CN115028631A - Triazole pyrimidine derivative, and pharmaceutical composition and application thereof - Google Patents

Abstract

The present disclosure relates to a bag made ofCompounds represented by the general formula, pharmaceutically acceptable salts, enantiomers, diastereomers, atropisomers, racemates, polymorphs, solvates or isotopically labeled compounds (including deuterium substitutions) thereof, pharmaceutical compositions containing the same and uses thereof. The partial compounds according to the present disclosure have significantly higher molecular and cellular activities than the positive control compound, EED226, and can be used for the preparation of a medicament for the treatment of diseases or conditions mediated by EED and/or PRC 2.

Description

Triazole pyrimidine derivative, and pharmaceutical composition and application thereof

Technical Field

The present disclosure relates to the field of pharmaceutical chemistry and pharmacotherapeutics, and generally to a class of triazolopyrimidine compounds, pharmaceutical compositions containing the same, and uses thereof. Specifically, the disclosure relates to a triazolopyrimidine compound, a pharmaceutically acceptable salt, enantiomer, diastereomer, atropisomer, racemate, polymorph, solvate or isotopically labeled compound (including deuterium substitution), a pharmaceutical composition and application thereof in treatment of diseases or disorders mediated by EED and/or PRC2, especially tumor diseases.

Background

The Polycomb inhibition Complex PRC2(Polycomb regenerative complete 2) is a core member of Polycomb family protein (Polycomb Group), has histone methyltransferase activity, and can specifically catalyze trimethylation modification of 27 th lysine (H3K27me3) of histone H3, thereby inhibiting the expression of specific genes. The methyltransferase activity of PRC2 is derived from its catalytic member EZH2, whereas EZH2 alone is not catalytically active, requiring at least the formation of complexes with the other two members of PRC2, EED and SUZ12, to catalyze methylation modifications. Thus, EZH2, EED and SUZ12 are considered core components of the PRC2 complex. Recent studies have found that the core component of PRC2 is overexpressed in various tumor cells, and the abnormal activity thereof is a direct cause of the onset and exacerbation of various malignant tumors. Meanwhile, recent gene sequencing results of lymphoma patients show that EZH2 has activating mutation in germinal center B cell lymphoma (GCB-DLBCL) patients, and the mutated EZH2 changes the substrate specificity of PRC2, thereby increasing the level of H3K27me3 in cells. Downregulating the expression of EZH2 or other core components by siRNA approach will significantly inhibit the proliferation of lymphoma cells, indicating that the development of GCB-DLBCL is closely related to the over-activation of PRC 2. Thus, PRC2 is a very promising target for anti-cancer drug development, and the discovery of inhibitors targeting PRC2 is a hotspot of current research in the pharmaceutical community. Recently, two major pharmaceutical companies, Nowa and Erberweiwei, invented a class of small molecules that inhibit PRC2 activity by targeting EEDs (references: EED226 of Nowa, US2016/0176882, J.Med.chem.2017,60,2215-. In conclusion, the PRC2 complex is considered as a key driver for the development of various malignant tumors, and the development of inhibitors for inhibiting the activity of PRC2 by targeting EED is currently very hot in the industry, which is beneficial for the development of new drugs related thereto.

Disclosure of Invention

One object of the present disclosure is to provide a triazolopyrimidine compound represented by general formula I, a pharmaceutically acceptable salt, enantiomer, diastereomer, atropisomer, racemate, polymorph, solvate or isotopically labeled compound thereof,

wherein R is 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 method for preparing the above compound.

It is a further object of the present disclosure to provide a pharmaceutical composition comprising a therapeutically effective amount of one or more of the above compounds or pharmaceutically acceptable salts, enantiomers, diastereomers, atropisomers, racemates, polymorphs, solvates or isotopically labeled compounds thereof.

It is a further object of the present disclosure to provide the use of the above compound, or a pharmaceutically acceptable salt, enantiomer, diastereomer, atropisomer, racemate, polymorph, solvate or isotopically labeled compound thereof, in the manufacture of a medicament for the treatment of a disease or disorder mediated by PRC 2.

It is a further object of the present disclosure to provide a method of treating a disease or disorder mediated by EED and/or PRC2 characterized by administering to a subject a therapeutically effective amount of one or more of the above compounds or pharmaceutically acceptable salts, enantiomers, diastereomers, atropisomers, racemates, polymorphs, solvates or isotopically labeled compounds thereof.

According to one aspect of the present disclosure, there is provided a compound represented by formula I, a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, an atropisomer, a racemate, a polymorph, a solvate, or an isotopically labeled compound thereof:

wherein the content of the first and second substances,

(1)

is a single or double bond;

(2)R ¹ 、R ² and R ³ Each independently is hydrogen, halogen、C ₁ -C ₄ Alkyl radical, C ₁ -C ₄ Haloalkyl, -O- (C) ₁ -C ₄ Alkyl group), C ₁ -C ₄ Haloalkoxy or 3-6 membered cycloalkyl;

R ⁴ is hydrogen, OH, ═ O, or C ₁ -C ₄ An alkyl group;

R ⁵ is hydrogen, halogen, C ₁ -C ₄ Alkyl, or C ₁ -C ₄ A haloalkyl group;

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

R ⁶ independently selected from halogen, CN, by 0-1R ^A1 Substituted C ₁ -C ₆ Alkyl, 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 groups);

R ^A2 selected from hydrogen, C ₁ -C ₄ Alkyl, 3-6 membered cycloalkyl, and C ₁ -C ₄ A haloalkyl group;

R ^A3 independently at each occurrence is selected from C ₁ -C ₄ Alkyl, 3-6 membered cycloalkyl, 6-10 membered aromatic ring group, and 5-10 membered heteroaromatic ring group;

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

R ^a and R ^b Independently at each occurrence, selected from hydrogen, C ₁ -C ₄ Alkyl, and 3-6 membered ringAn alkyl group;

(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 )-；

R ^c And R ^d Independently at each occurrence, selected from hydrogen, halogen, OH, C ₁ -C ₄ Alkyl radical, C ₁ -C ₄ Haloalkyl, C ₁ -C ₄ Alkoxy, and C ₁ -C ₄ A haloalkoxy group; or R ^c And R ^d Are linked to each other to form-R ^c’ -L-R ^d’ -and is bonded to the carbon atom to which it is attached to form a ring, wherein L is absent or-O-; r ^c ' and R ^d ' independently of one another are absent, C ₁ -C ₄ Alkylene radical, C ₁ -C ₃ Haloalkylene, or substituted by 1-2 halogens, OH, or C ₁ -C ₄ Alkyl substituted C ₁ -C ₃ An alkylene group;

n is an integer selected from 1 to 4.

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

the method comprises the following steps:

preparing lactone analogue according to the way described in the reaction formula I, specifically, starting from A-1, carrying out suzuki reaction with boric acid ester A-2 to generate A-3, hydrolyzing A-3 to obtain acid A-4, and carrying out a trans-kavalactonization reaction with A-5 to generate lactone A-6, wherein R is ¹ 、R ⁶ M, Z and n are as defined above;

the second method comprises the following steps:

according to the reaction formulaThe method described in the second paragraph prepares lactam analogues, specifically, starting from A-1, the lactam analogues are subjected to a suzuki reaction with Boc-protected amino borate A-7 to generate A-8, then Boc is removed to obtain A-9, and finally intramolecular amine transesterification is carried out under the action of potassium tert-butoxide to obtain A-10, wherein R is ¹ 、R ⁶ 、R ^A4 M, Z and n are as defined above;

the third method comprises the following steps:

preparing lactam analog according to the route described in the reaction formula III, specifically, carrying out suzuki reaction on A-1 and boric acid ester A-11 to generate A-12, then carrying out intramolecular ester condensation under the condition of tert-butyl alcohol to obtain A-13, and finally obtaining A-14 under the condition of trifluoroacetic acid, wherein R is R ¹ 、R ⁶ M, Z and n are as defined above.

According to another aspect of the present disclosure, there is provided a pharmaceutical composition comprising one or more selected from the group consisting of the compounds described above, pharmaceutically acceptable salts thereof, enantiomers, diastereomers, atropisomers, racemates, polymorphs, solvates or isotopically labeled compounds, and at least one pharmaceutically acceptable carrier, diluent or excipient.

According to another aspect of the present disclosure, there is provided the use of a compound as described above, a pharmaceutically acceptable salt, enantiomer, diastereomer, atropisomer, racemate, polymorph, solvate or isotopically labeled compound thereof, or a pharmaceutical composition as described above, in the manufacture of a medicament for the treatment of a disease or condition mediated by EED and/or PRC 2.

Advantageous effects

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 novel medicaments for treating diseases or conditions mediated by EED and/or PRC 2.

Detailed Description

To make the features and effects of the present invention comprehensible to those having ordinary knowledge in the art, general description and definitions are made with respect to terms and phrases mentioned in the specification and claims. 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.

In this document, the terms "comprising," "including," "having," "containing," or any other similar term, are intended to be open-ended franslational phrase (open-ended franslational phrase) and are intended to cover non-exclusive inclusions. For example, a composition or article comprising a plurality of elements is not limited to only those elements recited herein, but may include other elements not expressly listed but generally inherent to such composition or article. In addition, unless expressly stated to the contrary, the term "or" is intended to mean an inclusive "or" rather than an exclusive "or". For example, the condition "a or B" is satisfied in any of the following cases: a is true (or present) and B is false (or not present), a is false (or not present) and B is true (or present), both a and B are true (or present). Furthermore, in this document, the terms "comprising," including, "" having, "" containing, "and" containing "are to be construed as specifically disclosed and to cover both closed and semi-closed conjunctions, such as" consisting of … "and" consisting essentially of ….

All features or conditions defined herein 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 have covered and specifically disclosed all possible subranges and individual numerical values within the ranges, particularly integer numerical values. For example, a description of a range of "1 to 8" should be considered 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, and so on, particularly subranges bounded by all integer values, and should be considered to have specifically disclosed individual values such as 1,2, 3, 4,5, 6, 7, 8, and so on, within the range. Unless otherwise indicated, the foregoing explanatory methods apply to all matters contained in the entire disclosure, whether broad or not.

If an amount or other value or parameter is expressed as a range, preferred range, or a list of upper and lower limits, it is to be understood that all ranges subsumed therein for any pair of that range's upper or preferred value and that range's lower or preferred value, whether or not such ranges are separately disclosed, are specifically disclosed herein. Further, when a range of numerical values is recited herein, unless otherwise stated, the range is intended to include the endpoints thereof, and all integers and fractions within the range.

In this context, numerical values should be understood to have the precision of the number of significant digits of the value, provided that the object of the invention is achieved. For example, the number 40.0 should be understood to encompass a range from 39.50 to 40.49.

In this document, where Markush group (Markush group) or Option language is used to describe features or examples of the invention, those skilled in the art will recognize that a sub-group of all elements or any individual element within a Markush group or list of options may also be used to describe the invention. For example, if X is described as "selected from the group consisting of ₁ 、X ₂ And X ₃ The group "also indicates that X has been fully described as X ₁ Is claimed with X ₁ And/or X ₂ Claim (5). Furthermore, where Markush group or option terms are used to describe features or examples of the invention, those skilled in the art will recognize that any combination of sub-groups of all elements or individual elements within the Markush group or option list can also be used to describe the invention. Accordingly, for example, if X is described as being "selected from" X ₁ 、X ₂ And X ₃ Group consisting of "and Y is described as" selected from Y ₁ 、Y ₂ And Y ₃ Group of X is X ₁ Or X ₂ Or X ₃ And Y is Y ₁ Or Y ₂ Or Y ₃ Claim (5).

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

According to one embodiment of the present disclosure, there is provided a compound represented by formula I, a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, an atropisomer, a racemate, a polymorph, a solvate, or an isotopically labeled compound thereof:

wherein, the first and the second end of the pipe are connected with each other,

(1)

is a single or double bond;

(2)R ¹ 、R ² and R ³ Each independently of the others is hydrogen, halogen, C ₁ -C ₄ Alkyl radical, C ₁ -C ₄ Haloalkyl, -O- (C) ₁ -C ₄ Alkyl group), C ₁ -C ₄ Haloalkoxy or 3-6 membered cycloalkyl;

R ⁴ is hydrogen, OH, ═ O or C ₁ -C ₄ An alkyl group;

R ⁵ is hydrogen, halogen, C ₁ -C ₄ Alkyl or C ₁ -C ₄ A haloalkyl group;

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

R ⁶ independently selected from halogen, CN, by 0-1R ^A1 Substituted C ₁ -C ₆ Alkyl, 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 selected from hydrogen, C ₁ -C ₄ Alkyl, 3-6 membered cycloalkyl and C ₁ -C ₄ A haloalkyl group;

R ^A3 independently at each occurrence is selected from C ₁ -C ₄ Alkyl, 3-6 membered cycloalkyl, 6-10 membered aromatic ring group and 5-10 membered heteroaromatic ring group;

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

R ^a and R ^b Independently at each occurrence, 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 )-；

R ^c And R ^d Independently at each occurrence, selected from hydrogen, halogen, OH, C ₁ -C ₄ Alkyl radical, C ₁ -C ₄ Haloalkyl, C ₁ -C ₄ Alkoxy and C ₁ -C ₄ A haloalkoxy group; or R ^c And R ^d Are linked to each other to form-R ^c’ -L-R ^d’ -and is bonded to the carbon atom to which it is attached to form a ring, wherein L is absent or-O-; r is ^c’ And R ^d’ Each independently is absent, C ₁ -C ₄ Alkylene radical, C ₁ -C ₃ Haloalkylene, or substituted by 1-2 halogens, OH, or C ₁ -C ₄ Alkyl substituted C ₁ -C ₃ An alkylene group;

n is an integer selected from 1 to 4.

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

wherein the content of the first and second substances,

(1)R ¹ is hydrogen, halogen, C ₁ -C ₄ Alkyl radical, C ₁ -C ₄ Haloalkyl or 3-6 membered cycloalkyl;

(2)R ⁴ is hydrogen, OH or ═ O;

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

R ⁶ independently selected from halogen, by 0-1R ^A1 Substituted C ₁ -C ₆ Alkyl, 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 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 Independently at each occurrence, selected from hydrogen, C ₁ -C ₄ Alkyl and 3-6 membered cycloalkyl;

R ^a and R ^b Independently at each occurrence, 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 )-；

R ^c And R ^d Independently at each occurrence is selected from hydrogen, halogen, OH, C ₁ -C ₄ Alkyl radical, C ₁ -C ₄ Haloalkyl, C ₁ -C ₄ Alkoxy, and C ₁ -C ₄ A haloalkoxy group; or R ^c And R ^d Are linked to each other to form-R ^c’ -L-R ^d’ -and is bonded to the carbon atom to which it is attached to form a ring, wherein L is absent or-O-; r is ^c’ And R ^d’ Each independently is absent, C ₁ -C ₄ Alkylene radical, C ₁ -C ₃ Haloalkylene, or substituted by 1-2 halogens, OH, or C ₁ -C ₄ Alkyl substituted C ₁ -C ₃ An alkylene group;

n is an integer selected from 1 to 3.

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

wherein the content of the first and second substances,

(1)R ¹ is hydrogen or halogen;

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

R ⁶ independently selected from halogen, by 0-1R ^A1 Substituted C ₁ -C ₆ Alkyl, 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 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 Independently at each occurrence selected from hydrogen, C ₁ -C ₄ Alkyl and 3-6 membered cycloalkyl;

R ^a and R ^b Independently at each occurrence 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 Independently at each occurrence, selected from hydrogen, halogen, OH, C ₁ -C ₄ Alkyl and C ₁ -C ₄ A haloalkyl group;

n is 1 or 2.

According to another embodiment of the present disclosure, the compound of formula I is selected from the group consisting of compounds of formulae I-3a and I-3 b:

wherein the content of the first and second substances,

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

R ^A4 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 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 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 the context of the present disclosure, it is,

halogen means fluorine, chlorine, bromine or iodine.

Alkyl refers to a straight or branched monovalent saturated hydrocarbon group; such as 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 means a cyclic, monovalent, saturated radical containing on the ring an atom selected from oxygen, nitrogen, sulfur; such as pyrrolidinyl, tetrahydrofuranyl, tetrahydrothienyl, and the like.

The aromatic ring means an aromatic hydrocarbon such as benzene, naphthalene, anthracene, or the like.

The heteroaromatic ring is a ring having aromaticity including an atom selected from oxygen, nitrogen and sulfur in the ring, and examples thereof include furan, pyrrole, thiophene and imidazole.

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

Alkylene means a linear or branched divalent saturated hydrocarbon group; for example, methylene, ethylene, 1, 3-propylene, 1, 2-propylene, 1, 4-butylene, 1, 3-butylene, and the like.

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

According to one embodiment of the present disclosure, there is provided a pharmaceutical composition comprising one or more selected from the group consisting of the above-mentioned compounds, pharmaceutically acceptable salts, enantiomers, diastereomers, atropisomers, racemates, polymorphs, solvates or isotopically labeled compounds thereof, 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 the group consisting of other anti-cancer agents, immune modulators, anti-allergic agents, antiemetics, pain relievers, cytoprotective agents, and combinations thereof. Here, the other anticancer agents refer to anticancer agents other than the above-mentioned compounds of the present disclosure, pharmaceutically acceptable salts, enantiomers, diastereomers, atropisomers, racemates, polymorphs, solvates and isotopically labeled compounds thereof.

According to one embodiment of the present disclosure there is provided the use of the compound, a pharmaceutically acceptable salt, enantiomer, diastereomer, atropisomer, racemate, polymorph, solvate or isotopically labeled compound thereof, or the pharmaceutical composition in the manufacture of a medicament for the treatment of a disease or condition mediated by EED and/or PRC 2.

According to another embodiment of the present disclosure, wherein the disease or condition mediated by EED and/or PRC2 includes diffuse large B-cell lymphoma, follicular lymphoma, leukemia, multiple myeloma, mesothelioma, gastric cancer, malignant rhabdoid tumor, hepatocellular carcinoma, prostate cancer, breast cancer, cholangiocarcinoma and gallbladder cancer, bladder cancer; including neuroblastoma, schwannomas, 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, renal cell carcinoma, rectal cancer, thyroid cancer, parathyroid tumor, uterine tumor, and soft tissue sarcoma.

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

the method comprises the following steps:

preparation of lactone analogs according to the route described in equation one, in particular starting from A-1 with boronic acidsEster A-2 is subjected to suzuki reaction to generate A-3, A-3 is hydrolyzed to obtain acid A-4, and then the acid A-4 and A-5 are subjected to a mountain-mouth lactonization reaction to generate lactone A-6, wherein R ¹ 、R ⁶ M, Z or n are as defined above;

the second method comprises the following steps:

the lactam analogs were prepared according to the route described in scheme two, specifically starting from A-1 by suzuki reaction with Boc protected aminoboronic ester A-7 to form A-8, followed by removal of Boc to give A-9 and finally by intramolecular amine transesterification under the action of potassium tert-butoxide to give A-10, where R is ¹ 、R ⁶ 、R ^A4 M, Z and n are as defined above;

the third method comprises the following steps:

the lactam analog is prepared according to the pathway described in the reaction formula III, specifically, starting from A-1, the lactam analog reacts with boric acid ester A-11 in a suzuki reaction to generate A-12, then intramolecular ester is condensed under the condition of tert-butyl alcohol to obtain A-13, and finally, A-14 is obtained under the condition of trifluoroacetic acid, wherein R is R ¹ 、R ⁶ M, Z and n are as defined above.

Compound a-1 can be prepared according to CN109790166A or according to conventional methods known in the art.

The optimal reaction conditions and reaction times for each individual step may vary depending on the particular reactants used and the substituents present in all of the reactants. Solvents, temperatures and other reaction conditions can be readily selected by one skilled in the art unless otherwise specified. Specific steps are provided in the synthesis examples section. The reaction may be further processed in a conventional manner, for example by removing the solvent from the residue and further purified 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.

The starting materials, if not commercially available, may be prepared by a procedure selected from standard organic chemistry techniques, techniques analogous to the synthesis of known structural analogs, or techniques analogous to the procedures described in the schemes or synthetic 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 (e.g., prepared by asymmetric induction of an appropriate reaction step), or by resolving a mixture of stereoisomers of the compound or intermediate using standard procedures (e.g., chromatographic separation, recrystallization, or enzymatic resolution).

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 a starting material, or by resolving a mixture of geometric isomers of the compound or intermediate using standard procedures, such as chromatographic separation.

In preparing polymorphs of the compounds of the present disclosure, crystals are obtained by using a recrystallization method and the crystal structure thereof is obtained by X-ray diffraction.

When preparing the isotopically labeled compounds of the present disclosure, the reaction may be carried out using isotopically labeled starting materials as needed.

Examples

The following examples were prepared, isolated and characterized using the methods disclosed herein. The following examples illustrate some of the scope of the disclosure and are not meant to be all of the scope of the disclosure.

Example 1 Synthesis of ZB-PY-E18

To compound 1(100mg, 0.230mmol), (2-aminophenyl) boronic acid (63.0mg, 0.460mmol) and potassium carbonate (K) ₂ CO ₃ 95.2mg, 0.690mmol) of the resulting solution was purged with nitrogen in a mixed solvent of 1, 4-dioxane (5mL) and water (1mL) for 1 minute, and Pd (dppf) Cl was added ₂ (17.4mg,0.0230mmol) and the sealed tube was heated in a Microwave (MW) reactor at 125 ℃ for 1 hour. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate (same drying conditions as in the examples below unless otherwise specified), concentrated, and purified by silica gel column chromatography (same silica gel column chromatography conditions as in the examples below as an eluent) to give ZB-PY-E18(26.0mg, 28%) as a yellow solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ(ppm):9.51(s,1H),8.67(s,1H),8.53(t,J＝5.1Hz,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(100mg, 0.230mmol), (2-aminopyridin-3-yl) boronic acid (95.2mg, 0.690mmol) and potassium carbonate (95.2mg, 0.690mmol) in a mixed solvent of 1, 4-dioxane (5mL) and water (1mL) was bubbled nitrogen for 1 minute, PdCl was added ₂ (dppf) (17.4mg,0.0230mmol) and the sealed tube was heated in a microwave reactor at 125 ℃ for 1 hour. Water was added to the reaction solution, 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(17.0mg, 18.4%) as a white solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ(ppm):9.30(s,1H),8.70-8.65(m,2H),8.32(d,J＝7.8Hz,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(100mg, 0.230mmol), (2- (hydroxymethyl) phenyl) boronic acid (105mg, 0.690mmol), tetrakis (triphenylphosphine) palladium (Pd (PPh) ₃ ) ₄ 26.6mg,0.0230mmol) and potassium carbonate (95.2mg, 0.690mmol) were added to a mixed solvent of 1, 4-dioxane (5mL) and water (1mL) to displace nitrogen and heated at 110 ℃ for 4 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated and chromatographed on silica gel (5% methanol in dichloromethane as eluent) to give the title compound 3.2(60.0mg, 56%) as a pale yellow solid. LCMS: [ M + H ]] ⁺ ＝463.1。

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

Synthesis of ZB-PY-E16: triethylamine (TEA, 62.4mg, 0.618mmol) and 2,4, 6-trichlorobenzoyl chloride (125mg, 0.517mmol) were added to a solution of compound 3.3(45.0mg, 0.103mmol) in toluene (5mL) and reacted at 25 ℃ for 4 hours. The reaction solution was then diluted with toluene (5mL) and added dropwise slowly to a solution of 4-dimethylaminopyridine (DMAP, 17.7mg, 0.144mmol) in toluene (10mL) at 80 ℃ 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(15.0mg, 35%) as a white solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ8.87(s,1H)，8.73(t,J＝5.1Hz,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＝4.9Hz,2H)，4.56(t,J＝8.7Hz,2H)，3.35(t,J＝8.8Hz,2H).LCMS:[M+H] ⁺ ＝417.3。

Example 4 Synthesis of ZB-PY-E21

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

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

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

Synthesis of ZB-PY-E21: triethylamine (60.6mg, 0.600mmol) and 2,4, 6-trichlorobenzoyl chloride (121mg, 0.500mmol) were added to a solution of compound 4.4(43.6mg, 0.1mmol) in toluene (5mL) and reacted at 25 ℃ for 4 hours. Then the reaction solution was treated with tolueneBenzene (5mL) was diluted and added dropwise slowly to a solution of 4-dimethylaminopyridine (17.2mg, 0.14mmol) in toluene (10mL) at 80 ℃ 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(13.0mg, 31%) as a white solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ8.90(s,1H),8.84(t,J＝5.1Hz,1H),8.64(dd,J＝4.7,1.6Hz,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(100mg, 0.230mmol), (2- (2-hydroxyethyl) phenyl) boronic acid (74.7mg, 0.461mmol), potassium carbonate (95.2mg, 0.690mmol) and tetrakis (triphenylphosphine) palladium (26.6mg,0.0230mmol) were added to a mixed solvent of 1, 4-dioxane (5mL) and water (1mL), nitrogen was replaced and the mixture was heated at 100 ℃ for 4 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated and chromatographed on silica gel (5% methanol in dichloromethane as eluent) to give compound 5.2(80.0mg, 73%) as a pale yellow solid. LCMS: [ M + H] ⁺ ＝477.2。

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

Synthesis of ZB-PY-E19: triethylamine (67.7mg, 0.670mmol) and 2,4, 6-trichlorobenzoyl chloride (243mg, 0.558mmol) were added to compound 5.3(50.0mg, 0.112mmol) in toluene (5mL) and reacted at 25 ℃ for 4 hours. The reaction solution was then diluted with toluene (5mL) and added dropwise slowly to a solution of 4-dimethylaminopyridine (19.3mg, 0.157mmol) in toluene (10mL) at 80 ℃ 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(13.0mg, 31%) as a white solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ8.81(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(100mg, 0.230mmol), (2- (((tert-butoxycarbonyl) amino) methyl) phenyl) boronic acid (154mg, 0.461mmol), potassium carbonate (95.2mg, 0.690mmol) and tetrakis (triphenylphosphine) palladium (26.6mg,0.023mmol) were added to a mixed solvent of 1, 4-dioxane (5mL) and water (1mL), and heated at 100 ℃ for 4 hours while displacing nitrogen. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated and chromatographed on silica gel (3% methanol in dichloromethane as eluent) to give compound 6.2(52mg, 40%) as a pale yellow solid. LCMS: [ M + H] ⁺ ＝562.3。

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

Synthesis of ZB-PY-E20: to a solution of compound 6.3(42.0mg, 0.091mmol) in N, N-Dimethylformamide (DMF) was added potassium tert-butoxide (t-BuOK, 30.6mg, 0.273mmol) at 0 ℃ and the reaction was carried out for 2 hours at 0 ℃. AddingQuench with water and extract with ethyl acetate. The organic phase was washed with brine, dried, concentrated and purified by silica gel column chromatography to give ZB-PY-E20(15.0mg, 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(200mg, 0.461mmol), tert-butyl 2- (2- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenyl) acetate (293mg, 0.922mmol), tetrakis (triphenylphosphine) palladium (53.2mg,0.0461mmol) and potassium carbonate (191mg, 1.38mmol) were added to a mixed solvent of 1, 4-dioxane (10mL) and water (2mL), and heated at 100 ℃ for 4 hours while displacing nitrogen. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated and chromatographed on silica gel (3% methanol in dichloromethane as eluent) to give compound 7.2(210mg, 83%) as a white solid. LCMS: [ M + H ]] ⁺ ＝547.2。

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

Synthesis of compound 7.4: potassium tert-butoxide (111mg, 0.989mmol) was added to a solution of compound 7.3(213mg, 0.330mmol) in toluene (10mL) at 0 ℃ and stirred at 0 ℃ for 2 hours. Saturated aqueous ammonium chloride solution was added thereto, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated and chromatographed on silica gel (3% methanol in dichloromethane as eluent) to give compound 7.4(110mg, 56%) as a pale yellow solid. LCMS: [ M + H] ⁺ ＝601.2。

Synthesis of ZB-PY-E22: compound 7.4(110mg, 0.183mmol) was dissolved in dichloromethane (10mL) and trifluoroacetic acid (2mL) 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(15.0mg, 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(200mg, 0.461mmol), 4,4,4',4',5,5,5',5' -octamethyl-2, 2' -bis (1,3, 2-dioxaborolane) (234mg, 0.922mmol), potassium acetate (KOAC, 1.36g, 1.38mmol) and PdCl ₂ (dppf) (34.9mg,0.0461mmol) was added to 1, 4-dioxane (10mL), nitrogen was replaced, and the mixture was heated at 100 ℃ for 2 hours. The reaction mixture was concentrated, and subjected to silica gel column chromatography to give compound 8.2(65mg, 29.2%). LCMS: [ MH] ⁺ ＝483.2。

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

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

Synthesis of Compound ZB-PY-E23 to a solution of Compound 8.4(25.7mg, 0.0528mmol) in N, N-dimethylformamide (5mL) was added potassium tert-butoxide (15.2mg, 0.158mmol), and the reaction was carried out at 0 ℃ for 2 hours. 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 objective compound ZB-PY-E23(8mg, 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(200mg, 0.460mmol), (2- (((tert-butoxycarbonyl) amino) methyl) pyridin-3-yl) boronic acid (174mg, 0.692mmol), potassium carbonate (191mg, 1.38mmol) and tetrakis (triphenylphosphine) palladium (34.9mg,0.0461mmol) were added to a mixed solvent of 1, 4-dioxane (5mL) and water (1mL), replaced with nitrogen, and heated at 100 ℃ for 4 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated and chromatographed on silica gel (3% methanol in dichloromethane as eluent) to give compound 9.2(120mg, 46%) as a pale yellow solid. LCMS: [ M + H] ⁺ ＝563.3。

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

Synthesis of Compound ZB-PY-E24: to a solution of compound 9.3(98.2mg,0.213 mmol) in N, N-dimethylformamide (5mL) at 0 ℃ was added potassium tert-butoxide (78.0mg, 0.639mmol), and the reaction was carried out at 0 ℃ for 2 hours. Quenched by addition of water and extracted with ethyl acetate. Organic phaseWashed with brine, dried, concentrated and purified by silica gel column chromatography to give ZB-PY-E24(25.0mg, 28%) as a white solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ8.80(s,1H),8.64(t,J＝5.1Hz,1H),8.52(dd,J＝4.8,1.6Hz,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＝14.2,6.0Hz,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(174mg, 0.402mmol), tert-butyl 3- (2- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenyl) propionate (200mg, 0.602mmol), tetrakis (triphenylphosphine) palladium (46.4mg,0.0602mmol) and potassium carbonate (222mg, 1.61mmol) were added to a mixed solvent of 1, 4-dioxane (10mL) and water (2mL), nitrogen was replaced, and the mixture was heated at 100 ℃ for 4 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated and chromatographed on silica gel (3% methanol in dichloromethane as eluent) to give compound 10.2(110mg, 49%) as a white solid. LCMS: [ M + H] ⁺ ＝561.2。

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

Synthesis of compound 10.4: potassium tert-butoxide (61.1mg, 0.546mmol) was added to a solution of compound 10.3(120mg, 0.182mmol) in toluene (10mL) at 0 ℃ and stirred at 0 ℃ for 2 hours. Saturated aqueous ammonium chloride solution was added to quench, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated and chromatographed on silica gel (5% methanol in methylene chloride)Alkane solution as eluent) to give compound 10.4(45.0mg, 40%) as a light yellow solid. LCMS: [ M + H] ⁺ ＝617.3。

Synthesis of Compound ZB-PY-E25: compound 10.4(45.0mg, 0.0730mmol) was dissolved in dichloromethane (10mL) and trifluoroacetic acid (2mL) 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(15.0mg, 50%) as a yellow solid. ¹ H NMR (400MHz, chloroform- _d )δ8.27(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＝8.7Hz,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(100mg, 0.230mmol), (2- (((tert-butoxycarbonyl) (methyl) amino) methyl) pyridin-3-yl) boronic acid (123mg, 0.461mmol), potassium carbonate (95.2mg, 0.690mmol) and tetrakis (triphenylphosphine) palladium (26.6mg,0.0230mmol) were added to a mixed solvent of 1, 4-dioxane (5mL) and water (1mL), nitrogen was replaced, and the mixture was heated at 100 ℃ for 4 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated and chromatographed on silica gel (5% methanol in dichloromethane as eluent) to give compound 11.2(73mg, 40%) as a pale yellow solid. LCMS: [ M + H] ⁺ ＝577.3。

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

Synthesis of Compound ZB-PY-E32: to a solution of compound 11.3(60.1mg, 0.126mmol) in N, N-dimethylformamide at 0 ℃ was added potassium tert-butoxide (42.3mg, 0.378mmol), and the reaction was carried out at 0 ℃ for 2 hours. Quenching with water and ethyl acetateAnd (4) extracting. The organic phase was washed with brine, dried, concentrated and purified by silica gel column chromatography to give ZB-PY-E32(17.0mg, 31%) as a white solid. LCMS: [ M + H] ⁺ ＝431.2。 ¹ H NMR(400MHz,DMSO-d ₆ )δ8.81(s,1H),8.67(t,J＝5.1Hz,1H),8.53(dd,J＝4.9,1.6Hz,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,0.0460mmol) and potassium carbonate (190mg, 1.38mmol) were added to a mixed solvent of 1, 4-dioxane (5mL) and water (1mL), nitrogen was replaced and the mixture was heated at 110 ℃ for 4 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated and chromatographed on silica gel (5% methanol in dichloromethane as eluent) to give compound 12.2(130mg, 59%) as a pale yellow solid. LCMS: [ M + H ]] ⁺ ＝477.1。

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

Synthesis of ZB-PY-E26: triethylamine (94.7mg, 0.938mmol) and 2,4, 6-trichlorobenzoyl chloride (189mg, 0.781mmol) were added to a solution of compound 12.3(70mg, 0.156mmol) in toluene (5mL) and reacted at 25 ℃ for 4 hours. The reaction was then diluted with toluene (5mL) and added dropwise slowly to a solution of 4-dimethylaminopyridine (26.8mg, 0.218mmol) in toluene (10mL)To the solution, the solution was added dropwise at 80 ℃ 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(15.0mg, 22%) as a white solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ8.84(s,1H),8.72(t,J＝5.1Hz,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＝8.8Hz,2H),3.33–3.35(m,2H)，1.65(d,J＝6.4Hz,3H).LCMS:[M+H] ⁺ ＝431.3。

Example Synthesis of 13 ZB-PY-E27

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

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

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

Synthesis of Compound ZB-PY-E27: triethylamine (75.5mg, 0.748mmol) and 2,4, 6-trichlorobenzoyl chloride (151mg, 0.624mmol) were added to a solution of compound 13.4(56mg, 0.125mmol) in toluene (5mL) and reacted at 25 ℃ for 4 hours. The reaction was then diluted with toluene (5mL) and added dropwise slowly to a solution of 4-dimethylaminopyridine (21.5mg, 0.175mmol) in toluene (10mL) at 80 ℃ for 4 h. 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(17.0mg, 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(289mg, 0.667mmol), (2- (((tert-butyldimethylsilyl) oxo) methyl) -6-cyclopropylpyridin-3-yl) boronic acid (307mg, 1.00mmol), potassium carbonate (276mg, 2.00mmol) and palladium tetratriphenylphosphine (77.0mg,0.0667mmol) were added to a mixed solvent of 1, 4-dioxane (5mL) and water (1mL), replaced with nitrogen and heated at 100 ℃ for 4 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated and chromatographed on silica gel (5% methanol in dichloromethane as eluent) to give compound 14.2(170mg, 41%) as a pale yellow solid. LCMS: [ M + H] ⁺ ＝618.3。

Compound 14.3The synthesis of (2): tetrabutylammonium fluoride (1M, 0.413mL) was added to a solution of compound 14.2(170mg, 0.275mmol) in tetrahydrofuran (5mL), and the resulting solution was stirred at room temperature for 4 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated and chromatographed on silica gel (5% methanol in dichloromethane as eluent) to give compound 14.3(130.0mg, 94%) as a pale yellow solid. LCMS: [ M + H] ⁺ ＝504.10。

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

Synthesis of Compound ZB-PY-E28: triethylamine (60.6mg, 0.600mmol) and 2,4, 6-trichlorobenzoyl chloride (121mg, 0.500mmol) were added to a solution of compound 14.4(47.5mg, 0.100mmol) in toluene (5mL) and reacted at 25 ℃ for 4 hours. The reaction was then diluted with toluene (5mL) and added dropwise slowly to a solution of 4-dimethylaminopyridine (17.2mg, 0.14mmol) in toluene (10mL) at 80 ℃ for 4 h. 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(17.0mg, 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＝12.4,8.0,5.2Hz,1H),0.90–1.10(m,4H).LCMS:[M+H] ⁺ ＝458.2。

Example 15 Synthesis of ZB-PY-E31

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

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

Synthesis of Compound ZB-PY-E31: to a solution of compound 15.3(60.1mg, 0.126mmol) in N, N-dimethylformamide at 0 ℃ was added potassium tert-butoxide (42.3mg, 0.378mmol), and the reaction was carried out at 0 ℃ for 2 hours. Quenched by addition of water 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(13.0mg, 24%) as a white solid. LCMS: [ M + H ]] ⁺ ＝430.1。 ¹ H NMR(400MHz,DMSO-d ₆ )δ8.80(s,1H),8.57(d,J＝5.0Hz,1H),7.32–7.55(m,5H),6.96(dd,J＝10.3,8.6Hz,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＝8.7Hz,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(321mg, 0.741mmol), (3- (((tert-butyldimethylsilyl) oxy) methyl) -1-methyl-1H-pyrazol-4-yl) boronic acid (600mg, 2.22mmol), potassium carbonate (306mg, 2.22mmol) and tetrakis (triphenylphosphine) palladium (85.6mg,0.0741mmol) were added to 1, 4-dioxane (5mL) and water(1mL) was added to the mixed solvent, and the mixture was heated at 100 ℃ for 4 hours while replacing nitrogen. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated and chromatographed on silica gel (5% methanol in dichloromethane as eluent) to give compound 16.2(210mg, 49%) as a pale yellow solid. LCMS: [ M + H ]] ⁺ ＝581.3。

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

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

Synthesis of Compound ZB-PY-E30: triethylamine (104mg, 1.02mmol) and 2,4, 6-trichlorobenzoyl chloride (207mg, 0.850mmol) were added to a solution of compound 16.4(75.0mg, 0.171mmol) in toluene (5mL) and reacted at 25 ℃ for 4 hours. The reaction solution was then diluted with toluene (5mL) and added dropwise slowly to a solution of 4-dimethylaminopyridine (25.2mg, 0.205mmol) in toluene (10mL) at 80 ℃ 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(30.0mg, 37%) as a white solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ8.83(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＝8.7Hz,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(321mg, 0.741mmol), (5- (((tert-butyldimethylsilyl) oxo) methyl) -1-methyl-1H-pyrazol-4-yl) boronic acid (600mg, 2.22mmol), potassium carbonate (306mg, 2.22mmol) and tetrakis (triphenylphosphine) palladium (85.6mg,0.0741mmol) were added to a mixed solvent of 1, 4-dioxane (5mL) and water (1mL), nitrogen was replaced, and the mixture was heated at 100 ℃ for 4 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated and chromatographed on silica gel (5% methanol in dichloromethane as eluent) to give compound 17.2(300mg, 70%) as a pale yellow solid. LCMS: [ M + H] ⁺ ＝581.3。

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

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

Synthesis of Compound ZB-PY-E29: triethylamine (194mg, 0.510mmol) and 2,4, 6-trichlorobenzoyl chloride (387mg, 1.60mmol) were added to a solution of compound 17.4(140mg, 0.320mmol) in toluene (5mL) and reacted at 25 ℃ for 4 hours. The reaction mixture was then diluted with toluene (5mL) and added dropwise slowly to a solution of 4-dimethylaminopyridine (47.2mg, 0.384mmol) in toluene (10mL) at 80 deg.CAfter 4 hours addition, the reaction was cooled, quenched with saturated aqueous sodium bicarbonate 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(30.0mg, 37%) as a white solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ8.85(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 inhibition complex 2(PRC2) enzyme activity assay

The activity of the PRC2 enzyme was detected using a time-resolved fluorescence energy transfer (TR-FRET) method. First, the enzyme was mixed with different concentrations of the compound and incubated for 30 minutes at room temperature. The enzymatic reaction was initiated by the addition of a biotin-labeled histone H3 polypeptide substrate and the cofactor S-adenosylmethionine (SAM). After 4 hours of reaction at room temperature, the recipient Acceptor and Donor Donor were added and incubated for half an hour. The fluorescence signal was detected with a multifunctional microplate reader EnVision (Perkin Elmer Corp.). Data were analyzed using GraphPad Prism 5.0 software to obtain IC ₅₀ The value is obtained.

The compounds described in Table 1 can be prepared by the methods described in the above examples, EED226 being a positive control compound (nat. chem. biol.2017,13, 381-388).

Compound (I)	Enzyme activity IC 50 (nM)	Compound (I)	Enzyme activity IC 50 (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: long-term growth inhibition experiment of cells

Pfeiffer cells in exponential growth phase were seeded in 24-well plates at a cell density of 1 × 10E5 cells/mL. Cells were treated the same day with different concentrations of compound added. Fresh media and compounds were replaced at 4 and 7 days of compound treatment. After 11 days of compound treatment, cell viability was determined using CellTiter-Glo reagent (Promega). Data were analyzed using GraphPad Prism 5.0 software to obtain IC ₅₀ The value is obtained.

The compounds described 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).

As can be seen from the data in tables 1 and 2 above, the IC of some of the compounds of the present disclosure for the PRC2 enzyme ₅₀ Value of<10nM, significantly higher than the positive control compound EED 226; similarly, IC of some compounds of the disclosure for the long-term growth inhibition assay of Pfeiffer cells ₅₀ Value of<1nM, significantly higher than the positive control compound EED 226.

The above embodiments are merely exemplary in nature and are not intended to limit the claimed embodiments or the application or uses of such embodiments. In this document, the term "exemplary" represents "as an example, instance, or illustration. Any exemplary embodiment herein is not necessarily to be construed as preferred or advantageous over other embodiments.

In addition, while at least one exemplary embodiment or comparative example has been presented in the foregoing detailed description, it should be appreciated that a vast number of variations are possible. It should also be appreciated that the embodiments described herein are not intended to limit the scope, applicability, or configuration of the claimed subject matter in any way. Rather, the foregoing implementations will provide those of ordinary skill in the art with a convenient road map for implementing the described embodiment or embodiments. Further, various changes may be made in the function and arrangement of elements without departing from the scope defined in the claims, which includes known equivalents and all foreseeable equivalents at the time of filing this patent application.

Claims (10)

1. A compound represented by the general formula I, a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, an atropisomer, a racemate, a polymorph, a solvate, or an isotopically labeled compound thereof:

wherein the content of the first and second substances,

(1)

is a single or double bond;

(2)R ¹ 、R ² and R ³ Each independently of the others is hydrogen, halogen, C ₁ -C ₄ Alkyl radical, C ₁ -C ₄ Haloalkyl, -O- (C) ₁ -C ₄ Alkyl group), C ₁ -C ₄ Haloalkoxy or 3-6 membered cycloalkyl;

R ⁴ is hydrogen, OH, ═ O or C ₁ -C ₄ An alkyl group;

R ⁵ is hydrogen, halogen, C ₁ -C ₄ Alkyl or C ₁ -C ₄ A haloalkyl group;

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

R ⁶ independently selected from halogen, CN, by 0-1R ^A1 Substituted C ₁ -C ₆ Alkyl, 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 groups);

R ^A2 selected from hydrogen, C ₁ -C ₄ Alkyl, 3-6 membered cycloalkyl, and C ₁ -C ₄ A haloalkyl group;

R ^A3 independently at each occurrence is selected from C ₁ -C ₄ Alkyl, 3-6 membered cycloalkyl, 6-10 membered aromatic ring group, and 5-10 membered heteroaromatic ring group;

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

R ^a and R ^b Independently at each occurrence, 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 )-；

R ^c And R ^d Independently at each occurrence, selected from hydrogen, halogen, OH, C ₁ -C ₄ Alkyl radical, C ₁ -C ₄ Haloalkyl, C ₁ -C ₄ Alkoxy, and C ₁ -C ₄ A haloalkoxy group; or R ^c And R ^d Are linked to each other to form-R ^c’ -L-R ^d’ -and is bonded to the carbon atom to which it is attached to form a ring, wherein L is absent or-O-; r ^c’ And R ^d’ Each independently is absent, C ₁ -C ₄ Alkylene radical, C ₁ -C ₃ Haloalkylene, or substituted by 1-2 halogens, OH, or C ₁ -C ₄ Alkyl substituted C ₁ -C ₃ An alkylene group;

n is an integer selected from 1 to 4.

2. The compound according to claim 1, a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, an atropisomer, a racemate, a polymorph, a solvate or an isotopically labeled compound thereof, wherein the compound represented by the general formula I is selected from the group consisting of compounds represented by the general formula I-1:

wherein the content of the first and second substances,

(1)R ¹ is hydrogen, halogen, C ₁ -C ₄ Alkyl radical, C ₁ -C ₄ Haloalkyl, or 3-6 membered cycloalkyl;

(2)R ⁴ is hydrogen, OH, or ═ O;

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

R ⁶ independently selected from halogen, by 0-1R ^A1 Substituted C ₁ -C ₆ Alkyl, 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 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 Independently at each occurrence, selected from hydrogen, C ₁ -C ₄ Alkyl and 3-6 membered cycloalkyl;

R ^a and R ^b Independently at each occurrence, 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 )-；

R ^c And R ^d Independently at each occurrence, selected from hydrogen, halogen, OH, C ₁ -C ₄ Alkyl radical, C ₁ -C ₄ Haloalkyl, C ₁ -C ₄ Alkoxy, and C ₁ -C ₄ A haloalkoxy group; or R ^c And R ^d Are linked to each other to form-R ^c’ -L-R ^d’ -and is bonded to the carbon atom to which it is attached to form a ring, wherein L is absent or-O-; r ^c’ And R ^d’ Each independently is absent, C ₁ -C ₄ Alkylene radical, C ₁ -C ₃ Haloalkylene, or substituted by 1-2 halogens, OH, or C ₁ -C ₄ Alkyl substituted C ₁ -C ₃ An alkylene group;

n is an integer selected from 1 to 3.

3. The compound according to claim 1, a pharmaceutically acceptable salt, enantiomer, diastereomer, atropisomer, racemate, polymorph, solvate or isotopically labeled compound thereof, wherein the compound represented by formula I is selected from the group consisting of compounds represented by formula I-2:

wherein the content of the first and second substances,

(1)R ¹ is hydrogen or halogen;

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

R ⁶ independently selected from halogen, by 0-1R ^A1 Substituted C ₁ -C ₆ Alkyl, 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 Independently at each occurrence, selected from hydrogen, C ₁ -C ₄ Alkyl and 3-6 membered cycloalkyl;

R ^a and R ^b Independently at each occurrence, 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 Independently at each occurrence, selected from hydrogen, halogen, OH, C ₁ -C ₄ Alkyl, and C ₁ -C ₄ A haloalkyl group;

n is 1 or 2.

4. The compound according to claim 1, a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, an atropisomer, a racemate, a polymorph, a solvate or an isotopically labeled compound thereof, wherein the compound represented by the general formula I is selected from the group consisting of compounds represented by general formulae I-3a and I-3 b:

wherein

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

R ^A4 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 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 Independently selected from C ₁ -C ₄ Alkyl and 3-6 membered cycloalkyl;

R ^A4 and R ^A5 Independently selected from hydrogen, C ₁ -C ₄ Alkyl and 3-6 membered cycloalkyl.

5. The compound according to any one of claims 1 to 4, a pharmaceutically acceptable salt, enantiomer, diastereomer, atropisomer, racemate, polymorph, solvate or isotopically labeled compound thereof, wherein the compound represented by formula I is selected from the group consisting of:

6. a pharmaceutical composition comprising one or more selected from the group consisting of a compound according to any one of claims 1 to 5, pharmaceutically acceptable salts, enantiomers, diastereomers and racemates thereof, 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 anti-cancer agents, immunomodulators, anti-allergic agents, antiemetics, pain relievers, cytoprotective agents, and combinations thereof.

9. Use of a compound according to any one of claims 1 to 5, a pharmaceutically acceptable salt, enantiomer, diastereomer or racemate thereof, or a pharmaceutical composition according to any one of claims 6 to 8, in the manufacture of a medicament for the treatment of a disease or condition mediated by EED and/or PRC 2.

10. Use according to claim 9, wherein

The diseases or disorders 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, esophageal cancer, head and neck cancer, lung cancer, nasopharyngeal cancer, ovarian cancer, pancreatic cancer, renal cell carcinoma, rectal cancer, thyroid cancer, parathyroid tumor, uterine tumor, and soft tissue sarcoma.