CN115028631A - Triazolopyrimidine derivatives, pharmaceutical compositions and uses thereof - Google Patents

Abstract

The present disclosure relates to a compound represented by the following general formula, pharmaceutically acceptable salts, enantiomers, diastereomers, atropisomers, racemates, polymorphs, solvents thereof Compounds or isotopically-labeled compounds (including deuterium substituted), pharmaceutical compositions comprising the same and uses thereof. The molecular activity and cellular activity of some compounds according to the present disclosure are significantly higher than that of the positive control compound EED226, and can be used for the preparation of medicaments for the treatment of diseases or conditions mediated by EED and/or PRC2.

Description

Triazolopyrimidine derivatives, pharmaceutical compositions and uses thereof

technical field

The present disclosure relates to the fields of medicinal chemistry and pharmacotherapeutics, and generally relates to a class of triazolopyrimidine compounds, pharmaceutical compositions comprising the same, and uses thereof. In particular, the present disclosure relates to a triazolopyrimidine compound, its pharmaceutically acceptable salts, enantiomers, diastereomers, atropisomers, racemates, polymorphs compounds, solvates or isotopically-labeled compounds (including deuterium substitutions), as well as pharmaceutical compositions and their use in the treatment of diseases or disorders mediated by EED and/or PRC2, in particular neoplastic diseases.

Background technique

Polycomb Repressive Complex 2 (Polycomb Repressive Complex 2) is a core member of the Polycomb family of proteins. It has histone methyltransferase activity and can specifically catalyze the trimethylation of lysine at position 27 of histone H3. Modification (H3K27me3), thereby inhibiting the expression of specific genes. The methyltransferase activity of PRC2 is derived from its catalytic member EZH2. However, EZH2 has no catalytic activity when it exists alone. It needs to form a complex with at least two other members of PRC2, EED and SUZ12, to catalyze methylation modification. Thus, EZH2, EED and SUZ12 are considered to be the core components of the PRC2 complex. Recent studies have found that the core components of PRC2 are overexpressed in various tumor cells, and its abnormal activity is the direct cause of the onset and aggravation of various malignant tumors. At the same time, recent gene sequencing results of lymphoma patients have shown that EZH2 has an activating mutation in germinal center B-cell lymphoma (GCB-DLBCL) patients, and the mutated EZH2 alters the substrate specificity of PRC2, thereby increasing H3K27me3 in cells Level. Down-regulating the expression of EZH2 or other core components by siRNA method will significantly inhibit the proliferation of lymphoma cells, which indicates that the occurrence and development of GCB-DLBCL is closely related to the over-activation of PRC2. Therefore, PRC2 is a very promising target for anticancer drug development, and the discovery of inhibitors targeting PRC2 is currently a hot research topic in the pharmaceutical industry. Recently, two major pharmaceutical companies, Novartis and AbbVie, have invented a class of small molecules that inhibit the activity of PRC2 by targeting EED (reference: Novartis' EED226, US2016/0176882, J.Med.Chem.2017,60,2215 -2226, J.Med.Chem.2017,60,415-427, Nat.Chem.Biol.2017,13,381-388; AbbVie's A-395, Nat.Chem.Biol.2017,13,389-395), the class The compounds showed strong inhibitory activities at the molecular level, cellular level and animal experiments. In summary, the PRC2 complex is considered to be a key driver of the occurrence and development of various malignant tumors, and the development of inhibitors that inhibit the activity of PRC2 by targeting EED is currently very hot in the industry, which is beneficial for related new drug development.

SUMMARY OF THE INVENTION

An object of the present disclosure is to provide a triazolopyrimidine compound represented by the general formula I, its pharmaceutically acceptable salts, enantiomers, diastereomers, atropisomers, racemates form, polymorph, solvate or isotopically labeled compound,

wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , M, Y, Z and n are as defined herein.

Another object of the present disclosure is to provide a preparation method of the above compound.

Yet another object of the present disclosure is to provide a compound comprising a therapeutically effective amount of one or more of the above compounds or their pharmaceutically acceptable salts, enantiomers, diastereomers, atropisomers, exoisomers Pharmaceutical compositions of racemates, polymorphs, solvates or isotopically labeled compounds.

Yet another object of the present disclosure is to provide the above-mentioned compounds or their pharmaceutically acceptable salts, enantiomers, diastereomers, atropisomers, racemates, polymorphs, solvates or the use of an isotopically labeled compound in the manufacture of a medicament for the treatment of a disease or condition mediated by PRC2.

Yet another object of the present disclosure is to provide a method of treating a disease or condition mediated by EED and/or PRC2, characterized by administering to a subject a therapeutically effective amount of one or more of the above-mentioned compounds or medicaments thereof The salts, enantiomers, diastereomers, atropisomers, racemates, polymorphs, solvates or isotopically labeled compounds used.

According to one aspect of the present disclosure, there is provided a compound represented by general formula I, pharmaceutically acceptable salts, enantiomers, diastereomers, atropisomers, racemates thereof , polymorphs, solvates or isotopically labeled compounds:

in,

为单键或双键；(1)

is a single bond or a double bond;

(2) R ¹ , R ² and R ³ are each independently hydrogen, halogen, C ₁ -C ₄ alkyl, C ₁ -C ₄ haloalkyl, -O-(C ₁ -C ₄ alkyl), C ₁ -C ₄ haloalkoxy or 3-6 membered cycloalkyl;

R ⁴ is hydrogen, OH, =O, or C ₁ -C ₄ alkyl;

R ⁵ is hydrogen, halogen, C ₁ -C ₄ alkyl, or C ₁ -C ₄ haloalkyl;

(3) M ring is a 6-10-membered aromatic ring or a 5-10-membered heteroaromatic ring, and M is substituted by 0-4 R ⁶ ;

R ⁶ is independently selected from halogen, CN, C ₁ -C ₆ alkyl substituted with 0-1 R ^A1 , 3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl, -OR ^A2 , -C (=O)R ^A3 , -NR ^A4 R ^A5 , -NHC(=O)R ^A3 , -S(=O) ₂ R ^A3 , -S(=O) ₂ NR ^A4 R ^A5 , -NHS(=O) ₂ (C ₁ -C ₄ alkyl), and -C(=O)NR ^A4 R ^A5 ;

R ^A1 is selected from C ₁ -C ₆ alkyl, 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, OH, -NR ^a R ^b , C ₁ -C ₄ alkoxy, -C(= O) NR ^a R ^b , -S(=O) ₂ (C ₁ -C ₄ alkyl), and -NHC(=O) (C ₁ -C ₄ alkyl);

R ^A2 is selected from hydrogen, C ₁ -C ₄ alkyl, 3-6 membered cycloalkyl, and C ₁ -C ₄ haloalkyl;

R ^A3 at each occurrence is independently selected from C ₁ -C ₄ alkyl, 3-6 membered cycloalkyl, 6-10 membered aryl, and 5-10 membered heteroaryl;

R ^A4 and R ^A5 are at each occurrence independently selected from hydrogen, C ₁ -C ₄ alkyl and 3-6 membered cycloalkyl;

R ^a and R ^b are at each occurrence independently selected from hydrogen, C ₁ -C ₄ alkyl, and 3-6 membered cycloalkyl;

(4) Y is -C(O)-, -SO ₂ -, -C(R ^c )(R ^d )-, or -S(O)-;

(5) each Z is independently -O-, -N(R ^A4 )-, -S- or -C(R ^c )(R ^d )-;

^Rc and ^Rd are at each occurrence independently selected from hydrogen, halogen, OH, _C1 - _C4 alkyl, _C1 - _C4 haloalkyl, _C1 - _C4 alkoxy, and _C1 -C ₄ haloalkoxy; or R ^c and R ^d are attached to each other to form -R ^c' -LR ^d' - and to the carbon atom attached to it to form a ring, wherein L is absent or -O-; R ^c ' and R ^d ' are each independently absent, C ₁ -C ₄ alkylene, C ₁ -C ₃ haloalkylene, or replaced by 1-2 halogens, OH, or C ₁ -C ₄ alkyl Substituted C ₁ -C ₃ alkylene;

N is an integer selected from 1-4.

According to another aspect of the present disclosure, there is provided a preparation method of the compound, wherein the method is one of the following methods:

method one:

The lactone analogs were prepared according to the route described in Reaction Formula 1. Specifically, starting from A-1, suzuki reaction with boronate A-2 was carried out to generate A-3, A-3 was hydrolyzed to obtain acid A-4, and then reacted with boronate A-2. A-5 undergoes a Yamaguchi lactonization reaction to form a lactone A-6, wherein the definitions of R ¹ , R ⁶ , M, Z and n are consistent with the above;

Method Two:

The lactam analogs were prepared according to the approach described in Reaction Formula 2. Specifically, starting from A-1, suzuki reaction with Boc-protected amino boronate ester A-7 occurred to generate A-8, and then Boc was removed to obtain A-9, Finally, A-10 is obtained by intramolecular amine transesterification under the action of potassium tert-butoxide, wherein, the definitions of R ¹ , R ⁶ , R ^A4 , M, Z and n are the same as the above;

Method three:

The lactam analogs were prepared according to the route described in the reaction formula 3. Specifically, starting from A-1, suzuki reaction with boronate ester A-11 was carried out to generate A-12, and then the molecular lactone was condensed under the condition of tert-butanol to obtain A-13, A-14 is finally obtained under the condition of trifluoroacetic acid, wherein the definitions of R ¹ , R ⁶ , M, Z and n are the same as the above.

According to another aspect of the present disclosure, there is provided a pharmaceutical composition comprising a compound selected from the group consisting of the above-mentioned compounds, pharmaceutically acceptable salts thereof, enantiomers, diastereomers, atrop One or more of an isomer, racemate, polymorph, solvate, or isotopically-labeled compound, and at least one pharmaceutically acceptable carrier, diluent, or excipient.

According to another aspect of the present disclosure, there is provided the above compound, pharmaceutically acceptable salts, enantiomers, diastereomers, atropisomers, racemates, polymorphs thereof , solvate or isotopically-labeled compound or pharmaceutical composition as described above in the manufacture of a medicament for the treatment of a disease or condition mediated by EED and/or PRC2.

beneficial effect

The molecular activity and cellular activity of some compounds according to the present disclosure are significantly higher than that of the positive control compound EED226, and can be used to prepare new drugs for the treatment of diseases or conditions mediated by EED and/or PRC2.

Detailed ways

In order for those with ordinary knowledge in the art to understand the features and effects of the present invention, general descriptions and definitions of terms and terms mentioned in the specification and the scope of the patent application are provided below. Unless otherwise specified, all technical and scientific terms used herein have the ordinary meanings understood by those skilled in the art to the present invention, and in case of conflict, the definitions in this specification shall prevail.

As used herein, the terms "comprising", "including", "having", "containing" or any other similar terms are open-ended transitional phrases intended to cover non-exclusive inclusions. For example, a composition or article of manufacture containing a plurality of elements is not limited to only those elements listed herein, but can also include other elements not expressly listed, but which are generally inherent to the composition or article of manufacture. Otherwise, unless expressly stated to the contrary, the term "or" refers to an inclusive "or" rather than an exclusive "or". For example, the condition "A or B" is satisfied by any of the following: A is true (or present) and B is false (or absent), A is false (or absent) and B is true (or present), Both A and B are true (or exist). In addition, in this document, the terms "comprising", "including", "having", "containing" should be interpreted as having specifically disclosed and encompassing both "consisting of" and "substantially consisting of" and other closures Formal or semi-closed connectives.

In this document, all features or conditions defined as numerical ranges or percentage ranges are for brevity and convenience only. Accordingly, the description of numerical ranges or percentage ranges should be considered to encompass and specifically disclose all possible sub-ranges and individual numerical values within the range, particularly integer numerical values. For example, a range description of "1 to 8" should be deemed to have specifically disclosed all subranges such as 1 to 7, 2 to 8, 2 to 6, 3 to 6, 4 to 8, 3 to 8, etc., particularly are sub-ranges bounded by all integer values, and should be deemed to have specifically disclosed individual values such as 1, 2, 3, 4, 5, 6, 7, 8, etc. within the range. Unless otherwise indicated, the foregoing method of interpretation applies to all content throughout this disclosure, whether broad or not.

If a quantity or other value or parameter is expressed as a range, a preferred range, or a series of upper and lower limits, it should be understood that any upper or preferred value of the range and the lower limit of the range have been specifically disclosed herein or all ranges of preferred values, whether or not those ranges are disclosed separately. Furthermore, when a range of values is referred to herein, unless otherwise indicated, the range shall include its endpoints and all integers and fractions within the range.

As used herein, numerical values should be understood to have an accuracy of significant digits of the numerical value, provided that the purpose of the invention can be achieved. For example, the number 40.0 should be understood to cover the range from 39.50 to 40.49.

Where Markush group or alternative terminology is used herein to describe features or examples of the invention, those skilled in the art will understand the Markush group or subgroups of all elements within a list of options Groups or any individual element may also be used to describe the invention. For example, if X is described as "selected from the group consisting of X ₁ , X ₂ and X ₃ ", it also means that the claim that X is X ₁ and that X is X ₁ and/or X ₂ have been fully described claim. Furthermore, where Markush group or option terminology is used to describe features or instances of the invention, those skilled in the art will recognize subgroups or individual elements of all elements within the Markush group or option list Any combination of , can also be used to describe the invention. Accordingly, for example, if X is described as "selected from the group consisting of X ₁ , X ₂ and X ₃ " and Y is described as "selected from the group consisting of Y ₁ , Y ₂ and Y ₃ group of "," means that the claim that X is X ₁ or X ₂ or X ₃ and Y is Y ₁ or Y ₂ or Y ₃ has been fully described.

The following detailed description is merely exemplary in nature and is not intended to limit the invention and its uses. Furthermore, there is no intention to be bound by any theory presented in the preceding prior art or this summary or in the following detailed description or examples.

According to one embodiment of the present disclosure, there is provided a compound represented by general formula I, pharmaceutically acceptable salts, enantiomers, diastereomers, atropisomers, racemates thereof Forms, polymorphs, solvates or isotopically labeled compounds:

in,

为单键或双键；(1)

is a single bond or a double bond;

(2) R ¹ , R ² and R ³ are each independently hydrogen, halogen, C ₁ -C ₄ alkyl, C ₁ -C ₄ haloalkyl, -O-(C ₁ -C ₄ alkyl), C ₁ -C ₄ haloalkoxy or 3-6 membered cycloalkyl;

R ⁴ is hydrogen, OH, =O or C ₁ -C ₄ alkyl;

R ⁵ is hydrogen, halogen, C ₁ -C ₄ alkyl or C ₁ -C ₄ haloalkyl;

(3) M ring is a 6-10-membered aromatic ring or a 5-10-membered heteroaromatic ring, and M is substituted by 0-4 R ⁶ ;

R ⁶ is independently selected from halogen, CN, C ₁ -C ₆ alkyl substituted with 0-1 R ^A1 , 3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl, -OR ^A2 , -C (=O)R ^A3 , -NR ^A4 R ^A5 , -NHC(=O)R ^A3 , -S(=O) ₂ R ^A3 , -S(=O) ₂ NR ^A4 R ^A5 , -NHS(=O) ₂ (C ₁ -C ₄ alkyl) and -C(=O)NR ^A4 R ^A5 ;

R ^A1 is selected from C ₁ -C ₆ alkyl, 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, OH, -NR ^a R ^b , C ₁ -C ₄ alkoxy, -C(= O) NR ^a R ^b , -S(=O) ₂ (C ₁ -C ₄ alkyl) and -NHC(=O) (C ₁ -C ₄ alkyl);

R ^A2 is selected from hydrogen, C ₁ -C ₄ alkyl, 3-6 membered cycloalkyl and C ₁ -C ₄ haloalkyl;

R ^A3 at each occurrence is independently selected from C ₁ -C ₄ alkyl, 3-6 membered cycloalkyl, 6-10 membered aryl and 5-10 membered heteroaryl;

R ^A4 and R ^A5 are at each occurrence independently selected from hydrogen, C ₁ -C ₄ alkyl and 3-6 membered cycloalkyl;

R ^a and R ^b are at each occurrence independently selected from hydrogen, C ₁ -C ₄ alkyl, and 3-6 membered cycloalkyl;

(4) Y is -C(O)-, -SO ₂ -, -C(R ^c )(R ^d )- or -S(O)-;

(5) each Z is independently -O-, -N(R ^A4 )-, -S- or -C(R ^c )(R ^d )-;

^Rc and ^Rd are at each occurrence independently selected from hydrogen, halogen, OH, _C1 -C4alkyl, _C1 - _C4haloalkyl , _C1 - _C4alkoxy , and _{C1 - C4} haloalkoxy; or R ^c and R ^d are attached to each other to form -R ^c' -LR ^d' - and to the carbon atom to which they are attached to form a ring, where L is absent or -O-; R ^c' and R ^d' are each independently absent, C ₁ -C ₄ alkylene, C ₁ -C ₃ haloalkylene, or substituted with 1-2 halo, OH, or C ₁ -C ₄ alkyl C ₁ -C ₃ alkylene;

N is an integer selected from 1-4.

According to another embodiment of the present disclosure, the compound represented by the general formula I is selected from the compounds represented by the general formula I-1:

in,

(1) R ¹ is hydrogen, halogen, C ₁ -C ₄ alkyl, C ₁ -C ₄ haloalkyl or 3-6 membered cycloalkyl;

(2) R ⁴ is hydrogen, OH or =O;

(3) M ring is a benzene ring or a 5-membered heteroaromatic ring, and M is substituted by 0-2 R ⁶ ;

R ⁶ is independently selected from halogen, C ₁ -C ₆ alkyl substituted with 0-1 R ^A1 , 3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl, -OR ^A2 , -S(= O) ₂ R ^A3 , -S(=O) ₂ NR ^A4 R ^A5 and -C(=O)NR ^A4 R ^A5 ;

R ^A1 is selected from C ₁ -C ₆ alkyl, 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, OH and -NR ^a R ^b ;

R ^A2 is selected from hydrogen, C ₁ -C ₄ alkyl and 3-6 membered cycloalkyl;

R ^A3 is selected from C ₁ -C ₄ alkyl and 3-6 membered cycloalkyl;

R ^A4 and R ^A5 are at each occurrence independently selected from hydrogen, C ₁ -C ₄ alkyl and 3-6 membered cycloalkyl;

R ^a and R ^b are at each occurrence independently selected from hydrogen, C ₁ -C ₄ alkyl and 3-6 membered cycloalkyl;

(4) Y is -C(O)- or -SO ₂ -;

(5) each Z is independently -O-, -N(R ^A4 )-, -S- or -C(R ^c )(R ^d )-;

^Rc and ^Rd are at each occurrence independently selected from hydrogen, halogen, OH, _C1 - _C4 alkyl, _C1 - _C4 haloalkyl, _C1 - _C4 alkoxy, and _C1 -C ₄ haloalkoxy; or R ^c and R ^d are attached to each other to form -R ^c' -LR ^d' - and to the carbon atom attached to it to form a ring, wherein L is absent or -O-; R ^{c '} and R ^d' are each independently absent, C ₁ -C ₄ alkylene, C ₁ -C ₃ haloalkylene, or replaced by 1-2 halogens, OH, or C ₁ -C ₄ alkane substituted C ₁ -C ₃ alkylene;

n is an integer selected from 1-3.

According to another embodiment of the present disclosure, the compound represented by the general formula I is selected from the compounds represented by the general formula I-2:

in,

(1) R ¹ is hydrogen or halogen;

(2) M ring is a benzene ring, and M is unsubstituted or substituted by 1-2 R ⁶ ;

R ⁶ is independently selected from halogen, C ₁ -C ₆ alkyl substituted with 0-1 R ^A1 , 3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl, -OR ^A2 , -S(= O) ₂ R ^A3 , -S(=O) ₂ NR ^A4 R ^A5 and -C(=O)NR ^A4 R ^A5 ;

R ^A1 is selected from C ₁ -C ₆ alkyl, 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, OH and -NR ^a R ^b ;

R ^A2 is selected from hydrogen, C ₁ -C ₄ alkyl and 3-6 membered cycloalkyl;

R ^A3 is selected from C ₁ -C ₄ alkyl and 3-6 membered cycloalkyl;

R ^A4 and R ^A5 are at each occurrence independently selected from hydrogen, C ₁ -C ₄ alkyl and 3-6 membered cycloalkyl;

R ^a and R ^b are at each occurrence independently selected from hydrogen, C ₁ -C ₄ alkyl and 3-6 membered cycloalkyl;

(3) each Z is independently -O-, -N(R ^A4 )- or -C(R ^c )(R ^d )-;

R ^c and R ^d are independently selected at each occurrence from hydrogen, halogen, OH, C ₁ -C ₄ alkyl and C ₁ -C ₄ haloalkyl;

n is 1 or 2.

According to another embodiment of the present disclosure, the compound represented by the general formula I is selected from the compounds represented by the general formula I-3a and I-3b:

in,

(1) each Z is independently -O- or -N(R ^A4 )-;

R ^A4 is selected from hydrogen, C ₁ -C ₄ alkyl and 3-6 membered cycloalkyl;

(2) A ¹ , A ² , A ³ and A ⁴ independently represent N or CR ^x ;

R ^x is independently selected from hydrogen, halogen, C ₁ -C ₄ alkyl, 3-6 membered cycloalkyl, -S(=O) ₂ R ^A3 , -S(=O) ₂ NR ^A4 R ^A5 and -C (=O)NR ^A4 R ^A5 ;

R ^A3 is selected from C ₁ -C ₄ alkyl and 3-6 membered cycloalkyl;

R ^A4 and R ^A5 are independently selected from hydrogen, C ₁ -C ₄ alkyl and 3-6 membered cycloalkyl.

According to another embodiment of the present disclosure, the compound represented by the general formula I is selected from the following compounds:

In this disclosure,

Halogen means fluorine, chlorine, bromine or iodine.

Alkyl refers to a linear or branched monovalent saturated hydrocarbon group; for example, methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tert-butyl, and the like.

Cycloalkyl refers to a cyclic monovalent saturated hydrocarbon group; for example, cyclopropyl, cyclobutyl, cyclopentyl, and the like.

Heterocycloalkyl refers to a cyclic monovalent saturated group containing atoms selected from oxygen, nitrogen, and sulfur in the ring; for example, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothienyl, and the like.

Aromatic ring refers to aromatic hydrocarbons such as benzene, naphthalene, anthracene, and the like.

The heteroaromatic ring refers to an aromatic ring containing an atom selected from oxygen, nitrogen and sulfur in the ring, such as furan, pyrrole, thiophene, imidazole and the like.

For example, the numbers in "3-6 yuan" and "6-10 yuan" represent the number of ring atoms.

Alkylene refers to a linear or branched divalent saturated hydrocarbon group; for example, methylene, ethylene, 1,3-propylene, 1,2-propylene, 1,4-butylene, 1 , 3-Butylene etc.

Isotopically-labeled compounds are, for example, deuterium-substituted compounds.

According to one embodiment of the present disclosure, there is provided a pharmaceutical composition comprising a compound selected from the group consisting of the above-mentioned compounds, their pharmaceutically acceptable salts, enantiomers, diastereomers, atrop One or more of an isomer, racemate, polymorph, solvate, or isotopically-labeled compound, and at least one pharmaceutically acceptable carrier, diluent, or excipient.

According to another embodiment of the present disclosure, wherein the pharmaceutical composition may further comprise at least one additional therapeutic agent.

According to another embodiment of the present disclosure, wherein the additional therapeutic agent is selected from other anticancer agents, immunomodulatory agents, antiallergic agents, antiemetic agents, pain relief agents, cytoprotective agents, and combinations thereof. Here, the other anticancer agent refers to compounds other than the above-mentioned compounds of the present disclosure, their pharmaceutically acceptable salts, enantiomers, diastereomers, atropisomers, racemates, polyisomers Anticancer agents other than crystalline forms, solvates and isotopically labeled compounds.

According to one embodiment of the present disclosure, there is provided the compound, pharmaceutically acceptable salts, enantiomers, diastereomers, atropisomers, racemates, polymorphs thereof , solvate or isotopically labeled compound, or the use of said pharmaceutical composition in the manufacture of a medicament for the treatment of a disease or condition mediated by EED and/or PRC2.

According to another embodiment of the present disclosure, wherein the disease or disorder mediated by EED and/or PRC2 comprises diffuse large B-cell lymphoma, follicular lymphoma, leukemia, multiple myeloma, mesothelioma , gastric cancer, malignant rhabdoid tumor, hepatocellular carcinoma, prostate cancer, breast cancer, bile duct and gallbladder cancer, bladder cancer; including neuroblastoma, schwannoma, glioma, glioblastoma and astrocytes cancer; cervical cancer, colon cancer, melanoma, endometrial cancer, esophageal cancer, head and neck cancer, lung cancer, nasopharyngeal cancer, ovarian cancer, pancreatic cancer, renal cell cancer, rectal cancer, thyroid cancer, parathyroid tumor, uterus Tumors and soft tissue sarcomas.

According to an embodiment of the present disclosure, there is provided a preparation method of the compound, wherein the method is one of the following methods:

method one:

The lactone analogs were prepared according to the route described in Reaction Formula 1. Specifically, starting from A-1, suzuki reaction with boronate A-2 was carried out to generate A-3, A-3 was hydrolyzed to obtain acid A-4, and then reacted with boronate A-2. A-5 undergoes Yamaguchi lactonization reaction to generate lactone A-6, wherein the definitions of R ¹ , R ⁶ , M, Z or n are the same as the above;

Method Two:

The lactam analogs were prepared according to the approach described in Reaction Formula 2. Specifically, starting from A-1, suzuki reaction with Boc-protected amino boronate ester A-7 occurred to generate A-8, and then Boc was removed to obtain A-9, Finally, A-10 is obtained by intramolecular amine transesterification under the action of potassium tert-butoxide, wherein the definitions of R ¹ , R ⁶ , R ^A4 , M, Z and n are the same as the above;

Method three:

The lactam analogs were prepared according to the route described in the reaction formula 3. Specifically, starting from A-1, suzuki reaction with boronate ester A-11 was carried out to generate A-12, and then the molecular lactone was condensed under the condition of tert-butanol to obtain A-13, A-14 is finally obtained under the condition of trifluoroacetic acid, wherein the definitions of R ¹ , R ⁶ , M, Z and n are the same as the above.

For the preparation of compound A-1, see CN109790166A, or it can be prepared according to conventional methods known in the art.

Optimal reaction conditions and reaction times for each individual step can vary depending on the particular reactants used and the substituents present in all reactants. Unless otherwise specified, solvents, temperatures and other reaction conditions can be readily selected by those skilled in the art. Specific procedures are provided in the Synthetic Examples section. The reaction can be further processed in a conventional manner, eg, by removing the solvent from the residue and further purifying according to methods generally known in the art such as, but not limited to, crystallization, distillation, extraction, trituration, and chromatography. Unless otherwise stated, starting materials and reactants are commercially available or can be prepared by one skilled in the art from commercially available materials using methods described in the chemical literature.

Starting materials, if not commercially available, can be prepared by procedures selected from standard organic chemistry techniques, techniques analogous to the synthesis of analogs of known structures, or techniques analogous to those described in the Schemes or Synthesis Examples section above . When an optically active form of a compound of the present disclosure is desired, it can be obtained by performing one of the steps described herein using an optically active starting material (eg, by asymmetric induction of an appropriate reaction procedure), or by using standard procedures (eg, chromatographic separation) , recrystallization or enzymatic resolution) to obtain a mixture of stereoisomers from compounds or intermediates.

Similarly, when a pure geometric isomer of a compound of the present disclosure is desired, it can be obtained by performing one of the above steps using the pure geometric isomer as the starting material, or by resolving the compound or intermediate using standard procedures such as chromatographic separation obtained as a mixture of geometric isomers.

When preparing the polymorphs of the compounds of the present disclosure, crystals are obtained by adopting a recrystallization method, and their crystal structures are obtained according to X-ray diffraction.

In the preparation of isotopically-labeled compounds of the compounds of the present disclosure, isotopically-labeled starting materials can be used for the reaction as desired.

Example

The following examples were prepared, isolated and characterized using the methods disclosed herein. The following examples illustrate some of the scope of the disclosure, but do not imply the entire scope of the disclosure.

Example 1 Synthesis of ZB-PY-E18

To compound 1 (100 mg, 0.230 mmol), (2-aminophenyl)boronic acid (63.0 mg, 0.460 mmol) and potassium carbonate (K ₂ CO ₃ , 95.2 mg, 0.690 mmol) in 1,4-dioxane ( A mixture of 5 mL) and water (1 mL) was bubbled with nitrogen for 1 min, Pd(dppf)Cl ₂ (17.4 mg, 0.0230 mmol) was added and the sealed tube was heated in a microwave (MW) reactor at 125 °C for 1 h . Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate (unless explicitly stated otherwise, the drying conditions were the same in the following examples), concentrated, and purified by silica gel column chromatography (8% methanol in dichloromethane as eluent) ( Unless explicitly stated otherwise, silica gel column chromatography conditions were the same in the following examples) to give ZB-PY-E18 (26.0 mg, 28%) as a yellow solid. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 9.51 (s, 1H), 8.67 (s, 1H), 8.53 (t, J=5.1 Hz, 1H), 7.98 (s, 1H), 7.84 (d,J＝8.1Hz,1H),7.15-7.18(m,2H),7.02(ddd,J＝8.1,6.1,2.1Hz,1H),6.95(dd,J＝10.3,8.7Hz,1H), 6.71(dd,J＝8.7,3.8Hz,1H),4.72(d,J＝5.1Hz,2H),4.55(t,J＝8.8Hz,2H),3.30(t,J＝8.8Hz,2H). LCMS: [M+H] ⁺ =402.2.

Example 2 Synthesis of ZB-PY-E14.

To compound 1 (100 mg, 0.230 mmol), (2-aminopyridin-3-yl)boronic acid (95.2 mg, 0.690 mmol) and potassium carbonate (95.2 mg, 0.690 mmol) in 1,4-dioxane (5 mL) Nitrogen was bubbled through a mixed solvent of water (1 mL) for 1 min, PdCl ₂ (dppf) (17.4 mg, 0.0230 mmol) was added and the sealed tube was heated at 125° C. for 1 h in a microwave reactor. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated, and purified by silica gel column chromatography to give ZB-PY-E14 (17.0 mg, 18.4%) as a white solid. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 9.30 (s, 1H), 8.70-8.65 (m, 2H), 8.32 (d, J=7.8 Hz, 1H), 8.19 (dd, J= 4.6,1.6Hz,1H),8.07(s,1H),7.09(dd,J=7.8,4.6Hz,1H),6.95(dd,J=10.3,8.6Hz,1H),6.71(dd,J=8.6 ,3.8Hz,1H),4.73(d,J＝4.9Hz,2H),4.55(t,J＝8.8Hz,2H),3.30(t,J＝8.8Hz,2H).LCMS:[M+H] ⁺ = 403.2.

Example 3 Synthesis of ZB-PY-E16

Synthesis of compound 3.2: compound 1 (100 mg, 0.230 mmol), (2-(hydroxymethyl)phenyl)boronic acid (105 mg, 0.690 mmol), tetrakis(triphenylphosphine)palladium (Pd(PPh ₃ ) ₄ , 26.6 mg, 0.0230 mmol) and potassium carbonate (95.2 mg, 0.690 mmol) were added to a mixed solvent of 1,4-dioxane (5 mL) and water (1 mL) to replace nitrogen and heated at 110° C. for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated, and subjected to column chromatography on silica gel (5% methanol in dichloromethane as eluent) to give the title compound 3.2 (60.0 mg, 56%) as a pale yellow solid. LCMS: [M+H] ⁺ =463.1.

Synthesis of compound 3.3: A mixture of compound 3.2 (60.0 mg, 0.130 mmol) and lithium hydroxide (LiOH, 62.3 mg, 2.60 mmol) in tetrahydrofuran (THF, 7 mL) and water (3 mL) was heated to 60 °C for 12 hours . The pH was adjusted to 2-3 with 3N aqueous hydrochloric acid at 0°C. The reaction was extracted with ethyl acetate. The organic phase was washed with brine, dried, and concentrated to obtain the title compound 3.3 (45 mg, 80%) as a white solid. LCMS: [M+H] ⁺ =435.0.

Synthesis of ZB-PY-E16: Triethylamine (TEA, 62.4 mg, 0.618 mmol) and 2,4,6-trichlorobenzoyl chloride (125 mg, 0.517 mmol) were added to compound 3.3 (45.0 mg, 0.103 mmol) In toluene (5 mL) solution, the reaction was carried out at 25 °C for 4 hours. Then, the reaction solution was diluted with toluene (5 mL) and slowly added dropwise to a solution of 4-dimethylaminopyridine (DMAP, 17.7 mg, 0.144 mmol) in toluene (10 mL) at 80° C. for 4 hours. The reaction was cooled, quenched with saturated sodium bicarbonate solution, and extracted with ethyl acetate. The organic phase was washed with brine, dried, concentrated and purified by silica gel column chromatography to give ZB-PY-E16 (15.0 mg, 35%) as a white solid. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.87 (s, 1H), 8.73 (t, J=5.1 Hz, 1H), 7.67 (s, 1H), 7.37-7.60 (m, 4H), 6.96 ( t, J=9.4Hz, 1H), 6.72 (dd, J=8.7, 3.8Hz, 1H), 5.78 (d, J=12.4Hz, 1H), 5.03 (d, J=12.4Hz, 1H), 4.76 ( d, J=4.9 Hz, 2H), 4.56 (t, J=8.7 Hz, 2H), 3.35 (t, J=8.8 Hz, 2H). LCMS: [M+H] ⁺ =417.3.

Example 4 Synthesis of ZB-PY-E21

Synthesis of compound 4.2: compound 1 (100 mg, 0.230 mmol), (2-(((tert-butyldimethylsilyl)oxy)methyl)pyridin-3-yl)boronic acid (123 mg, 0.690 mmol), carbonic acid Potassium (95.2 mg, 0.690 mmol) and tetrakis(triphenylphosphine)palladium (26.6 mg, 0.0230 mmol) were added to a mixed solvent of 1,4-dioxane (5 mL) and water (1 mL), nitrogen was replaced, and Heated at 100°C for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated, and subjected to silica gel column chromatography (5% methanol in dichloromethane as eluent) to give compound 4.2 (67.7 mg, 51%) as a pale yellow solid. LCMS: [M+H] ⁺ = 578.26.

Synthesis of compound 4.3: To a solution of compound 4.2 (67.7 mg, 0.117 mmol) in tetrahydrofuran (5 mL) was added tetrabutylammonium fluoride (TBAF, 1 M in tetrahydrofuran, 0.351 mL). The resulting solution was stirred at room temperature (rt) for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated, and subjected to silica gel column chromatography (5% methanol in dichloromethane as eluent) to give compound 4.3 (50.0 mg, 92%) as a pale yellow solid. LCMS: [M+H] ⁺ =464.10.

Synthesis of compound 4.4: A mixture of compound 4.3 (50.0 mg, 0.108 mmol) and lithium hydroxide (51.8 mg, 2.16 mmol) in tetrahydrofuran (7 mL) and water (3 mL) was heated to 60°C for 12 hours. The pH was adjusted to 2-3 with 3N aqueous hydrochloric acid at 0°C, and extracted with ethyl acetate. The organic phase was washed with brine, dried, and concentrated to give compound 4.4 (43.6 mg, 93%) as a white solid. LCMS: [M+H] ⁺ =436.0.

Synthesis of ZB-PY-E21: Triethylamine (60.6 mg, 0.600 mmol) and 2,4,6-trichlorobenzoyl chloride (121 mg, 0.500 mmol) were added to compound 4.4 (43.6 mg, 0.1 mmol) in toluene (5 mL) solution, reacted at 25°C for 4 hours. Then, the reaction solution was diluted with toluene (5 mL) and slowly added dropwise to a solution of 4-dimethylaminopyridine (17.2 mg, 0.14 mmol) in toluene (10 mL), and added dropwise at 80° C. for 4 hours. The reaction was cooled, quenched with saturated sodium bicarbonate solution, and extracted with ethyl acetate. The organic phase was washed with brine, dried, concentrated and purified by silica gel column chromatography to give ZB-PY-E21 (13.0 mg, 31%) as a white solid. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.90 (s, 1H), 8.84 (t, J=5.1 Hz, 1H), 8.64 (dd, J=4.7, 1.6 Hz, 1H), 8.03 (dd, J=8.0, 1.6Hz, 1H), 7.73 (s, 1H), 7.56 (dd, J=7.9, 4.8Hz, 1H), 6.93–6.99 (m, 1H), 6.72 (dd, J=8.7, 3.9Hz) ,1H),5.95(d,J＝11.5Hz,1H),4.98(d,J＝11.5Hz,1H),4.77(d,J＝4.7Hz,2H),4.56(t,J＝8.7Hz,2H) ), 3.30-3.31 (m, 2H). LCMS: [M+H] ⁺ =418.2.

Example 5 Synthesis of ZB-PY-E19:

Synthesis of compound 5.2: compound 1 (100 mg, 0.230 mmol), (2-(2-hydroxyethyl)phenyl)boronic acid (74.7 mg, 0.461 mmol), potassium carbonate (95.2 mg, 0.690 mmol) and tetrakis(triphenylene) phosphine)palladium (26.6 mg, 0.0230 mmol) was added to a mixed solvent of 1,4-dioxane (5 mL) and water (1 mL), replaced with nitrogen and heated at 100° C. for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated, and subjected to silica gel column chromatography (5% methanol in dichloromethane as eluent) to give compound 5.2 (80.0 mg, 73%) as a pale yellow solid. LCMS: [M+H] ⁺ =477.2.

Synthesis of compound 5.3: A mixture of compound 5.2 (80.0 mg, 0.168 mmol) and lithium hydroxide (80.6 mg, 3.36 mmol) in tetrahydrofuran (7 mL) and water (3 mL) was heated to 60°C for 12 hours. The pH was adjusted to 2-3 with 3N aqueous hydrochloric acid at 0°C, and extracted with ethyl acetate. The organic phase was washed with brine, dried, and concentrated to give compound 5.3 (50.0 mg, 93%) as a white solid. LCMS: [M+H] ⁺ =450.20.

Synthesis of ZB-PY-E19: Triethylamine (67.7 mg, 0.670 mmol) and 2,4,6-trichlorobenzoyl chloride (243 mg, 0.558 mmol) were added to compound 5.3 (50.0 mg, 0.112 mmol) in toluene (5 mL) solution, reacted at 25°C for 4 hours. Then, the reaction solution was diluted with toluene (5 mL) and slowly added dropwise to a solution of 4-dimethylaminopyridine (19.3 mg, 0.157 mmol) in toluene (10 mL), and added dropwise at 80° C. for 4 hours. The reaction was cooled, quenched with saturated sodium bicarbonate solution, and extracted with ethyl acetate. The organic phase was washed with brine, dried, concentrated and purified by silica gel column chromatography to give ZB-PY-E19 (13.0 mg, 31%) as a white solid. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.81 (s, 1H), 8.58 (t, J=5, 6Hz, 1H) 7.37-7.42 (m, 1H), 7.29-7.37 (m, 2H), 7.21-7.27(m, 2H), 6.93-7.00(m, 1H), 6.72(dd, J=8.7, 3.7Hz, 1H), 4.72-4.73(m, 2H), 4.57(t, J=8.7Hz, 2H),4.41-4.51(m,1H),4.27-4.37(m,1H),3.35(t,J=8.9Hz,2H),3.03(t,J=7.5Hz,2H).LCMS:[M+ H] ⁺ =431.20.

Example 6 Synthesis of ZB-PY-E20:

Synthesis of compound 6.2: compound 1 (100 mg, 0.230 mmol), (2-(((tert-butoxycarbonyl)amino)methyl)phenyl)boronic acid (154 mg, 0.461 mmol), potassium carbonate (95.2 mg, 0.690 mmol) and tetrakis(triphenylphosphine)palladium (26.6 mg, 0.023 mmol) were added to a mixed solvent of 1,4-dioxane (5 mL) and water (1 mL), and the mixture was heated at 100° C. for 4 hours by replacing nitrogen. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated, and subjected to column chromatography on silica gel (3% methanol in dichloromethane as eluent) to give compound 6.2 (52 mg, 40%) as a pale yellow solid. LCMS: [M+H] ⁺ =562.3.

Synthesis of compound 6.3: at 0°C, compound 6.2 (52.0 mg, 0.0925 mmol) was dissolved in a 1,4-dioxane solution of hydrochloric acid (4 M, 5 mL), reacted for 1 hour, and concentrated to obtain compound 6.3 (42.0 mg, 98%) as a yellow solid. LCMS: [M+H] ⁺ =462.3.

Synthesis of ZB-PY-E20: To a solution of compound 6.3 (42.0 mg, 0.091 mmol) in N,N-dimethylformamide (DMF) at 0°C was added potassium tert-butoxide (t-BuOK, 30.6 mg, 0.273 mmol), and reacted at 0 °C for 2 hours. Water was added to quench and extracted with ethyl acetate. The organic phase was washed with brine, dried, concentrated and purified by silica gel column chromatography to give ZB-PY-E20 (15.0 mg, 40%) as a white solid. ¹ H NMR (400MHz, DMSO-d ₆ )δ8.78(s,1H),8.52(t,J=5.1Hz,1H),8.21-8.24(m,1H),7.36-7.49(m,4H), 7.24–7.32 (m, 1H), 6.93–7.00 (m, 1H), 6.71 (dd, J=8.6, 3.9Hz, 1H), 4.70–4.86 (m, 3H), 4.56 (t, J=8.8Hz, 2H), 3.91-4.05 (m, 1H), 3.29-3.33 (m, 2H). LCMS: [M+H] ⁺ =416.2.

Example 7 Synthesis of ZB-PY-E22

Synthesis of compound 7.2: compound 1 (200 mg, 0.461 mmol), tert-butyl 2-(2-(4,4,5,5-tetramethyl-1,3,2-dioxaboropenan-2-yl) )phenyl)acetate (293 mg, 0.922 mmol), tetrakis(triphenylphosphine)palladium (53.2 mg, 0.0461 mmol) and potassium carbonate (191 mg, 1.38 mmol) were added to 1,4-dioxane (10 mL) ) and water (2 mL) in a mixed solvent of substituted nitrogen and heated at 100° C. for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated, and subjected to column chromatography on silica gel (3% methanol in dichloromethane as eluent) to give compound 7.2 (210 mg, 83%) as a white solid. LCMS: [M+H] ⁺ =547.2.

Synthesis of compound 7.3: A solution of compound 7.2 (210 mg, 0.385 mmol) in tetrahydrofuran (5 mL) was stirred at 25°C overnight. The reaction solution was concentrated and purified by silica gel column chromatography to obtain compound 7.3 (213 mg, 86%) as a pale yellow solid. LCMS: [M+H] ⁺ =647.2.

Synthesis of compound 7.4: Potassium tert-butoxide (111 mg, 0.989 mmol) was added to a solution of compound 7.3 (213 mg, 0.330 mmol) in toluene (10 mL) at 0°C, and stirred at 0°C for 2 hours. It was quenched by addition of saturated aqueous ammonium chloride, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated, and subjected to silica gel column chromatography (3% methanol in dichloromethane as eluent) to give compound 7.4 (110 mg, 56%) as a pale yellow solid. LCMS: [M+H] ⁺ =601.2.

Synthesis of ZB-PY-E22: Compound 7.4 (110 mg, 0.183 mmol) was dissolved in dichloromethane (10 mL), then trifluoroacetic acid (2 mL) was added. The resulting solution was stirred at room temperature for 2 hours, then concentrated and purified by silica gel column chromatography to give ZB-PY-E22 (15.0 mg, 40%) as a yellow solid. ¹ H NMR (400MHz, DMSO-d ₆ )δ8.87(t, J=5.1Hz, 1H), 8.79(s, 1H), 8.32(s, 1H), 7.80-7.93(m, 1H), 7.31- 7.49(m, 3H), 6.97(dd, J=10.3, 8.7Hz, 1H), 6.73(dd, J=8.7, 3.9Hz, 1H), 4.79(d, J=4.7Hz, 2H), 4.57(t , J=8.7Hz, 2H), 3.85 (s, 2H), 3.29-3.33 (m, 2H). LCMS: [M+H] ⁺ =401.20.

Example 8 Synthesis of ZB-PY-E23

Synthesis of compound 8.2: compound 1 (200 mg, 0.461 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2 - Dioxaborolane) (234 mg, 0.922 mmol), potassium acetate (KOAC, 1.36 g, 1.38 mmol) and _PdCl2 (dppf) (34.9 mg, 0.0461 mmol) were added to 1,4-dioxane (10 mL) ), nitrogen was replaced, and then heated at 100°C for 2 hours. The reaction solution was concentrated and separated by silica gel column chromatography to obtain compound 8.2 (65 mg, 29.2%). LCMS: [MH] ⁺ =483.2.

Synthesis of compound 8.3: compound 8.2 (65 mg, 0.135 mmol), tert-butyl(1-(2-bromophenyl)cyclopropyl)carbamate (84.0 mg, 0.270 mmol), tetrakis(triphenylphosphine) Palladium (15.6 mg, 0.0135 mmol) and potassium carbonate (55.9 mg, 0.405 mmol) were added to a mixed solvent of 1,4-dioxane (6 mL) and water (1.5 mL), replaced with nitrogen, and heated at 100°C for 2 Hour. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated, and subjected to column chromatography on silica gel (5% methanol in dichloromethane as eluent) to give compound 8.3 (31.0 mg, 39%) as a white solid. LCMS: [M+H] ⁺ =588.4.

Synthesis of compound 8.4: Compound 8.3 (31 mg, 0.0528 mmol) was added to a solution of hydrochloric acid (HCl) in 1,4-dioxane (4N, 5 mL) and stirred at room temperature for 1 hour. The reaction solution was concentrated to obtain compound 8.4 (25.7 mg, 100%) LCMS: [M+H] ⁺ =488.4.

Synthesis of compound ZB-PY-E23: To a solution of compound 8.4 (25.7 mg, 0.0528 mmol) in N,N-dimethylformamide (5 mL), potassium tert-butoxide (15.2 mg, 0.158 mmol) was added, and the reaction was carried out at 0°C 2 hours. It was quenched by addition of saturated aqueous ammonium chloride solution and extracted with ethyl acetate. The organic phase was washed with brine, dried, concentrated and purified by silica gel column chromatography to give the target compound ZB-PY-E23 (8 mg, 34%) as a white solid. ¹ H NMR (400MHz, DMSO-d ₆ )δ8.73(s,1H), 8.62(s,1H), 8.51(s,1H), 7,28-7.62(m,4H), 6.91-7.00(m ,1H),6.72(dd,J＝8.6,3.8Hz,1H),4.65-4.80(m,2H),4.57(t,J＝8.7Hz,2H),3.29-3.33(m,2H),1.05- 1.12 (m, 2H), 0.79-0.86 (m, 1H), 0.66-0.72 (m, 1H). LCMS: [M+H] ⁺ =442.2.

Example 9 Synthesis of ZB-PY-E24

Synthesis of compound 9.2: compound 1 (200 mg, 0.460 mmol), (2-(((tert-butoxycarbonyl)amino)methyl)pyridin-3-yl)boronic acid (174 mg, 0.692 mmol), potassium carbonate (191 mg) , 1.38 mmol) and tetrakis(triphenylphosphine)palladium (34.9 mg, 0.0461 mmol) were added to a mixed solvent of 1,4-dioxane (5 mL) and water (1 mL), replaced with nitrogen, and heated at 100 °C 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated, and subjected to column chromatography on silica gel (3% methanol in dichloromethane as eluent) to give compound 9.2 (120 mg, 46%) as a pale yellow solid. LCMS: [M+H] ⁺ =563.3.

Synthesis of compound 9.3: At 0°C, compound 9.2 (120 mg, 0.213 mmol) was dissolved in a solution of hydrochloric acid in 1,4-dioxane (4N, 5 mL) and reacted for 1 hour. The reaction solution was concentrated to obtain compound 9.3 (98.2 mg, 100%) as a yellow solid. LCMS: [M+H] ⁺ =463.3.

Synthesis of compound ZB-PY-E24: To a solution of compound 9.3 (98.2 mg, 0.213 mmol mmol) in N,N-dimethylformamide (5 mL) was added potassium tert-butoxide (78.0 mg, 0.639 mmol) at 0°C ) at 0°C for 2 hours. Water was added to quench and extracted with ethyl acetate. The organic phase was washed with brine, dried, concentrated and purified by silica gel column chromatography to give ZB-PY-E24 (25.0 mg, 28%) as a white solid. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.80 (s, 1H), 8.64 (t, J=5.1 Hz, 1H), 8.52 (dd, J=4.8, 1.6 Hz, 1H), 8.46 (dd, J=8.4, 6.0Hz, 1H), 7.89 (dd, J=7.8, 1.7Hz, 1H), 7.40–7.48 (m, 2H), 6.96 (dd, J=10.3, 8.7Hz, 1H), 6.71 (dd , J=8.6, 3.9Hz, 1H), 4.95 (dd, J=14.2, 8.5Hz, 1H), 4.74 (d, J=5.3Hz, 2H), 4.56 (t, J=8.7Hz, 2H), 4.01 (dd, J=14.2, 6.0 Hz, 1H) 3.31 - 3.34 (m, 2H). LCMS: [M+H] ⁺ =417.2.

Example 10 Synthesis of ZB-PY-E25

Synthesis of compound 10.2: compound 1 (174 mg, 0.402 mmol), tert-butyl 3-(2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2- (200 mg, 0.602 mmol), tetrakis(triphenylphosphine)palladium (46.4 mg, 0.0602 mmol) and potassium carbonate (222 mg, 1.61 mmol) were added to 1,4-dioxane ( In a mixed solvent of 10 mL) and water (2 mL), nitrogen was replaced, and the mixture was heated at 100° C. for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated, and subjected to column chromatography on silica gel (3% methanol in dichloromethane as eluent) to give compound 10.2 (110 mg, 49%) as a white solid. LCMS: [M+H] ⁺ =561.2.

Synthesis of compound 10.3: (Boc) ₂ O (85.6 mg, 0.393 mmol) and 4-dimethylaminopyridine (2.41 mg, 0.0196 mmol) were added to a solution of compound 10.2 (110 mg, 0.196 mmol) in tetrahydrofuran (5 mL), and stirred at room temperature overnight. The reaction solution was concentrated and purified by silica gel column chromatography to obtain compound 10.3 (120 mg, 93%) as a pale yellow solid. LCMS: [M+H] ⁺ =661.2.

Synthesis of compound 10.4: Potassium tert-butoxide (61.1 mg, 0.546 mmol) was added to a solution of compound 10.3 (120 mg, 0.182 mmol) in toluene (10 mL) at 0°C, and stirred at 0°C for 2 hours. It was quenched by addition of saturated aqueous ammonium chloride, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated, and subjected to column chromatography on silica gel (5% methanol in dichloromethane as eluent) to give compound 10.4 (45.0 mg, 40%) as a pale yellow solid. LCMS: [M+H] ⁺ =617.3.

Synthesis of compound ZB-PY-E25: Compound 10.4 (45.0 mg, 0.0730 mmol) was dissolved in dichloromethane (10 mL), then trifluoroacetic acid (2 mL) was added. The resulting solution was stirred at room temperature for 2 hours, then concentrated, and purified by silica gel column chromatography to give ZB-PY-E25 (15.0 mg, 50%) as a yellow solid. ¹ H NMR (400 MHz, chloroform- _d ) δ 8.27 (s, 1H), 7.61 (s, 1H), 7.36–7.42 (m, 1H), 7.27–7.32 (m, 3H), 6.83 (t, J= 9.4Hz, 1H), 6.65 (dd, J=8.7, 3.9Hz, 1H), 5.96 (s, 1H), 4.85 (d, J=5.2Hz, 2H), 4.61 (t, J=8.7Hz, 2H) , 3.43 (t, J=8.7 Hz, 2H), 2.86-3.17 (m, 4H). LCMS: [M+H] ⁺ =415.20.

Example 11 Synthesis of ZB-PY-E32

Synthesis of compound 11.2: compound 1 (100 mg, 0.230 mmol), (2-(((tert-butoxycarbonyl)(methyl)amino)methyl)pyridin-3-yl)boronic acid (123 mg, 0.461 mmol), carbonic acid Potassium (95.2 mg, 0.690 mmol) and tetrakis(triphenylphosphine)palladium (26.6 mg, 0.0230 mmol) were added to a mixed solvent of 1,4-dioxane (5 mL) and water (1 mL) to replace nitrogen gas, Heated at 100°C for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated, and subjected to column chromatography on silica gel (5% methanol in dichloromethane as eluent) to give compound 11.2 (73 mg, 40%) as a pale yellow solid. LCMS: [M+H] ⁺ =577.3.

Synthesis of compound 11.3: Compound 11.2 (73.0 mg, 0.126 mmol) was dissolved in a solution of hydrochloric acid in 1,4-dioxane (4M, 5 mL) at 0°C, and reacted for 1 hour. The reaction solution was concentrated to obtain compound 11.3 (60.1 mg, 98%) as a yellow solid. LCMS: [M+H] ⁺ =477.3.

Synthesis of compound ZB-PY-E32: Add potassium tert-butoxide (42.3 mg, 0.378 mmol) to a solution of compound 11.3 (60.1 mg, 0.126 mmol) in N,N-dimethylformamide at 0 °C, 0 °C React for 2 hours. Water was added to quench and extracted with ethyl acetate. The organic phase was washed with brine, dried, concentrated and purified by silica gel column chromatography to give ZB-PY-E32 (17.0 mg, 31%) as a white solid. LCMS: [M+H] ⁺ =431.2. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.81 (s, 1H), 8.67 (t, J=5.1 Hz, 1H), 8.53 (dd, J=4.9, 1.6 Hz, 1H), 7.94 (dd, J=7.9, 1.7Hz, 1H), 7.61–7.42 (m, 2H), 6.96 (dd, J=10.3, 8.6Hz, 1H), 6.71 (dd, J=8.6, 3.8Hz, 1H), 5.41 (d , J=14.5Hz, 1H), 4.74(d, J=4.7Hz, 2H), 4.56(t, J=8.7Hz, 2H), 4.07(d, J=14.4Hz, 1H), 3.34(t, J =8.7Hz,2H),3.05(s,3H).

Example 12 Synthesis of ZB-PY-E26

Synthesis of compound 12.2: compound 1 (200mg, 0.460mmol), (2-(1-hydroxyethyl)phenyl)boronic acid pinacol ester (228mg, 0.920mmol), tetrakis(triphenylphosphine)palladium (53.2mg) , 0.0460 mmol) and potassium carbonate (190 mg, 1.38 mmol) were added to a mixed solvent of 1,4-dioxane (5 mL) and water (1 mL), replaced with nitrogen and heated at 110° C. for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated, and subjected to column chromatography on silica gel (5% methanol in dichloromethane as eluent) to give compound 12.2 (130 mg, 59%) as a pale yellow solid. LCMS: [M+H] ⁺ =477.1.

Synthesis of compound 12.3: A mixture of compound 12.2 (130 mg, 0.273 mmol) and lithium hydroxide (131 mg, 5.46 mmol) in tetrahydrofuran (7 mL) and water (3 mL) was heated to 60°C for 12 hours. The pH was adjusted to 2-3 with 3N aqueous hydrochloric acid at 0°C, and extracted with ethyl acetate. The organic phase was washed with brine, dried, and concentrated to give compound 12.3 (70 mg, 57%) as a white solid. LCMS: [M+H] ⁺ =449.2.

Synthesis of ZB-PY-E26: Triethylamine (94.7 mg, 0.938 mmol) and 2,4,6-trichlorobenzoyl chloride (189 mg, 0.781 mmol) were added to compound 12.3 (70 mg, 0.156 mmol) in toluene ( 5mL) solution, react at 25°C for 4 hours. Then, the reaction solution was diluted with toluene (5 mL) and slowly added dropwise to a solution of 4-dimethylaminopyridine (26.8 mg, 0.218 mmol) in toluene (10 mL), and added dropwise at 80° C. for 4 hours. The reaction solution was cooled, quenched with saturated aqueous sodium bicarbonate solution, and extracted with ethyl acetate. The organic phase was washed with brine, dried, concentrated and purified by silica gel column chromatography to give ZB-PY-E26 (15.0 mg, 22%) as a white solid. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.84 (s, 1H), 8.72 (t, J=5.1 Hz, 1H), 7.66 (s, 1H), 7.75–7.69 (m, 1H), 7.48– 7.51 (m, 3H), 6.96 (dd, J=10.3, 8.7Hz, 1H), 6.72 (dd, J=8.7, 3.9Hz, 1H), 6.29 (q, J=6.4Hz, 1H), 4.67–4.89 (m, 2H), 4.56 (t, J=8.8 Hz, 2H), 3.33-3.35 (m, 2H), 1.65 (d, J=6.4 Hz, 3H). LCMS: [M+H] ⁺ =431.3.

Example 13 Synthesis of ZB-PY-E27

Synthesis of compound 13.2: compound 1 (200 mg, 0.460 mmol), (2-(((tert-butyldimethylsilyl)oxo)methyl)-6-methylpyridin-3-yl)boronic acid ( 244 mg, 0.920 mmol), potassium carbonate (190 mg, 1.38 mmol) and tetrakis(triphenylphosphine)palladium (53.2 mg, 0.0460 mmol) were added to a mixture of 1,4-dioxane (5 mL) and water (1 mL) In the solvent, nitrogen was replaced and heated at 100°C for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated, and subjected to silica gel column chromatography (5% methanol in dichloromethane as eluent) to give compound 13.2 (170 mg, 62%) as a pale yellow solid. LCMS: [M+H] ⁺ = 592.26.

Synthesis of compound 13.3: To a solution of compound 13.2 (170 mg, 0.287 mmol) in tetrahydrofuran (5 mL) was added tetrabutylammonium fluoride (1 M, 0.431 mL) and the resulting solution was stirred at room temperature for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated, and subjected to column chromatography on silica gel (5% methanol in dichloromethane as eluent) to give compound 13.3 (130.0 mg, 92%) as a pale yellow solid. LCMS: [M+H] ⁺ =478.10.

Synthesis of compound 13.4: A mixture of compound 13.3 (130 mg, 0.273 mmol) and lithium hydroxide (131 mg, 5.46 mmol) in tetrahydrofuran (7 mL) and water (3 mL) was heated to 60°C for 12 hours. The pH was adjusted to 2-3 with 3N aqueous hydrochloric acid at 0°C, and extracted with ethyl acetate. The organic phase was washed with brine, dried, and concentrated to give compound 13.4 (56 mg, 46%) as a white solid. LCMS: [M+H] ⁺ =450.0.

Synthesis of compound ZB-PY-E27: Triethylamine (75.5 mg, 0.748 mmol) and 2,4,6-trichlorobenzoyl chloride (151 mg, 0.624 mmol) were added to compound 13.4 (56 mg, 0.125 mmol) in toluene (5 mL) solution, reacted at 25°C for 4 hours. Then, the reaction solution was diluted with toluene (5 mL) and slowly added dropwise to a solution of 4-dimethylaminopyridine (21.5 mg, 0.175 mmol) in toluene (10 mL) at 80° C. for 4 hours. The reaction solution was cooled, quenched with saturated aqueous sodium bicarbonate solution, and extracted with ethyl acetate. The organic phase was washed with brine, dried, concentrated and purified by silica gel column chromatography to give ZB-PY-E27 (17.0 mg, 32%) as a white solid. ¹ H NMR (400MHz, DMSO-d ₆ )δ8.90(s,1H),8.84(t,J=5.1Hz,1H),7.99(d,J=8.3Hz,1H),7.71(s,1H) ,7.48(d,J＝8.1Hz,1H),6.96(dd,J＝10.2,8.7Hz,1H),6.71(dd,J＝8.6,3.8Hz,1H),5.91(d,J＝12.3Hz, 1H), 4.96(d, J＝12.5Hz, 1H), 4.76(d, J＝4.7Hz, 2H). 4.56(t, J＝8.7Hz, 2H), 3.35(t, J＝8.7Hz, 2H) , 2.57(s,3H).LCMS: [M+H] ⁺ =432.2.

Example 14 Synthesis of ZB-PY-E28

Synthesis of compound 14.2: compound 1 (289 mg, 0.667 mmol), (2-(((tert-butyldimethylsilyl)oxo)methyl)-6-cyclopropylpyridin-3-yl)boronic acid (307 mg, 1.00 mmol), potassium carbonate (276 mg, 2.00 mmol) and tetrakistriphenylphosphine palladium (77.0 mg, 0.0667 mmol) were added to a mixed solvent of 1,4-dioxane (5 mL) and water (1 mL) , nitrogen was replaced and heated at 100°C for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated, and subjected to column chromatography on silica gel (5% methanol in dichloromethane as eluent) to give compound 14.2 (170 mg, 41%) as a pale yellow solid. LCMS: [M+H] ⁺ = 618.3.

Synthesis of compound 14.3: To a solution of compound 14.2 (170 mg, 0.275 mmol) in tetrahydrofuran (5 mL) was added tetrabutylamine fluoride (1 M, 0.413 mL) and the resulting solution was stirred at room temperature for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated, and subjected to silica gel column chromatography (5% methanol in dichloromethane as eluent) to give compound 14.3 (130.0 mg, 94%) as a pale yellow solid. LCMS: [M+H] ⁺ =504.10.

Synthesis of compound 14.4: A mixture of compound 14.3 (130 mg, 0.258 mmol) and lithium hydroxide (216 mg, 5.16 mmol) in tetrahydrofuran (7 mL) and water (3 mL) was heated to 60°C for 12 hours. The pH was adjusted to 2-3 with 3N aqueous hydrochloric acid at 0°C, and extracted with ethyl acetate. The organic phase was washed with brine, dried, and concentrated to give compound 14.4 (47.5 mg, 39%) as a white solid. LCMS: [M+H] ⁺ =476.0.

Synthesis of compound ZB-PY-E28: Triethylamine (60.6 mg, 0.600 mmol) and 2,4,6-trichlorobenzoyl chloride (121 mg, 0.500 mmol) were added to compound 14.4 (47.5 mg, 0.100 mmol) In a toluene (5 mL) solution, the reaction was carried out at 25°C for 4 hours. Then, the reaction solution was diluted with toluene (5 mL) and slowly added dropwise to a solution of 4-dimethylaminopyridine (17.2 mg, 0.14 mmol) in toluene (10 mL), and added dropwise at 80° C. for 4 hours. The reaction solution was cooled, quenched with saturated aqueous sodium bicarbonate solution, and extracted with ethyl acetate. The organic phase was washed with brine, dried, concentrated and purified by silica gel column chromatography to give ZB-PY-E28 (17.0 mg, 37%) as a white solid. ¹ H NMR(400MHz,DMSO-d6)δ8.88(s,1H),8.80(t,J=5.1Hz,1H),7.88(d,J=8.1Hz,1H),7.68(s,1H), 7.43(d,J＝8.1Hz,1H),6.96(dd,J＝10.3,8.7Hz,1H),6.71(dd,J＝8.6,3.8Hz,1H),5.87(d,J＝12.2Hz,1H) ), 4.86(d, J＝12.2Hz, 1H), 4.75(d, J＝4.7Hz, 2H), 4.56(t, J＝8.7Hz, 2H), 3.35(t, J＝8.7Hz, 2H), 2.20 (ddd, J=12.4, 8.0, 5.2 Hz, 1H), 0.90-1.10 (m, 4H). LCMS: [M+H] ⁺ =458.2.

Example 15 Synthesis of ZB-PY-E31

Synthesis of compound 15.2: compound 1 (100 mg, 0.230 mmol), (2-(((tert-butoxycarbonyl)(methyl)amino)methyl)pyridin-3-yl)boronic acid (123 mg, 0.461 mmol), Potassium carbonate (95.2 mg, 0.690 mmol) and tetrakistriphenylphosphine palladium (26.6 mg, 0.0230 mmol) were added to a mixed solvent of 1,4-dioxane (5 mL) and water (1 mL), and nitrogen was replaced at 100 ℃ heated for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated, and subjected to column chromatography on silica gel (5% methanol in dichloromethane as eluent) to give compound 15.2 (73 mg, 40%) as a pale yellow solid. LCMS: [M+H] ⁺ =576.3.

Synthesis of compound 15.3: Compound 15.2 (73.0 mg, 0.126 mmol) was dissolved in a solution of hydrochloric acid in 1,4-dioxane (4M, 5 mL) at 0°C, and reacted for 1 hour. The reaction solution was concentrated to obtain compound 15.3 (60.1 mg, 98%) as a yellow solid. LCMS: [M+H] ⁺ =476.3.

Synthesis of compound ZB-PY-E31: Add potassium tert-butoxide (42.3 mg, 0.378 mmol) to a solution of compound 15.3 (60.1 mg, 0.126 mmol) in N,N-dimethylformamide at 0 °C, 0 °C React for 2 hours. Water was added to quench and extracted with ethyl acetate. The organic phase was washed with brine, dried, concentrated and purified by silica gel column chromatography to give ZB-PY-E31 (13.0 mg, 24%) as a white solid. LCMS: [M+H] ⁺ =430.1. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.80 (s, 1H), 8.57 (d, J=5.0 Hz, 1H), 7.32-7.55 (m, 5H), 6.96 (dd, J=10.3, 8.6 Hz, 1H), 6.71(dd, J＝8.6, 3.8Hz, 1H), 5.24(d, J＝14.8Hz, 1H), 4.74(d, J＝4.7Hz, 2H), 4.56(t, J＝8.7 Hz, 2H), 4.05(d, J=14.7Hz, 1H), 3.34(t, J=8.7Hz, 2H), 2.99(s, 3H).

Example 16 Synthesis of ZB-PY-E30

Synthesis of compound 16.2: compound 1 (321 mg, 0.741 mmol), (3-(((tert-butyldimethylsilyl)oxy)methyl)-1-methyl-1H-pyrazole-4- yl)boronic acid (600 mg, 2.22 mmol), potassium carbonate (306 mg, 2.22 mmol) and tetrakis(triphenylphosphine)palladium (85.6 mg, 0.0741 mmol) were added to 1,4-dioxane (5 mL) and water ( 1 mL) of a mixed solvent, replaced with nitrogen, and heated at 100° C. for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated, and subjected to column chromatography on silica gel (5% methanol in dichloromethane as eluent) to give compound 16.2 (210 mg, 49%) as a pale yellow solid. LCMS: [M+H] ⁺ =581.3.

Synthesis of compound 16.3: To a solution of compound 16.2 (210 mg, 0.361 mmol) in tetrahydrofuran (5 mL) was added tetrabutylammonium fluoride (1 M, 0.722 mL) and the resulting solution was stirred at room temperature for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated, and subjected to column chromatography on silica gel (5% methanol in dichloromethane as eluent) to give compound 16.3 (130.0 mg, 77%) as a pale yellow solid. LCMS: [M+H] ⁺ =467.10.

Synthesis of compound 16.4: A mixture of compound 16.3 (130 mg, 0.278 mmol) and lithium hydroxide (233 mg, 5.56 mmol) in tetrahydrofuran (7 mL) and water (3 mL) was heated to 60°C for 12 hours. The pH was adjusted to 2-3 with 3N aqueous hydrochloric acid at 0°C, and extracted with ethyl acetate. The organic phase was washed with brine, dried, and concentrated to give the title compound (75 mg, 61%) as a white solid. LCMS: [M+H] ⁺ =439.0.

Synthesis of compound ZB-PY-E30: Triethylamine (104 mg, 1.02 mmol) and 2,4,6-trichlorobenzoyl chloride (207 mg, 0.850 mmol) were added to compound 16.4 (75.0 mg, 0.171 mmol) in toluene (5 mL) solution, reacted at 25°C for 4 hours. Then, the reaction solution was diluted with toluene (5 mL) and slowly added dropwise to a solution of 4-dimethylaminopyridine (25.2 mg, 0.205 mmol) in toluene (10 mL) at 80° C. for 4 hours. The reaction solution was cooled, quenched with saturated aqueous sodium bicarbonate solution, and extracted with ethyl acetate. The organic phase was washed with brine, dried, concentrated and purified by silica gel column chromatography to give ZB-PY-E30 (30.0 mg, 37%) as a white solid. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.83(s, 1H), 8.52(t, J=5.1Hz, 1H), 8.08(s, 1H), 7.59(s, 1H), 6.94(dd, J=10.3, 8.7Hz, 1H), 6.69 (dd, J=8.7, 3.8Hz, 1H), 5.23 (s, 2H), 4.72 (d, J=5.0Hz, 2H), 4.54 (t, J=8.7 Hz, 2H), 3.89 (s, 3H), 3.27-3.32 (m, 2H). LCMS: [M+H] ⁺ =421.2.

Example 17 Synthesis of ZB-PY-E29

Synthesis of compound 17.2: compound 1 (321 mg, 0.741 mmol), (5-(((tert-butyldimethylsilyl)oxo)methyl)-1-methyl-1H-pyrazole-4- yl)boronic acid (600 mg, 2.22 mmol), potassium carbonate (306 mg, 2.22 mmol) and tetrakis(triphenylphosphine)palladium (85.6 mg, 0.0741 mmol) were added to 1,4-dioxane (5 mL) and water ( 1 mL) of a mixed solvent, replaced with nitrogen, and heated at 100° C. for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated, and subjected to column chromatography on silica gel (5% methanol in dichloromethane as eluent) to give compound 17.2 (300 mg, 70%) as a pale yellow solid. LCMS: [M+H] ⁺ =581.3.

Synthesis of compound 17.3: To a solution of compound 17.2 (300 mg, 0.516 mmol) in tetrahydrofuran (5 mL) was added tetrabutylammonium fluoride (1 M, 1.03 mL) and the resulting solution was stirred at room temperature for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated, and subjected to silica gel column chromatography (5% methanol in dichloromethane as eluent) to give compound 17.3 (203.0 mg, 84%) as a pale yellow solid. LCMS: [M+H] ⁺ =467.10.

Synthesis of compound 17.4: A mixture of compound 17.3 (203 mg, 0.435 mmol) and lithium hydroxide (365 mg, 8.69 mmol) in tetrahydrofuran (7 mL) and water (3 mL) was heated to 60°C for 12 hours. The pH was adjusted to 2-3 with 3N aqueous hydrochloric acid at 0°C, and extracted with ethyl acetate. The organic phase was washed with brine. Drying and concentration gave the title compound (140 mg, 73%) as a white solid. LCMS: [M+H] ⁺ =439.0.

Synthesis of compound ZB-PY-E29: Triethylamine (194 mg, 0.510 mmol) and 2,4,6-trichlorobenzoyl chloride (387 mg, 1.60 mmol) were added to compound 17.4 (140 mg, 0.320 mmol) in toluene ( 5mL) solution, react at 25°C for 4 hours. Then the reaction solution was diluted with toluene (5 mL) and slowly added dropwise to a solution of 4-dimethylaminopyridine (47.2 mg, 0.384 mmol) in toluene (10 mL), and added dropwise at 80°C for 4 hours. Quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was washed with brine, dried, concentrated and purified by silica gel column chromatography to give ZB-PY-E29 (30.0 mg, 37%) as a white solid. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.85(s, 1H), 8.61(t, J=5.1Hz, 1H), 7.79(s, 1H), 7.68(s, 1H), 6.95(t, J=9.5Hz, 1H), 6.70(dd, J=8.6, 3.8Hz, 1H), 5.49(s, 2H), 4.72(d, J=4.9Hz, 2H), 4.55(t, J=8.7Hz, 2H), 3.95 (s, 3H), 3.30-3.34 (m, 2H). LCMS: [M+H] ⁺ =421.2.

Example 18: Polycomb inhibitory complex 2 (PRC2) enzyme activity assay

The activity of the PRC2 enzyme was detected using the time-resolved fluorescence energy transfer (TR-FRET) method. First, enzymes are mixed with different concentrations of compounds and incubated for 30 minutes at room temperature. The enzymatic reaction is initiated by addition of biotinylated histone H3 polypeptide substrate and cofactor S-adenosylmethionine (SAM). After 4 hours of reaction at room temperature, acceptor and donor Donor were added and incubated for half an hour. Fluorescence signals were detected with a multi-plate reader EnVision (Perkin Elmer). Data were analyzed with GraphPad Prism 5.0 software to obtain _IC50 values.

The compounds in Table 1 can be prepared by the methods described in the above examples, and EED226 is a positive control compound (Nat. Chem. Biol. 2017, 13, 381-388).

compound Enzyme activity IC₅₀(nM) compound Enzyme activity IC₅₀(nM) ZB-PY-E14 16.6 ZB-PY-E25 <50 ZB-PY-E16 7.8 ZB-PY-E26 <50 ZB-PY-E18 22.5 ZB-PY-E27 <50 ZB-PY-E19 7.0 ZB-PY-E28 <50 ZB-PY-E20 18.8 ZB-PY-E29 <50 ZB-PY-E21 55.0 ZB-PY-E30 <50 ZB-PY-E22 19.1 ZB-PY-E31 <50 ZB-PY-E23 <50 ZB-PY-E32 <50 ZB-PY-E24 <50 EED226 104

Example 19: Cell long-term growth inhibition experiment

Pfeiffer cells in exponential growth phase were seeded in 24-well plates at a cell density of 1*10E5 cells/mL. Cells were treated with different concentrations of compounds on the day. On days 4 and 7 of compound treatment, fresh medium and compounds were replaced. After 11 days of compound treatment, cell viability was determined using CellTiter-Glo reagent (Promega). Data were analyzed with GraphPad Prism 5.0 software to obtain _IC50 values.

The compounds in Table 2 can be prepared by the methods described in the above examples, and EED226 is a positive compound (Nat. Chem. Biol. 2017, 13, 381-388).

From the data in Table 1 and Table 2 above, it can be seen that the _IC50 value of some compounds of the present disclosure to PRC2 enzyme is less than 10 nM, which is significantly higher than that of the positive control compound EED226; similarly, for the long-term growth inhibition experiment of Pfeiffer cells, The IC ₅₀ values of some compounds of the present disclosure are <1 nM, which is significantly higher than that of the positive control compound EED226.

The above embodiments are merely auxiliary descriptions in nature, and are not intended to limit the embodiments of the application target or the applications or uses of these embodiments. As used herein, the term "exemplary" means "serving as an instance, instance, or illustration." Any exemplary embodiment herein is not necessarily to be construed as preferred or advantageous over other embodiments.

Furthermore, while at least one illustrative or comparative example has been presented in the foregoing embodiments, it should be understood that the present invention is capable of numerous variations. It should also be appreciated that the embodiments described herein are not intended to limit the scope, utility, or configuration of the claimed object in any way. Rather, the foregoing embodiments will provide those of ordinary skill in the art with a convenient guide for implementing the described embodiment or embodiments. Furthermore, various changes may be made in the function and arrangement of elements without departing from the scope of the claimed scope, which includes known equivalents and all foreseeable equivalents at the time of filing this patent application .

Claims (10)

1. A compound represented by general formula I, its pharmaceutically acceptable salt, enantiomer, diastereomer, atropisomer, racemate, polymorph, solvate substances or isotopically labeled compounds:

in, (1)

is a single bond or a double bond; (2) R ¹ , R ² and R ³ are each independently hydrogen, halogen, C ₁ -C ₄ alkyl, C ₁ -C ₄ haloalkyl, -O-(C ₁ -C ₄ alkyl), C ₁ -C ₄ haloalkoxy or 3-6 membered cycloalkyl; R ⁴ is hydrogen, OH, =O or C ₁ -C ₄ alkyl; R ⁵ is hydrogen, halogen, C ₁ -C ₄ alkyl or C ₁ -C ₄ haloalkyl; (3) M ring is a 6-10-membered aromatic ring or a 5-10-membered heteroaromatic ring, and M is substituted by 0-4 R ⁶ ; R ⁶ is independently selected from halogen, CN, C ₁ -C ₆ alkyl substituted with 0-1 R ^A1 , 3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl, -OR ^A2 , -C (=O)R ^A3 , -NR ^A4 R ^A5 , -NHC(=O)R ^A3 , -S(=O) ₂ R ^A3 , -S(=O) ₂ NR ^A4 R ^A5 , -NHS(=O) ₂ (C ₁ -C ₄ alkyl), and -C(=O)NR ^A4 R ^A5 ; R ^A1 is selected from C ₁ -C ₆ alkyl, 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, OH, -NR ^a R ^b , C ₁ -C ₄ alkoxy, -C(= O) NR ^a R ^b , -S(=O) ₂ (C ₁ -C ₄ alkyl), and -NHC(=O) (C ₁ -C ₄ alkyl); R ^A2 is selected from hydrogen, C ₁ -C ₄ alkyl, 3-6 membered cycloalkyl, and C ₁ -C ₄ haloalkyl; R ^A3 at each occurrence is independently selected from C ₁ -C ₄ alkyl, 3-6 membered cycloalkyl, 6-10 membered aryl, and 5-10 membered heteroaryl; R ^A4 and R ^A5 are at each occurrence independently selected from hydrogen, C ₁ -C ₄ alkyl and 3-6 membered cycloalkyl; R ^a and R ^b are at each occurrence independently selected from hydrogen, C ₁ -C ₄ alkyl, and 3-6 membered cycloalkyl; (4) Y is -C(O)-, -SO ₂ -, -C(R ^c )(R ^d )-, or -S(O)-; (5) each Z is independently -O-, -N(R ^A4 )-, -S- or -C(R ^c )(R ^d )-; ^Rc and ^Rd are at each occurrence independently selected from hydrogen, halogen, OH, _C1 - _C4 alkyl, _C1 - _C4 haloalkyl, _C1 - _C4 alkoxy, and _C1 -C ₄ haloalkoxy; or R ^c and R ^d are attached to each other to form -R ^c' -LR ^d' - and to the carbon atom attached to it to form a ring, wherein L is absent or -O-; R ^{c '} and R ^d' are each independently absent, C ₁ -C ₄ alkylene, C ₁ -C ₃ haloalkylene, or replaced by 1-2 halogens, OH, or C ₁ -C ₄ alkyl Substituted C ₁ -C ₃ alkylene; n is an integer selected from 1-4. 2. The compound according to claim 1, its pharmaceutically acceptable salts, enantiomers, diastereomers, atropisomers, racemates, polymorphs, solvates Or an isotopically-labeled compound, wherein the compound represented by the general formula I is selected from the compounds represented by the general formula I-1:

in, (1) R ¹ is hydrogen, halogen, C ₁ -C ₄ alkyl, C ₁ -C ₄ haloalkyl, or 3-6 membered cycloalkyl; (2) R ⁴ is hydrogen, OH, or =O; (3) M ring is a benzene ring or a 5-membered heteroaromatic ring, and M is substituted by 0-2 R ⁶ ; R ⁶ is independently selected from halogen, C ₁ -C ₆ alkyl substituted with 0-1 R ^A1 , 3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl, -OR ^A2 , -S(= O) ₂ R ^A3 , and -S(=O) ₂ NR ^A4 R ^A5 , -C(=O)NR ^A4 R ^A5 ; R ^A1 is selected from C ₁ -C ₆ alkyl, 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, OH, and -NR ^a R ^b ; R ^A2 is selected from hydrogen, C ₁ -C ₄ alkyl, and 3-6 membered cycloalkyl; R ^A3 is selected from C ₁ -C ₄ alkyl, and 3-6 membered cycloalkyl; R ^A4 and R ^A5 are independently selected at each occurrence from hydrogen, C ₁ -C ₄ alkyl and 3-6 membered cycloalkyl; R ^a and R ^b are at each occurrence independently selected from hydrogen, C ₁ -C ₄ alkyl, and 3-6 membered cycloalkyl; (4) Y is -C(O)- or -SO ₂ -; (5) each Z is independently -O-, -N(R ^A4 )-, -S- or -C(R ^c )(R ^d )-; ^Rc and ^Rd are at each occurrence independently selected from hydrogen, halogen, OH, _C1 - _C4 alkyl, _C1 - _C4 haloalkyl, _C1 - _C4 alkoxy, and _C1 -C ₄ haloalkoxy; or R ^c and R ^d are attached to each other to form -R ^c' -LR ^d' - and to the carbon atom attached to it to form a ring, wherein L is absent or -O-; R ^{c '} and R ^d' are each independently absent, C ₁ -C ₄ alkylene, C ₁ -C ₃ haloalkylene, or replaced by 1-2 halogens, OH, or C ₁ -C ₄ alkyl Substituted C ₁ -C ₃ alkylene; n is an integer selected from 1-3. 3. The compound of claim 1, its pharmaceutically acceptable salt, enantiomer, diastereomer, atropisomer, racemate, polymorph, solvate Or an isotopically-labeled compound, wherein the compound represented by the general formula I is selected from the compounds represented by the general formula I-2:

in, (1) R ¹ is hydrogen or halogen; (2) M ring is a benzene ring, and M is unsubstituted or substituted by 1-2 R ⁶ ; R ⁶ is independently selected from halogen, C ₁ -C ₆ alkyl substituted with 0-1 R ^A1 , 3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl, -OR ^A2 , -S(= O) ₂ R ^A3 , -S(=O) ₂ NR ^A4 R ^A5 and -C(=O)NR ^A4 R ^A5 ; R ^A1 is C ₁ -C ₆ alkyl, 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, OH or -NR ^a R ^b ; R ^A2 is hydrogen, C ₁ -C ₄ alkyl or 3-6 membered cycloalkyl; R ^A3 is C ₁ -C ₄ alkyl or 3-6 membered cycloalkyl; R ^A4 and R ^A5 are at each occurrence independently selected from hydrogen, C ₁ -C ₄ alkyl and 3-6 membered cycloalkyl; R ^a and R ^b are at each occurrence independently selected from hydrogen, C ₁ -C ₄ alkyl and 3-6 membered cycloalkyl; (3) each Z is independently -O-, -N(R ^A4 )-, or -C(R ^c )(R ^d )-; ^Rc and ^Rd are at each occurrence independently selected from hydrogen, halogen, OH, _C1 - _C4 alkyl, and _C1 - _C4 haloalkyl; n is 1 or 2. 4. The compound of claim 1, its pharmaceutically acceptable salt, enantiomer, diastereomer, atropisomer, racemate, polymorph, solvate Or an isotopically-labeled compound, wherein the compound represented by the general formula I is selected from the compounds represented by the general formula I-3a and I-3b:

in (1) each Z is independently -O- or -N(R ^A4 )-; R ^A4 is selected from hydrogen, C ₁ -C ₄ alkyl and 3-6 membered cycloalkyl; (2) A ¹ , A ² , A ³ and A ⁴ independently represent N or CR ^x ; R ^x is independently selected from hydrogen, halogen, C ₁ -C ₄ alkyl, 3-6 membered cycloalkyl, -S(=O) ₂ R ^A3 , -S(=O) ₂ NR ^A4 R ^A5 , and - C(=O)NR ^A4 R ^A5 ; R ^A3 is independently selected from C ₁ -C ₄ alkyl and 3-6 membered cycloalkyl; R ^A4 and R ^A5 are independently selected from hydrogen, C ₁ -C ₄ alkyl and 3-6 membered cycloalkyl. 5. The compound of any one of claims 1-4, pharmaceutically acceptable salts, enantiomers, diastereomers, atropisomers, racemates, polymorphs thereof Forms, solvates or isotopically labeled compounds, wherein the compound represented by the general formula I is selected from the following compounds:

6. A pharmaceutical composition comprising a compound selected from the group consisting of compounds according to any one of claims 1-5, pharmaceutically acceptable salts thereof, enantiomers, diastereomers and racemates one or more of the body, and at least one pharmaceutically acceptable carrier, diluent or excipient. 7. The pharmaceutical composition of claim 6, wherein the pharmaceutical composition further comprises at least one additional therapeutic agent. 8. The pharmaceutical composition of claim 7, wherein the additional therapeutic agent is selected from the group consisting of other anticancer agents, immunomodulatory agents, antiallergic agents, antiemetics, pain relief agents, cytoprotective agents, and combinations thereof. 9. The compound according to any one of claims 1-5, a pharmaceutically acceptable salt, enantiomer, diastereomer or racemate thereof, or according to claim 6-8 Use of any one of the pharmaceutical compositions in the manufacture of a medicament for the treatment of a disease or condition mediated by EED and/or PRC2. 10. The use according to claim 9, wherein The diseases or conditions mediated by EED and/or PRC2 include lymphoma, leukemia, multiple myeloma, mesothelioma, gastric cancer, malignant rhabdoid tumor, hepatocellular carcinoma, prostate cancer, breast cancer, bile duct and gallbladder cancer , bladder cancer, brain tumor, cervical cancer, colon cancer, melanoma, endometrial cancer, esophagus cancer, head and neck cancer, lung cancer, nasopharyngeal cancer, ovarian cancer, pancreatic cancer, renal cell cancer, rectal cancer, thyroid cancer, Parathyroid tumors, uterine tumors and soft tissue sarcomas.