WO2023016511A1

WO2023016511A1 - Azaaryl compound, and preparation method therefor and use thereof

Info

Publication number: WO2023016511A1
Application number: PCT/CN2022/111668
Authority: WO
Inventors: 马世超
Original assignee: 上海青煜医药科技有限公司
Priority date: 2021-08-11
Filing date: 2022-08-11
Publication date: 2023-02-16
Also published as: TW202312995A; CN115703799A

Abstract

Provided are an azaaryl compound as represented by formula (I), and a pharmaceutically acceptable salt, stereoisomer or solvate thereof, or an isotope labeled compound thereof. In addition, provided are a method for preparing the compound, a composition containing the compound and the use of the compound in the preparation of a drug for treating diseases or disorders associated with the action mechanism of EED protein and/or PRC2 protein complexes.

Description

Azaaryl compound, its preparation method and application

This application claims the priority of the Chinese patent application 2021109212000 with the filing date of 2021/8/11. This application cites the full text of the above-mentioned Chinese patent application.

technical field

The invention relates to an azaaryl compound, its preparation method and application.

Background technique

PcG (Polycomb Group) proteins are an important class of chromatin modifying enzymes. It regulates the transcription of genes by modifying chromatin, and plays an important role in the growth, differentiation and long-term cell memory of stem cells. In mammalian cells, PcG proteins are mainly divided into two types of transcriptional repression complexes, namely PRC1 (Polycomb Repressive Complex 1) and PRC2 (Polycomb Repressive Complex 2). Among them, PRC2 inhibits the expression of related genes by methylating the 27th lysine (H3K27) of histone 3 in chromatin. The PRC2 protein complex is mainly composed of core proteins such as EZH2 (Enhancer of Zeste Homolog 2) (or its very similar homologous protein EZH1), EED (Embryonic ectoderm Development) and SUZ12 (Suppressor of Zeste 12). Among them, EZH2 has enzyme catalytic activity, and can transfer the methyl group of the substrate SAM (S-adenosyl-L-methionine) to H3K27 through the SET (Su(var), E(Z), and Trithorax) protein domain, thereby One- to three-methylation modification of H3K27 is achieved. The enzymatic activity of EZH2 also depends on other components of PRC2, such as EED proteins belonging to the WD40 repeat structural protein family. The combination of EED and trimethylated H3K27Me3 can greatly promote the enzymatic function of EZH2 on the one hand, and on the other hand, it can also position the PCR2 complex on the chromatin that needs to be modified. Abnormal function of PRC2, such as EZH2 overexpression or gain-of-function mutation, is associated with many clinical tumor diseases, including lung cancer, breast cancer, rectal cancer, prostate cancer, bladder cancer, pancreatic cancer, sarcoma, and lymphoma. PRC2 is also associated with a variety of cellular immune functions, such as EZH2 involved in the regulation of lymphocyte activation, and can also cooperate with glycolysis to promote the response of T cells to tumor cells. Therefore, the development of small molecule inhibitors of PRC2 has important and broad drug development value.

The research and development around PRC2 inhibitors mainly involves the development of EZH2 inhibitors and EED inhibitors. EZH2 inhibitors currently entering the clinic include EPZ-6438 (Epizyme, clinical phase II), GSK2816126 (GSK, clinical phase I), and CPI-1205 (Constellation, clinical phase I) and so on. Although several EZH2 inhibitors have entered the clinical research stage, these inhibitors all contain a common pharmacophore of 2-pyridone. Moreover, in clinical treatment with existing EZH2 inhibitors, secondary mutations have begun to appear. EED inhibitors have an allosteric inhibitory effect on the enzyme function of EZH2, and can achieve the same or similar biological functions as EZH2. Moreover, on the one hand, EED inhibitors can overcome the drug resistance problem of EZH2 well, and on the other hand, EED inhibitors can be used in combination with EZH2 inhibitors to achieve a better synergistic effect. Therefore, it is very important to develop new EED inhibitors. meaning.

Contents of the invention

The technical problem to be solved by the present invention is that the EED inhibitors in the prior art have a single structure. Therefore, the present invention provides an azaaryl compound, its preparation method and application. The azaaryl compound of the invention exhibits good inhibitory activity on tumor cells and has broad prospects for drug development.

The present invention provides a compound represented by formula (I), its pharmaceutically acceptable salt, stereoisomer, solvate or their isotope-labeled compounds:

where A is independently

X is N or CR ² ;

R ¹ , R ² and _R ³ are independently H, halogen, cyano, R ^5b substituted or unsubstituted amino, R ^5b -O-, R ⁵ substituted or unsubstituted C _1-4 alkyl, R ⁵ substituted Or unsubstituted C ₃ - ₆ cycloalkyl, R ⁵ substituted or unsubstituted C ₁ -C ₄ haloalkyl, -C(=O)R ⁶ , -CO ₂ R ⁶ , -C(=O)NR ⁶ R ⁷ , -SO ₂ R ⁶ or -SO ₂ NR ⁶ R ⁷ ; when there are multiple substituents, they are the same or different;

R ⁴ is independently H, halogen, cyano, C _1-4 haloalkyl, -C(=O)NR ^4a1 R ^4a2 , -S(=O) ₂ R ^4b or -C(=O)R ^4c ;

R ^4a1 , R ^4a2 and R ^4b are independently H or C _1-4 alkyl;

R ^4c is independently H, C _1-4 alkyl or -OC _1-4 alkyl;

R ⁵ is independently C _1-4 alkyl-O-, C _1-4 alkyl, C _1-4 haloalkyl, -N(R ^5b R ^5b ), amino protected by N protecting group, fluorine, hydroxyl;

R ^5b , R ⁶ and R ⁷ are independently H, C _1-4 alkyl, C _1-4 haloalkyl;

"*" indicates that when the carbon atom with "*" is a chiral carbon atom, it is in R configuration, S configuration or a mixture thereof.

In some preferred embodiments of the present invention, certain groups in the compounds shown in formula (I), their pharmaceutically acceptable salts, stereoisomers, solvates or their isotope-labeled compounds As defined below, the unmentioned groups are described in any scheme of the application (referred to as "in some preferred embodiments of the present invention"):

The compound shown in formula (I) is selected from the compounds shown below:

In the above compounds, X, R ¹ , R ³ and R ⁴ are as defined above.

In certain preferred embodiments of the invention:

for

or a mixture thereof.

In certain preferred embodiments of the invention:

When R ¹ , R ² , R ³ and R ⁴ are independently halogen, said halogen is fluorine, chlorine, bromine or iodine, preferably chlorine.

In certain preferred embodiments of the invention:

When R ¹ , R ² and R ³ are independently R ⁵ substituted or unsubstituted C _1-4 alkyl, said C _1-4 alkyl is methyl, ethyl, n-propyl _, isopropyl , n-butyl, isobutyl or tert-butyl, preferably methyl or ethyl.

In certain preferred embodiments of the invention:

When R ¹ , R ² and R ³ are independently R ⁵ substituted or unsubstituted C _3-6 cycloalkyl groups, the C _3-6 cycloalkyl groups are cyclopropyl, cyclobutyl _, _cyclo Pentyl or cyclohexyl.

In certain preferred embodiments of the invention:

When R ¹ , R ² and R ³ are independently R ⁵ substituted or unsubstituted C ₁ -C ₄ haloalkyl, the halogen is fluorine, chlorine, bromine or iodine, preferably fluorine.

In certain preferred embodiments of the invention:

When R ¹ , R ² and R ³ are independently R ⁵ substituted or unsubstituted C ₁ -C ₄ haloalkyl, said C ₁ -C ₄ alkyl is methyl, ethyl, n-propyl, iso Propyl, n-butyl, isobutyl or tert-butyl, preferably methyl or ethyl.

In certain preferred embodiments of the invention:

When R ¹ , R ² and R ³ are independently R ⁵ substituted or unsubstituted C ₁ -C ₄ haloalkyl groups, the number of halogenated groups is 1, 2, 3, 4 or 5; for example, 3 indivual.

In certain preferred embodiments of the invention:

When R ¹ , R ² and R ³ are independently R ⁵ substituted or unsubstituted C ₁ -C ₄ haloalkyl, the C ₁ -C ₄ haloalkyl is trifluoromethyl.

In certain preferred embodiments of the invention:

When R ⁴ , R ⁵ , R ^5b , R ⁶ and R ⁷ are independently C ₁ -C ₄ haloalkyl, the halogen is fluorine, chlorine, bromine or iodine, preferably fluorine.

In certain preferred embodiments of the invention:

When R ⁴ , R ⁵ , R ^5b , R ⁶ and R ⁷ are independently C ₁ -C ₄ haloalkyl, wherein the C ₁ -C ₄ alkyl is methyl, ethyl, n-propyl, isopropyl , n-butyl, isobutyl or tert-butyl, preferably methyl or ethyl.

In certain preferred embodiments of the invention:

When R ⁵ is independently a C ₁ -C ₄ haloalkyl group, wherein the C ₁ -C ₄ alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl , preferably methyl or ethyl.

When R ⁴ , R ^5b , R ⁶ and R ⁷ are independently C ₁ -C ₄ haloalkyl, wherein the C ₁ -C ₄ alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl group, isobutyl or tert-butyl, preferably methyl or ethyl.

In certain preferred embodiments of the invention:

When R ⁴ , R ⁵ , R ^5b , R ⁶ and R ⁷ are independently C ₁ -C ₄ haloalkyl, the number of said halo is 1, 2, 3, 4 or 5; for example, 3 .

In certain preferred embodiments of the invention:

When R ⁴ , R ⁵ , R ^5b , R ⁶ and R ⁷ are independently C ₁ -C ₄ haloalkyl, said C ₁ -C ₄ haloalkyl is trifluoromethyl.

In certain preferred embodiments of the invention:

When R ^4a1 , R ^4a2 , R ^4b , R ^5b , R ⁶ and R ⁷ are independently C _1-4 alkyl, the C ₁ -C ₄ alkyl is methyl, ethyl, n-propyl, Isopropyl, n-butyl, isobutyl or tert-butyl, preferably methyl or ethyl.

In certain preferred embodiments of the invention:

When R ^4c is independently C _1-4 alkyl or -OC _1-4 alkyl, said C ₁ -C ₄ alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl , isobutyl or tert-butyl, preferably methyl or ethyl.

In certain preferred embodiments of the invention:

When R ⁵ is independently C _1-4 alkyl, C _1-4 alkyl-O-, C _1-4 haloalkyl, said C _1-4 alkyl, C _1-4 alkyl-O- and C _1-4 alkyl in C _1-4 haloalkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, preferably methyl.

In certain preferred embodiments of the invention:

When R ⁵ is independently an amino group protected by an N protecting group, the protecting group can be

R ^5a _is C _1-4 alkyl or C _1-4 alkyl-O-; the C _1-4 alkyl in C 1-4 alkyl and C _1-4 alkyl-O- can be independently methyl , ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl;

For example

May be tert-butoxycarbonyl.

In certain preferred embodiments of the invention:

X is N.

In certain preferred embodiments of the invention:

^R is selected from methyl, cyano, trifluoromethyl, difluoromethyl,

In certain preferred embodiments of the invention:

^R2 is H.

In certain preferred embodiments of the invention:

^R3 is selected from H, methyl, halo or cyano.

In certain preferred embodiments of the invention:

R ⁴ is selected from cyano, -C(=O)NR ^4a1 R ^4a2 , -S(=O) ₂ R ^4b or -C(=O)R ^4c .

In certain preferred embodiments of the invention:

R ^4a1 , R ^4a2 , R ^4b , R ^4c , R ^5b , R ⁶ and R ⁷ are independently methyl.

In certain preferred embodiments of the invention:

X is N;

^R is selected from methyl, cyano, trifluoromethyl, difluoromethyl,

^R is selected from H, methyl, halogen or cyano;

R ⁴ is selected from cyano, -C(=O)NR ^4a1 R ^4a2 , -S(=O) ₂ R ^4b or -C(=O)R ^4c ;

R ^4a1 , R ^4a2 , R ^4b and R ^4c are as defined above.

In certain preferred embodiments of the invention:

X is N;

R ¹ is cyano, R ^5b substituted _or unsubstituted amino, R ⁵ substituted or unsubstituted C _1-4 _alkyl ; for example R ⁵ substituted or unsubstituted amino, R ⁵ substituted or unsubstituted C _1-4 alkyl;

R ³ is H, R ⁵ substituted or unsubstituted C _1-4 alkyl, halogen or cyano _; for example H;

R ⁵ is independently C _1-4 alkyl, -N(R ^5b R ^5b ), hydroxyl;

R ⁴ is selected from cyano, -C(=O)NR ^4a1 R ^4a2 , -S(=O) ₂ R ^4b or -C(=O)R ^4c ; for example R ⁴ is selected from cyano or -S(=O ) ₂ R ^4b ; For example, R ^4b is independently C _1-4 alkyl;

R ^5b is independently H or C _1-4 alkyl.

In certain preferred embodiments of the invention:

X is N;

R ¹ is R ⁵ substituted or unsubstituted C _1-4 alkyl _;

^R3 is H;

R ⁵ is independently C _1-4 alkyl, -N(R ^5b R ^5b ), hydroxyl;

R ⁴ is selected from cyano or -S(=O) ₂ R ^4b ;

R ^4b is independently C _1-4 alkyl;

R ^5b is independently H or C _1-4 alkyl.

In some preferred embodiments of the present invention, the compound represented by the formula (I) is selected from the compounds shown below:

In the above compounds, R ¹ is as defined above.

In some preferred embodiments of the present invention, the compound described in formula (I) is selected from the compounds shown below:

The present invention also provides isotope-labeled compounds of the compound represented by formula (I), its pharmaceutically acceptable salt, stereoisomer or solvate thereof. The isotope in the isotope-labeled compound is selected from ² H, ³ H, ¹¹ C, ¹³ C, ¹⁴ C ^, ¹⁵ N, ¹⁷ O, 18 O, ¹⁸ F, ³¹ P, ³² P, ³⁵ S, ³⁶ Cl and ¹²⁵ I. The atoms that can be labeled with isotopes in the compound of formula (I) include but are not limited to hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine and iodine, etc., which can be respectively labeled with isotopes ² H, ³ H, ¹¹ C, ¹³ C , ¹⁴ C, ¹⁵ N, ¹⁷ O, ¹⁸ O, ¹⁸ F, ³¹ P, ³² P, ³⁵ S, ³⁶ Cl and ¹²⁵ I etc. instead.

The present invention also provides the preparation method of the compound shown in formula (I), which comprises the following steps:

Halogenated intermediate B ₀ and intermediate E ₀ are subjected to a coupling reaction to obtain a compound shown in formula (I);

Among them, W represents halogen (such as Br); R ^x is -B (OH) ₂ or

A, R ¹ , R ³ , R ⁴ , and X are as defined above.

The present invention also provides a preparation method of intermediate compound B ₀ , which can be any of the following schemes:

Option 1 includes the following steps:

Step (1) Compound SM-5 is ring-closed under the action of a catalyst (such as phosphorus oxychloride) to obtain compound SM-6,

Step (2) Compound SM-6 is reacted under the action of an oxidizing agent (such as m-chloroperoxybenzoic acid) to obtain compound SM-7,

Step (3) Reaction of compound SM-7 with H ₂ N-CH ₂ -A in the presence of a base (such as sodium carbonate) to obtain compound B ₀ ;

Scheme two, it is as shown in step (2) to step (3) in scheme one;

Scheme three, it is as shown in step (3) in scheme one;

The reaction equation is as follows:

Wherein, W, A, R ⁴ are as defined above.

In some preferred embodiments of the present invention: the solvate is a hydrate.

The solvent involved in the present invention can be selected from, for example, methanol, ethanol, isopropanol, toluene, xylene, chlorobenzene, water, methylene chloride, chloroform, carbon tetrachloride, 1,2-dichloroethane, Dioxane, DMF, acetonitrile, DMSO, NMP, THF or combinations thereof; for example selected from dichloromethane, chloroform, 1,2-dichloroethane, dioxane, DMF, acetonitrile, DMSO, NMP, THF or a combination thereof.

The bases involved in the present invention may include organic bases and inorganic bases.

The organic base involved in the present invention can be selected from, for example, TEA, DIPEA or a combination thereof.

The inorganic base involved in the present invention can be selected from, for example, sodium hydride, sodium methoxide, potassium carbonate, sodium carbonate, cesium carbonate, potassium tert-butoxide, sodium tert-butoxide, lithium tert-butoxide, LiHMDS, LDA, butyllithium , potassium hydroxide, potassium acetate, lithium aluminum hydride or combinations thereof; for example selected from sodium hydride, potassium carbonate, sodium carbonate, cesium carbonate, potassium tert-butoxide, sodium tert-butoxide, LiHMDS, LDA, butyllithium or combinations thereof .

The isotope-labeled compound of the compound shown in formula (I) according to the present invention can be prepared by a synthetic method similar to that of the unlabeled compound, the difference is that the unlabeled starting material and/or reagent are replaced by isotope-labeled Starting materials and/or reagents.

The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of the above-mentioned compound represented by formula (I), its pharmaceutically acceptable salt, stereoisomer, its solvate or their Isotope-labeled compounds and pharmaceutically acceptable auxiliary materials. The pharmaceutically acceptable auxiliary material can be one or more of diluents, absorbents, wetting agents, binders, disintegrants and lubricants.

The present invention also provides the above-mentioned compound represented by formula (I), its pharmaceutically acceptable salt, stereoisomer, its solvate or their isotope-labeled compound, or the above-mentioned pharmaceutical composition in Use in the preparation of medicines. The medicine is preferably a medicine for treating cancer.

The present invention also provides the above-mentioned compound represented by formula (I), its pharmaceutically acceptable salt, stereoisomer, its solvate or their isotope-labeled compound, or the above-mentioned pharmaceutical composition in Use of the invention in the preparation of cancer drugs mediated by EED protein and/or PRC2 protein complexes.

Preferably, the cancer includes but is not limited to diffuse large B-cell lymphoma, follicular lymphoma, non-Hodgkinson lymphoma and other lymphoma, leukemia, multiple myeloma, mesothelioma, gastric cancer, malignant rhabdoid tumors, liver cancer, prostate cancer, breast cancer, brain tumors including neuroblastoma, glioma, glioblastoma and astrocytoma, cervical cancer, colon cancer, melanoma, endometrial cancer, esophageal cancer, Head and neck cancer, lung cancer, nasopharyngeal cancer, ovarian cancer, pancreatic cancer, kidney cancer, rectal cancer, thyroid cancer, parathyroid tumors, uterine tumors and soft tissue sarcomas, etc.

Preferably, the compounds, their pharmaceutically acceptable salts, stereoisomers, solvates or their isotope-labeled compounds are used in combination with other drugs; more preferably, the other drugs are selected from anticancer drugs, tumor Immunological drugs, antiallergic drugs, antiemetics, analgesics, or cytoprotective drugs.

The present invention also provides a pharmaceutical preparation, which comprises the above-mentioned compound represented by formula (I), its pharmaceutically acceptable salt, stereoisomer, its solvate or their isotope-labeled compound, or A pharmaceutical composition as described above. The pharmaceutical preparation is preferably a tablet, capsule (e.g. sustained release or timed release capsule), pill, powder, granule (e.g. granule), elixir, tincture, suspension (e.g. nanosuspension, microsuspension) and spray-dried dispersions and other forms of suspensions, syrups, emulsions, solutions and the like. The preferred mode of administration of the pharmaceutical preparation is oral administration, sublingual administration, injection including subcutaneous injection, intravenous injection, intramuscular injection, intrasternal injection, injection, etc., nasal administration (such as nasal membrane inhalation), topical ( Such as creams and ointments), rectal administration (such as suppositories) and so on. The compounds disclosed in the present invention can be administered alone or together with appropriate pharmaceutical carriers.

The present invention also provides that the aforesaid pharmaceutical preparation can be formulated into an appropriate drug dose to facilitate and control the dosage of the drug. Dosage regimens of the compounds disclosed in the present invention vary according to specific factors, such as pharmacodynamics and mode of administration, subject, gender, age, health status and body weight of the subject, disease characteristics, other simultaneous drug-taking conditions, and the duration of drug administration. Frequency, liver and kidney function, desired effect, etc. The compounds disclosed in the present invention can be taken in a single dose per day, or the total dose can be divided into multiple doses (such as two to four times per day).

The present invention also provides a method for treating cancer, which comprises administering a therapeutically effective amount of the above-mentioned compound represented by formula (I), its pharmaceutically acceptable salt, stereoisomer or their solvates or their isotope-labeled compounds, or pharmaceutical compositions as described above. The cancers include, but are not limited to, diffuse large B-cell lymphoma, follicular lymphoma, non-Hodgkinson lymphoma and other lymphomas, leukemia, multiple myeloma, mesothelioma, gastric cancer, malignant rhabdoid tumor, Liver cancer, prostate cancer, breast cancer, brain tumors including neuroblastoma, glioma, glioblastoma and astrocytoma, cervical cancer, colon cancer, melanoma, endometrial cancer, esophageal cancer, head and neck cancer , lung cancer, nasopharyngeal cancer, ovarian cancer, pancreatic cancer, kidney cancer, rectal cancer, thyroid cancer, parathyroid tumors, uterine tumors and soft tissue sarcoma, etc.

The present invention also provides the compound represented by formula (I), its pharmaceutically acceptable salt, stereoisomer or solvate thereof, or the aforementioned isotope-labeled compound used in combination with other drugs, and the other drugs are selected from: anticancer Drugs, tumor immune drugs, anti-allergic drugs, antiemetic drugs, analgesics and cell protection drugs, etc., have a better effect when used together.

In the present invention, the cancer is preferably a cancer mediated by a complex with EED protein and/or PRC2 protein.

The present invention also provides a method for inhibiting the activity of EED protein and/or PRC2 protein complex, which comprises administering to the subject a therapeutically effective amount of the above-mentioned compound represented by formula (I), its pharmaceutically acceptable salts, stereoisomers or solvates thereof, or their isotope-labeled compounds, or pharmaceutical compositions as described above.

The present invention also provides a method for blocking the combination of EED and H3K27 (such as H3K27me3), which includes administering to the subject a therapeutically effective amount of the above-mentioned compound represented by formula (I), its pharmaceutically acceptable Salts, stereoisomers or solvates thereof or their isotopically labeled compounds, or pharmaceutical compositions as described above.

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.

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 "about" when used in reference to a specifically recited value means that the value may vary by no more than 1% from the recited value. For example, as used herein, the expression "about 100" includes all values between 99 and 101 and in between (eg, 99.1, 99.2, 99.3, 99.4, etc.).

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".

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 indicated, conventional methods within the skill of the art, such as mass spectroscopy, NMR, IR and UV/VIS spectroscopy and pharmacological methods are employed. 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 theses, 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, C _1-6 alkyl refers to an alkyl group as defined below having a total of 1 to 6 carbon atoms (eg, 1, 2, 3, 4, 5, 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.

When a substituent is listed without specifying which atom it is attached to in a general chemical formula (including but not specifically mentioned compounds), such substituent may be bonded via any atom thereof. Combinations of substituents and/or variations thereof are permissible only if such combinations result in stable compounds.

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 each part of this specification, the substituents of the compounds disclosed in the present invention are disclosed according to the type or range of the group. It is specifically intended that the invention includes each individual subcombination of individual members of these radical classes and ranges. The term " _Cx - _Cyalkyl " or " _Cxyalkyl " refers to a straight or branched chain saturated hydrocarbon containing x to y carbon atoms. For example, the term "C ₁ -C ₆ alkyl" or "C _1-6 alkyl" specifically refers to independently disclosed methyl, ethyl, C ₃ alkyl, C ₄ alkyl, C ₅ alkyl and C ₆ alkane "C _1-4 alkyl" or "C ₁ -C ₄ alkyl" specifically refers to independently disclosed methyl, ethyl, C ₃ alkyl (ie propyl, including n-propyl and isopropyl), _C4 alkyl (ie, butyl, including n-butyl, isobutyl, sec-butyl and tert-butyl).

When a substituent is not clearly indicated in the enumerated group, this group only means unsubstituted. For example, when there is no limitation of "substituted or unsubstituted" before "C ₁ -C ₄ alkyl", it only refers to "C ₁ -C ₄ alkyl" itself or "unsubstituted C ₁ -C ₄ alkyl".

In various sections of the invention linking substituents are described. When the structure clearly requires a linking group, the Markush variables recited for that group are to be understood as linking groups. For example, if the structure requires a linking group and the Markush group definition for that variable recites "alkyl," it is understood that "alkyl" represents a linking alkylene group.

In some specific structures, when an alkyl group is clearly indicated as a linking group, then the alkyl group represents a linked alkylene group, for example, the group "halo-C ₁ -C ₆ alkane C ₁ -C ₆ alkyl in "group" should be understood as C ₁ -C ₆ alkylene.

In addition, it should be noted that, unless otherwise clearly stated, the description method "...independently" used in the present invention should be interpreted in a broad sense, which means that the described individuals are independent of each other and can be independent are the same or different specific groups. In more detail, the description "...independently" can mean that in different groups, the specific options expressed by the same symbols do not affect each other; it can also mean that in the same group, the same symbols The specific options expressed between them do not affect each other.

It should be understood that as used herein, a singular form such as "a" includes plural referents unless otherwise specified.

The term "one (one) or more (ones)" or "one (one) or two (more than one)" means that 1, 2, 3, 4, 5, 6, 7, 8, 9 or More.

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

When indicating a single key, it means

the mix of.

Those skilled in the art can understand that, according to the practice used in this field, the application describes the structural formula used in the group

and

means that the corresponding group R is connected to other fragments and groups in the compound through this site.

"Hydroxy" means an -OH group.

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

When R in is a carbonyl group,

for

"Cyano" means -CN.

"Amino" refers to _-NH2 .

"R ^5b substituted amino group", R ^5b can be one or two, when R ^5b is two, they can be the same group or different groups.

"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, Consisting solely of carbon and hydrogen atoms, having for example 1 to 12 (preferably 1 to 8, more preferably 1 to 6, more preferably 1 to 4) carbon atoms, and attached to the remainder of the molecule by a single bond, 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.

In this application, the term "haloalkyl", as a group or part of another group, means that one or more hydrogen atoms in an alkyl group (as defined in this invention) are replaced by a halogen (as defined in this invention) ), the number of halogens can be one or more; when the number of halogens is multiple, the halogens are the same or different. For example, fluoroalkyl refers to an alkyl group substituted with one or more fluorines. Examples of haloalkyl include, but are not limited to, trifluoromethyl, difluoromethyl, and monofluoromethyl.

In the present application, preferably, the number of carbon atoms in the "heterocycloalkyl" is 3, 4, 5 or 6, the heteroatoms are selected from N, O and S, and the number of heteroatoms is 1, 2, 3 or 4, Further preferably, the number of carbon atoms is 4 or 5, the heteroatoms are selected from N and O, and the number of heteroatoms is 1 or 2; for example

In the present application, preferably, the number of carbon atoms in the "heterospirocycloalkyl" is 4, 5 or 6, the heteroatoms are selected from N, O and S, and the number of heteroatoms is 1, 2, 3 or 4, further Preferably, the heteroatoms are selected from N and O, and the number of heteroatoms is 1 or 2; for example

In this application, the term "cyclohydrocarbyl", as a group or part of another group, means a stable non-aromatic monocyclic or polycyclic hydrocarbon group composed only of carbon and hydrogen atoms, which may include fused rings system, bridged ring system or spiro ring system, has 3 to 15 carbon atoms, preferably has 3 to 10 carbon atoms, more preferably has 3 to 8 carbon atoms, and it is saturated or unsaturated and can be changed via any suitable The carbon atoms are connected to the rest of the molecule by single bonds. Unless specifically stated otherwise in this specification, carbon atoms in a cycloalkyl group may be optionally oxidized. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclooctyl, 1H-indene Base, 2,3-indanyl, 1,2,3,4-tetrahydro-naphthyl, 5,6,7,8-tetrahydro-naphthyl, 8,9-dihydro-7H-benzo Cyclohepten-6-yl, 6,7,8,9-tetrahydro-5H-benzocycloheptenyl, 5,6,7,8,9,10-hexahydro-benzocyclooctenyl, Fluorenyl, bicyclo[2.2.1]heptyl, 7,7-dimethyl-bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, bicyclo[2.2.2]octyl Base, bicyclo[3.1.1]heptyl, bicyclo[3.2.1]octyl, bicyclo[2.2.2]octenyl, bicyclo[3.2.1]octenyl, adamantyl, octahydro -4,7-methylene-1H-indenyl and octahydro-2,5-methylene-pentalenyl, etc.

In this application, the term "cycloalkyl", as a group or part of another group, means a saturated cyclic hydrocarbon group.

In this application, the term "cycloalkenyl", as a group or part of another group, means a cyclic hydrocarbon group having at least one double bond (eg, a carbon-carbon double bond). A cycloalkenyl group can be attached to the rest of the molecule through an atom of a double bond within it.

In this application, the term "heterocyclyl", as a group or part of another group, means a group consisting of 2 to 14 carbon atoms and 1 to 6 heteroatoms selected from nitrogen, phosphorus, oxygen and sulfur 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 by a single bond. 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 , more preferably a stable 4- to 8-membered non-aromatic monocyclic, bicyclic, bridged or spirocyclic group comprising 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur. Examples of heterocyclic groups include, but are not limited to: pyrrolidinyl, morpholinyl, piperazinyl, homopiperazinyl, piperidinyl, thiomorpholinyl, 2,7-diaza-spiro[3.5]nonyl Alkane-7-yl, 2-oxa-6-aza-spiro[3.3]heptane-6-yl, 2,5-diaza-bicyclo[2.2.1]heptane-2-yl, aza Cyclobutanyl, pyranyl, tetrahydropyranyl, thiopyranyl, tetrahydrofuryl, oxazinyl, dioxolyl, tetrahydroisoquinolyl, decahydroisoquinolyl, imidazolinyl, Imidazolidinyl, Quinazinyl, Thiazolidinyl, Isothiazolidinyl, Isoxazolidinyl, Indolinyl, Octahydroindolyl, Octahydroisoindolyl, Pyrrolidinyl, Pyrazolidinyl , Phthalimide, etc.

In the present application, the term "aryl", as a group or part of another group, means a conjugated hydrocarbon ring system group having 6 to 18 carbon atoms, preferably having 6 to 10 carbon atoms. 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 cycloalkyl or heterocyclyl as defined above, provided that the aryl is via The atoms on the aromatic ring are connected to the rest of the molecule by 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 be fused to a cycloalkyl or heterocyclyl as defined above, provided that hetero An aryl group is connected to the rest of the molecule by a single bond through an atom on the aromatic 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 comprising 1 to 5 heteroatoms selected from nitrogen, oxygen and sulfur, more preferably 1 to 4 heteroatoms selected from nitrogen , a stable 5- to 10-membered aromatic group containing heteroatoms of nitrogen, oxygen and sulfur or a 5- to 6-membered aromatic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur (for example heteroaryl is a C ₁ -C ₅ heteroaryl group, wherein the heteroatoms are selected from N, O and S, and the number of heteroatoms is 1, 2, 3 or 4). 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 this application, "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes both instances where the event or circumstance occurs and instances where it does not occur. For example, "optionally substituted aryl" means that the aryl is substituted or unsubstituted, and the description includes both substituted and unsubstituted aryl. The "optional" substituents described in the claims and description of the present invention are selected from the group consisting of alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, cyano, nitro , optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl.

In the present invention, the term "substituted" or "substituent" means that one or more hydrogen atoms are replaced by the specified group. When the number of substituents is not specified, there may be one or more substituents; when the substitution position is not specified, the substitution may be at any position, but only when a stable or chemically feasible chemical compound is formed is considered Allowed.

When any variable (eg, R) occurs more than once in the composition or structure of a compound, its definition is independent at each occurrence. Thus, for example, if a group is substituted with 0-2 R, said group may optionally be substituted with up to two R, with independent options for each occurrence of R. For example, in

In , when n is 2, it means that the benzene ring is replaced by 2 R, and each R has an independent option, that is, the 2 R can be the same or different. Also, combinations of substituents and/or variations thereof are permissible only if such combinations result in stable compounds.

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.

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. Stereoisomers of the compounds of the present invention may exist as (R)- or (S)-isomers.

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.

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

In this application, "pharmaceutically acceptable excipients" include, but are not limited to, any adjuvants, carriers, excipients, glidants, sweeteners approved by the relevant government regulatory agencies 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.

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.

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, etc. Suitable protecting groups for amino, amidino and guanidino include tert-butoxycarbonyl, benzyloxycarbonyl and the like. 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.

On the basis of not violating common knowledge in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain preferred examples of the present invention.

The reagents and raw materials used in the present invention are all commercially available.

The positive progress effect of the present invention lies in: with US20160176882A1, compound N-((5-fluoro-2,3-dihydrobenzofuran-4-yl)methyl)-8-(2-methylpyridine- Compared with 3-yl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine, the anti-cell proliferation activity of the compound of the present invention is increased by about 10 times. When the compound disclosed in the present invention is combined with the EED protein, the bicyclic structure outside the combined "pocket" can make the compound have better metabolic stability.

Detailed ways

The present invention is further illustrated below by means of examples, but the present invention is not limited to the scope of the examples. For the experimental methods that do not specify specific conditions in the following examples, select according to conventional methods and conditions, or according to the product instructions.

The starting materials used in the following examples can be purchased from chemical distributors such as Aldrich, TCI, Alfa Aesar, Bi De, Anaiji, etc., or can be synthesized by known methods.

In the following examples, the ice bath refers to -5 degrees Celsius to 0 degrees Celsius, the room temperature refers to 10 degrees Celsius to 30 degrees Celsius, and the reflux temperature generally refers to the solvent reflux temperature under normal pressure. Reaction overnight means that the time is 8-15 hours. In the following examples, the specific operating temperature is not limited, all carried out at room temperature.

In the following examples, the separation and purification of intermediates and final products are carried out by normal phase or reverse phase chromatographic column separation or other suitable methods. The normal-phase flash chromatography column uses ethyl acetate and n-hexane or methanol and methylene chloride as mobile phases. Reversed-phase preparative high-pressure liquid chromatography (HPLC) uses C18 column and detects with UV 214nm and 254nm, and its mobile phase is A (water and 0.1% formic acid), B (acetonitrile) or mobile phase A (water and 0.1% carbonic acid ammonium hydrogen), B (acetonitrile).

In each embodiment:

LCMS instrument: Pump Agilent 1260UV detector: Agilent 1260 DAD

Mass Spectrometer API 3000

Chromatographic column: Waters sunfire C18, 4.6×50mm, 5μm

Mobile phase: AH ₂ O (0.1% HCOOH); B-acetonitrile

NMR instrument: Bruker Ascend 400M ( ¹ H NMR: 400 MHz; ¹³ C NMR: 100 MHz).

Example 1

Intermediate 4-(aminomethyl)-5-fluoro-2,3-dihydrobenzofuran-3-ol (A):

Step 1: Intermediate 2-bromo-3,6-difluorobenzaldehyde (A-2):

Add A-1 (22 g, 114 mmol) and dry THF (200 mL) into a dry 500 mL three-neck flask, and cool to -70°C. Lithium diisopropylamide (2M, 68.4 mL) was slowly added dropwise to the reaction solution. After the reaction solution was stirred at the same temperature for 45 minutes, DMF (17.8 mL, 228 mmol) was added. The reaction solution was stirred at the same temperature for two hours and then raised to 0°C. Saturated ammonium chloride (200 mL) was added to the reaction solution. The reaction solution was extracted with EtOAc (200mL×2). The combined organic phases were washed once with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified on silica gel (petroleum ether:ethyl acetate=100:1) to obtain A-2 (20 g, 79.4% yield) as a pale yellow solid.

¹ H NMR (400MHz, CDCl ₃ ) δ10.34 (s, 1H), 7.34 (ddd, J = 9.2, 7.4, 4.5Hz, 1H), 7.16 (td, J = 9.3, 4.0Hz, 1H) ppm.

Step 2: Intermediate 2-bromo-3-fluoro-6-methoxybenzaldehyde (A-3):

A-2 (20 g, 90.5 mmol) was added into a 2 L three-necked flask, and stirred and dissolved with anhydrous THF (1000 ml) and MeOH (200 ml). Sodium methoxide (5.87 g, 108.6 mmol) was added, and the reaction solution was stirred at 60° C. for 18 hours. Concentrate under reduced pressure to remove most of the solvent, and add 500 mL of water. The suspension was stirred for 30 minutes and filtered to collect the solid. The solid was slurried with a mixture of petroleum ether and ethyl acetate (5:1), filtered to obtain a solid, and dried under reduced pressure to obtain A-3 (18 g, 85% yield) as a yellow solid.

LC-MS: m/z 233.1[M+H] ⁺ .

Step 3: Intermediate 2-bromo-3-fluoro-6-hydroxybenzaldehyde (A-4):

A-3 (16.8 g, 72.1 mmol) and dichloromethane (300 mL) were added to a 1 L one-necked flask. Boron tribromide (21.7 g, 86.5 mmol) was slowly added dropwise at -78°C, and the reaction solution was raised to room temperature and stirred for 18 hours. The reaction solution was diluted with dichloromethane (300 mL), and saturated sodium bicarbonate (300 mL) was added slowly. The organic phase was washed twice with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified on silica gel (petroleum ether:ethyl acetate=50:1) to afford A-4 (10 g, 63.3% yield) as a pale yellow solid.

¹ H NMR (400MHz, CDCl ₃ ) δ11.77(s, 1H), 10.34(s, 1H), 7.29(dt, J=12.7, 6.3Hz, 1H), 6.94(dd, J=9.3, 4.1Hz, 1H)ppm.

Step 4: Intermediate 4-bromo-5-fluoro-2,3-dihydrobenzofuran-3-ol (A-5):

Add trimethylsulfoxide iodide (9.73 g, 44.2 mmol) and DMSO (50 ml) into a dry three-necked flask. Sodium tert-butoxide (4.25 g, 44.2 mmol) was added under an ice-water bath. The reaction solution was stirred at room temperature for 2 hours, and A-4 (8.8 g, 40.2 mmol) was added. After the reaction solution was stirred at room temperature for 18 hours, ethyl acetate (250ml) and water (250ml) were added, and extracted with ethyl acetate (250ml×2). The organic phase was washed once with water and brine respectively, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified on silica gel (petroleum ether:ethyl acetate=10:1) to afford A-5 (6.2 g, 66.2% yield) as a white solid.

¹ H NMR (400MHz, CDCl ₃ ) δ7.03(t, J=8.7Hz, 1H), 6.76(dd, J=8.8, 3.5Hz, 1H), 5.51-5.40(m, 1H), 4.58(ddd, J=13.4,10.8,4.6Hz,2H),2.33(d,J=4.9Hz,1H)ppm.

Step 5: Intermediate 5-fluoro-3-hydroxy-2,3-dihydrobenzofuran-4-carbonitrile (A-6):

A-5 (2.7g, 11.6mmol), zinc cyanide (2.04g, 17.4mmol), DMF (50ml) and palladium tetrakistriphenylphosphine (1.34g, 1.16mmol) were successively added into a dry one-necked flask. The reaction solution was heated to 120° C. under nitrogen protection and stirred for 18 hours. The reaction solution was cooled to room temperature, extracted with ethyl acetate (200mL×3) and water (200ml). The organic phase was washed once with water and brine respectively, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified on silica gel (petroleum ether:ethyl acetate=5:1) to afford A-6 (1.6 g, 77% yield) as a white solid.

¹ H NMR (400MHz, CDCl ₃ ) δ7.20-6.99 (m, 2H), 5.63 (dd, J=6.9, 2.9Hz, 1H), 4.69 (dd, J=10.8, 7.1Hz, 1H), 4.55( dd,J＝10.8,3.1Hz,1H),2.74(s,1H)ppm.

Step 6: Intermediate (5-fluoro-2,3-dihydrobenzofuran-4-yl)methanamine (A2):

A-6 (1.55g, 8.65mmol), trifluoroacetic acid (1.09g, 8.65mmol), methanol (20mL) and 10% palladium carbon (2g, containing 50% water) were sequentially added into a 100mL single-necked flask. The reaction mixture was sparged with hydrogen for 5 minutes, purged three times with a hydrogen balloon, and stirred at 60° C. for 48 hours under a hydrogen balloon. The mixture was filtered through diatomaceous earth, washed with methanol (50mL X 2), the filtrate was concentrated under reduced pressure, dioxane (10mL) and 10M aqueous sodium hydroxide solution (1mL) were added, extracted with dioxane (10mL X 2), The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Compound A2 (1.4 g, 90% purity, 87% yield) was obtained and used directly in the next step.

¹ H NMR (400MHz, CD ₃ OD) δ3.27(t,2H),3.77(s,2H),4.56(t,2H),6.59(dd 1H),6.81(dd,1H)ppm; LC-MS :m/z 168.1[M+H] ⁺ .

Example 2

8-(5-((Dimethylamino)methyl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-5-(((5-fluoro-2, 3-Dihydrobenzofuran-4-yl)methyl)amino)imidazo[1,5-c]pyrimidine-1-carbonitrile (BR-1)

Step 1: 2-(5-bromo-2-(methylthio)pyrimidin-4-yl)-2-((dibenzylidene)amino)acetonitrile (2)

To a solution of sodium hydride (3.63 g, 90.8 mmol, 60% purity) in N,N-dimethylformamide (60.0 mL) was added N-(dibenzylidene)aminoacetonitrile 1 (10.0 g, 45.4mmol) in N,N-dimethylformamide (50.0mL) solution, after 2 minutes, 5-bromo-4-chloro-2-(methylthio)pyrimidine (10.8g, 45.4mmol) was added under zero temperature N, N-dimethylformamide (30.0 mL) was reacted at 40°C for 2 hours. LCMS monitors that the reaction is completed, quenched by ammonium chloride aqueous solution, extracted with ethyl acetate and spin-dried to obtain crude product 2-(5-bromo-2-(methylthio)pyrimidin-4-yl)-2-((dibenzylidene ) amino) acetonitrile 2 (15.0 g, crude product), used directly in the next step.

¹ H NMR (400MHz,DMSO-d ₆ )δ7.58-7.52(m,7H),7.44-7.40(m,3H),7.26-7.23(m,2H),4.37(s,3H)ppm; LCMS m /z 424.7[M+H] ⁺ .

Step 2: 2-amino-2-(5-bromo-2-(methylthio)pyrimidin-4-yl)acetonitrile (3)

2-(5-bromo-2-(methylthio)pyrimidin-4-yl)-2-((dibenzylidene)amino)acetonitrile 2 (15.0 g, crude product) was dissolved in tetrahydrofuran at zero degree (200mL) and aqueous solution (150mL) were added with concentrated hydrochloric acid (12M, 23.6mL) and reacted at 20°C for 2 hours. TLC monitors the end of the reaction. After the reaction solution is spun off from tetrahydrofuran, aqueous sodium hydroxide solution (1.00M) is added to adjust the pH to about 8.0, extracted with ethyl acetate, dried, and dried to obtain the crude product 2-amino-2-(5-bromo-2 -(Methylthio)pyrimidin-4-yl)acetonitrile 3 (9.00 g, crude) was used directly in the next step.

¹ H NMR (400MHz,DMSO-d ₆ )δ8.86(s,1H),5.28(s,1H),2.89(s,2H),2.57(s,3H)ppm; LCMS m/z 260.9[M+ H] ⁺ .

Step 3: N-((5-bromo-2-(methylthio)pyrimidin-4-yl)(cyano)methyl)formamide (4)

Formic acid (39.0g, 848mmol) and acetic anhydride (34.8g, 341mmol) were mixed and reacted at 50°C for 1 hour, then added to 2-amino-2-(5-bromo-2-(methylthio) after cooling to 0°C ) pyrimidin-4-yl) acetonitrile 3 (9.00 g, crude product) in dichloromethane (100 mL) solution. Reaction at 20 degrees for 2h. TLC monitors the end of the reaction, adjusts it to 8 with aqueous sodium hydroxide solution (1.00M) at zero degrees, extracts with ethyl acetate, dries, and selects dry to obtain the N-((5-bromo-2-( Methylthio)pyrimidin-4-yl)(cyano)methyl)formamide 4 (7.00 g, 22.6 mmol, 73.2% yield).

Step 4: 8-bromo-5-(methylthio)imidazo[1,5-c]pyrimidine-1-carbonitrile (5)

To N-((5-bromo-2-(methylthio)pyrimidin-4-yl)(cyano)methyl)formamide 4 (7.00 g, 24.3 mmol) was added phosphorus oxychloride (28.8 g, 188 mmol) , reacted at 110 degrees for 1 hour. TLC monitors the end of the reaction, selects a part of phosphorus oxychloride from the reaction solution, then quenches it with water at <30°C, and finally adjusts it to 8 with NaOH aqueous solution (6.00M), extracts with ethyl acetate, and dries to obtain the crude product. Petroleum ether/ethyl acetate (5:1, 100mL) beating, the solid was filtered out to obtain a pure product, and a small amount of product was obtained by column chromatography of the filtrate, which was finally combined to obtain 8-bromo-5-(methylthio)imidazo[1,5 -c] Pyrimidine-1-carbonitrile 5 (3.20 g, 11.7 mmol, 48.1% yield).

¹ H NMR (400MHz, DMSO-d ₆ ) δ8.81(s,1H), 8.06(s,1H), 2.76(s,3H)ppm; LCMS m/z 270.9[M+H] ⁺ .

Step 5: 8-bromo-5-(methylsulfonyl)imidazo[1,5-c]pyrimidine-1-carbonitrile (6)

8-Bromo-5-(methylthio)imidazo[1,5-c]pyrimidine-1-carbonitrile 5 (0.60 g, 2.23 mmol) was dissolved in dichloromethane (5 mL), and 85% m-Chloroperbenzoic acid (1.40 g, 6.91 mmol) was stirred at 20°C for 12 hours. The completion of the reaction was monitored by LCMS, and the reaction solution was directly used in the next step.

LCMS m/z 302.9[M+H] ⁺ .

Step 6: 8-bromo-5-(((5-fluoro-2,3-dihydrobenzofuran-4-yl)methyl)amino)imidazo[1,5-c]pyrimidine-1-carbonitrile ( 7)

8-Bromo-5-(methylsulfonyl)imidazo[1,5-c]pyrimidine-1-carbonitrile 6 (671mg, 2.23mmol) and (5-fluoro-2,3-dihydrobenzofuran-4 -yl)methylamine A (453mg, 2.23mmol) was dissolved in dichloromethane (5mL), sodium carbonate (1.18g, 11.1mmol) was added, and stirred at 20°C for 4 hours. LCMS monitors the end of the reaction, and the reaction solution is diluted with dichloromethane, washed with brine, and the organic phase is dried and spin-dried to obtain the brown solid product 8-bromo-5-(((5-fluoro-2,3-dihydrobenzene furan-4-yl)methyl)amino)imidazo[1,5-c]pyrimidine-1-carbonitrile 7 (400 mg, 46.2% yield).

¹ H NMR (400MHz, DMSO-d ₆ ) δ8.90-8.77 (m, 2H), 7.75 (s, 1H), 6.93 (t, J = 9.4Hz, 1H), 6.69 (dd, J = 4.0, 8.8 Hz, 1H), 4.67(s, 2H), 4.53(t, J=8.8Hz, 2H), 3.29-3.24(m, 3H) ppm; LCMS m/z 388.0[M+H] ⁺ .

Step 7: 8-bromo-5-(bromomethyl)-[1,2,4]triazolo[1,5-a]pyridine (9)

Add 8-bromo-5-methyl-[1,2,4]triazolo[1,5-a]pyridine 8 (2g, 9.43mmol) and carbon tetrachloride (20ml) in a 100mL round bottom flask, NBS (2 g, 11.3 mmol) and AIBN (310 mg, 0.2 mmol) were added under ice water bath. After the addition was complete, the reaction temperature was raised to 80 °C, and the reaction was maintained for 16 hours overnight. The reaction solution was washed with aqueous sodium sulfite solution and spin-dried to obtain a reddish-brown solid, which was purified with a flash separation column (eluent: petroleum ether: ethyl acetate = 3:1) to obtain a reddish-brown solid. Pure 8-bromo-5-(bromomethyl)-[1,2,4]triazolo[1,5-a]pyridine 9 was obtained after vacuum drying (1.7 g, 62% yield).

¹ H NMR (400MHz, CDCl ₃ ) δ8.48 (s, 1H), 7.77 (d, J = 7.7Hz, 1H), 7.07 (d, J = 7.7Hz, 1H), 4.90 (s, 2H) ppm; LCMS: m/z 291.9[M+H] ⁺ .

Step 8: 1-(8-bromo-[1,2,4]triazolo[1,5-a]pyridin-5-yl)-N,N-dimethylmethylamine (10)

8-Bromo-5-(bromomethyl)-[1,2,4]triazolo[1,5-a]pyridine 9 (1 g, 3.44 mmol) and tetrahydrofuran (15 ml) were added to a 100 mL round bottom flask, A 1M solution of dimethylamine tetrahydrofuran (17ml, 17.2mmol) and potassium carbonate (0.59g, 1.43mmol) were added under an ice-water bath. After the addition was completed, the reaction temperature was raised to 80° C., and the reaction was maintained for 3 h. After the reaction was completed, the liquid was spin-dried to obtain a yellow-brown solid, which was eluted and purified with a flash separation column (petroleum ether: ethyl acetate = 5:1) to obtain a yellow solid 1-(8-bromo-[1,2,4] Triazolo[1,5-a]pyridin-5-yl)-N,N-dimethylmethanamine 10, (0.6 g, 68.3% yield).

¹ H NMR (400MHz,DMSO-d ₆ )δ8.74-8.38(m,1H),8.14-7.85(m,1H),7.31-6.90(m,1H),4.03-3.78(m,2H),2.38 -2.20(m,6H)ppm; LCMS: m/z 255.0,257.0[M+H] ⁺ .

Step 9: (5-((Dimethylamino)methyl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)boronic acid (11)

1-(8-Bromo-[1,2,4]triazolo[1,5-a]pyridin-5-yl)-N,N-dimethylmethylamine 10 (100mg, 391μmol), diboronic acid Pinacol ester (159 mg, 627 μmol), potassium acetate (115 mg, 1.18 mmol), [1,1-bis(diphenylphosphino) ferrocene] dichloropalladium dichloromethane (64.0 mg, 78.4 μmol) in Dissolve in dioxane (6 mL) in a glove box and react at 125°C for 5 hours. The completion of the reaction was monitored by LCMS, and the reaction solution was directly used for the next reaction.

LCMS m/z 221.1[M+H] ⁺ .

Step 10: 8-(5-((dimethylamino)methyl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-5-(((5-fluoro -2,3-dihydrobenzofuran-4-yl)methyl)amino)imidazo[1,5-c]pyrimidine-1-carbonitrile (BR-1)

Add 8-bromo-5-(((5-fluoro-2,3-dihydrobenzofuran-4-yl)methyl)amino)imidazo[1,5-c]pyrimidine to the boric acid reaction solution in the previous step -1-carbonitrile 7 (151 mg, 390 μmol), potassium carbonate (108 mg, 781 μmol), [1,1-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane (31.9 mg, 39.0 μmol), Water (1 mL), dioxane (3 mL), react at 100°C for 2 hours. LCMS monitors the end of the reaction, and after the reaction solution is concentrated, the solution obtained by column chromatography purification and perp-HPLC is adjusted to pH 8 with saturated aqueous sodium bicarbonate solution, concentrated to remove the organic solvent, and filtered to obtain a white solid 8-(5-((di Methylamino)methyl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-5-(((5-fluoro-2,3-dihydrobenzofuran- 4-yl)methyl)amino)imidazo[1,5-c]pyrimidine-1-carbonitrile BR-1 (70.0 mg, 36.8% yield).

¹ H NMR (400MHz, DMSO-d ₆ )δ=8.93(s,1H),8.83(s,1H),8.50(s,1H),7.85-7.76(m,2H),7.32(d,J=7.6 Hz,1H),6.96(t,J=9.6Hz,1H),6.72(dd,J=4.0,8.8Hz,1H),4.77(s,2H),4.56(t,J=8.8Hz,2H), 4.03(s,2H),3.35-3.35(m,2H),2.34(s,6H)ppm; LCMS m/z 484.4[M+H] ⁺ .

Example 3

5-(((5-fluoro-2,3-dihydrobenzofuran-4-yl)methyl)amino)-8-(5-(2-hydroxypropan-2-yl)-[1,2, 4] Triazolo[1,5-a]pyridin-8-yl)imidazo[1,5-c]pyrimidine-1-carbonitrile (BR-2)

Step 1: (E)-5-Bromo-6-(((dimethylamino)methylene)amino)-pyridine-2-carboxylic acid methyl ester E-2

(E)-5-bromo-6-(((dimethylamino)methylene)amino)-pyridine-2-carboxylic acid methyl ester (5.0g, 21.6mmol), 1,1-dimethoxy- N,N-Dimethylmethylamine (5.16g, 43.3mmol) was dissolved in toluene (100mL), the temperature of the reaction solution was raised to 110°C and the reaction was carried out for 4 hours. No raw material was detected by TLC. The reaction solution was concentrated to obtain the crude product (E)-5 -Bromo-6-(((dimethylamino)methylene)amino)-pyridine-2-carboxylic acid methyl ester E-2 (6.00 g).

LCMS: m/z 585.7[M+H] ⁺ .

Step 2: (E)-5-bromo-6-(((hydroxylamino)methylene)amino)-pyridine-2-carboxylic acid methyl ester E-3

Add (E)-5-bromo-6-(((dimethylamino)methylene)amino)-pyridine-2-carboxylic acid methyl ester E-2 (6.0g, 20.97mmol), hydrochloric acid Hydroxylamine (2.91g, 41.94mmol), sodium acetate (3.44g, 41.9mmol), ethanol (60mL). The reaction solution was heated to 50°C under a nitrogen atmosphere and reacted for 4 hours. LCMS monitored the completion of the reaction, cooled to room temperature, and filtered to obtain (E)-5-bromo-6-(((hydroxylamino)methylene)amino)-pyridine- 2-Methyl carboxylate E-3 (5.9 g, 85.0% yield).

¹ H NMR (400MHz, CDCl ₃ ) δ8.27 (br d, J = 8.5Hz, 1H), 8.19-8.13 (m, 1H), 7.85 (d, J = 7.9Hz, 1H), 7.50 (d, J =7.9Hz,1H), 3.90(s,3H)ppm; LCMS: m/z 273.7[M+H] ⁺ .

Step 3: Methyl 8-bromo-[1,2,4]triazolo[1,5-a]pyridine-5-carboxylate E-4

Add (E)-5-bromo-6-(((hydroxylamino)methylene)amino)-pyridine-2-carboxylic acid methyl ester E-3 (10g, 36.49mmol) in tetrahydrofuran (100mL) at room temperature Trifluoroacetic anhydride (23.0 g, 109 mmol) was added dropwise. Then slowly warm to 75°C and stir the reaction for 3 hours. After the reaction was completed, the mixture was quenched by adding saturated aqueous sodium bicarbonate solution (100 mL) at 0° C., extracted with ethyl acetate (100 mL*3), and the combined organic phases were dried and concentrated. The concentrate was purified by silica gel column (petroleum ether: ethyl acetate = 5:1 to 4:1) to obtain the white solid product 8-bromo-[1,2,4]triazolo[1,5-a]pyridine-5 - Methyl formate E-4 (5.00 g, yield 53.5%).

Step 4: 2-(8-Bromo-[1,2,4]triazolo[1,5-a]pyridin-5-yl)propan-2-ol E-5

At -30°C, add 8-bromo-[1,2,4]triazolo[1,5-a]pyridine-5-carboxylic acid methyl ester E-4 (500mg, 1.95mmol) in THF solution (10mL ) was added methylmagnesium bromide (3M, 2.60 mL) dropwise and stirred at this temperature for three hours. After the reaction, add water (10mL) to the reaction solution to quench, extract with ethyl acetate (10mL*3), combine the organic phases to dry, concentrate and purify with a flash silica gel column to obtain 2-(8-bromo -[1,2,4]triazolo[1,5-a]pyridin-5-yl)propan-2-ol E-5 (400 mg, 80.0% yield).

¹ H NMR (400MHz,DMSO-d ₆ )δ8.61(s,1H),8.05(d,J=8.0Hz,1H),7.28(d,J=8.0Hz,1H),5.91(s,1H) ,1.72(s,6H)ppm; LCMS: m/z 255.9[M+H] ⁺ .

Step 5: (5-(2-Hydroxypropan-2-yl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)boronic acid (12)

2-(8-Bromo-[1,2,4]triazolo[1,5-a]pyridin-5-yl) tertiary alcohol E-5 (100mg, 390μmol), pinacol diborate (158mg , 624μmol), potassium acetate (114mg, 1.17mmol), [1,1-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane (63.7mg, 78.0μmol) were dissolved in dichloromethane in glove box Hexane (6 mL) was reacted at 125°C for 6 hours. The completion of the reaction was monitored by LCMS, and the reaction solution was directly used for the next reaction.

LCMS m/z 222.2[M+H] ⁺ .

Step 6: 5-(((5-fluoro-2,3-dihydrobenzofuran-4-yl)methyl)amino)-8-(5-(2-hydroxypropan-2-yl)-[1 ,2,4]triazolo[1,5-a]pyridin-8-yl)imidazo[1,5-c]pyrimidine-1-carbonitrile (BR-2)

Add 8-bromo-5-(((5-fluoro-2,3-dihydrobenzofuran-4-yl)methyl)amino)imidazo[1,5-c to the reaction solution of boronic acid 12 in the previous step ]pyrimidine-1-carbonitrile 7 (151 mg, 389 μmol), potassium carbonate (107 mg, 778 μmol), [1,1-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane (31.7 mg, 38.9 μmol ), water (1 mL), dioxane (3 mL), and react at 100° C. for 2 hours. LCMS monitors the end of the reaction, and after the reaction solution is concentrated, the solution obtained by column chromatography purification and perp-HPLC is adjusted to pH 8 with saturated aqueous sodium bicarbonate solution, and the organic solvent is removed by concentration, and the white solid 5-(((5-fluoro -2,3-Dihydrobenzofuran-4-yl)methyl)amino)-8-(5-(2-hydroxypropan-2-yl)-[1,2,4]triazolo[1, 5-a]pyridin-8-yl)imidazo[1,5-c]pyrimidine-1-carbonitrile (50.0 mg, 26.0% yield).

¹ H NMR (400MHz, DMSO-d ₆ )δ=8.90(s,1H),8.84(s,1H),8.51(s,1H),7.83-7.78(m,2H),7.44(d,J=7.6 Hz,1H),6.95(t,J=9.4Hz,1H),6.75-6.67(m,1H),5.86(s,1H),4.77(s,2H),4.56(t,J=8.6Hz,2H ), 3.37-3.32(m,2H), 1.77(s,6H)ppm; LCMS m/z 485.4[M+H] ⁺ .

Example 4

8-(5-(Dimethylamino)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-5-(((5-fluoro-2,3-dihydro Benzofuran-4-yl)methyl)amino)imidazo[1,5-c]pyrimidine-1-carbonitrile (BR-3)

Step 1: 8-iodo-N,N-dimethyl-[1,2,4]triazolo[1,5-a]pyridin-5-amine 14

To a solution of dimethylamine in tetrahydrofuran (4 mL, 2M/L) was added 5-chloro-8-iodo-[1,2,4]triazolo[1,5-a]pyridine 13 (200 mg, 0.72 mmol). The mixture was stirred at 90 °C for 16 h. LCMS showed that the reaction was complete, and it was concentrated and purified (petroleum ether: ethyl acetate = 1:1) to obtain 8-iodo-N,N-dimethyl-[1,2,4]triazolo[1,5-a ] Pyridin-5-amine 14 (190 mg, 90%).

LCMS: m/z 288.9[M+H] ⁺ .

Step 2: (5-(Dimethylamino)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)boronic acid (15)

8-iodo-N,N-dimethyl-[1,2,4]triazolo[1,5-a]pyridin-5-amine 14 (100mg, 414μmol), pinacol diborate (168mg , 663μmol), potassium acetate (122mg, 1.24mmol), [1,1-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane (67.7mg, 82.9μmol) were dissolved in dichloromethane in glove box Hexane (5 mL) was reacted at 125°C for 6 hours. The completion of the reaction was monitored by LCMS, and the reaction solution was directly used for the next reaction.

LCMS m/z 207.2[M+H] ⁺ .

Step 3: 8-(5-(Dimethylamino)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-5-(((5-fluoro-2,3 -Dihydrobenzofuran-4-yl)methyl)amino)imidazo[1,5-c]pyrimidine-1-carbonitrile

Add 8-bromo-5-(((5-fluoro-2,3-dihydrobenzofuran-4-yl)methyl)amino)imidazo[1,5-c] to the boric acid 15 reaction liquid in the previous step Pyrimidine-1-carbonitrile (160 mg, 412 μmol), potassium carbonate (114 mg, 825 μmol), [1,1-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane (33.6 mg, 41.2 μmol), Water (1 mL), dioxane (3 mL) were reacted at 100° C. for 2 hours. LCMS monitors the end of the reaction. After the reaction solution is concentrated, the solution obtained by column chromatography purification and perp-HPLC is adjusted to pH 8 with saturated aqueous sodium bicarbonate solution, concentrated to remove the organic solvent, and filtered to obtain a white solid 8-(5-(dimethyl Amino)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-5-(((5-fluoro-2,3-dihydrobenzofuran-4-yl) Methyl)amino)imidazo[1,5-c]pyrimidine-1-carbonitrile BR-3 (13.0 mg, 6.61% yield).

¹ H NMR (400MHz, DMSO-d ₆ )δ=8.90-8.68(m,2H),8.40(s,1H),7.70-7.61(m,2H),6.94(t,J=9.2Hz,1H), 6.74-6.66(m,1H),6.56(d,J=8.0Hz,1H),4.74(s,2H),4.55(t,J=8.6Hz,2H),3.37-3.33(m,2H),3.17 (s,6H)ppm; LCMS m/z 470.4[M+H] ⁺ .

Example 5

8-(5-((Dimethylamino)methyl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-N-((5-fluoro-2,3 -Dihydrobenzofuran-4-yl)methyl)-1-(methylsulfonyl)imidazo[1,5-c]pyrimidin-5-amine (BR-4)

Step 1: 8-bromo-N-((5-fluoro-2,3-dihydrobenzofuran-4-yl)methyl)imidazo[1,5-c]pyrimidin-5-amine (17)

Dissolve 8-bromo-5-(methylthio)imidazo[1,5-c]pyrimidine 16 (4.0g, 16.4mmol) in dichloromethane (60ml), add m-chloroperoxybenzoic acid at 0°C (4.99g, 24.6mmol, 85% purity) and reacted at 0°C for 55 minutes, then added triethylamine (6.63g, 65.5mmol) dropwise at 0°C and stirred for 5 minutes, and finally 5-fluoro-2,3 -Dihydrobenzofuran-4-yl)methylamine A (3.34g, 16.4mmol, HCl) was added to the reaction solution, heated to 20°C and stirred for 3 hours. TLC showed that no raw material remained, and water was added to the reaction solution to quench , extracted with dichloromethane, the organic phase was washed, dried, concentrated, and the crude product was purified by a flash silica gel column to obtain 8-bromo-N-((5-fluoro-2,3-dihydrobenzofuran-4-yl)methyl base) imidazo[1,5-c]pyrimidin-5-amine 17 (4.0 g, 40.3% yield).

¹ H NMR (400MHz, CDCl ₃ )δ8.23(s,1H),7.41-7.30(m,2H),6.87-6.77(m,1H),6.71-6.58(m,1H),6.10(br s, 1H), 4.79-4.71(m, 2H), 4.60(t, J=8.8Hz, 2H), 3.37(t, J=8.8Hz, 2H) ppm; LCMS: m/z 363.0[M+H] ⁺

Step 2: Using the method of Step 10 of Example 2, 8-bromo-N-((5-fluoro-2,3-dihydrobenzofuran-4-yl)methyl)imidazo[1,5-c ]pyrimidin-5-amine (17) and (5-((dimethylamino)methyl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)boronic acid (11 ) as raw material can be obtained: 8-(5-((dimethylamine) methyl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-N-(( 5-fluoro-2,3-dihydrobenzofuran-4-yl)methyl)imidazo[1,5-c]pyrimidin-5-amine (18):

¹ H NMR (400MHz, CDCl ₃ )δ8.22(s,1H),8.03(s,1H),7.78(s,1H),7.71(d,J=7.4Hz,1H),7.30-7.27(m, 1H), 7.07(s, 1H), 6.87-6.72(m, 1H), 6.65(dd, J=3.9, 8.6Hz, 1H), 4.82(s, 2H), 4.60(t, J=8.8Hz, 2H ), 4.16(s, 2H), 3.67(t, J=6.0Hz, 1H), 3.43(br t, J=8.6Hz, 2H), 2.52(s, 6H)) ppm; LCMS: m/z 459.2[ M+H] ⁺

Step 3: 8-(5-((dimethylamine)methyl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-N-((5-fluoro -2,3-Dihydrobenzofuran-4-yl)methyl)-1-iodoimidazo[1,5-c]pyrimidin-5-amine (19)

8-(5-((dimethylamine)methyl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-N-((5-fluoro-2 ,3-dihydrobenzofuran-4-yl)methyl)imidazo[1,5-c]pyrimidin-5-amine (400mg, 0.782mmol) was dissolved in acetic acid (2ml), and N-iodine was added at 0°C Subsuccinimide (137mg, 0.62mmol), heated to 20°C and reacted for 0.25 hours, TLC and LCMS monitored the completion of the reaction, adjusted the pH of the reaction solution to 7, extracted with ethyl acetate, washed the organic phase, dried and concentrated, The crude product was purified by flash silica gel column to obtain 8-(5-((dimethylamine)methyl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-N-( (5-Fluoro-2,3-dihydrobenzofuran-4-yl)methyl)-1-iodoimidazo[1,5-c]pyrimidin-5-amine (190 mg, 33.9% yield).

¹ H NMR (400MHz,DMSO-d ₆ )δ8.73(s,1H),8.53-8.47(m,1H),8.47-8.43(m,1H),7.64(d,J=7.3Hz,1H), 7.33-7.26(m, 2H), 6.95(t, J=9.4Hz, 1H), 6.74-6.66(m, 1H), 4.74(d, J=4.9Hz, 2H), 4.55(t, J=8.8Hz ,2H), 4.03(s,2H), 3.30(br s,2H), 2.35(s,6H)ppm; LCMS: m/z 585.5[M+H] ⁺

Step 4: 8-(5-((dimethylamine)methyl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-N-((5-fluoro -2,3-Dihydrobenzofuran-4-yl)methyl)-1-(methylsulfonyl)imidazo[1,5-c]pyrimidin-5-amine (BR-4)

8-(5-((dimethylamine)methyl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-N-((5-fluoro-2 ,3-dihydrobenzofuran-4-yl)methyl)-1-iodoimidazo[1,5-c]pyrimidin-5-amine (100mg, 0.172mmol), cuprous iodide (98mg, 0.513 mmol), sodium methyl sulfinate (52.4mg, 0.513mmol) was dissolved in dimethyl sulfoxide (1ml), nitrogen was blown for 5 minutes, microwaved at 120°C for 20 minutes, and then lowered to 100°C for 3 hours to monitor the reaction by LCMS At the end, the reaction solution was concentrated, and the crude product was purified by Pre-HPLC to obtain 8-(5-((dimethylamine)methyl)-[1,2,4]triazolo[1,5-a]pyridine-8 -yl)-N-((5-fluoro-2,3-dihydrobenzofuran-4-yl)methyl)-1-(methylsulfonyl)imidazo[1,5-c]pyrimidine-5- Amine (30 mg, 32.3% yield).

¹ H NMR (400MHz,DMSO-d ₆ )δ8.94(s,1H),8.89-8.80(m,1H),8.36(br s,1H),7.66-7.56(m,2H),7.26-7.15( m,1H),6.96(t,J=9.4Hz,1H),6.75-6.67(m,1H),4.82-4.72(m,2H),4.56(t,J=8.7Hz,2H),3.99(br s,2H),3.31-3.31(m,2H),2.87(s,3H),2.33(br s,6H)ppm; LCMS: m/z 537.2[M+H] ⁺ .

Example 6

According to the method of Example 5, replace with (5-(2-hydroxypropan-2-yl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)boronic acid (12) (5-((Dimethylamino)methyl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)boronic acid (11) can give the following compound:

2-(8-(5-(((5-fluoro-2,3-dihydrobenzofuran-4-yl)methyl)amino)-1-(methylsulfonyl)imidazo[1,5-c ]pyrimidin-8-yl)-[1,2,4]triazolo[1,5-a]pyridin-5-yl)propan-2-ol (BR-5) ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.94(s,1H),8.88-8.76(m,1H),8.38(s,1H),7.65-7.57(m,2H),7.34(d,J＝7.4Hz,1H),6.96( t,J=9.4Hz,1H),6.80-6.62(m,1H),5.82(s,1H),4.85-4.68(m,2H),4.56(t,J=8.7Hz,2H),3.32-3.31 (m,2H),2.88(s,3H),1.76(s,6H)ppm; LCMS: m/z 538.2[M+H] ⁺

Pharmacology and Application

As one of the main components of the PRC2 protein complex, EED plays an important role in the overall function of PRC2 although it does not have enzymatic activity. The effect of EED on PRC2 is specifically manifested in two aspects: 1) EED directly binds to trimethylated H3K27Me3, which can locate the PCR2 complex on the chromatin that needs to be modified; 2) EED has a positive effect on the enzymatic function of EZH2. Great allosteric boost. Therefore, the development of compounds that are targets of EED as an allosteric protein provides a new strategy for providing inhibition of EZH2 enzymatic activity. Moreover, such inhibitors have better or complementary advantages than EZH2 enzyme catalytic site inhibitors. For example, when patients develop resistance to EZH2 enzyme inhibitors, EED inhibitors can also inhibit EZH2 enzyme activity. The invention discloses that the compound can be used as an EED target inhibitor, and has a therapeutic effect on diseases related to the mechanism of action of EED and/or PRC2.

The biological functions of the compounds disclosed in the present invention have been proved in biochemical and cell level tests. For example, in a biochemical test, the compound disclosed in the present invention can have a strong competitive binding effect with the H3K27Me3 polypeptide binding to the EED protein (IC ₅₀ can reach <1nM). At the cellular level, the compounds disclosed in the present invention can not only inhibit the methylation level of histone H3K27 but also inhibit the proliferation of cancer cells through this effect (IC ₅₀ can reach <1nM).

Example 7

ELISA (H3K27 trimethylation) analysis

Representative compounds of the disclosure were serially diluted in 3-fold in DMSO, and 10 concentration gradients were tested per compound, up to a maximum assay concentration of 10 μΜ. Compounds were diluted 200-fold into G401 cells cultured in 96-well plates (DMSO final concentration 0.5%). After the cells were cultured for 72 hours, the trimethylation level of histone H3K27 was detected by ELISA method.

Histone extraction: Compound-treated cells in 96-well plates were washed with 1x PBS (10x PBS buffer (80g NaCl (Sigma, product number S3014), 2g KCl (Sigma, product number 60128), _14.4g Na2HP04 (Sigma, Product No. S5136), 2.4g KH ₂ P04 (Sigma, Product No. P9791) into 1L water, pH to 7.4), wash three times, add 100 μL 0.4N HCl to each well, place at 4°C, gently shake for 2 hours to lyse the cells. The lysate was then neutralized with 80 μL of neutralization buffer (0.5 M disodium hydrogen phosphate, pH 12.5, 2.5 mM DTT; 1% cocktail (Sigma, product number P8340) (mix the cell lysate and neutralization buffer thoroughly).

ELISA detection method: transfer the cell lysate in parallel to two 384-well detection plates (PerkinElmer, OptiPlate-384HB, product number 6007290), one plate is used to detect the trimethylation level of H3K27, and the other plate is used to measure H3 adjusted to a final volume of 50 μL/well with PBS, and coated overnight at 4°C. The next day, discard the solution in the well, and use TBST buffer (lxTBS (10x TBS: 24.2g Tris (Sigma, product number T6066), 80g NaCl (Sigma, product number S3014) in 1L water, adjust the pH to 7.6 with HCl) ,0.1% Tween-20) washed 5 times, and dried on absorbent paper. Add 70 μL of blocking buffer (TBST, 5% BSA) to the coated reaction wells and incubate at room temperature for 1 hour. Discard the blocking buffer and add the primary antibody (30 μL/well). All required primary antibodies were diluted with blocking buffer, and the dilution ratios were as follows: anti-H3K27me3 antibody (Cell Signaling Technology, product number 9733), 1:2000 dilution; anti-H3 antibody (Cell Signaling Technology, product number 4499), 1:10000 dilution. After adding the primary antibody, incubate at room temperature for 1 hour. After washing with TBST for 5 times, the water was drained, and the secondary antibody (30 μL/well) was added to each reaction well, and incubated at room temperature for 1 hour. The secondary antibody (anti-rabbit antibody (Jackson ImmunoResearch, product number 111-035-003)) was diluted 2000 times with blocking buffer before use. After 1 hour, wash with TBST and dry. Add 30 μL of ECL substrate (Pierce, product number 34080) to each well, and centrifuge at 2000 rpm for 30 seconds. The signal of each sample was detected by using Molecular Devices, SpectraMax. Data processing: H3K27 methylation reads were first normalized by H3 signal, 0.5% DMSO-treated samples were used as a control, and the percentage inhibition of the compound was calculated. The data were fitted to a dose-response curve using the GraphPad prisim5 program to obtain the _IC50 values of the test compounds.

Table 1 below shows the _IC50 values of some compounds.

Table 1

Example 8

Cell Proliferation Assay

Human B-cell non-Hodgkin's lymphoma cells KARPAS-422 S were cultured in culture flasks using standard cell culture conditions. The culture medium is containing 15% fetal bovine serum (FBS, Invitrogen, product number 10099-141), 1% penicillin/streptomycin solution (P/S) RPMI-1640 (Invitrogen, product number 11875), and the culture bottle is placed at temperature Cultured in a sterile incubator at 37°C, 95% relative humidity, 5% _CO2 . In order to detect the effect of PRC2 inhibitors on cell proliferation, cells in the exponential growth phase were seeded into 96-well plates (Corning, product number 3904) at a density of 1×10 ⁴ cells/well, and 100 μL of medium was added to each well. Subsequently, the compounds disclosed in this paper at different concentrations were added to the wells in which the cells had been inoculated (9 concentration gradients were set for each compound, the highest detection concentration was 10 μM, and 3-fold serial dilutions), and 2 wells were set for each treatment concentration. Repeat in parallel with a final DMSO concentration of 0.5%. Afterwards, the number of surviving cells was measured with Vi-CELL (Beckman Coulter) every 3 to 4 days. The cells counted each time were inoculated into a new 96-well plate at the same density (1×10 ⁴ cells/well), supplemented with fresh medium to 100 μL, and added different concentrations of compounds at the same time. After culturing until the 13th day, add 100 μL of CellTiter-Glo (CellTiter-GloCellTiter-GloCellTiter-GloCTG) (Promega, Product No. G7573) to each well, place it in the dark at room temperature for 10-20 minutes, and use Molecular Devices, SpectraMax i3X to read the luminescent signal . The data were fitted to dose-response curves using GraphPad prisim5 to obtain _IC50 values for test compounds.

Table 5 below shows the _IC50 values of some compounds.

Table 2

The compounds disclosed in the present invention above can be used to treat cancers related to the mechanism of action of EED protein and/or PRC2 protein complex, including but not limited to diffuse large B-cell lymphoma, follicular lymphoma, non-Hodgkinson lymphoma, etc. Isolymphoma, leukemia, multiple myeloma, mesothelioma, gastric cancer, malignant rhabdoid tumor, liver cancer, prostate cancer, breast cancer, brain tumors including neuroblastoma, glioma, glioblastoma, and astrocytoma Cytoma, cervical cancer, colon cancer, melanoma, endometrial cancer, esophageal cancer, head and neck cancer, lung cancer, nasopharyngeal cancer, ovarian cancer, pancreatic cancer, kidney cancer, rectal cancer, thyroid cancer, parathyroid tumors, uterus Tumors and soft tissue sarcomas, among others.

Claims

A compound as shown in formula (I), its pharmaceutically acceptable salt, stereoisomer, solvate or their isotope-labeled compound, is characterized in that:

where A is independently

X is N or CR 2 ;

R 1 , R 2 and R 3 are independently H, halogen, cyano, R 5b substituted or unsubstituted amino, R 5b -O-, R 5 substituted or unsubstituted C 1-4 alkyl, R 5 substituted or unsubstituted C 3 - 6 cycloalkyl, R 5 substituted or unsubstituted C 1 - 4 haloalkyl, -C(=O)R 6 , -CO 2 R 6 , -C(=O)NR 6 R 7 , -SO 2 R 6 or -SO 2 NR 6 R 7 ; when there are multiple substituents, they are the same or different;

R 4 is independently H, halogen, cyano, C 1-4 haloalkyl, -C(=O)NR 4a1 R 4a2 , -S(=O) 2 R 4b or -C(=O)R 4c ;

R 4a1 , R 4a2 and R 4b are independently H or C 1-4 alkyl;

R 4c is independently H, C 1-4 alkyl or -OC 1-4 alkyl;

R 5 is independently C 1-4 alkyl-O-, C 1-4 alkyl, C 1-4 haloalkyl, -N(R 5b R 5b ), amino protected by N protecting group, fluorine, hydroxyl;

R 5b , R 6 and R 7 are independently H, C 1-4 alkyl or C 1-4 haloalkyl;

"*" indicates that when the carbon atom with "*" is a chiral carbon atom, it is in R configuration, S configuration or a mixture thereof.
The compound shown in formula (I), its pharmaceutically acceptable salt, stereoisomer, solvate or their isotope-labeled compound as claimed in claim 1, is characterized in that:

The compound shown in the formula (I) is selected from the compounds shown below:

And/or, when R 1 , R 2 , R 3 and R 4 are independently halogen, said halogen is fluorine, chlorine, bromine or iodine;

And/or, when R 1 , R 2 and R 3 are independently R 5 substituted or unsubstituted C 1-4 alkyl, said C 1-4 alkyl is methyl, ethyl, n-propyl , isopropyl, n-butyl, isobutyl or tert-butyl;

And/or, when R 1 , R 2 and R 3 are independently R 5 substituted or unsubstituted C 3-6 cycloalkyl, said C 3-6 cycloalkyl is cyclopropyl, cyclo Butyl, cyclopentyl or cyclohexyl;

And/or, when R 1 , R 2 and R 3 are independently R 5 substituted or unsubstituted C 1-4 haloalkyl, said halogen is fluorine, chlorine, bromine or iodine;

And/or, when R 1 , R 2 and R 3 are independently R 5 substituted or unsubstituted C 1-4 haloalkyl, said C 1-4 alkyl is methyl, ethyl, n-propyl , isopropyl, n-butyl, isobutyl or tert-butyl;

And/or, when R 1 , R 2 and R 3 are independently R 5 substituted or unsubstituted C 1-4 haloalkyl groups, the number of said halogens is 1, 2, 3, 4 or 5 ;

And/or, when R 4 , R 5 , R 5b , R 6 and R 7 are independently C 1-4 haloalkyl, the halogen is fluorine, chlorine, bromine or iodine;

And/or, when R 4 , R 5 , R 5b , R 6 and R 7 are independently C 1-4 haloalkyl, wherein C 1-4 alkyl is methyl, ethyl, n-propyl, iso Propyl, n-butyl, isobutyl or tert-butyl;

And/or, when R 4 , R 5 , R 5b , R 6 and R 7 are independently C 1-4 haloalkyl, the number of said halogen is 1, 2, 3, 4 or 5;

And/or, when R 4a1 , R 4a2 , R 4b , R 5b , R 6 and R 7 are independently C 1-4 alkyl, said C 1-4 alkyl is methyl, ethyl, normal Propyl, isopropyl, n-butyl, isobutyl or tert-butyl;

And/or, when R 4c is independently C 1-4 alkyl or -OC 1-4 alkyl, said C 1-4 alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl;

And/or, when R 5 is independently C 1-4 alkyl, C 1-4 alkyl-O-, C 1-4 haloalkyl, said C 1-4 alkyl, C 1-4 alkane The C 1-4 alkyl in the group -O- and C 1-4 haloalkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl;

And/or, when R 5 is independently an amino group protected by an N protecting group, the protecting group is
R 5a is C 1-4 alkyl or C 1-4 alkyl-O-.
The compound shown in formula (I), its pharmaceutically acceptable salt, stereoisomer, solvate or their isotope-labeled compound as claimed in claim 1 or 2, is characterized in that:

When R 5 is independently an amino group protected by an N protecting group, the protecting group is
When R 5a is C 1-4 alkyl or C 1-4 alkyl-O-, C 1-4 alkyl and C 1-4 alkyl in C 1-4 alkyl-O- are independently methyl , ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl;

And/or, X is N;

And/or, R 1 is selected from methyl, cyano, trifluoromethyl, difluoromethyl,

And/or, R 2 is H;

And/or, R 3 is selected from H, methyl, halogen or cyano;

And/or, R 4 is selected from cyano, -C(=O)NR 4a1 R 4a2 , -S(=O) 2 R 4b or -C(=O)R 4c ;

And/or, R 4a1 , R 4a2 , R 4b , R 4c , R 5b , R 6 and R 7 are independently methyl.
The compound shown as formula (I), its pharmaceutically acceptable salt, stereoisomer, solvate or their isotope-labeled compound as claimed in claim 1, is characterized in that: wherein, said formula ( I) described compound is selected from the compound shown below:

In the above-mentioned compound, the definition of R is as shown in claim 1;

and / or,
for
or a mixture thereof.
The compound represented by formula (I), its pharmaceutically acceptable salt, stereoisomer, solvate or their isotope-labeled compound according to any one of claims 1 to 4, characterized in that: The compound shown in formula (I) is selected from the compounds shown below:
A compound shown in formula (I) as described in any one of claims 1-5, its pharmaceutically acceptable salt, stereoisomer, solvate or their isotope-labeled compound, it is characterized in that: The isotopes are selected from 2 H, 3 H, 11 C, 13 C, 14 C, 15 N, 18 F, 31 P, 32 P, 35 S, 36 Cl and 125 I.
A method for preparing the compound shown in formula (I) as described in any one of claims 1-5, characterized in that: it comprises the steps of:

Halogenated intermediate B 0 and intermediate E 0 are subjected to a coupling reaction to obtain a compound shown in formula (I);

Among them, W represents a halogen, such as Br; R x is -B (OH) 2 or
A, R 1 , R 3 , R 4 , and X are as defined in any one of claims 1-5.
A preparation method of intermediate compound B 0 , characterized in that: it is any of the following schemes:

Option 1 includes the following steps:

Step (1) Compound SM-5 is ring-closed under the action of a catalyst to obtain compound SM-6,

Step (2) Compound SM-6 is reacted under the action of an oxidizing agent to obtain compound SM-7,

Step (3) Reaction of compound SM-7 with H 2 N-CH 2 -A in the presence of a base to obtain compound B 0 ;

Scheme two, it is as shown in step (2) to step (3) in scheme one;

Scheme three, it is as shown in step (3) in scheme one;

The reaction equation is as follows:

Wherein, W, A, R 4 are as defined in any one of claims 1-5.
A pharmaceutical composition, characterized in that: it comprises a therapeutically effective amount of the compound shown in formula (I) as described in any one of claims 1-6, its pharmaceutically acceptable salt, stereoisomer body, its solvate or their isotope-labeled compounds and pharmaceutically acceptable excipients.
A kind of application of substance A in the preparation of medicine, it is characterized in that: described substance A is the compound shown in formula (I) as described in any one in claim 1-6, its pharmaceutically acceptable Salt, stereoisomer, its solvate or their isotope labeling compound or pharmaceutical composition as claimed in claim 9; Described medicine is preferably the medicine for treating cancer; Described medicine is more preferably treatment and EED protein and/or drugs for cancers mediated by PRC2 protein complexes.
The application according to claim 10, characterized in that:

The cancer is selected from diffuse large B cell lymphoma, follicular lymphoma and other lymphoma, leukemia, multiple myeloma, mesothelioma, gastric cancer, malignant rhabdoid tumor, liver cancer, prostate cancer, breast cancer, brain tumor including Neuroblastoma, glioma, glioblastoma and astrocytoma, cervical cancer, colon cancer, melanoma, endometrial cancer, esophageal cancer, head and neck cancer, lung cancer, nasopharyngeal cancer, ovarian cancer, pancreatic cancer Carcinoma, kidney cancer, rectal cancer, thyroid cancer, parathyroid tumor, uterine tumor and soft tissue sarcoma; lymphoma such as diffuse large B-cell lymphoma and follicular lymphoma are preferred;

And/or, the substance A is used in combination with other drugs; preferably, the other drugs are selected from anticancer drugs, tumor immune drugs, antiallergic drugs, antiemetics, analgesics and cell protection drugs.
A pharmaceutical preparation, characterized in that: it comprises the compound shown in formula (I) as described in any one of claims 1-6, its pharmaceutically acceptable salt, stereoisomer, and its solvate Or their isotope-labeled compounds or pharmaceutical compositions as claimed in claim 9, said pharmaceutical preparations are preferably tablets, capsules, pills, powders, granules, elixirs, tinctures, suspensions, syrups, emulsions and solutions; more preferably Preferably, the administration mode of the pharmaceutical preparation is selected from oral administration, sublingual administration, subcutaneous injection, intravenous injection, intramuscular injection, intrasternal injection, nasal administration, local topical administration and rectal administration;

And/or, the pharmaceutical preparation is taken once or multiple times a day.