are each independently hydrogen or C1-C3 alkyl, or R ^1a and R ^1b together form oxo (=O), or R ^1a and R ^1b form a 3-5 membered group with the carbon atom to which they are attached Carbocyclyl; wherein, the alkyl or carbocyclyl may be optionally substituted by halogen or C1-C3 alkyl;

R ^2a and R ^2b are each independently hydrogen or C1-C3 alkyl, or R ^2a and R ^2b together form oxo (=O); wherein, the alkyl can optionally be replaced by halogen or C1-C3 alkyl replace;

q is 1 or 2;

R ³ is selected from: C6-C10 aryl-L- or 5-10 membered heteroaryl-L-, wherein the aryl and heteroaryl can be optionally substituted by 1-3 R';

Each R' is independently selected from: halogen, amino, cyano, oxo (=O), C1-C3 hydroxyalkyl, -Y-C1-C4 alkyl, 3-10 membered carbocyclyl-L' -Y-, 3-10 membered heterocyclyl-L'-Y-, C6-C10 aryl-L'-Y- or 5-10 membered heteroaryl-L'-Y-, wherein the alkyl Base, aryl, heteroaryl, carbocyclyl, heterocyclyl can be optionally substituted by 1-3 R";

Each R" is independently selected from: cyano, oxo (=O), halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, or 3-8 membered heterocyclyl;

Each Y is independently selected from: absent, -O-, -S-, -NR ^c -, -NR ^c CO-, -SO-, -SO ₂ -, -CH(OH)-, or -CO- ;

each L and L' is independently selected from: absent or -CR ^d R ^e -;

R ^c , R ^d and R ^e are each independently selected from: hydrogen or C1-C3 alkyl; wherein, the alkyl may be optionally substituted by halogen or C1-C3 alkyl;

R ⁴ is H, halogen, C1-C6 alkyl, C1-C6 haloalkyl or C1-C6 alkoxy;

X ¹ , X ² and X ³ are each independently CR ⁵ or N, and at most two of X ¹ , X ² and X ³ are N at the same time;

^Each R is independently selected from: H, halogen, hydroxyl, amino, cyano, -C1-C3 alkyl, -C1-C3 hydroxyalkyl, C2-C4 alkenyl or C2-C4 alkynyl; wherein, The alkyl, C2-C4 alkenyl or C2-C4 alkynyl may be optionally substituted by halogen or C1-C3 alkyl.

In another preferred embodiment, each R' is independently selected from: halogen, amino, cyano, oxo (=O), C1-C3 hydroxyalkyl, -Y-C1-C3 alkyl, 3-10 Carbocyclyl-L'-Y-, 3-10-membered heterocyclyl-L'-Y-, C6-C10 aryl-L'-Y- Or 5-10 membered heteroaryl-L'-Y-, wherein said alkyl, aryl, heteroaryl, carbocyclyl and heterocyclyl may be optionally substituted by 1-3 R".

In another preferred example, R ^1a and R ^1b are each independently hydrogen or R ^1a and R ^1b together form oxo (=O).

In another preferred example, q is 1.

In another preferred example, R ^2a and R ^2b are each independently hydrogen or R ^2a and R ^2b together form oxo (=O); q is 1.

In another preference, ^R3 is selected from:

in,

Each ^R6 is independently selected from: halogen, amino, cyano, oxo (=O), C1-C3 hydroxyalkyl, -Y-C1-C4 alkyl, 3-10 membered carbocyclyl-L' -Y or -3-10 membered heterocyclyl-L'-Y-, wherein the alkyl, carbocyclyl and heterocyclyl can be optionally substituted by 1-3 R";

each L and L' is independently selected from: absent or -CR ^d R ^e -;

R ^c , R ^d and R ^e are each independently selected from: hydrogen or C1-C3 alkyl;

m is 0, 1, 2, 3 or 4;

R ⁷ is H or C1-C3 alkyl.

In another preferred embodiment, each ^R6 is independently selected from: halogen, amino, cyano, oxo (=O), C1-C3 hydroxyalkyl, -Y-C1-C3 alkyl, 3-10 Membered carbocyclyl-L'-Y or -3-10 membered heterocyclyl-L'-Y-, wherein the alkane A radical, a carbocyclyl, a heterocyclyl can be optionally substituted by 1-3 R".

In another preferred embodiment, ^R4 is H, halogen, C1-C3 alkyl, C1-C3 haloalkyl or C1-C3 alkoxy; preferably, ^R4 is H, chlorine, fluorine, methyl, ethyl , methoxy or trifluoromethyl.

In another preferred embodiment, X ¹ is CR ⁵ ; X ² is N; X ³ is CR ⁵ ; each R ⁵ is independently H, halogen, hydroxyl, amino, cyano, -C1-C3 hydroxyalkyl , C2-C4 alkenyl or C2-C4 alkynyl;

Preferably, X ¹ is CH; X ² is N; X ³ is CH.

In another preferred example, each R ⁶ is independently selected from: halogen, amino, cyano, oxo (=O), C1-C3 hydroxyalkyl, C1-C3 alkyl, C1-C3 haloalkyl, C1-C4 alkyl-O-, C3-C6 cycloalkyl, C3-C6 cycloalkyl-O-, C3-C6 cycloalkyl CH ₂ -, C3-C6 cycloalkyl CH ₂ -O-, -CONR ^c C3-C6 cycloalkyl, CONR ^c C1-C3 alkyl, CONR ^c C1-C3 haloalkyl.

In another preferred example, each R ⁶ is independently selected from: halogen, amino, cyano, oxo (=O), C1-C3 hydroxyalkyl, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkyl-O-, C3-C6 cycloalkyl, C3-C6 cycloalkyl-O-, C3-C6 cycloalkyl CH ₂ -, C3-C6 cycloalkyl CH ₂ -O-, -CONR ^c C3-C6 cycloalkyl, CONR ^c C1-C3 alkyl, CONR ^c C1-C3 haloalkyl.

In another preferred example, R ^1a , R ^1b , R 2a , R ^2b , R ³ , R ⁴ , R ⁵ , R ⁶ , q, X ¹ , X ² , X ³ , Y, ^L and L' are all The corresponding groups of each specific compound in the example.

In another preferred embodiment, the compound is selected from the following group:

In another preferred example, the compound is the compound shown in the examples.

In a second aspect, the present invention provides a pharmaceutical composition, wherein the pharmaceutical composition comprises:

(1) A therapeutically effective amount selected from the compound described in the first aspect, its pharmaceutically acceptable salt, enantiomer, diastereoisomer, tautomer, cis-trans isomer, One or more of solvates, polymorphs and deuterated compounds as active ingredients; and

(2) Optionally, a pharmaceutically acceptable carrier.

9. The compound as described in the first aspect, its pharmaceutically acceptable salt, enantiomer, diastereomer, tautomer, cis-trans isomer, solvate, polymorph The use of the substance or deuterated substance or the pharmaceutical composition described in the second aspect in the preparation of a medicament for preventing or treating MTAP ^-/- related cancers.

In another preferred example, the cancer is selected from: liver cancer, breast cancer, skin cancer, pancreatic cancer, head and neck cancer, intestinal cancer, lung cancer, gastric cancer, esophageal cancer, kidney cancer, bladder cancer, urethral cancer, prostate cancer, testicular cancer Carcinoma, Uterine Cancer, Ovarian Cancer, Vaginal Cancer, Fallopian Tube Cancer, Cholangiocarcinoma, Multiple Myeloma, Spinal Neurofibroma, Astrocytoma, Glioma, Acute Lymphocytic Leukemia, Chronic Lymphocytic Leukemia, Hodgkin Lymphoma, non-Hodgkin's lymphoma, sarcoma.

It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described in the following (such as embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, we will not repeat them here.

Detailed ways

After extensive and in-depth research, the inventors unexpectedly developed for the first time a phthalazidinone compound that can effectively inhibit the MTA synergistic PRMT5, which can be used to prepare drugs for treating cancers related to MTAP ^-/- . On this basis, the present invention has been accomplished.

Glossary

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

As used herein, the term "comprises" or "includes (comprising)" can be open, semi-closed and closed. In other words, the term also includes "consisting essentially of", or "consisting of".

Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. For the experimental methods without specific conditions indicated in the following examples, the conventional conditions or the conditions suggested by the manufacturer are usually followed. Percentages and parts are by weight unless otherwise indicated.

Group definition

Definitions of standard chemical terms can be found in references, including Carey and Sundberg "ADVANCED ORGANIC CHEMISTRY 4TH ED." Vols. A (2000) and B (2001), Plenum Press, New York. Unless otherwise stated, conventional methods within the skill of the art, such as mass spectrometry, NMR, IR and UV/VIS Spectroscopic and pharmacological methods. Unless specific definitions are set forth, terms employed herein in the relevant descriptions of analytical chemistry, synthetic organic chemistry, and pharmaceutical and medicinal chemistry are those that are known in the art. Standard techniques can be used in chemical syntheses, chemical analyses, pharmaceutical preparation, formulation and delivery, and treatment of patients. For example, reactions and purifications can be carried out using the manufacturer's instructions for the kit, or by methods known in the art or as described herein. The techniques and methods described above can generally be performed according to conventional methods well known in the art as described in various general and more specific documents that are cited and discussed in this specification. In this specification, groups and substituents thereof can be selected by those skilled in the art to provide stable moieties and compounds.

When a substituent is described by a conventional chemical formula written from left to right, the substituent also includes chemically equivalent substituents obtained when the structural formula is written from right to left. For example, -CH ₂ O- is equivalent to -OCH ₂ -.

The section headings used herein are for the purpose of organizing the article only and should not be construed as limitations on the subject matter described. All documents or portions of documents cited in this application, including but not limited to patents, patent applications, articles, books, manuals, and treatises, are hereby incorporated by reference in their entirety.

Certain chemical groups defined herein are preceded by abbreviated symbols to indicate the total number of carbon atoms present in the group. For example, C1-C6 alkyl refers to an alkyl group as defined below having a total of 1 to 6 carbon atoms. The total number of carbon atoms in the abbreviated notation does not include carbons that may be present in substituents of the stated group.

In addition to the foregoing, when used in the specification and claims of the present application, the following terms have the meanings shown below unless otherwise specified.

In this application, the term "halogen" means fluorine, chlorine, bromine or iodine.

"Hydroxy" means an -OH group.

"Hydroxyalkyl" means an alkyl group as defined below substituted with a hydroxyl group (-OH).

"Carbonyl" means a -C(=O)- group.

"Nitro" means _-NO2 .

"Cyano" means -CN.

"Amino" refers to _-NH2 .

"Substituted amino" means an amino group substituted by one or two alkyl, alkylcarbonyl, arylalkyl, heteroarylalkyl groups as defined below, for example, monoalkylamino, dialkylamino, Alkylamido, arylalkylamino, heteroarylalkylamino.

"Carboxy" means -COOH.

In this application, as a group or a part of other groups (such as used in groups such as halogen-substituted alkyl groups), the term "alkyl" refers to a fully saturated straight-chain or branched hydrocarbon chain group, Consists solely of carbon and hydrogen atoms, has for example 1 to 12 (preferably 1 to 8, more preferably 1 to 6) carbon atoms, and is connected to the rest of the molecule by 1 or more single bonds, Examples include but are not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-methylbutyl, 2,2- Dimethylpropyl, n-hexyl, heptyl, 2-methylhexyl, 3-methylhexyl, octyl, nonyl, decyl, etc. For the purposes of the present invention, the term "alkyl" preferably refers to an alkyl group containing 1 to 6 carbon atoms.

In this application, the term "alkenyl", as a group or part of another group, means consisting only of carbon atoms and hydrogen atoms, containing at least one double bond, having, for example, 2 to 14 (preferably 2 to 10 2, more preferably 2 to 6) carbon atoms and is connected to the rest of the molecule by 1 or more single bonds, such as but not limited to vinyl, propenyl, alkenyl Propyl, but-1-enyl, but-2-enyl, pent-1-enyl, pent-1,4-dienyl and the like.

The term "alkynyl" herein, as a group or part of another group, means consisting solely of carbon and hydrogen atoms, containing at least one carbon-carbon triple bond, having, for example, 2 to 14 (preferably 2 to 10 2, more preferably 2 to 6) carbon atoms and connected to the rest of the molecule by 1 or more single bonds, straight or branched hydrocarbon chain groups, such as but not limited to ethynyl, 1-propyne base, 1-butynyl, heptynyl, octynyl, etc.

In this application, as a group or a part of other groups, the term "carbocycle (group)" means a stable non-aromatic monocyclic or polycyclic hydrocarbon group composed only of carbon atoms and hydrogen atoms, which may include A fused ring system, a bridged ring system or a spiro ring system having 3 to 15 carbon atoms, preferably 3 to 10 carbon atoms, more preferably 3 to 8 carbon atoms, more preferably 3 to 6 carbon atoms (i.e. C3-C6), and it is a saturated or unsaturated ring (ie cycloalkyl, cycloalkenyl, etc.) and can be connected to the rest of the molecule by 1 or more single bonds via any suitable carbon atom. Unless specifically stated otherwise in this specification, the carbon atoms in the carbocyclyl group can be optionally oxidized. Examples of carbocyclyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, adamantyl, 2,3-indanyl, octahydro-4, 7-methylene-1H-indenyl, 1,2,3,4-tetrahydro-naphthyl, 5,6,7,8-tetrahydro-naphthyl, cyclopentenyl, cyclohexenyl, cyclo Hexadienyl, 1H-indenyl, 8,9-dihydro-7H-benzocyclohepten-6-yl, 6,7,8,9-tetrahydro-5H-benzocycloheptenyl, 5 ,6,7,8,9,10-hexahydro-benzocyclooctenyl, fluorenyl, bicyclo[1.1.1]pentane, bicyclo[2.2.1]heptyl, 7,7-dimethyl Base-bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, bicyclo[2.2.2]octyl, bicyclo[3.1.1]heptyl, bicyclo[3.2.1] Octyl, bicyclo[2.2.2]octenyl, bicyclo[3.2.1]octenyl, octahydro-2,5-methylene-pentalenyl, etc.

In this application, as a group or part of other groups, the term "cycloalkyl" refers to the above-mentioned fully saturated carbocyclic (group), typical cycloalkyl groups include but not limited to cyclopropyl, cyclobutyl , cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, adamantyl, etc.

In this application, as a group or part of other groups, the term "cycloalkenyl" refers to a partially unsaturated carbocyclic (group), typical cycloalkenyl groups include but not limited to cyclobutenyl, cyclopentenyl alkenyl, cyclohexenyl, etc.

In this application, the term "heterocycle (group)", as a group or part of another group, means 2 to 14 carbon atoms and 1 to 6 heteroatoms selected from nitrogen, phosphorus, oxygen and sulfur Composed of stable 3- to 20-membered non-aromatic cyclic groups. Unless otherwise specified in this specification, the heterocyclic group may be a monocyclic, bicyclic, tricyclic or multicyclic ring system, which may include a fused ring system, a bridged ring system or a spiro ring system; The nitrogen, carbon, or sulfur atoms of can be optionally oxidized; the nitrogen atoms can be optionally quaternized; and the heterocyclyl can be partially or fully saturated. A heterocyclyl group can be attached to the rest of the molecule via a carbon atom or a heteroatom and via 1 or more single bonds. In heterocyclyl groups comprising fused rings, one or more rings may be aryl or heteroaryl as defined below, provided that the point of attachment to the rest of the molecule is a non-aromatic ring atom. For the purposes of the present invention, heterocyclyl is preferably a stable 4- to 11-membered non-aromatic monocyclic, bicyclic, bridged or spirocyclic group comprising 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur group, more preferably a stable 4- to 8-membered non-aromatic monocyclic, bicyclic, bridged or spirocyclic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur. Heterocycle Examples of radicals include, but are not limited to: pyrrolidinyl, morpholinyl, piperazinyl, homopiperazinyl, piperidinyl, thiomorpholinyl, 2-azabicyclo[2.2.2]octanyl, 2 ,7-diaza-spiro[3.5]nonan-7-yl, 2-oxa-6-aza-spiro[3.3]heptane-6-yl, 2,5-diaza-bicyclo[2.2 .1] Heptan-2-yl, azetidinyl, pyranyl, tetrahydropyranyl, thiopyranyl, tetrahydrofuranyl, oxazinyl, dioxolyl, tetrahydroisoquinolinyl , decahydroisoquinolinyl, imidazolinyl, imidazolidinyl, quinazinyl, thiazolidinyl, isothiazolidinyl, isoxazolidinyl, dihydroindolyl, octahydroindolyl, octahydroiso Indolyl, pyrrolidinyl, pyrazolidinyl, phthalimidyl, etc.

In this application, the term "aryl" as a group or part of another group means a conjugated Hydrocarbon ring system group. For the purposes of the present invention, aryl can be a monocyclic, bicyclic, tricyclic or multicyclic ring system and can also be fused to a carbocyclyl or heterocyclyl as defined above, provided that the aryl is via The atoms in the aromatic ring are connected to the rest of the molecule by 1 or more single bonds. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, phenanthrenyl, fluorenyl, 2,3-dihydro-1H-isoindolyl, 2-benzoxazolinone, 2H-1, 4-Benzoxazin-3(4H)-on-7-yl and the like.

In the present application, the term "arylalkyl" refers to an alkyl group as defined above substituted with an aryl group as defined above.

In this application, the term "heteroaryl", as a group or part of another group, means having 1 to 15 carbon atoms (preferably 1 to 10 carbon atoms) and 1 to 6 carbon atoms selected from the group consisting of nitrogen A 5- to 16-membered conjugated ring system group of heteroatoms of oxygen and sulfur. Unless specifically stated otherwise in this specification, heteroaryl may be a monocyclic, bicyclic, tricyclic or multicyclic ring system, and may also be fused to a carbocyclyl or heterocyclyl as defined above, provided that the heteroaryl An aryl group is attached to the rest of the molecule by one or more single bonds through atoms on the heteroaromatic ring. A nitrogen, carbon or sulfur atom in a heteroaryl can be optionally oxidized; the nitrogen atom can be optionally quaternized. For the purposes of the present invention, heteroaryl is preferably a stable 5- to 12-membered aromatic group containing 1 to 5 heteroatoms selected from nitrogen, oxygen and sulfur, more preferably 1 to 4 heteroatoms selected from A stable 5- to 10-membered aromatic group of heteroatoms selected from nitrogen, oxygen and sulfur or a 5- to 6-membered aromatic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur. Examples of heteroaryl groups include, but are not limited to, thienyl, imidazolyl, pyrazolyl, thiazolyl, oxazolyl, oxadiazolyl, isoxazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, Benzimidazolyl, benzopyrazolyl, indolyl, furyl, pyrrolyl, triazolyl, tetrazolyl, triazinyl, indolyl, isoindolyl, indazolyl, isoindazolyl , Purinyl, quinolinyl, isoquinolinyl, diazinyl, naphthyridinyl, quinoxalinyl, pteridinyl, carbazolyl, carbolinyl, phenanthridinyl, phenanthrolinyl, acridine Base, phenazinyl, isothiazolyl, benzothiazolyl, benzothienyl, oxatriazolyl, cinnolinyl, quinazolinyl, phenylthio, indolizyl, o-phenanthrenyl, Isoxazolyl, phenoxazinyl, phenothiazinyl, 4,5,6,7-tetrahydrobenzo[b]thienyl, naphthopyridyl, [1,2,4]triazolo[4 ,3-b]pyridazine, [1,2,4]triazolo[4,3-a]pyrazine, [1,2,4]triazolo[4,3-c]pyrimidine, [1, 2,4]triazolo[4,3-a]pyridine, imidazo[1,2-a]pyridine, imidazo[1,2-b]pyridazine, imidazo[1,2-a]pyrazine wait.

In the present application, the term "heteroarylalkyl" refers to an alkyl group as defined above substituted by a heteroaryl group as defined above.

In the present application, the term "absent" means that the two sides of the group defined above are directly connected by a chemical bond. For example, "B is absent in A-B-C" means "A-C".

In this application, middle Indicates the attachment position of the group R.

In this application, except for the special instructions in the claims, "optionally" and "optionally" mean that the subsequently described event or situation may or may not occur, and the description includes both the occurrence and non-occurrence of the event or situation Case. For example, "optionally substituted aryl" means that the hydrogens on the aryl are substituted or unsubstituted, and the description includes both substituted and unsubstituted aryls. For example, the term "optionally substituted", "substituted" or "substituted by" as used herein, where no substituent is explicitly listed, means that one or more of the given atom or group hydrogen atoms are independently substituted by one or more, such as 1, 2, 3 or 4, substituents independently selected from: deuterium (D), halogen, -OH, oxo (=O), Mercapto, cyano, -CD ₃ , -C1-C6 alkyl (preferably -C1-3 alkyl), C2-C6 alkenyl, C2-C6 alkynyl, cycloalkyl (preferably C3-C8 cycloalkyl), Aryl, heterocyclic group (preferably 3-8 membered heterocyclic group), heteroaryl, aryl-C1-C6 alkyl-, heteroaryl-C1-C6 alkyl-, C1-C6 haloalkyl-,- OC1-C6 alkyl (preferably -OC1-C3 alkyl), -OC2-C6 alkenyl, -O cycloalkyl, -O heterocyclyl, -O aryl, -O heteroaryl, -OC1-C6 alkane phenyl, -C1-C6 alkyl-OH (preferably -C1-C4 alkyl-OH), -C1-C6 alkyl-SH, -C1-C6 alkyl-O-C1-C6 alkyl, -OC1 -C6 haloalkyl, -NH ₂ , -C1-C6 alkyl-NH ₂ (preferably -C1-C3 alkyl-NH ₂ ), -N(C1-C6 alkyl) ₂ (preferably -N(C1-C3 alkane base) ₂ ), -NH(C1-C6 alkyl) (preferably -NH(C1-C3 alkyl)), -N(C1-C6 alkyl) (C1-C6 alkylphenyl), -NH(C1 -C6alkylphenyl), -N(C1-C6alkyl)(aryl), -NH(aryl), nitro, -C(O)-OH, -C(O)OC1-C6alkyl (preferably -C(O)OC1-C3 alkyl), -CONR ⁱ R ⁱⁱ (wherein R ⁱ and R ⁱⁱ are H, D and C1-6 alkyl, preferably C1-3 alkyl), -NHC (O) (C1-C6 alkyl), -NHC (O) (phenyl), -N (C1-C6 alkyl) C (O) (C1-C6 alkyl), -N (C1-C6 alkyl) C ( O)(phenyl), -C(O)C1-C6 alkyl, -C(O)heteroaryl (preferably -C(O)-5-7 membered heteroaryl), -C(O)C1- C6 alkylphenyl, -C(O)C1-C6 haloalkyl, -OC(O)C1-C6 alkyl (preferably -OC(O)C1-C3 alkyl), -S(O) ₂ -C1- C6 alkyl, -S(O)-C1-C6 alkyl, -S(O) ₂ -phenyl, -S(O) ₂ -C1-C6 haloalkyl, -S(O) ₂ NH ₂ , -S (O) ₂ NH(C1-C6 alkyl), -S(O) ₂ NH (phenyl), -NHS(O) ₂ (C1-C6 alkyl), -NHS(O) ₂ (phenyl) and -NHS(O) ₂ (C1-C6 haloalkyl), wherein each of said alkyl, alkenyl, alkynyl, cycloalkyl, phenyl, aryl, heterocyclyl and heteroaryl is optionally Further substituted with one or more substituents selected from the group consisting of halogen, -OH, oxo (=O), -NH ₂ , cycloalkyl, 3-8 membered heterocyclyl, C1-C4 alkyl, C1- C4 haloalkyl-, -OC1-C4 alkyl, -C1-C4 alkyl-OH, -C1-C4 alkyl-O-C1-C4 alkyl, -OC1-C4 haloalkyl, cyano, nitro, - C(O)-OH, -C(O)OC1-C6 alkyl, -CON(C1-C6 alkyl) ₂ , -CONH(C1-C6 alkyl), -CONH ₂ , -NHC(O)(C1 -C6 alkyl), -NH(C1-C6 alkyl)C(O)(C1-C6 alkyl), -SO ₂ (C1-C6 alkyl), -SO ₂ (phenyl), -SO ₂ ( C1-C6 haloalkyl), -SO ₂ NH ₂ , -SO ₂ NH (C1-C6 alkyl), -SO ₂ NH (phenyl), -NHSO ₂ (C1-C6 alkyl), -NHSO ₂ (benzene radical) and -NHSO ₂ (C1-C6 haloalkyl). When an atom or group is substituted by multiple substituents, the substituents may be the same or different. As used herein, the terms "moiety", "structural moiety", "chemical moiety", "group", "chemical group" refer to a specific segment or functional group in a molecule. Chemical moieties are generally considered to be chemical entities embedded or attached to molecules.

In the present invention, (C1-C4 alkyl) ₂ amino represents 2 C1-C4 alkyl substituted amines, such as wait.

"Multiple" in the present invention means 2, 3 or 4.

active ingredient

As used herein, "compound of the present invention" or "active ingredient" refers to the compound shown in formula I, and also includes its pharmaceutically acceptable salt, enantiomer, diastereoisomer, tautomer isomers, cis-trans isomers, solvates, polymorphs, deuteriums, or combinations thereof.

"Stereoisomer" refers to compounds composed of the same atoms, bonded by the same bonds, but having different three-dimensional structures. The present invention will encompass each stereoisomer and mixtures thereof.

When olefinic double bonds are contained in the compounds of the present invention, unless otherwise stated, the compounds of the present invention are intended to include both E- and Z-geometric isomers.

"Tautomer" refers to isomers formed by the transfer of a proton from one atom of a molecule to another atom of the same molecule. All tautomeric forms of the compounds of the invention are also intended to be within the scope of the invention.

The compounds of the present invention, or pharmaceutically acceptable salts thereof, may contain one or more chiral carbon atoms, and thus may give rise to enantiomers, diastereoisomers and other stereoisomeric forms. Each chiral carbon atom can be defined as (R)- or (S)- based on stereochemistry. The present invention is intended to include all possible isomers, as well as their racemates and optically pure forms. The preparation of the compounds of the present invention can select racemates, diastereomers or enantiomers as starting materials or intermediates. Optically active isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, such as crystallization and chiral chromatography.

Conventional techniques for the preparation/isolation of individual isomers include chiral synthesis from suitable optically pure precursors, or resolution of racemates (or racemates of salts or derivatives) using, for example, chiral high performance liquid chromatography. ), see, for example, Gerald Gübitz and Martin G. Schmid (Eds.), Chiral Separations, Methods and Protocols, Methods in Molecular Biology, Vol.243, 2004; A.M.Stalcup, Chiral Separations, Annu.Rev.Anal.Chem.3 :341-63, 2010; Fumiss et al. (eds.), VOGEL'S ENCYCLOPEDIA OF PRACTICAL ORGANIC CHEMISTRY 5.sup.TH ED., Longman Scientific and Technical Ltd., Essex, 1991, 809-816; Heller, Acc.Chem . Res. 1990, 23, 128.

In this application, the term "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

"Pharmaceutically acceptable acid addition salt" refers to a salt formed with an inorganic or organic acid that retains the biological effectiveness of the free base without other side effects. Inorganic acid salts include but not limited to hydrochloride, hydrobromide, sulfate, nitrate, phosphate, etc.; organic acid salts include but not limited to formate, acetate, 2,2-dichloroacetate , Trifluoroacetate, Propionate, Caproate, Caprylate, Caprate, Undecylenate, Glycolate, Gluconate, Lactate, Sebacate, Hexanoate glutarate, malonate, oxalate, maleate, succinate, fumarate, tartrate, citrate, palmitate, stearate, oleate , cinnamate, laurate, malate, glutamate, pyroglutamate, aspartate, benzoate, mesylate, benzenesulfonate, p-toluenesulfonate , alginate, ascorbate, salicylate, 4-amino salicylate, naphthalene disulfonate, etc. These salts can be prepared by methods known in the art.

"Pharmaceutically acceptable base addition salt" refers to a salt formed with an inorganic base or an organic base that can maintain the biological effectiveness of the free acid without other side effects. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like. Preferred inorganic salts are ammonium, sodium, potassium, calcium and magnesium salts. Salts derived from organic bases include, but are not limited to, those of primary, secondary, and tertiary amines, substituted amines, including natural substituted amines, cyclic amines, and basic ion exchange resins , such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, triethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, bicyclic Hexylamine, Lysine, Arginine, Histidine, Caffeine, Procaine, Choline, Betaine, Ethylenediamine, Glucosamine, Methylglucamine, Theobromine, Purine, Piperazine, Piperazine Pyridine, N-ethylpiperidine, polyamine resin, etc. Preferred organic bases include isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline and caffeine. These salts can be prepared by methods known in the art.

Pharmaceutical compositions and methods of administration

In this application, a "pharmaceutical composition" refers to a formulation of a compound of the present invention with a vehicle generally accepted in the art for the delivery of a biologically active compound to a mammal (eg, a human). The medium includes a pharmaceutically acceptable carrier. The purpose of the pharmaceutical composition is to promote the administration of the organism, facilitate the absorption of the active ingredient and thus exert its biological activity.

The compound of general formula (I) can be used in combination with other known drugs for treating or improving similar diseases. In the case of combined administration, the administration method and dose of the original drug can be kept unchanged, and the compound of formula I can be administered simultaneously or subsequently. When the compound of formula I is taken together with one or several other drugs, the pharmaceutical composition containing one or several known drugs and the compound of formula I can be preferably used. Drug combinations also include administration of a compound of formula I and one or more other known drugs for overlapping periods of time. When the compound of formula I is used in combination with one or several other drugs, the dosage of the compound of formula I or known drugs may be lower than that of their single administration.

Drugs or active ingredients that can be combined with the compound described in general formula (I) include but are not limited to: PD-1 inhibitors (such as nivolumab, pembrolizumab, etc.), PD-L1 inhibitors ( Such as durvalumab, atezolizumab, etc.), CD47 antibodies (such as Hu5F9-G4, CC-90002, etc.), CD20 antibodies (such as rituximab, ibrutinib, etc.), KRAS inhibitors ( Such as AMG510, etc.), ALK inhibitors (such as ceritinib, alectinib, brigatinib, lorlatinib, ocatinib), EGFR inhibitors (such as afatinib, gefitinib , erlotinib, lapatinib, dacomitinib, icotinib, osimertinib, etc.), VEGFR inhibitors (such as sorafenib, pazopanib, regorafenib, carbo Tini, sunitinib, etc.), PI3K inhibitors (such as Dactolisib, Taselisib, etc.), BTK inhibitors (such as ibrutinib, tirabrutinib, acabrutinib, zanubrutinib, Vicat Butinib, etc.), HDAC inhibitors (such as Vorinostat, Fimepinostat, Givinostat, Tucidinostat, etc.), CDK inhibitors (such as Palbociclib, Ribociclib, Abemaciclib, etc.), MEK inhibitors (such as Selumetinib (AZD6244), Trametinib, etc.), ERK inhibitors (such as BVD523, HH2710, etc.), mTOR inhibitors (such as Vistusertib, etc.), SHP2 inhibitors (such as RMC-4630, JAB-3068), SOS1 inhibitors (such as BI1701963, etc.), PRMT1 inhibitors , a MAT2A inhibitor, or a combination thereof.

The term "pharmaceutically acceptable" as used herein refers to a substance that does not affect the biological activity or properties of the compounds of the present invention (such as a carrier or diluent), and is relatively nontoxic, ie, the substance can be administered to an individual without causing an adverse biological response or interacting in an undesirable manner with any of the components contained in the composition.

In this application, "pharmaceutically acceptable carrier" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetener approved by the relevant government regulatory agency as acceptable for human or livestock use , diluent, preservative, dye/colorant, flavoring agent, surfactant, wetting agent, dispersing agent, suspending agent, stabilizing agent, isotonic agent, solvent or emulsifying agent.

The mode of administration of the compound or pharmaceutical composition of the present invention is not particularly limited, and representative modes of administration include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration .

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is admixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with (a) fillers or extenders, for example, Starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders such as hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) humectants, For example, glycerol; (d) disintegrants, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) slow agents, such as paraffin; (f) Absorption accelerators such as quaternary ammonium compounds; (g) wetting agents such as cetyl alcohol and glyceryl monostearate; (h) adsorbents such as kaolin; and (i) lubricants such as talc, hard Calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage form may also contain buffering agents.

Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shell materials, such as enteric coatings and others well known in the art. They may contain opacifying agents and, in such compositions, the release of the active compound or compounds may be in a certain part of the alimentary canal in a delayed manner. Examples of usable embedding components are polymeric substances and waxy substances. The active compounds can also be in microencapsulated form, if desired, with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances, etc.

Besides such inert diluents, the compositions can also contain adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Suspensions, in addition to the active compounds, may contain suspending agents, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances, and the like.

Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols, and suitable mixtures thereof.

The "tumor" mentioned in the present invention includes but not limited to lung cancer, pancreatic cancer, colorectal cancer, leukemia, Ewing's sarcoma, breast cancer, prostate cancer, T-cell lymphoma, B-cell lymphoma, malignant rhabdomyoma, synovial sarcoma, Endometrioma, stomach cancer, liver cancer, kidney cancer, melanoma, ovarian cancer, glioma, cholangiocarcinoma, nasopharyngeal cancer, cervical cancer, head and neck cancer, esophageal cancer, thyroid cancer and bladder cancer and other diseases. As used herein, the terms "prophylactic", "prevention" and "prevention" include reducing the likelihood of a disease or condition occurring or worsening in a patient.

As used herein, the term "treatment" and other similar synonyms include the following meanings:

(i) preventing the occurrence of a disease or condition in a mammal, especially when such mammal is susceptible to the disease or condition but has not been diagnosed as having the disease or condition;

(ii) inhibiting a disease or condition, i.e. arresting its development;

(iii) ameliorating a disease or condition, i.e., causing regression of the state of the disease or condition; or

(iv) Alleviating the symptoms caused by the disease or condition.

The term "effective amount", "therapeutically effective amount" or "pharmaceutically effective amount" as used herein refers to at least one agent or compound which, when administered, is sufficient to relieve to some extent one or more symptoms of the disease or condition being treated amount. The result may be a reduction and/or alleviation of a sign, symptom or cause, or any other desired change in a biological system. For example, a therapeutically "effective amount" is the amount of a composition comprising a compound disclosed herein required to provide a clinically significant disease-modifying effect. Effective amounts suitable for any individual case can be determined using techniques such as dose escalation assays.

As used herein, the terms "administering", "administering", "administering" and the like refer to methods capable of delivering a compound or composition to the desired site of biological action. These methods include, but are not limited to, oral routes, duodenal routes, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intraarterial injection or infusion), topical administration and rectal administration. Administration techniques useful for the compounds and methods described herein are well known to those skilled in the art, as described, for example, in Goodman and Gilman, The Pharmacological Basis of Therapeutics, current ed.; Pergamon; and Remington's, Pharmaceutical Sciences (current edition), Mack Publishing Co., Those discussed in Easton, Pa. In preferred embodiments, the compounds and compositions discussed herein are administered orally.

As used herein, the terms "pharmaceutical combination", "drug combination", "combination", "administration of other treatments", "administration of other therapeutic agents" and the like refer to drug treatments obtained by mixing or combining more than one active ingredient, It includes fixed and non-fixed combinations of active ingredients. The term "fixed combination" refers to the simultaneous administration to a patient of at least one compound described herein and at least one co-agent in the form of a single entity or single dosage form. The term "variable combination" refers to simultaneous, concomitant or sequential administration at variable intervals of at least one compound described herein and at least one synergistic agent as separate entities to a patient. These also apply to cocktail therapy, eg the administration of three or more active ingredients.

The pharmaceutical composition of the present invention comprises the compound of the present invention or a pharmacologically acceptable salt thereof within a safe and effective amount range and a pharmaceutically acceptable excipient or carrier. Wherein, "safe and effective dose" refers to: the amount of the compound is sufficient to obviously improve the condition without causing severe side effects. Usually, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, more preferably 10-1000 mg of the compound of the present invention per dose. Preferably, the "one dose" is a capsule or tablet.

Compound preparation method

Methods for preparing compounds of formula I are described in the following schemes. In some cases, the order in which the steps of a reaction scheme are performed can be altered to facilitate a reaction or to avoid undesired side reaction products. The compounds of the present invention can also be conveniently prepared by combining various synthetic methods described in the specification or known in the art, and such combinations can be easily performed by those skilled in the art to which the present invention belongs.

Those skilled in the art will also understand that in the methods described below, the functional groups of intermediate compounds may need to be protected by appropriate protecting groups. Such functional groups include hydroxyl, amino, mercapto and carboxylic acid. Suitable hydroxy protecting groups include trialkylsilyl or diarylalkylsilyl groups (eg tert-butyldimethylsilyl, tert-butyldiphenylsilyl or trimethylsilyl) , tetrahydropyranyl, benzyl, allyl, etc. Suitable protecting groups for amino, amidino and guanidino include tert-butoxycarbonyl, benzyloxycarbonyl, 9-fluorenylmethoxycarbonyl, benzyl, p-methoxybenzyl, allyl, allyloxycarbonyl , p-toluenesulfonyl, pivaloyl, trifluoroacetyl, etc. Suitable protecting groups for mercapto include -C(O)-R" (where R" is alkyl, aryl or aralkyl), p-methoxybenzyl, trityl and the like. Suitable carboxy protecting groups include alkyl, aryl or aralkyl esters.

Protecting groups can be introduced and removed according to standard techniques known to those skilled in the art and as described herein. The use of protecting groups is described in detail in Greene, T.W. and P.G.M. Wuts, Protective Groups in Organi Synthesis, (1999), 4th Ed., Wiley. The protecting group can also be a polymeric resin.

Usually, in the preparation process, each reaction is usually carried out in an inert solvent at room temperature to reflux temperature (such as 0, in the preparation flow, preferably 10 or 100 in the preparation flow. The reaction time is usually 0.1 hour-60 hours, relatively Preferably it is 0.5-48 hours.

Preferably, the compound of formula I can be prepared by the following method:

(1) Reductive amination reaction between compound a and b or formation of imine first and then affinity addition reaction to obtain compound c;

(2) Compound c undergoes a condensation reaction in the presence of a condensing agent or undergoes an ammonolysis reaction in the presence of a base to generate compound d;

(3) Suzuki reaction occurs between compound d and compound e to generate compound I;

Preferably, compound d can also directly generate compound d through compound f and compound b in the presence of a base.

Preferably, the condensing agent is selected from the group consisting of dicyclohexylcarbodiimide, diisopropylcarbodiimide, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide salt salt, 1-hydroxy-7-azobenzotriazole, 1-hydroxybenzotriazole, N-hydroxysuccinimide, N,N,N,N′-tetramethyl-O-( 7-Azabenzotriazol-1-yl)urea hexafluorophosphate, benzotriazole-N,N,N,N'-tetramethyluronium hexafluorophosphate, 1H-benzotriazole-1 -Oxotris(dimethylamino)phosphonium hexafluorophosphate, propyl phosphoric anhydride, 2-hydroxypyridine nitrogen nitride, 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate or its combination;

The base is selected from the group consisting of sodium hydride, sodium methoxide, sodium ethoxide, potassium tert-butoxide, sodium tert-butoxide, sodium hydroxide, potassium hydroxide, n-butyllithium, lithium diisopropylamide, hexamethyl Lithium disilazide, sodium hexamethyldisilazide, potassium hexamethyldisilamide, potassium carbonate, cesium carbonate, sodium carbonate, triethylamine, diisopropylethylamine, 1.8-diazepine Bicyclo[5.4.0]undec-7-ene, or a combination thereof;

Wherein, R ^1a , R ^1b , R ^2a , R ^2b , R ³ , R ⁴ , X ¹ , X ² and X ³ are as defined above.

Compared with the prior art, the present invention has the following beneficial effects:

(1) The present invention provides a compound as shown in formula I or a pharmaceutically acceptable salt thereof with a novel structure;

(2) The compound of the present invention can inhibit PRMT5 synergistically with MTA, and it can be used to prepare drugs for treating cancers related to MTAP ^-/- .

The technical solutions of the present invention will be further described below through specific embodiments. It should be clear to those skilled in the art that the examples are only for helping to understand the present invention, and should not be regarded as specific limitations on the present invention.

For the experimental methods without specific conditions indicated in the following examples, the conventional conditions or the conditions suggested by the manufacturer are usually followed. Percentages and parts are by weight unless otherwise indicated.

The experimental materials and reagents used in the following examples can be obtained from commercially available channels unless otherwise specified.

In each embodiment, ¹ H NMR is recorded by a BRUKER AVANCE NEO 400MHz nuclear magnetic resonance instrument, and the chemical shift is expressed in δ (ppm); liquid mass spectrometry (LCMS) is recorded by Shimadzu LC-20AD, SIL-20A, CTO-20AC, SPD - M20A, CBM-20A, LCMS-2020 type mass spectrometer records; preparative HPLC separation using Gilson-281 type liquid chromatograph.

Example

1. Preparation of Intermediate A

(1) For the synthesis of compound A-1, refer to patent WO2021050915A1.

MS-ESI [M+H] ⁺ , calculated 354, found 354.

¹ H NMR (400MHz, DMSO-d6) δ12.69(s, 1H), 8.26(s, 1H), 8.17(d, J=8.4Hz, 1H), 8.02(dd, J=1.6, 8.4Hz, 1H ), 4.41 (d, J=6.0Hz, 2H), 1.40 (s, 9H).

(2) To a solution of compound A-1 (200mg, 565μmol) in dioxane (5.0mL), add pinacol borate (175mg, 689μmol), [1,1'-bis(diphenylphosphine ) ferrocene] palladium dichloride (40.0 mg, 54.7 μmol) and potassium acetate (115 mg, 1.17 mmol). The reaction solution was stirred at 80° C. for 1 hour under the protection of nitrogen. Add water (10.0mL) to the reaction solution, extract with ethyl acetate (10.0mL×2), combine the organic phases, dry over anhydrous sodium sulfate, filter, concentrate under reduced pressure, and the crude product is subjected to silica gel column chromatography (petroleum ether/ethyl acetate Esters=10:1) Compound A was isolated.

MS-ESI [M+H] ⁺ , calculated 320, found 320.

2. Preparation of Intermediate B

(1) To a solution of compound B-1 (100 g, 399 mmol) in tetrahydrofuran (1000 mL) was added dropwise a solution of isopropylmagnesium chloride (2.0 mol/L, 240 mL) in tetrahydrofuran at 0°C. The reaction solution was stirred at 25° C. for 4 hours under nitrogen protection. Dry ice (3.0 kg) was added to the reaction solution at -70°C, and stirred at 25°C for 4 hours. Water (300 mL) was added to the reaction solution, and concentrated under reduced pressure to remove tetrahydrofuran. The aqueous phase was washed with ethyl acetate (200 mL×3), and the pH value of the aqueous phase was adjusted to 1 with hydrochloric acid (4.0 mol/L). Solids were precipitated and filtered to obtain a filter cake as compound B-2.

¹ H NMR (400 MHz, DMSO-d6) δ 8.82 (d, J=6.8 Hz, 2H), 2.60 (s, 3H).

(2) To a methanol (120.mL) solution of compound B-2 (10.0 g, 46.3 mmol) was added thionyl chloride (10.0 mL, 138 mmol). The reaction solution was stirred at 80° C. for 12 hours. Add water (50.0 mL) to the reaction solution, adjust the pH to 7 with saturated aqueous sodium bicarbonate, extract with ethyl acetate (100 mL×2), combine the organic phases, and wash with saturated aqueous sodium chloride (100 mL×1). Dry over anhydrous sodium sulfate, filter and concentrate under reduced pressure to obtain compound B-3.

MS-ESI [M+H] ⁺ , calculated 230, found 230.

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.86-8.92 (m, 1H), 8.74-8.79 (m, 1H), 3.94 (d, J=4.4Hz, 3H), 2.63-2.73 (m, 3H).

(3) To a solution of compound B-3 (1.0 g, 4.35 mmol) in acetic acid (15.0 mL), selenium dioxide (1.0 g, 9.01 mmol) was added. The reaction solution was stirred at 130° C. for 2 hours under the protection of nitrogen. The reaction solution was filtered and concentrated under reduced pressure to obtain compound B.

MS-ESI [M+H] ⁺ , calculated 246, found 246.

¹ H NMR (400 MHz, CDCl ₃ ) δ 10.25 (s, 1H), 9.07 (d, J=4.0 Hz, 2H), 3.90 (s, 1H)).

3. Preparation of Intermediate C

(1) To a solution of compound C-1 (4.00 g, 27.4 mmol) in ethanol (60.0 mL), methylhydrazine (4.74 g, 103 mmol, 5.42 mL) was added dropwise. The reaction solution was stirred at 70° C. for 4 hours under the protection of nitrogen. The reaction solution was filtered to obtain the filter cake as Compound C.

MS-ESI [M+H] ⁺ , calculated 173, found 173.

¹ H NMR (400 MHz, CDCl3) δ 7.41-7.47 (m, 1H), 7.33-7.40 (m, 2H), 3.88 (s, 3H).

4. Preparation of Intermediate D

(1) To a solution of compound D-1 (2.50 g, 17.1 mmol) in ethanol (40.0 mL), hydrazine hydrate (3.19 g, 62.5 mmol, 3.10 mL) was added dropwise. The reaction solution was stirred at 70° C. for 15 hours under the protection of nitrogen. The reaction solution was filtered to obtain the filter cake as compound D-2.

MS-ESI [M+H] ⁺ , calculated 159, found 159.

¹ H NMR (400MHz, DMSO-d ₆ )δ7.65(d, J=8.0Hz, 1H), 7.48(d, J=8.0Hz, 1H), 7.4-7.4(m, 1H), 5.10(s, 2H).

(2) Potassium carbonate (2.00 g, 14.5 mmol) and dimethyl sulfate (2.18 g, 17.3 mmol, 1.64 mL) were added to a solution of compound D-2 (2.08 mg, 13.2 mmol) in acetone (30.0 mL). The reaction solution was stirred at 25°C under nitrogen protection for 12 hours, dimethyl sulfate (2.50g, 19.8mmol, 1.88mL) was added, and the reaction solution was stirred at 25°C under nitrogen protection for 16 hours. Water (20.0 mL) was added to the reaction solution, extracted with dichloromethane (50.0 mL×3), and the combined organic phases were washed with saturated aqueous sodium chloride solution (20.0 mL×3). Dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The crude product is separated by silica gel column chromatography (dichloromethane/methanol=1:0 to 10:1) to obtain compound D.

MS-ESI [M+H] ⁺ , calculated 173, found 173.

¹ H NMR (400MHz, DMSO-d ₆ )δ7.60(d, J=8.0Hz, 1H), 7.31(d, J=8.0Hz, 1H), 7.12-7.24(m, 1H), 5.76(s, 2H), 3.88(s, 3H).

Embodiment 1 synthetic compound 1

(1) To a solution of intermediate B (1.20 g, 4.92 mmol) in dichloromethane (20.0 mL) was added compound C (650 mg, 3.77 mmol) and acetic acid (210 mg, 3.50 mmol). The reaction solution was stirred at 25°C under nitrogen protection for 12 hours, sodium cyanoborohydride (500 mg, 7.96 mmol) was added, and the reaction solution was stirred at 25°C under nitrogen protection for 2 hours. Water (10.0 mL) was added to the reaction solution, extracted with dichloromethane (10.0 mL×3), and the combined organic phases were washed with saturated aqueous sodium chloride solution (10.0 mL×3). Dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The crude product is separated by silica gel column chromatography (petroleum ether/ethyl acetate=1:0 to 3:1) to obtain compound 1-1.

MS-ESI [M+H] ⁺ , calculated 402, found 402.

¹ H NMR (400MHz, CDCl3) δ8.80 (d, J = 13.2Hz, 2H), 7.28-7.41 (m, 3H), 5.15 (s, 2H), 4.12 (s, 3H), 3.80 (s, 3H ).

(2) To a solution of compound 1-1 (170 mg, 425 μmol) in tetrahydrofuran (5.0 mL) was added sodium tert-butoxide (62.0 mg, 645 μmol). The reaction solution was stirred at 25°C for 2 hours. Water (10.0 mL) was added to the reaction solution, extracted with ethyl acetate (10.0 mL×3), the combined organic phases were washed with saturated aqueous sodium chloride solution (10.0 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=1:0 to 2:1) to obtain compound 1-2.

MS-ESI [M+H] ⁺ , calculated 370, found 370.

¹ H NMR (400MHz, CDCl3) δ9.16(s, 1H), 8.99(s, 1H), 7.74(d, J=8.4Hz, 1H), 7.60-7.66(m, 1H), 7.53-7.59(m ,1H), 5.13(s,2H), 4.17(s,3H).

(3) To a solution of compound 1-2 (15.0 mg, 40.7 μmol) in dioxane (3.0 mL), add compound A (150 mg, 118 μmol), water (1.0 mL), potassium carbonate (90 mg, 651 μmol) and Methanesulfonic acid (2-dicyclohexylphosphino-2',4',6'-tri-isopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl- 2-yl)palladium(II) (30 mg, 35.4 μmol). The reaction solution was stirred at 80°C under nitrogen protection for 5 hours, filtered, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane/methanol=1:0 to 25:1) to obtain compound 1-3.

MS-ESI [M+H] ⁺ , calculated 563, found 563.

¹ H NMR (400MHz, CDCl3) δ10.21(s,1H),9.33(s,1H),9.01(s,1H),8.60(d,J=7.2Hz,1H),8.27(s,1H), 7.99(d, J=6.0Hz, 1H), 7.70(d, J=8.4Hz, 1H), 7.61(d, J=7.2Hz, 1H), 7.49-7.56(m, 1H), 5.31(s, 2H ), 4.67(s,2H), 4.13(s,3H), 1.33(s,9H).

(4) To a solution of compound 1-3 (3.0 mg, 5.33 μmol) in dichloromethane (1.0 mL) was added a solution of hydrochloric acid in dioxane (4.0 mol/L, 0.50 mL). The reaction solution was stirred at 25°C for 20 minutes. The reaction solution was concentrated under reduced pressure to obtain the hydrochloride of compound 1.

MS-ESI [M+H] ⁺ , calculated 463, found 463.

¹ H NMR (400MHz, MeOD) δ9.07-9.15(m, 1H), 9.00(s, 1H), 8.43(d, J=8.4Hz, 1H), 8.04-8.13(m, 2H), 7.92(d ,J=8.8Hz,1H),7.70(d,J=7.2Hz,1H),7.55-7.62(m,1H),4.68(s,2H),4.13(s,2H),4.08(s,3H) .

Embodiment 2 synthetic compound 2

(1) To a solution of intermediate B (198 mg, 813 μmol) in toluene (10.0 mL), compound D (140 mg, 813 μmol) and tetraisopropyl titanate (462 mg, 1.63 mmol) were added. The reaction solution was stirred at 80°C under nitrogen protection for 12 hours, sodium cyanoborohydride (102mg, 1.63mmol) was added, the reaction solution was stirred at 80°C under nitrogen protection for 2 hours, and sodium cyanoborohydride (102mg, 1.63mmol) was added ), the reaction solution was stirred for 12 hours at 40° C. under nitrogen protection. Add water (20.0mL) to the reaction solution, extract with dichloromethane (50.0mL×3), combine the organic phases, dry over anhydrous sodium sulfate, filter, concentrate under reduced pressure, and the crude product is subjected to preparative high performance liquid chromatography (C18-6, 100mm × 30mm 5 μm, A: water (formic acid); B: acetonitrile, 26%-56%: 15 minutes) isolated to obtain compound 2-1.

MS-ESI [M+H] ⁺ , calculated 388, found 388.

¹ H NMR (400MHz,DMSO-d ₆ )δ8.90(s,1H),8.78(s,1H),7.65-7.75(m,1H),7.42-7.46(m,1H),7.21-7.27(m , 1H), 4.80 (d, J=6.0Hz, 2H), 4.15 (s, 1H), 3.81 (s, 3H).

(2) To a solution of compound 2-1 (25.0 mg, 64.7 μmol) in N,N-dimethylformamide (1.0 mL) was added propylphosphoric anhydride (1.8 mg, 97.1 μmol, 57.8 μL, 50.0%) and Diisopropylethylamine (25.1 mg, 194 μmol, 33.8 μL). The reaction solution was stirred at 25°C for 1 hour. Add water (20.0mL) to the reaction solution, and dichloromethane (50.0mL× 3) Extraction, the combined organic phases were washed with saturated aqueous sodium chloride solution (50.0 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain compound 2-2.

MS-ESI [M+H] ⁺ , calc. 368, found 368.

(3) To a solution of compound 2-2 (40.0 mg, 109 μmol) in dioxane (3.0 mL), add compound A (173 mg, 217 μmol), water (1.0 mL), potassium carbonate (45.0 mg, 326 μmol) and Methanesulfonic acid (2-dicyclohexylphosphino-2',4',6'-tri-isopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl- 2-yl)palladium(II) (9.2 mg, 10.9 μmol). The reaction solution was stirred at 80°C under nitrogen protection for 5 hours, water (20.0 mL) was added, extracted with ethyl acetate (50.0 mL×3), the combined organic phases were washed with saturated aqueous sodium chloride solution (50.0 mL×2), anhydrous Dry over sodium sulfate, filter and concentrate under reduced pressure. The crude product is separated by silica gel column chromatography (dichloromethane/methanol=1:0 to 10:1) to obtain compound 2-3.

MS-ESI [M+H] ⁺ , calculated 563, found 563.

(4) To a solution of compound 2-3 (20.0 mg, 35.6 μmol) in dichloromethane (1.0 mL) was added trifluoroacetic acid (0.20 mL). The reaction solution was stirred at 25°C for 60 minutes. The reaction solution was concentrated under reduced pressure, and the crude product was separated by preparative high-performance liquid chromatography (Welch Xtimate C18, 100mm×30mm 10 μm, A: water (formic acid); B: acetonitrile, 5%-35%: 10 minutes) to obtain compound 2. salt.

MS-ESI [M+H] ⁺ , calculated 463, found 463.

¹ H NMR (400MHz, MeOD) δ9.23-9.25 (m, 1H), 9.11-9.13 (m, 1H), 8.56-8.60 (m, 1H), 8.23-8.26 (m, 1H), 8.20-8.22 ( m,1H),7.99-8.03(m,1H),7.71(s,1H),7.45-7.50(m,1H),5.34-5.46(m,2H),4.54-4.60(m,2H),4.18- 4.20(m,3H).

Embodiment 3 synthetic compound 3

(1) To the solution of compound 3-1 (3.00g, 11.3mmol) in N-methylpyrrolidone (40.0mL), add Zinc cyanide (5.32g, 45.3mmol), tetrakis(triphenylphosphine)palladium (3.92g, 3.40mmol). The reaction solution was stirred at 110° C. for 12 hours under nitrogen protection. Add water (100mL), extract with ethyl acetate (150mL×3), combine the organic phases and wash with saturated aqueous sodium chloride solution (100mL×2), dry over anhydrous sodium sulfate, filter and concentrate under reduced pressure to obtain compound 3-2 .

MS-ESI [M+H] ⁺ , calculated 212, found 212.

(2) To compound 3-2 (6.5g, 12.3mmol, 40% purity) in tetrahydrofuran (65.0mL) solution, add sodium hydride (985mg, 24.6mmol, 60%), the reaction solution was stirred at 0 ℃ under nitrogen protection For 0.5 hours, iodomethane (5.24 g, 36.9 mmol, 2.30 mL) was added. The reaction solution was stirred at 0° C. for 2 hours under nitrogen protection. Add water (50.0mL), extract with ethyl acetate (150mL×2), wash the combined organic phase with saturated aqueous sodium chloride solution (100mL×2), dry over anhydrous sodium sulfate, filter and concentrate under reduced pressure, the crude product is passed through a silica gel column Compound 3-3 was isolated by chromatography (petroleum ether/ethyl acetate=1:0 to 2:1).

MS-ESI [M+H] ⁺ , calculated 226, found 226.

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.28 (s, 1H), 7.97 (s, 1H), 7.77 (s, 1H), 4.22 (s, 3H).

(3) Dibromohydantoin (933 mg, 3.26 mmol) was added to a solution of compound 3-3 (490 mg, 2.18 mmol) in acetic acid (35.0 mL), and the reaction solution was stirred at 40° C. for 48 hours under nitrogen protection. Water (5.0 mL) was added, extracted with dichloromethane (25 mL×2), the combined organic phases were washed with saturated aqueous sodium chloride solution (10.0 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure, the crude product was purified by silica gel Compound 3-4 was isolated by column chromatography (petroleum ether/ethyl acetate=1:0 to 4:1).

MS-ESI [M+H] ⁺ , calculated 306, found 306.

(4) To a solution of compound 3-4 (750 mg, 2.47 mmol) in toluene (40.0 mL), add benzophenone imine (11.34 g, 7.40 mmol), tris(dibenzylideneacetone) dipalladium (226 mg , 247 μmol), cesium carbonate (2.41 g, 7.40 mmol) and 4,5-bisdiphenylphosphine-9,9-dimethylxanthene (285 mg, 493 μmol). The reaction solution was stirred at 120° C. for 4 hours under nitrogen protection. Water (50.0 mL) was added to the reaction solution, extracted with dichloromethane (50.0 mL×3), the combined organic phases were washed with saturated aqueous sodium chloride solution (50.0 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=1:0 to 2:1) to obtain compound 3-5.

MS-ESI [M+H] ⁺ , calculated 405, found 405.

¹ H NMR (400MHz, CDCl ₃ ) δ7.92(d, J=7.2Hz, 1H), 7.80-7.84(m, 2H), 7.73(s, 1H), 7.58-7.61(m, 1H), 7.43- 7.56(m,5H),7.35-7.38(m,1H),7.29(br s,1H),3.95-4.00(m,3H)

(5) To a solution of compound 3-5 (690 mg, 1.71 mmol) in tetrahydrofuran (5.0 mL) was added hydrochloric acid (12 mol/L, 142 μL). The reaction solution was stirred at 25°C for 12 hours. Add sodium carbonate aqueous solution to the reaction solution to adjust the pH to neutral, extract with dichloromethane (50.0mL×3), combine the organic phases, dry over anhydrous sodium sulfate, filter and concentrate under reduced pressure, the crude product is subjected to silica gel column chromatography (dichloromethane Methane/methanol=1:0 to 10:1) isolated compound 3-6.

MS-ESI [M+H] ⁺ , calculated 241, found 241.

(6) To a solution of intermediate B (240 mg, 999 μmol) in toluene (6.0 mL), compound 3-6 (290 mg, 1.00 mmol) and tetraisopropyl titanate (568 mg, 2.00 mmol) were added. The reaction solution was stirred at 80°C under nitrogen protection After 12 hours, sodium cyanoborohydride (502 mg, 7.99 mmol) was added, and the reaction solution was stirred at 40° C. for 36 hours under the protection of nitrogen. Add water (20.0mL) to the reaction solution, extract with ethyl acetate (50.0mL×2), combine the organic phases, dry over anhydrous sodium sulfate, filter, concentrate under reduced pressure, and the crude product is subjected to preparative high performance liquid chromatography (C18-6, 100mm ×30mm 5 μm, A: water (0.1% hydrochloric acid); B: acetonitrile, 0%-35%: 15 minutes) were isolated to obtain compound 3-7.

MS-ESI [M+H] ⁺ , calculated 454, found 454.

¹ H NMR (400MHz, DMSO-d ₆ ) δ8.61-8.82(m, 2H), 8.34(br d, J=6.0Hz, 1H), 7.86(d, J=4.0Hz, 1H), 4.86(d , J=6.8Hz, 1H), 4.69 (br d, J=6.4Hz, 1H), 3.96 (d, J=13.2Hz, 3H).

(7) Add propyl phosphoric anhydride (841 mg, 1.32 mmol, 786 μL, 50.0%) and diisopropyl Ethylamine (341 mg, 2.64 mmol, 460 μL). The reaction solution was stirred at 25°C for 1 hour. Water (20.0 mL) was added to the reaction solution, extracted with ethyl acetate (20.0 mL×3), the combined organic phases were washed with saturated aqueous sodium chloride solution (50.0 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane/methanol=1:0 to 100:1) to obtain compound 3-8.

MS-ESI [M+H] ⁺ , calculated 436, found 436.

¹ H NMR (400MHz, CDCl ₃ )δ9.18(s,1H),8.93(s,1H),8.00(s,1H),7.82(s,1H),5.02(s,2H),4.22(s, 3H).

(8) To a solution of compound 3-8 (80mg, 183μmol) in dioxane (3.0mL), add compound A (89.0mg, 275μmol), water (0.5mL), potassium carbonate (36.0mg, 367μmol) and 1,1-Bis(diphenylphosphino)ferrocenepalladium dichloride (13.4 mg, 18.3 μmol). The reaction solution was stirred at 70° C. for 5 hours under nitrogen protection. Water (20.0 mL) was added, extracted with dichloromethane (50.0 mL×3), the combined organic phases were washed with saturated aqueous sodium chloride (50.0 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure, and the crude product was Compound 3-9 was isolated by silica gel column chromatography (dichloromethane/methanol=1:0 to 10:1).

MS-ESI [M+H] ⁺ , calculated 631, found 631.

(9) To a solution of compound 3-9 (40 mg, 63.4 μmol) in dichloromethane (1.0 mL) was added trifluoroacetic acid (0.20 mL). The reaction solution was stirred at 25°C for 60 minutes. The reaction solution was concentrated under reduced pressure, and the crude product was separated by preparative high-performance liquid chromatography (Welch Xtimate C18, 100mm×30mm 10 μm, A: water (formic acid); B: acetonitrile, 25%-55%: 10 minutes) to obtain compound 3. salt.

MS-ESI [M+H] ⁺ , calculated 531, found 531.

¹ H NMR (400MHz, MeOD) δ9.22(s, 1H), 9.09(s, 1H), 8.59(d, J=8.4Hz, 1H), 8.48(s, 1H), 8.27(d, J=8.4 Hz, 1H), 8.21(s, 1H), 7.99(s, 1H), 5.40(s, 2H), 4.65(s, 2H), 4.25(s, 3H).

Embodiment 4 synthetic compound 4

(1) Add sodium hydride (778mg, 19.5mmol, 60%) to compound 4-1 (4g, 17.7mmol) in tetrahydrofuran (32.0mL), stir the reaction solution at 0°C for 0.5 hours under nitrogen protection, add ring Propylmethanol (1.91 g, 26.5 mmol). The reaction solution was stirred at 0 °C for 1 hour under nitrogen protection. Add water (120mL), wash with methyl tert-butyl ether (60mL×2), adjust the pH value of the aqueous phase to 2 with aqueous hydrochloric acid, extract with ethyl acetate (50mL×2), combine the organic phases with saturated sodium chloride Washed with aqueous solution (100mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure, the crude product was separated by silica gel column chromatography (dichloromethane/methanol=1:0 to 10:1) to obtain compound 4-2.

¹ H NMR (400MHz, DMSO-d6) δ13.78(br s, 1H), 7.36(d, J=8.8Hz, 1H), 7.26(s, 1H), 4.03(d, J=7.2Hz, 2H) ,1.13-1.23(m,1H),0.53-0.59(m,2H),0.30-0.36(m,2H).

(2) To compound 4-2 (1.3g, 4.67mmol) in tetrahydrofuran (16.0mL), add borane tetrahydrofuran (1mol/L, 14.0mL), and stir the reaction solution at 15°C under nitrogen for 16 hours, then add Methanol (4.0 mL). Add water (40.0mL), extract with ethyl acetate (40mL×3), combine the organic phases and wash with saturated aqueous sodium chloride solution (100mL×2), dry over anhydrous sodium sulfate, filter and concentrate under reduced pressure, the crude product is passed through a silica gel column Compound 4-3 was isolated by chromatography (petroleum ether/ethyl acetate=1:0 to 10:1).

¹ H NMR (400MHz, CDCl ₃ ) δ6.98(d, J=8.8Hz, 1H), 6.87(s, 1H), 4.82(d, J=1.2Hz, 2H), 3.94(d, J=7.2Hz ,2H), 1.29-1.36(m,1H),0.67-0.76(m,2H),0.36-0.42(m,2H).

(3) To a solution of compound 4-3 (600mg, 2.27mmol) in tetrahydrofuran (2.0mL), add diphenylphosphoryl azide (933mg, 3.26mmol), 1,8-diazobispiro[5.4 .0] Undec-7-ene (691mg, 4.54mmol), the reaction solution was stirred at 20°C under nitrogen protection for 12 hours. Water (20.0 mL) was added, extracted with ethyl acetate (15 mL×3), the combined organic phases were washed with saturated aqueous sodium chloride solution (10.0 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure, the crude product was purified by silica gel Compound 4-4 was isolated by column chromatography (petroleum ether/ethyl acetate=1:0 to 10:1).

¹ H NMR (400MHz, DMSO-d6) δ7.33(d, J=9.2Hz, 1H), 7.24(s, 1H), 4.47(d, J=0.8Hz, 2H), 4.04(d, J=7.2 Hz, 2H), 1.21-1.30(m, 1H), 0.57-0.62(m, 2H), 0.35-0.39(m, 2H).

(4) To a solution of compound 4-4 (400mg, 1.38mmol) in tetrahydrofuran (20.0mL), add triphenyl Phosphine (1.09g, 4.15mmol), compound B (371mg, 1.52mmol), and the reaction solution was stirred at 20°C for 12 hours under nitrogen protection. The reaction solution was filtered and concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=1:0 to 6:1) to obtain compound 4-5.

¹ H NMR (400MHz, CDCl ₃ )δ8.83(s,1H),8.77(s,1H),8.61(s,1H),7.01(d,J＝8.8Hz,1H),6.89(s,1H) ,4.98(s,2H),3.90(d,J＝6.9Hz,2H),3.66(s,3H),1.29-1.33(m,1H),0.61-0.68(m,2H),0.33-0.39(m ,2H).

(5) Sodium cyanoborohydride (169.5 mg, 2.70 mmol) was added to a solution of compound 4-5 (440 mg, 899 μmol) in methanol (10.0 mL), and the reaction solution was stirred at 25° C. for 3 hours under nitrogen protection. Add water (20.0mL) to the reaction solution, extract with ethyl acetate (30.0mL×2), combine the organic phases, dry over anhydrous sodium sulfate, filter, concentrate under reduced pressure, and the crude product is subjected to silica gel column chromatography (dichloromethane/methanol =1:0 to 20:1) Compound 4-6 was isolated.

MS-ESI [M+H] ⁺ , calculated 461, found 461.

(6) To compound 4-6 (130mg, 283μmol) in dioxane (2.0mL) solution, add compound A (538mg, 708μmol, 42% purity), water (0.2mL), potassium carbonate (78.2mg, 566 μmol) and 1,1-bis(diphenylphosphino)ferrocenepalladium dichloride (31.0 mg, 42.4 μmol). The reaction solution was stirred at 80° C. for 2 hours under the protection of nitrogen. Water (20.0 mL) was added, extracted with dichloromethane (10.0 mL×3), the combined organic phases were washed with saturated aqueous sodium chloride (20.0 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure, and the crude product was Compound 4-7 was isolated by silica gel column chromatography (dichloromethane/methanol=1:0 to 50:1).

MS-ESI [M+H] ⁺ , calculated 654, found 654.

(7) To a solution of compound 4-7 (160 mg, 245 μmol) in dichloromethane (1.0 mL) was added trifluoroacetic acid (0.20 mL). The reaction solution was stirred at 25°C for 60 minutes. The reaction solution was concentrated under reduced pressure, and the crude product was separated by preparative high-performance liquid chromatography (Welch Xtimate C18, 100mm×30mm 10 μm, A: water (formic acid); B: acetonitrile, 25%-55%: 10 minutes) to obtain compound 4. salt.

MS-ESI [M+H] ⁺ , calculated 554, found 554.

¹ H NMR (400MHz, MeOD) δ9.04(s, 1H), 8.93(s, 1H), 8.53(d, J=8.4Hz, 1H), 8.16(dd, J=8.4, 1.2Hz, 1H), 8.12(s,1H),7.09(d,J=9.6Hz,1H),7.05(s,1H),4.99(s,2H),4.75(s,2H),4.56(s,2H),3.90(d , J=7.2Hz, 2H), 1.11-1.23(m, 1H), 0.37-0.46(m, 2H), 0.13-0.24(m, 2H).

Biological experiment example

I. PRMT5 inhibitory activity with or without MTA

Experimental principle: TR-FRET PRMT5 assay is based on PRMT5/MEP50 complex protein as enzyme, histone H4 (1-21) biotinylated peptide as substrate, Eu ³⁺ -nucleate-labeled methylated histone H4 arginine Acid 3 3H4R3 (me) antibody and ULight-streptavidin were used as detection reagents. The assay is divided into two steps: enzymatic and detection, and the TR-FRET signal is directly proportional to the concentration of methylated histone H4(1-21) biotinylated peptide. Testing was performed in the presence or absence of MTA to determine whether compounds exhibited synergistic effects of MTA.

Experimental materials: Bicine was purchased from Sigma, sodium chloride was purchased from Sigma, dithiothreitol was purchased from Sigma, pigskin gelatin was purchased from Sigma, Tween-20 was purchased from Sigma, SAM was purchased from NEB, Ultra Eu-Anti-Methylated Histone H4 Arginine 3 Antibody was purchased from PE, ULight-Streptavidin was purchased from PE, LANCE Detection Buffer was purchased from PE, Histone H4(1-21) Biotinylated Peptide Ac-SGRGKGGKGLGKGGAKRHRKVGG-K was purchased from GL Biochem, PRMT5/MEP50, FLAG-tag, His-tag (HEK293 source) were purchased from BPS, 384-well assay plates were purchased from Gerinier, 384-LDV plates were purchased from LABCYTE, and incubators were purchased from Grant -bio company, microplate reader purchased from TECAN.

experimental method:

(1). Preparation buffer: 20mM Bicine (pH7.6); 25mM sodium chloride; 2mM dithiothreitol (DTT); 0.005% pigskin gelatin; 0.01% Tween-20; no MTA or 0.0026 mM MTA.

(2). The compound to be tested was dissolved in DMSO to prepare a 10 mM stock solution. The compound to be tested was diluted with buffer to a maximum concentration of 10 μM, diluted 3 times, and tested in duplicate wells.

(3). Take an appropriate amount of PRMT5 protease (4.16 μM) mother solution and add buffer to prepare a 0.0076 μM PRMT5 enzyme reaction solution. Buffer solution (5 μL/well) was added to the positive control well, and 5 μL enzyme reaction solution was added to each well of the rest. Centrifuge at 1000r/min for 30s, and incubate in a room temperature incubator for 30min.

(4). Take an appropriate amount of 100% SAM (32000 μM) stock solution and 100 μM histone H4 (1-21) biotinylated peptide. Firstly, the SAM master solution was diluted 10 times with buffer, and the buffer solution was used to prepare a substrate mixture containing 0.32 μM histone H4 (1-21) biotinylated peptide and 2.6 μM SAM (10%). Add 5 μL of substrate mixture to each well, centrifuge at 1000 r/min for 30 s, and incubate in a room temperature incubator for 90 min.

(5). Take an appropriate amount of Eu-anti-methylated histone H4 arginine 3 antibody (625nM), ULight-streptavidin (10000nM) and 10×LANCE detection buffer. Dilute with ultrapure water to form a detection mixture containing 4nM Eu-anti-methylated histone H4 arginine 3 antibody, 53.3nM ULight-streptavidin and 1×LANCE detection buffer. Add 10 μL of detection mixture to each well, centrifuge at 1000 r/min for 30 s, and incubate in a room temperature incubator for 60 min. The detection signal was read with a PerkinElmer EnVision 2105 multi-mode microplate reader (excitation light is 320/340nm, emission light is 665nm), and the IC ₅₀ of the compound was calculated with GraphPad Prism 5.0 software, and the results are shown in Table 1.

Experimental results show that the compound of the invention can effectively inhibit the MTA synergistic PRMT5, and can be used to prepare medicines for treating cancers related to MTAP ^-/- .

II. HCT116 MTAP ^-/- and HCT 116 ^wt cell proliferation inhibitory activity

Experimental principle: HCT116 MTAP ^-/- cells lack MTAP, and the MTA level increases; HCT 116 ^wt cells do not lack MTAP, and the MTA level is normal. These two cell lines were used to determine whether compounds exhibited MTA synergistic inhibitory activity.

Experimental materials: HCT116 MTAP ^-/- and HCT 116 ^wt cells were purchased from Horizon; CellTiter-Glo reagent was purchased from Promega (Cat. No. G7571); RPMI-1640 was purchased from ATCC (Cat. No. 30-2001): fetal bovine serum (FBS) purchase From EXCELL (product number FND500); penicillin-streptomycin was purchased from Gibco (product number 15140-122); 0.25% trypsin-ethylenediaminetetraacetic acid digestion solution (Trypsin-EDTA) was purchased from Gibco (product number 25200-072) ); dimethyl sulfoxide (DMSO) was purchased from Sigma (Cat. No. D2650); 96-well plates were purchased from Corning (Cat. No. 3610); incubators were purchased from NuAire (model NU-5700E); TS-100); an automatic cell counter was purchased from Life technologies (model Countess II); a microplate reader was purchased from PerkinElmer (model Envision); the data processing software was GraphPad Prism 5.0.

Test method: HCT116 MTAP ^-/- or HCT116 ^wt cells in logarithmic growth phase were resuspended in growth medium (RPMI-1640+10% FBS) and diluted to the target density (5000/mL). Inoculate the above cell suspension into 96-well plate at 100 μL per well; incubate overnight at 37°C in a 5% CO ₂ incubator. The culture medium was used as the background control group. Cells were starved for 4 hours with 80 μL of serum-free medium. The compound to be tested was dissolved in DMSO to prepare a stock solution with a concentration of 10 mmol/L. Firstly, the stock solution was diluted to 2mmol/L (200X) with DMSO, and then serially diluted 3 times, with a total of 10 concentrations. Take 3 μL of the above solution at each concentration and dilute (2X) with 297 μL of growth medium. Then add 80 μL/well into the 96-well plate inoculated with cells. The cells added with the compound to be tested were placed in a 37°C, 5% _CO2 incubator and incubated for 120 hours. Equilibrate the 96-well plate at room temperature, add 40 μL CellTiter-Glo reagent to each well, mix on a vortexer for 2 minutes, incubate at room temperature for 60 minutes, read the luminescence value with an EnVision microplate reader, and calculate the IC ₅₀ of the compound with GraphPad Prism 5.0 software , and the results are shown in Table 2.

All documents mentioned in this application are incorporated by reference in this application as if each were individually incorporated by reference. In addition, it should be understood that after reading the above teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Claims

A compound represented by formula I, or a pharmaceutically acceptable salt, enantiomer, diastereomer, tautomer, cis-trans isomer, solvate, polymorph , deuterium or combinations thereof,

in,

R 1a and R 1b are each independently hydrogen or C1-C3 alkyl, or R 1a and R 1b together form oxo (=O), or R 1a and R 1b form a 3-5 membered group with the carbon atom to which they are attached Carbocyclyl; wherein, the alkyl or carbocyclyl may be optionally substituted by halogen or C1-C3 alkyl;

R 2a and R 2b are each independently hydrogen or C1-C3 alkyl, or R 2a and R 2b together form oxo (=O); wherein, the alkyl can optionally be replaced by halogen or C1-C3 alkyl replace;

q is 1 or 2;

R 3 is selected from: C6-C10 aryl-L- or 5-10 membered heteroaryl-L-, wherein the aryl and heteroaryl can be optionally substituted by 1-3 R';

Each R' is independently selected from: halogen, amino, cyano, oxo (=O), C1-C3 hydroxyalkyl, -Y-C1-C4 alkyl, 3-10 membered carbocyclyl-L' -Y-, 3-10 membered heterocyclyl-L'-Y-, C6-C10 aryl-L'-Y- or 5-10 membered heteroaryl-L'-Y-, wherein the alkyl Base, aryl, heteroaryl, carbocyclyl, heterocyclyl can be optionally substituted by 1-3 R";

Each R" is independently selected from: cyano, oxo (=O), halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, or 3-8 membered heterocyclyl;

Each Y is independently selected from: absent, -O-, -S-, -NR c -, -NR c CO-, -SO-, -SO 2 -, -CH(OH)-, or -CO- ;

each L and L' is independently selected from: absent or -CR d R e -;

R c , R d and R e are each independently selected from: hydrogen or C1-C3 alkyl; wherein, the alkyl may be optionally substituted by halogen or C1-C3 alkyl;

R 4 is H, halogen, C1-C6 alkyl, C1-C6 haloalkyl or C1-C6 alkoxy;

X 1 , X 2 and X 3 are each independently CR 5 or N, and at most two of X 1 , X 2 and X 3 are N at the same time;

Each R is independently selected from: H, halogen, hydroxyl, amino, cyano, -C1-C3 alkyl, -C1-C3 hydroxyalkyl, C2-C4 alkenyl or C2-C4 alkynyl; wherein, The alkyl, C2-C4 alkenyl or C2-C4 alkynyl may be optionally substituted by halogen or C1-C3 alkyl.
The compound as claimed in claim 1, or its pharmaceutically acceptable salt, enantiomer, diastereomer, tautomer, cis-trans isomer, solvate, polymorph , a deuterated substance or a combination thereof, characterized in that R 1a and R 1b are each independently hydrogen or R 1a and R 1b together form oxo (=O).
The compound according to any one of claims 1-2, or a pharmaceutically acceptable salt, enantiomer, diastereomer, tautomer, cis-trans isomer, solvent Compounds, polymorphs, deuteriums or combinations thereof, characterized in that R 2a and R 2b are each independently hydrogen or R 2a and R 2b together form oxo (=O); q is 1.
The compound according to any one of claims 1-3, or a pharmaceutically acceptable salt, enantiomer, diastereomer, tautomer, cis-trans isomer, solvent Compound, polymorph, deuterium or combination thereof, it is characterized in that, R 3 is selected from:

in,

Each R6 is independently selected from: halogen, amino, cyano, oxo (=O), C1-C3 hydroxyalkyl, -Y-C1-C4 alkyl, 3-10 membered carbocyclyl-L' -Y or -3-10 membered heterocyclyl-L'-Y-, wherein the alkyl, carbocyclyl and heterocyclyl can be optionally substituted by 1-3 R";

Each R" is independently selected from: cyano, oxo (=O), halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, or 3-8 membered heterocyclyl;

Each Y is independently selected from: absent, -O-, -S-, -NR c -, -NR c CO-, -SO-, -SO 2 -, -CH(OH)-, or -CO- ;

each L and L' is independently selected from: absent or -CR d R e -;

R c , R d and R e are each independently selected from: hydrogen or C1-C3 alkyl;

m is 0, 1, 2, 3 or 4;

R 7 is H or C1-C3 alkyl.
The compound according to any one of claims 1-3, or a pharmaceutically acceptable salt, enantiomer, diastereomer, tautomer, cis-trans isomer, solvent Compounds, polymorphs, deuteriums or combinations thereof, characterized in that R 4 is H, halogen, C1-C3 alkyl, C1-C3 haloalkyl or C1-C3 alkoxy; preferably, R 4 is H , chlorine, fluorine, methyl, ethyl, methoxy or trifluoromethyl.
The compound according to any one of claims 1-3, or a pharmaceutically acceptable salt, enantiomer, diastereomer, tautomer, cis-trans isomer, solvent Compounds, polymorphs, deuteriums or combinations thereof, characterized in that X 1 is CR 5 ; X 2 is N; X 3 is CR 5 ; each R 5 is independently H, halogen, hydroxyl, amino, Cyano, -C1-C3 hydroxyalkyl, C2-C4 alkenyl or C2-C4 alkynyl;

Preferably, X 1 is CH; X 2 is N; X 3 is CH.
The compound as claimed in claim 4, or its pharmaceutically acceptable salt, enantiomer, diastereomer, tautomer, cis-trans isomer, solvate, polymorph , a deuterated substance or a combination thereof, wherein each R 6 is independently selected from: halogen, amino, cyano, oxo (=O), C1-C3 hydroxyalkyl, C1-C3 alkyl, C1- C3 haloalkyl, C1-C4 alkyl-O-, C3-C6 cycloalkyl, C3-C6 cycloalkyl-O-, C3-C6 cycloalkyl CH 2 -, C3-C6 cycloalkyl CH 2 -O -, -CONR c C3-C6 cycloalkyl, CONR c C1-C3 alkyl, CONR c C1-C3 haloalkyl, wherein, R c is selected from: hydrogen or C1-C3 alkyl.
The compound according to any one of claims 1-6, or a pharmaceutically acceptable salt, enantiomer, diastereomer, tautomer, cis-trans isomer, solvent Compound, polymorph, deuterium or combination thereof, it is characterized in that, described compound is selected from the following group:
A kind of pharmaceutical composition, is characterized in that, described pharmaceutical composition comprises:

(1) A therapeutically effective amount selected from the compounds described in any one of claims 1 to 8, their pharmaceutically acceptable salts, enantiomers, diastereoisomers, and tautomers One or more of cis-trans isomers, solvates, polymorphs and deuterated substances as active ingredients; and

(2) Optionally, a pharmaceutically acceptable carrier.
The compound according to any one of claims 1-8, its pharmaceutically acceptable salt, enantiomer, diastereomer, tautomer, cis-trans isomer, solvate , polymorph or deuterium, or the pharmaceutical composition as claimed in claim 8 in the preparation of medicines for preventing or treating MTAP -/- related cancers.
The use according to claim 10, wherein the cancer is selected from the group consisting of: liver cancer, breast cancer, skin cancer, pancreatic cancer, head and neck cancer, colon cancer, lung cancer, gastric cancer, esophageal cancer, kidney cancer, bladder cancer, urethral cancer Cancer, Prostate Cancer, Testicular Cancer, Uterine Cancer, Ovarian Cancer, Vaginal Cancer, Fallopian Tube Cancer, Cholangiocarcinoma, Multiple Myeloma, Spinal Neurofibroma, Astrocytoma, Glioma, Acute Lymphoblastic Leukemia, Chronic Lymphoid Leukemia, Hodgkin's Lymphoma, Non-Hodgkin's Lymphoma, Sarcoma.