WO2022078403A1

WO2022078403A1 - Substituted pyridone compound and application

Info

Publication number: WO2022078403A1
Application number: PCT/CN2021/123577
Authority: WO
Inventors: 李桢; 唐锋; 刘乐; 赵春艳; 陈平; 唐任宏; 任晋生
Original assignee: 江苏先声药业有限公司
Priority date: 2020-10-15
Filing date: 2021-10-13
Publication date: 2022-04-21
Also published as: CN116782903A

Abstract

The present invention relates to a compound shown in general formula (A) or a pharmaceutically acceptable salt thereof, a pharmaceutical composition, a preparation method for the compound, and a use of the compound as an MAT2A inhibitor.

Description

Substituted pyridone compounds and applications

The present invention requires that it was submitted to the State Intellectual Property Office of China on October 15, 2020, the patent application number is 202011105645.3, and the name of the invention is "Substituted pyridone compounds and applications", submitted to the State Intellectual Property Office of China on February 10, 2021, The patent application number is 202110182410.2, the name of the invention is "Substituted pyridone compounds and their applications" and submitted to the State Intellectual Property Office of China on April 2, 2021, the patent application number is 202110361446.7, and the invention name is "Substituted pyridone compounds and their applications" ” of the priority of the earlier application. The entire contents of the aforementioned prior application are incorporated herein by reference.

technical field

The invention belongs to the field of medicine, and relates to a novel pyridone compound or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing them and the use as a MAT2A inhibitor.

Background technique

Methionine adenosyltransferase (MAT), also known as S-adenosylmethionine synthase, is able to catalyze the reaction of methionine (Met) with ATP to generate S-adenosylmethionine S-Adenosyl-L-methionine (SAM) class of enzymes. SAM is the main methyl donor in the body, which can regulate gene expression, transcription and translation through transmethylation reaction, and then have an important impact on cell growth, death and differentiation. Not only that, SAM is also involved in the biosynthesis of polyamines and glutathione.

There are three main subtypes of MAT enzymes, MAT1A, MAT2A and MAT2B. MAT1A mainly exists in normal hepatocytes, while MAT2A is widely distributed in extrahepatic cells. The two isoforms differ in catalytic efficiency and regulation. MAT2B does not have the ability to catalyze the synthesis of SAM, but as a regulatory subunit of MAT2A, after forming a complex with MAT2A, it regulates the catalytic activity of MAT2A.

Studies have shown that in liver cancer cells, the expression level of MAT1A is down-regulated and the expression of MAT2A is increased, which in turn promotes the proliferation of liver cancer cells. In addition, abnormally elevated expression levels of MAT2A also exist in many other tumors, and silencing the gene encoding MAT2A can lead to cancer cell death. Further, Marjon et al. (MTAP deletions in cancer create vulnerability to targeting of the MAT2A/PRMT5/RIOK1 axis. Marjon K, et al. Cell Reports. 2016, 15(3), 574–587) found MTAP-deficient cancer cell lines Sensitive to MAT2A inhibition. MTAP, also known as methylthioadenosine phosphorylase, is widely expressed in normal tissue cells. The enzyme catalyzes the conversion of methylthioadenosine (MTA) to 5-methylthioribose-1-phosphate and adenine. This process is also an important part of the methionine compensation pathway in the human body. When MTAP is depleted, the metabolic pathway of MTA is inhibited, which in turn leads to a large accumulation of MTA in the body, and finally makes cancer cells more sensitive to MAT2A inhibition.

The gene encoding human MTAP is located in the chromosome 9p21 region (chr9p21), and its frequency of homozygous deletion in all tumors is about 15%, and the deletion frequency varies in different tumors. Tumors with higher deletion frequencies include glioma, mesothelioma, melanoma, gastric cancer, esophageal cancer, bladder cancer, pancreatic cancer, non-small cell lung cancer, astrocytoma, osteosarcoma, head and neck cancer, and mucinous chondrosarcoma , ovarian cancer, endometrial cancer, breast cancer, soft tissue sarcoma, non-Hodgkin lymphoma, etc.

The 9p21 region of human chromosome contains not only the gene encoding MTAP, but also the tumor suppressor genes p16INK4A (also known as CDKN2A) and p15INK4B. MTAP is also absent in 80%-90% of CDKN2A-deficient tumors.

Given that the expression level of MAT2A is abnormally elevated in many types of tumors, including gastric cancer, colon cancer, liver cancer, and pancreatic cancer, and selective inhibition of MAT2A can reduce the proliferation activity of MTAP-deficient cancer cells. Therefore, selective inhibition of MAT2A can be used as an effective tumor therapy.

WO2018039972, WO2018045071 and WO2019191470 disclose MAT2A inhibitor heterocyclic compounds for the treatment of tumors.

SUMMARY OF THE INVENTION

The present invention provides a compound represented by general formula (A) or a pharmaceutically acceptable salt thereof:

Wherein, ring Q is a 5-6-membered heterocyclic group, a C ₆ -C ₁₀ -membered aryl group or a 5-10-membered heteroaryl group, and the 5-6-membered heterocyclic group, a C ₆ -C ₁₀ -membered aryl group or a 5-10 ^{A membered heteroaryl group is optionally substituted with R a} ^selected from F, Cl, Br, I, OH, CN, C ₂ -C ₃ alkynyl, C ₁ -C ₁₀ alkyl, C ₃ -C ₁₀ cycloalkyl or C ₁ -C ₁₀ alkoxy, said C ₁ -C ₁₀ alkyl, C ₃ -C ₁₀ cycloalkyl or C ₁ -C ₁₀ alkoxy optionally substituted by R ^b ;

R ^b is selected from F, Cl, Br, I, OH or CN;

Ring W is phenyl, pyridyl, pyrimidinyl, pyridazinyl, pyridinonyl, pyridazinonyl or

wherein Ring M is a 5-10 membered heteroaryl group or a 4-10 membered heterocyclic group, and said phenyl, pyridyl, pyrimidinyl, pyridazinyl, pyridinyl, pyridazinonyl or ring M is optionally replaced by R ^c is substituted, said R ^c is selected from OH, =O or the following groups optionally substituted by R ^c1 : C ₁ -C ₁₀ alkyl, C ₁ -C ₁₀ alkoxy, C ₃ -C ₁₀ cycloalkyl , P(O)(C ₁ -C ₃ alkyl) ₂ , 4-6 membered heterocycloalkyl, NHC(O)(C ₁ -C ₆ alkyl); X ₁ is selected from C or N, when X ₁ When selected from C,

Represents a double bond, when X ₁ is selected from N,

represents a single key;

R ^c1 is selected from deuterium, F, Cl, Br, I, CN, OH, NH ₂ or the following groups optionally substituted by R ^c2 : C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, NHC (O)O(C ₁ -C ₆ alkyl), (C ₁ -C ₆ alkyl) NHC(O)O, NH(C ₁ -C ₃ alkyl), S(O) ₂ (C ₁ -C ₃ alkyl), 4-10 membered heterocyclyl, 4-10 membered heterocyclyloxy, (C ₃ -C ₁₀ cycloalkyl) CH ₂ O, 5-6 membered heteroaryl, 5-6 membered heterocyclyl Aryloxy;

R ^c2 is selected from 4-6 membered heterocycloalkyl, =O, C ₁ -C ₃ alkyl, halogenated C ₁ -C ₃ alkyl, F, Cl, Br, I, CN, OH, CH ₂ OH or NH ₂ ;

X is selected from O, S or NR ² ;

L is selected from O, NH or a chemical bond;

R ¹ and R ² are independently selected from H, C ₁ -C ₁₀ alkyl, C ₃ -C ₁₀ cycloalkyl or 4-10 membered heterocyclic group, the C ₁ -C ₁₀ alkyl, C ₃ -C The ₁₀ -cycloalkyl or 4-10 membered heterocyclyl is optionally substituted ^{with Rd} ^selected from F, Cl, Br, I, OH, CN or _C1 _- C3 alkyl.

In some embodiments, Ring Q is 5-6 membered heterocyclyl, _C6 - _C10 aryl, or 5-10 membered heteroaryl, said 5-6 membered heterocyclyl, _C6 - _C10 aryl or 5-10 membered heteroaryl optionally substituted with R ^a selected from F, Cl, Br, I, OH, CN or the following groups ^optionally substituted with R ^: C ₁ -C ₁₀ alkanes base, C ₃ -C ₁₀ cycloalkyl, C ₁ -C ₁₀ alkoxy;

R ^b is selected from F, Cl, Br, I, OH or CN;

Ring W is phenyl, pyridyl, pyridinone, pyridazinone or

wherein ring M is a 5-6 membered heteroaryl group or a 4-10 membered heterocyclic group, and the phenyl, pyridyl, pyridinone, pyridazinonyl or ring M is optionally substituted with R ^c , the R ^c Selected from OH, =O or the following groups optionally substituted by R ^c1 : C ₁ -C ₁₀ alkyl, C ₁ -C ₁₀ alkoxy, C ₃ -C ₁₀ cycloalkyl, P(O)(C ₁ -C ₃ alkyl) ₂ , 4-6 membered heterocycloalkyl, NHC(O) (C ₁ -C ₆ alkyl); X ₁ is selected from C or N, when X ₁ is selected from C,

Represents a double bond, when X ₁ is selected from N,

represents a single key;

X is selected from O, S or NR ² ;

L is selected from O or NH;

R ¹ , R ² are independently selected from H, C ₁ -C ₁₀ alkyl or C ₃ -C ₁₀ cycloalkyl, said C ₁ -C ₁₀ alkyl or C ₃ -C ₁₀ cycloalkyl optionally being replaced by R ^d is substituted, and said R ^d is selected from F, Cl, Br, I, OH, CN or _C1 _- C3 alkyl.

R ^b is selected from F, Cl, Br, I, OH or CN;

Ring W is phenyl, pyridyl, pyridone or 9-10-membered heteroaryl, said phenyl, pyridyl, pyridone or 9-10-membered heteroaryl is optionally substituted by R ^c , said R ^c is selected from OH or the following groups optionally substituted by R ^c1 : C ₁ -C ₁₀ alkyl, C ₁ -C ₁₀ alkoxy, C ₃ -C ₁₀ cycloalkyl, P(O)(C ₁ - C ₃ alkyl) ₂ , 4-6 membered heterocycloalkyl;

R ^c1 is selected from F, Cl, Br, I, CN, OH, NH ₂ or the following groups optionally substituted by R ^c2 : C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, NHC(O )O(C ₁ -C ₆ alkyl), (C ₁ -C ₆ alkyl) NHC(O)O, NH(C ₁ -C ₃ alkyl), S(O) ₂ (C ₁ -C ₃ alkane) base), 4-10 membered heterocyclyl, 4-10 membered heterocyclyloxy, (C ₃ -C ₁₀ cycloalkyl) CH ₂ O, 5-6 membered heteroaryl, 5-6 membered heteroaryl Oxygen;

X is selected from O, S or NR ² ;

L is selected from O or NH;

In some embodiments, Ring Q is a 5-6 membered heterocyclyl, phenyl or 5-6 membered heteroaryl, optionally the 5-6 membered heterocyclyl, phenyl or 5-6 membered heteroaryl replaced by ^Ra .

In some embodiments, Ring Q is phenyl or 5-6 membered heteroaryl optionally substituted with ^Ra .

In some embodiments, Ring Q is piperidinyl, phenyl, or pyridyl, optionally substituted with ^Ra .

In some embodiments, Ring Q is phenyl or pyridyl optionally substituted with ^Ra .

In some embodiments, Ring Q is phenyl optionally substituted with ^Ra .

In some embodiments, ^Ra is F, Cl, Br, I, CN, C2 _- _C3alkynyl , C1 _- _C6alkyl , C3 _- _C6cycloalkyl , or optionally substituted with F C ₁ -C ₆ alkoxy.

In some embodiments, R ^a is F, Cl, Br, I, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, or C ₁ -C ₆ alkoxy optionally substituted with F. In some embodiments, ^Ra is F, Cl, Br, I, CN, methyl, cyclopropyl, ethynyl, prop-1-ynyl, or methoxy optionally substituted with F.

In some embodiments, ^Ra is F, Cl, Br, I, methyl, cyclopropyl, or methoxy optionally substituted with F.

In some embodiments, ^Ra is methoxy optionally substituted with F.

In some embodiments, ^Ra is F, Cl, Br, I, CN, methyl, cyclopropyl, ethynyl, prop-1-ynyl, difluoromethoxy, trifluoromethoxy, or methoxy base.

In some embodiments, ^Ra is difluoromethoxy.

In some embodiments, ring Q is selected from

In some embodiments, ring Q is selected from

In some embodiments, Ring W is phenyl, pyridyl, pyrimidinyl, pyridazinyl, 2-pyridinyl,

wherein ring M is a 5-10-membered heteroaryl group or a 5-10-membered heterocyclic group, the phenyl, pyridyl, pyrimidinyl, pyridazinyl, 2-pyridone,

or Ring M is optionally substituted with ^Rc . In some embodiments, Ring W is phenyl, pyridyl, 2-pyridone,

wherein ring M is a 5-6-membered heteroaryl group or a 5-10-membered heterocyclic group, and the phenyl, 2-pyridone, pyridyl,

or Ring M is optionally substituted with ^Rc .

In some embodiments, Ring W is phenyl, pyridinyl, 2-pyridonyl, or 9-10 membered heteroaryl, said phenyl, 2-pyridonyl, pyridyl, or 9-10 membered heteroaryl Optionally substituted with ^Rc .

In some embodiments, Ring W is the following groups ^optionally substituted with R:

Phenyl, 9-10 membered heteroaryl.

In some embodiments, Ring M is a 5-6 membered heteroaryl, 9-10 membered heteroaryl, or 5-9 membered heterocyclyl, which is optionally substituted with ^Rc .

In some embodiments, Ring M is a 5-6 membered heteroaryl or 6-8 membered heterocyclyl, which is optionally substituted with ^Rc .

In some embodiments, Ring W is

wherein Ring M is 5-6 membered heteroaryl, 9-10 membered heteroaryl or 5-9 membered heterocyclyl, and said Ring M is optionally substituted with ^Rc .

In some embodiments, Ring W is

wherein ring M is a 5-6 membered heteroaryl group or a 6-8 membered heterocyclyl group, and the ring M is optionally substituted with ^Rc .

In some embodiments, Ring W is

wherein Ring M is a 5-6 membered heteroaryl, optionally substituted with R ^c .

In some embodiments, Ring W is the following ^optionally substituted with R:

phenyl,

In some embodiments, Ring W is the following ^optionally substituted with R:

phenyl,

In some embodiments, Ring W is the following ^optionally substituted with R:

phenyl,

In some embodiments, Ring W is the following ^optionally substituted with R:

phenyl,

In some embodiments, R ^c is selected from =O or the following groups optionally substituted with R ^c1 : C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, NHC(O)(C ₁ -C ₆ alkyl).

In some embodiments, R ^c is selected from =O or the following groups optionally substituted with R ^c1 : methyl, methoxy,

NHC(O) _CH3 .

In some embodiments, R ^c is selected from the following groups optionally substituted with R ^c1 : C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy.

In some embodiments, R ^c is selected from the following groups optionally substituted with R ^c1 : methyl, methoxy,

In some embodiments, R ^c is selected from methyl optionally substituted with R ^c1 or methoxy optionally substituted with R ^c1 .

In some embodiments, R ^c1 is selected from deuterium atoms, F, Cl, Br, I, CN, OH, NH ₂ or ethoxy.

In some embodiments, R ^c1 is selected from deuterium atoms, F, Cl, Br, I, CN, OH.

In some embodiments, R ^c1 is selected from F, Cl, Br, I, CN, OH.

In some embodiments, R ^c is selected from CH ₂ OH, methyl, methoxy, CHF ₂ ,

CD ₃ , NHC(=O)CH ₃ , =O,

_CD3 , NHC(=O) _CH3 or =O.

In some embodiments, R ^c is selected from CH ₂ OH, methyl, methoxy, CHF ₂ or

In some embodiments, R ^c is selected from methyl optionally substituted with OH or methoxy optionally substituted with OH.

In some embodiments, Ring W is selected from

In some embodiments, Ring W is selected from

In some embodiments, Ring W is selected from

In some embodiments, Ring W is selected from

In some embodiments, Ring W is selected from

In some embodiments, X is selected from O, S or NH.

In some embodiments, X is selected from S or O.

In some embodiments, X is selected from S.

In some embodiments, R ¹ is selected from H, 4-6 membered heterocyclyl, C ₃ -C ₆ cycloalkyl optionally substituted with C ₁ -C ₃ alkyl, or C ₁ -C optionally substituted with F C ₆ alkyl.

In some embodiments, R ¹ is selected from H, C ₃ -C ₆ cycloalkyl optionally substituted with C ₁ -C ₃ alkyl, or C ₁ -C ₆ alkyl optionally substituted with F.

In some embodiments, R ¹ is selected from 4-6 membered heterocyclyl or C ₁ -C ₆ alkyl optionally substituted with F.

In some embodiments, R ¹ is selected from 4-membered heterocyclyl or C ₁ -C ₃ alkyl optionally substituted with F.

In some embodiments, R ¹ is selected from C ₁ -C ₆ alkyl.

In some embodiments, R ¹ is selected from methyl, ethyl, n-propyl,

or trifluoromethyl.

In some embodiments, R ¹ is selected from ethyl.

In some embodiments, L is selected from O or a bond.

In some embodiments, L is selected from O.

In some embodiments,

is selected from methoxy, ethoxy, n-propyloxy,

or trifluoromethyl.

In some embodiments, the compound represented by the general formula (A) or a pharmaceutically acceptable salt thereof is selected from the compound of formula (B) or a pharmaceutically acceptable salt thereof:

wherein rings W, R ^a , R ¹ are as defined above, and n is selected from 0, 1, 2, 3, 4 or 5.

In some embodiments, the compound represented by the general formula (A) or a pharmaceutically acceptable salt thereof is selected from the compound of formula (C) or a pharmaceutically acceptable salt thereof:

wherein Rings W, R ^a , R ¹ are as defined above.

In some embodiments, the compound of general formula (A) or a pharmaceutically acceptable salt thereof is selected from the compound of formula (D) or a pharmaceutically acceptable salt thereof:

wherein Ring W is as defined above.

In some embodiments, the compound or pharmaceutically acceptable salt of the general formula (A) is selected from the following compounds or pharmaceutically acceptable salts:

The present invention also provides a pharmaceutical composition comprising the compound represented by the general formula (A) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

The present invention relates to the use of a compound represented by general formula (A) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the preparation of a medicament for preventing or treating tumors.

Further, the present invention relates to the use of a compound represented by general formula (A) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the preparation of a medicament for preventing or treating tumors with reduced or absent MTAP activity.

The present invention relates to the use of a compound represented by the general formula (A) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in preventing or treating tumors.

Further, the present invention relates to the use of the compound represented by the general formula (A) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the prevention or treatment of tumors with reduced or absent MTAP activity.

The present invention relates to a compound of general formula (A) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for preventing or treating tumors.

Further, the present invention relates to a compound of general formula (A) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for preventing or treating tumors with reduced or absent MTAP activity.

The present invention also relates to a method of treating tumors comprising administering to a patient a therapeutically effective dose of a pharmaceutical formulation comprising a compound of general formula (A) or a pharmaceutically acceptable salt thereof described in the present invention.

Further, the present invention also relates to a method for treating tumors with reduced or absent MTAP activity, the method comprising administering to a patient a therapeutically effective dose of a compound comprising the general formula (A) of the present invention or a pharmaceutically acceptable salt thereof. pharmaceutical preparations.

In a preferred embodiment of the present invention, the tumors or tumors with reduced or absent MTAP activity include but are not limited to glioma, mesothelioma, melanoma, gastric cancer, esophageal cancer, bladder cancer, pancreatic cancer, non-small cell lung cancer, Astrocytoma, osteosarcoma, head and neck cancer, mucinous chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, soft tissue sarcoma, non-Hodgkin's lymphoma, etc.

Definition and Explanation of Terms

Unless otherwise stated, definitions of groups and terms set forth in the specification and claims of this application, including their definitions as examples, exemplary definitions, preferred definitions, definitions set forth in tables, and definitions of specific compounds in the examples etc., can be arbitrarily combined and combined with each other. Such combinations, group definitions and compound structures after the combination should fall within the scope described in the specification of the present application. A particular term should not be considered indeterminate or unclear unless specifically defined, but should be understood according to its ordinary meaning in the art. When a trade name appears herein, it is intended to refer to its corresponding commercial product or its active ingredient.

in this article

Indicates the attachment site.

The term "pharmaceutically acceptable salts" refers to pharmaceutically acceptable salts of non-toxic acids or bases, including salts of inorganic acids and bases, organic acids and bases.

A schematic representation of racemate or enantiomerically pure compounds herein is from Maehr, J. Chem. Ed. 1985, 62: 114-120. Use wedge and virtual wedge keys unless otherwise specified

Indicate the absolute configuration of a stereocenter, using black solid and virtual bonds

Indicates the relative configuration of a stereocenter (eg, cis-trans configuration of alicyclic compounds). When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, they include E, Z geometric isomers unless otherwise specified. Likewise, all tautomeric forms are included within the scope of the present invention.

The term "tautomer" refers to an isomer of a functional group resulting from the rapid movement of an atom in two positions in a molecule. The compounds of the present invention may exhibit tautomerism. Tautomeric compounds can exist as two or more interconvertible species. Tautomers generally exist in equilibrium, and attempts to separate individual tautomers usually result in a mixture whose physicochemical properties are consistent with a mixture of compounds. The position of equilibrium depends on the chemical properties within the molecule. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the ketone form predominates; in phenols, the enol form predominates. The present invention encompasses all tautomeric forms of the compounds.

The term "stereoisomer" refers to isomers resulting from different arrangements of atoms in a molecule in space, and includes cis-trans isomers, enantiomers, diastereomers and conformers.

The compounds of the present invention may have asymmetric atoms such as carbon atoms, sulfur atoms, nitrogen atoms, phosphorus atoms or asymmetric double bonds, and thus the compounds of the present invention may exist in specific geometric or stereoisomeric forms. Particular geometric or stereoisomeric forms may be cis and trans isomers, E and Z geometric isomers, (-)- and (+)-enantiomers, (R)- and (S) )-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic or other mixtures thereof, such as enantiomers or diastereomers Enriched mixtures, all of the above isomers and mixtures thereof are within the definition of the compounds of the present application. There may be additional asymmetric carbon atoms, asymmetric sulfur atoms, asymmetric nitrogen atoms or asymmetric phosphorus atoms in substituents such as alkyl groups, and these isomers and their mixtures involved in all substituent groups are also included in the Within the definition of the compounds of the present application. Compounds of the present application containing asymmetric atoms can be isolated in optically pure form or in racemic form, optically pure forms can be resolved from racemic mixtures, or synthesized by using chiral starting materials or chiral reagents .

The term "pharmaceutical composition" means a mixture of one or more compounds described herein, or a physiologically/pharmaceutically acceptable salt or prodrug thereof, and other chemical components, such as a physiologically/pharmaceutically acceptable carrier and excipients. The purpose of a pharmaceutical composition is to facilitate the administration of a compound to an organism.

The term "substituted" means that any one or more hydrogen atoms on a specified atom are replaced by a substituent, so long as the valence of the specified atom is normal and the compound after substitution is stable. When the substituent is oxo (ie =O), it means that two hydrogen atoms are substituted and oxo does not occur on an aromatic group.

The term "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes both the occurrence and the non-occurrence of said event or circumstance. For example, an ethyl group "optionally" substituted with halogen means that the ethyl group can be unsubstituted ( _CH2CH3 ₎ , monosubstituted ₍ eg, _CH2CH2F , _CH2CH2Cl , etc. ₎ , polysubstituted (eg _CHFCH2F , _CH2CHF2 , _CHFCH2Cl , _CH2CHCl2 , etc. ₎ or fully substituted ₍ _CF2CF3 , _CF2CCl3 , _CCl2CCl3 _, _etc. ₎ . It will be understood by those skilled in the art that for any group containing one or more substituents, no substitution or substitution pattern is introduced that is sterically impossible and/or cannot be synthesized.

When any variable (eg, ^Ra , ^Rb ) occurs more than once in the composition or structure of a compound, its definition in each case is independent. For example, if a group is substituted with 2 R ^bs , each R ^b has independent options.

When the number of a linking group is 0, such as -(CH ₂ ) ₀ -, it means that the linking group is a bond.

When one of the variables is selected from a chemical bond or does not exist, it means that the two groups to which it is connected are directly connected, for example, when L in A-L-Z represents a bond, it means that the structure is actually A-Z.

When a substituent's bond is cross-linked to two atoms on a ring, the substituent can bond to any atom on the ring. For example, structural unit

Indicates that R ⁵ can be substituted at any position on the benzene ring.

_Cm - _Cn herein means having an integer number of carbon atoms in the range mn. For example, "C ₁ -C ₁₀ " means that the group can have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, 9 carbon atoms or 10 carbon atoms.

The term "alkyl" refers to a hydrocarbon group of the general formula _CnH2n+1 _. The alkyl group can be straight or branched. For example, the term " _C1 - _C10 alkyl" is understood to mean a straight or branched chain saturated monovalent hydrocarbon radical having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. The alkyl group includes, but is not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl , 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methyl pentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl base, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl, etc.; "C ₁ -C ₆ alkane "Radical" should be understood to mean a linear or branched saturated monovalent hydrocarbon group having 1, 2, 3, 4, 5, 6 carbon atoms; "C ₁ -C ₃ alkyl" refers to methyl, ethyl, propyl ,Isopropyl.

The "C ₁ -C ₁₀ alkyl group" described herein may include "C ₁ -C ₆ alkyl group" or "C ₁ -C ₃ alkyl group", and the "C ₁ -C ₆ alkyl group" may further include "C 1 -C 6 alkyl group" ₁ _- C3 alkyl".

The term "alkoxy" can be understood as "alkyloxy" or "alkyl-O-", which refers to a monovalent group generated by the loss of a hydrogen atom on the hydroxyl group of linear or branched alcohols, such as the term " C ₁ -C ₁₀ alkoxy" can be understood as "C ₁ -C ₁₀ alkyloxy" or "C ₁ -C ₁₀ alkyl-O-", the term "C ₁ -C ₆ alkoxy" can be understood is "C ₁ -C ₆ alkyloxy" or "C ₁ -C ₆ alkyl-O-", preferably, "C ₁ -C ₁₀ alkoxy" may contain "C ₁ -C ₆ alkoxy"" and "C ₁ -C ₃ alkoxy" and other ranges.

The term "halogenated _C1 _- C3 alkyl" includes monohalogenated _C1 _- C3 alkyl or polyhalogenated _C1 _- C3 alkyl, examples including, but not limited to, trifluoromethyl, 2,2,2 - trichloroethyl or 3-fluoropropyl.

The term "alkynyl" refers to a straight or branched chain unsaturated aliphatic hydrocarbon group consisting of carbon atoms and hydrogen atoms having at least one triple bond. The term "C ₂ -C ₁₀ alkynyl" is understood to mean a straight-chain or branched unsaturated hydrocarbon group containing one or more triple bonds and having 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. Examples of "C ₂ -C ₁₀ alkynyl" include, but are not limited to, ethynyl (-C≡CH), propynyl (-C≡CCH _3, -CH ₂ C≡CH), but-1-ynyl, butanyl -2-alkynyl or but-3-ynyl. "C ₂ -C ₁₀ alkynyl" may include "C ₂ -C ₃ alkynyl", examples of "C ₂ -C ₃ alkynyl" include ethynyl (-C≡CH), prop-1-ynyl (-C _≡CCH3 ), prop- ₂ -ynyl (-CH2C≡CH).

The term "C ₃ -C ₁₀ cycloalkyl" is understood to mean a saturated monovalent monocyclic or bicyclic hydrocarbon ring having 3 to 10 carbon atoms. Such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl, or a bicyclic hydrocarbon group such as decalin. The term "C3 _- C6cycloalkyl" is understood to mean a saturated monovalent monocyclic or bicyclic hydrocarbon ring having 3 to ₆ carbon atoms.

The term "cycloalkoxy" is to be understood as "cycloalkyloxy" or "cycloalkyl-O-".

The term "heterocyclyl" refers to a fully saturated or partially saturated (non-aromatic heteroaromatic as a whole) monocyclic, paracyclic, spirocyclic or bridged ring group containing 1-5 ring atoms Heteroatoms or heteroatomic groups (ie, atomic groups containing heteroatoms), the "heteroatoms or heteroatomic groups" include but are not limited to nitrogen atoms (N), oxygen atoms (O), sulfur atoms (S), phosphorus atoms (P) , boron atom (B), -S(=O) ₂ -, -S(=O)-, -P(=O) ₂ -, -P(=O)-, -NH-, -S(=O )(=NH)-, -C(=O)NH- or -NHC(=O)NH-, etc.

The term "4-10 membered heterocyclyl" refers to a heterocyclyl with 4, 5, 6, 7, 8, 9 or 10 ring atoms, and its ring atoms contain 1-5 independently selected from the above-mentioned heteroatoms or heteroatoms. "4-10-membered heterocyclic group" includes "4-7-membered heterocyclic group", wherein specific examples of 4-membered heterocyclic group include but are not limited to azetidinyl or oxetanyl; 5-membered heterocyclic group Specific examples of heterocyclyl groups include, but are not limited to, tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl, 4,5-dihydrooxazolyl, or 2,5-dihydrooxazolyl. 5-dihydro-1H-pyrrolyl; specific examples of 6-membered heterocyclic groups include, but are not limited to, tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl , trithianyl, tetrahydropyridyl, or 4H-[1,3,4]thiadiazinyl; specific examples of 7-membered heterocyclyl include, but are not limited to, diazepanyl. The heterocyclic group may also be a bicyclic group, wherein specific examples of the 5,5-membered bicyclic group include but are not limited to hexahydrocyclopento[c]pyrrol-2(1H)-yl; Specific examples include, but are not limited to, hexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl, 5,6,7,8-tetrahydro-[1,2,4]triazolo[4 ,3-a]pyrazinyl or 5,6,7,8-tetrahydroimidazo[1,5-a]pyrazinyl. Optionally, the heterocyclic group may be a benzo-fused ring group of the above-mentioned 4-7 membered heterocyclic group, and specific examples include, but are not limited to, dihydroisoquinolinyl and the like. Preferably, "4-10 membered heterocyclyl" may include "5-10 membered heterocyclyl", "5-9 membered heterocyclyl", "4-6 membered heterocyclyl", "5-6 membered heterocyclyl" "Cyclyl", "6-9 membered heterocyclyl", "6-8 membered heterocyclyl", "5-10 membered heterocycloalkyl", "5-9 membered heterocycloalkyl", "4-6 membered heterocyclyl" membered heterocycloalkyl", "5-6 membered heterocycloalkyl", "6-8 membered heterocycloalkyl" and the like. According to the present invention, although some bicyclic heterocyclic groups partially contain a benzene ring or a heteroaromatic ring, the heterocyclic group as a whole is still non-aromatic.

The term "heterocyclyloxy" is to be understood as "heterocyclyl-O-".

The term "heterocycloalkyl" refers to a 4-10 membered cyclic group that is fully saturated and may exist as a monocyclic, paracyclic, bridged or spirocyclic ring. Unless otherwise indicated, the ring atoms of the heterocyclic ring contain 1-5 heteroatoms or heteroatomic groups (ie, atomic groups containing heteroatoms), and the "heteroatoms or heteroatomic groups" include but are not limited to nitrogen atoms (N), Oxygen atom (O), sulfur atom (S), phosphorus atom (P), boron atom (B), -S(=O) ₂ -, -S(=O)-, -NH-, -S(=O )(=NH)-, -C(=O)NH- or -NHC(=O)NH-, etc. The term "4-6 membered heterocycloalkyl" means a heterocycloalkyl group having 4, 5 or 6 ring atoms and containing 1-3 heteroatoms or heteroatomic groups independently selected from the above-mentioned ring atoms. , wherein, non-limiting examples of 4-membered heterocycloalkyl include but are not limited to azetidinyl, oxetanyl, thibutanyl, and examples of 5-membered heterocycloalkyl include but are not limited to tetrahydrofuranyl, tetrahydrothienyl , pyrrolidinyl, isoxazolidinyl, oxazolidinyl, isothiazolidinyl, thiazolidinyl, imidazolidinyl, tetrahydropyrazolidinyl, examples of 6-membered heterocycloalkyl include but are not limited to piperidinyl , tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl, piperazinyl, 1,4-thioxanyl, 1,4-dioxanyl, thiomorpholinyl, 1,3- Dithianyl, 1,4-Dithianyl.

The term "aryl" refers to an all-carbon monocyclic or fused polycyclic aromatic ring group having a conjugated pi electron system. For example, an aryl group can have 6-20 carbon atoms, 6-14 carbon atoms, or 6-12 carbon atoms. The term "C ₆ -C ₁₀ aryl" is to be understood as preferably denoting an aromatic or partially aromatic fully carbon monocyclic or bicyclic radical having 6 to 10 carbon atoms, especially a ring having 6 carbon atoms ("C6-C10 aryl"). C ₆ aryl"), such as phenyl; or a ring with 9 carbon atoms ("C ₉ aryl"), such as indanyl or indenyl, or a ring with 10 carbon atoms ("C ₁₀ aryl") ”), such as tetrahydronaphthyl, dihydronaphthyl or naphthyl.

The term "heteroaryl" refers to an aromatic monocyclic or fused polycyclic system containing at least one ring atom selected from N, O, S, and an aromatic ring group in which the remaining ring atoms are C. The term "5-10 membered heteroaryl" is understood to include monovalent monocyclic or bicyclic aromatic ring systems having 5, 6, 7, 8, 9 or 10 ring atoms, in particular 5 or 6 or 9 or 10 ring atoms and it contains 1-5, preferably 1-3 heteroatoms independently selected from N, O and S. And, additionally in each case may be benzo-fused. In particular, heteroaryl is selected from the group consisting of thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiazolyl Diazolyl and the like and their benzo derivatives such as benzofuranyl, benzothienyl, benzothiazolyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzotriazole base, indazolyl, indolyl, isoindolyl, etc.; or pyridyl, pyridazinyl, pyrimidyl, pyrazinyl, triazinyl, etc., and their benzo derivatives, such as quinolinyl, quinoline oxazolinyl, isoquinolinyl, etc; Naphthyridinyl, pteridyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl and the like. The term "5-6 membered heteroaryl" refers to an aromatic ring system having 5 or 6 ring atoms, and which contains 1-3, preferably 1-2, heteroatoms independently selected from N, O and S. The term "9-10 membered heteroaryl" refers to an aromatic ring system having 9 or 10 ring atoms, and which contains 1-5, preferably 1-3 heteroatoms independently selected from N, O and S.

The term "heteroaryloxy" is to be understood as "heteroaryl-O-".

The term "excipient" refers to a pharmaceutically acceptable inert ingredient.

Examples of classes of the term "excipient" include, without limitation, binders, disintegrants, lubricants, glidants, stabilizers, fillers, diluents, and the like. Excipients can enhance the handling characteristics of a pharmaceutical formulation, ie make the formulation more suitable for direct compression by increasing flowability and/or stickiness. Examples of typical "pharmaceutically acceptable carriers" suitable for the above-mentioned preparations are: carbohydrates, starches, cellulose and their derivatives and other commonly used adjuvants in pharmaceutical preparations.

The term "treating" means administering a compound or formulation described herein to prevent, ameliorate, or eliminate a disease or one or more symptoms associated with the disease, and includes:

(i) preventing the emergence of a disease or disease state in mammals, particularly when such mammals are susceptible to, but have not been diagnosed with, the disease state;

(ii) inhibiting the disease or disease state, i.e. arresting its development;

(iii) Alleviation of the disease or disease state, even if the disease or disease state resolves.

The term "therapeutically effective amount" means (i) treating or preventing a particular disease, condition or disorder, (ii) alleviating, ameliorating or eliminating one or more symptoms of a particular disease, condition or disorder, or (iii) preventing or delaying The amount of a compound of the invention to be used for the onset of one or more symptoms of a particular disease, condition or disorder described herein. The amount of a compound of the present invention that constitutes a "therapeutically effective amount" will vary depending on the compound, the disease state and its severity, the mode of administration, and the age of the mammal to be treated, but can be routinely determined by those skilled in the art according to its own knowledge and the present disclosure.

The term "pharmaceutically acceptable excipients" refers to those excipients which are not significantly irritating to the organism and which do not impair the biological activity and properties of the active compound. Suitable excipients are well known to those skilled in the art, such as carbohydrates, waxes, water-soluble and/or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, and the like.

The words "comprise", "comprise" or "comprise" and their English variants such as comprises or comprising are to be understood in an open, non-exclusive sense, ie, "including but not limited to".

The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments enumerated below, embodiments formed in combination with other chemical synthesis methods, and those well known to those skilled in the art Equivalent to alternatives, preferred embodiments include, but are not limited to, the embodiments of the present invention.

The present application also includes isotopically-labeled compounds of the present application which are the same as those described herein, but wherein one or more atoms have been replaced by an atom having an atomic weight or mass number different from that normally found in nature. Examples of isotopes that may be incorporated into the compounds of the present application include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as 2H, ^3H , ^11C , ^13C , ^14C , ¹³ , ^respectively N, ¹⁵ N, ¹⁵ O, ¹⁷ O, ¹⁸ O, ³¹ P, ³² P, ³⁵ S, ¹⁸ F, ¹²³ I, ¹²⁵ I and ³⁶ Cl and the like.

Certain isotopically-labeled compounds of the present application (eg, those labeled ^with3H ^and14C ) are useful in compound and/or substrate tissue distribution assays. Tritiated (ie ³ H) and carbon-14 (ie ¹⁴ C) isotopes are especially preferred for their ease of preparation and detectability. Positron emitting isotopes such ^as15O , ^13N , ^11C ^and18F can be used in positron emission tomography (PET) studies to determine substrate occupancy. Isotopically labeled compounds of the present application can generally be prepared by the following procedures analogous to those disclosed in the Schemes and/or Examples below, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

The pharmaceutical composition of the present application can be prepared by combining the compound of the present application with suitable pharmaceutically acceptable excipients, for example, it can be formulated into solid, semi-solid, liquid or gaseous preparations, such as tablets, pills, capsules, powders , granules, ointments, emulsions, suspensions, suppositories, injections, inhalants, gels, microspheres and aerosols, etc.

Typical routes of administration of a compound of the present application, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, Intramuscular, subcutaneous, intravenous administration.

The pharmaceutical composition of the present application can be manufactured by methods well known in the art, such as conventional mixing method, dissolving method, granulation method, sugar-coated pill method, grinding method, emulsification method, freeze-drying method and the like.

In some embodiments, the pharmaceutical composition is in oral form. For oral administration, the pharmaceutical compositions can be formulated by admixing the active compound with pharmaceutically acceptable excipients well known in the art. These excipients enable the compounds of the present application to be formulated into tablets, pills, lozenges, dragees, capsules, liquids, gels, slurries, suspensions, etc., for oral administration to patients.

Solid oral compositions can be prepared by conventional mixing, filling or tabletting methods. It can be obtained, for example, by mixing the active compound with solid excipients, optionally milling the resulting mixture, adding other suitable excipients if desired, and processing the mixture into granules to obtain tablets or icing core. Suitable adjuvants include, but are not limited to, binders, diluents, disintegrants, lubricants, glidants, sweeteners or flavoring agents, and the like.

The pharmaceutical compositions may also be suitable for parenteral administration as sterile solutions, suspensions or lyophilized products in suitable unit dosage forms.

In all methods of administration of the compounds of general formula I described herein, the doses administered per day are 0.01 to 100 mg/kg body weight, preferably 0.05 to 50 mg/kg body weight, more preferably 0.1 to 30 mg/kg body weight, in single or divided doses form.

The chemical reactions of specific embodiments of the present invention are carried out in suitable solvents suitable for the chemical changes of the present invention and their required reagents and materials. In order to obtain the compounds of the present invention, it is sometimes necessary for those skilled in the art to modify or select the synthetic steps or reaction schemes on the basis of the existing embodiments.

Detailed ways

The following examples illustrate the technical solutions of the invention in detail, but the protection scope of the present invention includes but is not limited thereto.

Unless otherwise specified, the ratio indicated by the mixed solvent is the volume mixing ratio.

Unless otherwise specified, % refers to weight percent wt %.

The eluent or mobile phase can be a mixed eluent or mobile phase composed of two or more solvents, and its ratio is the volume ratio of each solvent, such as "0-10% methanol/dichloromethane" means mixed eluent Or the volume ratio of methanol to dichloromethane in the mobile phase is 0:100 to 10:100.

Compounds are manually or

Software naming, commercially available compounds use supplier catalog names.

The structures of the compounds were determined by nuclear magnetic resonance (NMR) and/or mass spectrometry (MS). The units of NMR shifts are ^10-6 (ppm). The solvents for NMR measurement are deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol, etc., and the internal standard is tetramethylsilane (TMS). " _IC50 " refers to the half inhibitory concentration, the concentration at which half of the maximal inhibitory effect is achieved. "DCM" refers to dichloromethane, "PE" refers to petroleum ether, and "EA" refers to ethyl acetate.

Example 1, 4-(4-(difluoromethoxy)phenyl)-2-ethoxy-6-(2-(hydroxymethyl)-1-methyl-1H-benzo[d]imidazole -6-yl)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 1)

Step 1: Synthesis of N-((5-bromo-6-methoxypyridin-2-yl)carbamoyl)benzamide (Intermediate 1-2)

The starting material 1-1 (1 g, 4.93 mmol) was dissolved in tetrahydrofuran (15 mL), benzoyl isothiocyanate (763.59 mg, 4.68 mmol) was added at 0 °C, and the reaction was stirred at 0 °C for 1 hour, and then at 0 °C. The reaction was stirred at 25°C for 16 hours. LCMS detected that the reaction of the raw materials was complete, and concentrated under reduced pressure to remove the solvent to obtain the title compound (1.8 g), which was directly used in the next step.

MS m/z (ESI): 365.9 [M+H] ⁺ .

Step 2: Synthesis of 1-(5-bromo-6-methoxypyridin-2-yl)thiourea (Intermediate 1-3)

Intermediate 1-2 (1.8 g, 4.91 mmol) was dissolved in water (1.5 mL) and tetrahydrofuran (8 mL), sodium hydroxide (5 M, 1.18 mL) was added, heated to 80° C., and the reaction was stirred for 3 hours. LCMS detected the complete reaction of the raw materials, concentrated under reduced pressure to remove the solvent, added water (50 mL), ethyl acetate (50 mL), adjusted pH=1 with hydrochloric acid, and then added mixed solvent isopropanol: dichloromethane=1:3 (120 mL) for extraction , the organic phases were combined, dried over sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by flash column chromatography (

20g

Flash silica column, mobile phase 5-25% tetrahydrofuran/petroleum ether (5% DCM) gradient @ 30 mL/min) to give the title compound (1.1 g).

MS m/z (ESI): 261.8 [M+H] ⁺ .

Step 3: Synthesis of 6-bromo-5-methoxythiazolo[4,5-b]pyridin-2-amine (Intermediate 1-4)

Intermediate 1-3 (1 g, 3.81 mmol) was dissolved in chloroform (20 mL), cooled to 0°C, liquid bromine (621.86 mg, 3.89 mmol) was dissolved in chloroform (5 mL) and added dropwise to the reaction solution. The reaction solution was reacted at 0°C for 30 minutes and heated to 65°C for 2.5 hours. LCMS detected that the reaction of the raw materials was complete, the reaction solution was cooled to -20°C, filtered, and the filter cake was washed with n-pentane. The title compound (1.1 g) was obtained, which was directly used in the next reaction.

MS m/z (ESI): 260.0 [M+H] ⁺ .

Step 4: Synthesis of 2-amino-6-bromothiazolo[4,5-b]pyridin-5(4H)-one (Intermediate 1-5)

Intermediate 1-4 (600 mg, 2.31 mmol) was dissolved in 1,2-dichloroethane (15 mL) and boron tribromide (2.23 g, 8.90 mmol) was added, and the reaction solution was stirred at 50° C. for 16 hours. LCMS detected that the reaction of the raw materials was complete, the reaction solution was diluted with 1,2-dichloroethane, and methanol (15 mL) was added dropwise at 0°C under nitrogen protection. The reaction solution was concentrated to obtain a solid, methanol (15 mL) was added, ammonia water was added to adjust pH=9, and the mixture was filtered to obtain the title compound (100 mg).

MS m/z (ESI): 247.9 [M+H] ⁺ .

Step 5: Synthesis of 6-bromo-2-chlorothiazolo[4,5-b]pyridin-5(4H)-one (Intermediate 1-6)

Cupric chloride (327.82 mg, 2.44 mmol) and tert-butyl nitrite (399.00 mg, 3.87 mmol) were dissolved in acetonitrile (6 mL), followed by intermediate 1-5 (300 mg, 1.22 mmol) in acetonitrile (4 mL) ) was added dropwise to the reaction solution at 0°C. The reaction solution was stirred at 25°C for 16 hours. LCMS detected that the reaction of the raw materials was complete, 6M HCl (2 mL) was added dropwise to the reaction solution, stirred for 30 minutes, adjusted to pH 8 with sodium bicarbonate, extracted with a mixed solvent of methanol and dichloromethane (10:1), and the organic phases were combined and concentrated The title compound (310 mg) was obtained.

MS m/z (ESI): 264.9 [M+H] ⁺ .

Step 6: Synthesis of 6-bromo-2-ethoxythiazolo[4,5-b]pyridin-5(4H)-one (Intermediate 1-7)

Intermediate 1-6 (400 mg, 1.51 mmol) was dissolved in ethanol (15 mL), sodium ethoxide (512.59 mg, 7.53 mmol) was added, and the reaction was stirred at 25 ° C for 16 hours, then saturated ammonium chloride was added to the reaction solution, and stirred for 15 min, extracted with mixed organic solvent ethanol and dichloromethane (10:1, 100 mL*5), dried over sodium sulfate, and concentrated under reduced pressure to remove the solvent. The residue was purified by preparative thin layer chromatography (silica, petroleum ether:tetrahydrofuran=3:2) to give the title compound (130 mg).

MS m/z (ESI): 275 [M+H] ⁺ .

Step 7: 6-Bromo-4-(4-(difluoromethoxy)phenyl)-2-ethoxythiazolo[4,5-b]pyridin-5(4H)-one (Intermediate 1- 8) Synthesis

Intermediate 1-7 (120 mg, 436.17 μmol) was dissolved in dichloromethane (5 mL), (4-(difluoromethoxy)phenyl)boronic acid (163.94 mg, 872.34 μmol), copper acetate (87.15 mg, 479.79 μmol), pyridine (103.50 mg, 1.31 mmol), the reaction was stirred at 40° C. for 16 hours under an oxygen atmosphere, followed by addition of water (5 mL), ammonia (0.5 mL) and ethyl acetate (5 mL), extraction, and then the organic phases were combined and concentrate. The residue was purified by preparative thin layer chromatography (silica, petroleum ether:tetrahydrofuran=1:1). The title compound (110 mg).

MS m/z (ESI): 417 [M+H] ⁺ .

Step 8: 4-(4-(Difluoromethoxy)phenyl)-2-ethoxy-6-(2-(hydroxymethyl)-1-methyl-1H-benzo[d]imidazole- Synthesis of 6-yl)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 1)

Intermediate 1-8 (110 mg, 263.65 μmol) and Intermediate 1-9 (120 mg, 416.45 μmol) were dissolved in dioxane (2 mL) and water (0.5 mL), and cesium carbonate (171.80 mg) was added to the reaction system. , 527.29 μmol), under nitrogen atmosphere, Pd(dtbpf)Cl ₂ (17.18 mg, 26.36 μmol) was added. The reaction solution was stirred at 90°C for 16 hours. Water (15 mL) was added to the reaction solution, followed by extraction with ethyl acetate (10 mL). The organic phase was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by preparative high performance liquid chromatography (basic conditions, column: Phenomenex Luna C18 100*30mm*5μm; mobile phase: [water (0.05% ammonia v/v)-acetonitrile]; B%: 43%-63% , 9 minutes). The title compound (12 mg) was obtained.

MS m/z (ESI): 499.1 [M+H] ⁺ .

¹ H NMR (400MHz, METHANOL-d ₄ )δ8.18(s,1H),7.89(s,1H),7.65(d,J=8.4Hz,1H),7.54(d,J=8.4Hz,1H) ,7.45(d,J＝8.9Hz,2H),7.33(d,J＝8.8Hz,2H),6.96(t,J＝73.8Hz,1H),4.81(s,2H),4.39(q,J＝ 7.1Hz, 2H), 3.92 (s, 3H), 1.36 (t, J=7.1Hz, 3H).

Example 2, 4-[4-(difluoromethoxy)phenyl]-2-ethoxy-6-(4-methoxyphenyl)thiazolo[4,5-b]pyridine-5( 4H)-ketone (compound 2)

Step 1: 4-[4-(Difluoromethoxy)phenyl]-2-ethoxy-6-(4-methoxyphenyl)thiazolo[4,5-b]pyridine-5(4H )-ketone (compound 2) synthesis

Intermediate 1-8 (110 mg, 263.65 μmol) and reactant 2-1 (92.58 mg, 395.48 μmol) were dissolved in dioxane (3 mL) and water (0.75 mL), and cesium carbonate (171.80 mg, 527.30 μmol) was added ) and Pd(dtbpf)Cl ₂ (17.18 mg, 26.37 μmol), followed by heating to 90° C. under nitrogen atmosphere for 16 hours. The reaction was completed by LCMS detection. The reaction solution was diluted with water (5 mL), extracted twice with ethyl acetate (10 mL), and the organic phase was concentrated under reduced pressure to remove the solvent. The residue was purified by preparative thin layer plate (silica, tetrahydrofuran/petroleum ether=2/1), and purified by preparative high performance liquid phase (basic conditions, column: Boston Prime C18 150*30mm*5μm; mobile phase: [ A: water (0.05% ammonia v/v), B: acetonitrile]; B%: 50%-80%, 9 minutes). The title compound (2.6 mg) was obtained.

MS m/z (ESI): 445.1 [M+H] ⁺ .

¹ H NMR(400MHz,Methanol-d ₄ )δ8.05(s,1H),7.60(d,J=8.8Hz,2H),7.44-7.40(m,2H),7.35-7.29(m,2H), 6.96 (t, J=73 Hz, 1H), 6.94 (m, 2H), 4.38 (q, J=7.3 Hz, 2H), 3.82 (s, 3H), 1.35 (t, J=7.1 Hz, 3H).

Example 3, 4-[4-(difluoromethoxy)phenyl]-2-ethoxy-6-(1-methyl-6-oxo-1,6dihydropyridin-3-yl) Thiazolo[4,5-b]pyridin-5(4H)-one (Compound 3)

Intermediate 1-8 (60 mg, 143.81 μmol) and Intermediate 3-1 (50.71 mg, 215.71 μmol) were dissolved in dioxane (2 mL) and water (0.5 mL), and cesium carbonate (93.71 mg, 287.61 μmol) was added ) and Pd(dtbpf)Cl ₂ (9.37 mg, 14.38 μmol), followed by heating to 90° C. under nitrogen atmosphere for 16 hours. The reaction was completed by LCMS detection. The reaction solution was diluted with water (5 mL), extracted twice with ethyl acetate (10 mL), and the organic phase was concentrated under reduced pressure to remove the solvent. The residue was purified by preparative thin layer plate (silica, tetrahydrofuran/petroleum ether=2/1), and purified by preparative high performance liquid phase (basic conditions, column: Boston Prime C18 150*30mm*5μm; mobile phase: [ Water (0.05% ammonia v/v)-acetonitrile]; B%: 42%-72%, 9 minutes). The title compound (3.2 mg) was obtained.

MS m/z (ESI): 446.0 [M+H] ⁺ .

¹ H NMR(400MHz,Methanol-d ₄ )δ8.20(d,J=2.4Hz,1H),8.14(s,1H),7.90(dd,J=2.6,9.4Hz,1H),7.46-7.38( m,2H),7.36-7.28(m,2H),6.95(t,J=73.7Hz,1H),6.60(d,J=9.4Hz,1H),4.38(q,J=7.1Hz,2H), 3.62 (s, 3H), 1.35 (t, J=7.1 Hz, 3H).

Example 4, 4-(4-(difluoromethoxyphenyl)-2-ethoxy-6-(1-isobutyl-6-oxo-1,6-dihydropyridin-3-yl) )thiazolo[4,5-b]pyridin-5(4H)-one (Compound 4)

Intermediate 4-1 (53 mg, 191.74 μmol, which can be synthesized according to the method reported on page 48 of the patent WO2019102256 text) and intermediate 1-8 (40 mg, 95.87 μmol) were dissolved in dioxane (1 mL) and water (0.25 mL). ), cesium carbonate (62.47 mg, 191.74 μmol) and Pd(dtbpf)Cl ₂ (6.25 mg, 9.59 μmol) were added thereto, and the reaction solution was stirred at 100° C. for 2 h under nitrogen protection. LC-MS detected the completion of the reaction. The reaction solution was filtered, concentrated to dryness under reduced pressure, and purified by preparative high performance liquid chromatography (column: Gemini NX C18 5 μm*10*150mm; mobile phase: A: water (0.225% ammonium bicarbonate v/v), B: acetonitrile; B%: 30%-50%, 11 min) to obtain the title compound (10.0 mg).

MS m/z (ESI): 488.1 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.25-8.14 (m, 2H), 7.75-7.67 (m, 1H), 7.42-7.35 (m, 2H), 7.30 (t, J=72Hz 1H), 7.29–7.22 (m, 2H), 6.37 (d, J=9.5Hz, 1H), 4.31–4.19 (m, 2H), 3.67 (d, J=7.4Hz, 2H), 2.11–1.90 (m, 1H) ,1.24(t,J＝7.0Hz,3H),0.80(d,J＝6.7Hz,6H).

Example 5, 4-(4-(difluoromethoxy)phenyl)-2-ethoxy-6-(1-methyl-1H-benzo[d]imidazol-6-yl)thiazolo[ 4,5-b]pyridin-5(4H)-one (Compound 5)

Intermediate 1-8 (40 mg, 95.87 μmol) and Intermediate 5-1 (35 mg, 134.22 μmol) were dissolved in dioxane (1 mL) and water (0.2 mL), to which was added cesium carbonate (62.47 mg, 191.74 μmol) and Pd(dtbpf)Cl ₂ (6.25 mg, 9.59 μmol), the reaction solution was stirred at 90° C. for 2 h under nitrogen protection. LC-MS detected the completion of the reaction. The reaction solution was filtered, concentrated to dryness under reduced pressure, and purified by preparative high performance liquid chromatography (column: Gemini NX C18 5 μm*10*150mm; mobile phase: A: water (0.225% trifluoroacetic acid v/v), B: acetonitrile; B%: 30%-50%, 11 min) to obtain the title compound (23.0 mg).

MS m/z (ESI): 469.1 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ )δ9.23(s,1H), 8.31(s,1H), 8.19(s,1H), 7.91-7.79(m,2H), 7.47-7.40(m,2H) ), 7.37–7.30(m, 2H), 7.20(t, J=72.0Hz, 1H), 4.38–4.25(m, 2H), 4.01(s, 3H), 1.31(t, J=7.0Hz, 3H) .

Example 6, 4-(4-(difluoromethoxy)phenyl)-6-(1-(difluoromethyl)-6-oxo-1,6-dihydropyridin-3-yl)- 2-Ethoxythiazolo[4,5-b]pyridin-5(4H)-one (Compound 6)

Step 1: 4-(4-(Difluoromethoxy)phenyl)-2-ethoxy-6-(4,4,5,5-tetramethyl-1,3,2-dioxaboro Synthesis of Pentacyclo-2-yl)thiazolo[4,5-b]pyridin-5(4H)-one (Intermediate 6-1)

Compound 1-8 (4.2 g, 10 mmol) and bispinacol boronate (3.8 g, 15 mmol) were dissolved in anhydrous dioxane (42 mL), to which was added potassium acetate (1.9 g, 20 mmol) and Pd(dppf)Cl ₂ (1.1 g, 1.5 mmol), the reaction solution was stirred at 100° C. for 15 h under nitrogen protection. LC-MS detected the completion of the reaction. After the reaction was cooled to room temperature, the solvent was removed by rotary evaporation, water (40 mL) and ethyl acetate (50 mL) were added successively, the organic phase was washed with water (15 mL*2), and the washed organic phase was dried with an appropriate amount of anhydrous sodium sulfate. The title compound (3.2 g) was obtained by preparative thin layer chromatography (silica, petroleum ether:ethyl acetate=4:1).

MS m/z (ESI): 465.1 [M+H] ⁺ .

Step 2: 4-(4-(Difluoromethoxy)phenyl)-6-(1-(difluoromethyl)-6-oxo-1,6-dihydropyridin-3-yl)-2 - Synthesis of Ethoxythiazolo[4,5-b]pyridin-5(4H)-one (Compound 6)

Intermediate 6-2 (50 mg, 0.22 mmol) and intermediate 6-1 (124.5 mg, 0.268 mmol) were dissolved in anhydrous dioxane solution (0.5 mL), to which was added water (0.1 mL), carbonic acid Cesium (145.5 mg, 0.446 mmol) and Pd(dtbpf)Cl ₂ (14.6 mg, 0.022 mmol), the reaction solution was stirred at 100° C. for 2 h under nitrogen protection. LC-MS detected the completion of the reaction. The reaction solution was filtered, concentrated to dryness under reduced pressure, and purified by high pressure preparative [YMC-Actus Triart C18 column 5 μm silica, 30 mm diameter, 150 mm length; a mixture of water (containing 0.05% NH ₄ HCO ₃ ) and acetonitrile of decreasing polarity As eluent; acetonitrile gradient ratio 55%-80%, elution time 12 minutes] to obtain the title compound (15mg).

MS m/z (ESI): 482.0 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ )δ8.43(s,1H),8.36(s,1H),7.99-7.70(m,2H),7.49-7.12(m,5H),6.55(d,J =9.8Hz,1H),4.29-4.23(m,2H),1.24(t,J=7.0Hz,3H).

Example 7, 4-(4-(difluoromethoxy)phenyl)-2-ethoxy-6-(1-methyl-6-oxo-1,6-dihydropyridazine-3- yl)thiazolo[4,5-b]pyridin-5-(4H)-one (Compound 7)

Step 1: 4-(4-(Difluoromethoxy)phenyl)-2-ethoxy-6-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl ) Synthesis of thiazolo[4,5-b]pyridin-5-(4H)-one (compound)

Intermediate 1-8 (40 mg, 95.87 μmol) and Intermediate 7-1 (48.7 mg, 191.74 μmol) were dissolved in dioxane (2 mL), to which was added potassium acetate (28.2 mg, 287.61 μmol) and Pd (dppf)Cl ₂ (7.0 mg, 9.59 μmol), the reaction solution was stirred at 90° C. for 1 h under nitrogen protection. LC-MS detected the completion of the reaction. Intermediate 7-2 (36 mg, 191.74 μmol), cesium carbonate (62.6 mg, 191.74 μmol), Pd(dtbpf)Cl ₂ (6.3 mg, 9.5 μmol) and water (0.5 mL) were added to the reaction solution, and the mixture was heated under nitrogen. Stir under protection at 90 °C for 1 h. LC-MS detected the completion of the reaction. The reaction solution was filtered, concentrated to dryness under reduced pressure, and purified by preparative high performance liquid chromatography (column: Gemini NX C18 5 μm*10*150mm; mobile phase: A: water (0.225% ammonium bicarbonate v/v), B: acetonitrile; B%: 30%-50%, 11 min) to obtain the title compound (10.6 mg).

MS m/z(ESI): 447.1[M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.51 (s, 1H), 8.01 (d, J=9.7Hz, 1H), 7.48 (d, J=8.9Hz, 2H), 7.37 (t, J= 76.0Hz, 1H), 7.33(d, J=8.8Hz, 2H), 6.93(d, J=9.7Hz, 1H), 4.47–4.25(m, 2H), 3.72(s, 3H), 1.31(t, J=7.0Hz, 3H).

Example 8, 4-(4-(difluoromethoxy)phenyl)-2-ethoxy-6-(1-(methyl-d ₃ )-1H benzo[d]imidazol-6-yl )thiazolo[4,5-b]pyridin-5(4H)-one (Compound 8)

Step 1: Synthesis of 6-bromo-1-(methyl- _d3 )-1H-benzo[d]imidazole (Intermediate 8-2)

The reactant 8-1 (5.0 g, 25.38 mmol) was dissolved in tetrahydrofuran solution (50 mL), sodium hydride (60%) (1.12 g, 27.91 mmol) was added at 0-5 °C, and the reaction solution was stirred at 5 °C for 0.5 h , CD ₃ I (4.41 g, 30.45 mmol) was added to the reaction solution, and the mixture was stirred at room temperature for 12 h. LC-MS detected the end of the reaction, slowly poured the reaction solution into saturated aqueous ammonium chloride solution (50 mL), extracted with ethyl acetate (60 mL*2), combined the organic phases, washed with saturated brine (100 mL), and the washed organic Phase is dried with an appropriate amount of anhydrous sodium sulfate to obtain 5.0g of crude product, and 1.0g of crude product is prepared by supercritical liquid chromatography (column: DAICEL CHIRALPAK IG (250mm*50mm, 10 μm); mobile phase: [A is CO ₂ , B: ethanol solution containing 0.1% ammonia water]; B%: 35%) to obtain the title compound (400 mg).

MS m/z (ESI): 214.0 [M+H] ⁺ .

Step 2: 4-(4-(Difluoromethoxy)phenyl)-2-ethoxy-6-(1-(methyl- _d3 )-1H-benzo[d]imidazol-6-yl ) Synthesis of Thiazolo[4,5-b]pyridin-5(4H)-one (Compound 8)

Intermediate 6-1 (150 mg, 0.32 mmol) and intermediate 8-2 (83 mg, 0.39 mmol) were dissolved in anhydrous dioxane solution (2 mL), to which was added water (0.4 mL), cesium carbonate ( 211 mg, 0.65 mmol) and 1,1-bis(tert-butylphosphorus)ferrocene palladium chloride (Pd(dtbpf)Cl ₂ ) (21 mg, 0.03 mmol), the reaction solution was stirred at 100° C. for 2 h under nitrogen protection. LC-MS detected the completion of the reaction. After the reaction was cooled to room temperature, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and purified by preparative high performance liquid chromatography [YMC-Actus Triart C18 column 5 μm silica, 30 mm diameter, 150 mm length; water (containing 0.05% NH _{4 )} A mixture of decreasing polarity of HCO ₃ ) and acetonitrile as eluent; acetonitrile gradient ratio 55%-80%, elution time 13 minutes] to give the title compound (16 mg).

MS m/z (ESI): 472.0 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.34(s, 1H), 8.20(s, 1H), 7.97(d, J=1.6Hz, 1H), 7.66(d, J=8.5Hz, 1H) ,7.57–7.21(m,6H),4.36–4.31(m,2H),1.35–1.31(m,3H).

Example 9, N-(6-(4-(4-(difluoromethoxy)phenyl)-2-ethoxy-5-oxo-4,5-dihydrothiazolo[4,5- b]Pyridin-6-yl)-1-methyl-1H-benzo[d]imidazol-2-yl)acetamide (Compound 9)

Step 1: N-(6-(4-(4-(difluoromethoxy)phenyl)-2-ethoxy-5-oxo-4,5-dihydrothiazolo[4,5-b] ]pyridin-6-yl)-1-methyl-1H-benzo[d]imidazol-2-yl)acetamide (compound 9)

Intermediate 9-1 (67 mg, 0.25 mmol) and Intermediate 6-1 (115 mg, 0.30 mmol) were dissolved in dioxane (2 mL) and water (0.5 mL), to which was added cesium carbonate (164 mg, 0.50 mmol) and 1,1-bis(tert-butylphosphorus)ferrocene palladium chloride (Pd(dtbpf)Cl ₂ ) (24 mg, 0.04 mmol), the reaction solution was stirred at 80° C. for 16 h under nitrogen protection. LC-MS detected the completion of the reaction. The reaction solution was filtered, concentrated to dryness under reduced pressure, purified by preparative high performance liquid chromatography on YMC-Actus Triart C18 column 5μm silica, 30mm diameter, 150mm length; water (containing 0.05% NH ₄ HCO ₃ ) and acetonitrile with decreasing polarity The mixture was used as the eluent; the acetonitrile gradient ratio was 30%-50%, the elution time was 12 minutes) to obtain the title compound (11 mg).

MS m/z (ESI): 526.1 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ )δ10.62(s,1H), 8.31(s,1H), 7.87(s,1H), 7.60-7.43(m,4H), 7.39-7.30(m,3H) ), 4.35–4.30(m, 2H), 3.30(s, 3H), 2.16(s, 3H), 1.32(t, J=7.0Hz, 3H).

Example 10, 4-(4-(difluoromethoxy)phenyl)-2-ethoxy-6-(4-oxo-1,2,3,9b-tetrahydro-4λ ⁴ -benzo [c] Thieno[2,1-e]isothiazol-8-yl)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 10)

Intermediate 10-1 (200mg, 0.52mmol, can be synthesized according to the method reported in the literature "Tetrahydrobenzo[c]thieno-[2,1-e]isothiazole 4-Oxides:Three-Dimensional Heterocycles as Cross-Coupling Building Blocks") and Intermediate 6-1 (201 mg, 0.62 mmol) was dissolved in dioxane (2 mL) and water (0.4 mL), to which were added potassium phosphate (330 mg, 1.56 mmol) and 1,1-bis(tert-butylphosphorus) ) ferrocene palladium chloride (Pd(dtbpf)Cl ₂ ) (33 mg, 0.05 mmol), the reaction solution was stirred at 100° C. for 16 h under nitrogen protection. LC-MS detected the completion of the reaction. After the reaction was cooled to room temperature, water (30 mL) and ethyl acetate (40 mL) were added in sequence, the organic phase was washed with water (30 mL*2), and the washed organic phase was dried with an appropriate amount of anhydrous sodium sulfate. The crude product (210 mg) was obtained by thin layer chromatography (silica, dichloromethane:methanol = 5:1), and then purified by preparative high performance liquid chromatography (YMC-Actus Triart C18 column 5 μm silica, 30 mm diameter, 150 mm length; a mixture of water (containing 0.05% trifluoroacetic acid) and acetonitrile of decreasing polarity as eluent; acetonitrile gradient ratio 40%-60%, elution time 13 minutes) to give the title compound (30 mg).

MS m/z (ESI): 530.1 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ )δ8.16(s,1H),7.61(d,J=1.9Hz,1H),7.57-7.11(m,6H),6.76(d,J=8.3Hz, 1H), 4.91–4.84 (m, 1H), 4.36–4.25 (m, 2H), 3.77–3.67 (m, 1H), 3.62–3.49 (m, 1H), 2.79–2.67 (m, 1H), 2.22– 2.13 (m, 1H), 2.14–2.05 (m, 1H), 1.67–1.51 (m, 1H), 1.35–1.21 (m, 3H).

Example 11, 4-(4-(difluoromethoxy)phenyl)-6-(2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazole-7- yl)-2-ethoxythiazolo[4,5-b]pyridin-5(4H)-one (Compound 11)

Intermediate 11-1 (149mg, 628.03μmol, can be reported according to the method "Metal-Free Sequential C(sp2)-H/OH and C(sp3)-H Aminations of Nitrosoarenes and N-Heterocycles to Ring-Fused Imidazoles" synthesis) and intermediate 6-1 (200 mg, 523.36 μmol) were dissolved in dioxane (2 mL) and water (0.5 mL), to which was added cesium carbonate (341 mg, 1.05 mmol) and 1,1-bis(tert. Butylphosphorus) ferrocene palladium chloride (Pd(dtbpf)Cl ₂ ) (34 mg, 52.01 μmol), the reaction solution was stirred at 90° C. for 2 h under nitrogen protection. LC-MS detected the completion of the reaction. The reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (YMC-Actus Triart C18 column 5 μm silica, 30 mm diameter, 150 mm length; polar with water (containing 0.05% formic acid) and acetonitrile Decreasing mixture was used as eluent; acetonitrile gradient ratio 30%-50%, elution time 11 min) to give the title compound (57.0 mg).

MS m/z (ESI): 495.0 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ )δ8.29(s,1H),7.86(s,1H),7.56-7.19(m,6H),4.35-4.29(m,2H),4.09(t,J = 7.0Hz, 2H), 2.96–2.94 (m, 2H), 2.67–2.64 (m, 2H), 1.32 (t, J=7.0Hz, 3H).

Example 12, 4-(4-(difluoromethoxy)phenyl)-2-ethoxy-6-(2-methyl-2H-indazol-5-yl)thiazolo[4,5- b] Synthesis of Pyridin-5(4H)-one (Compound 12)

Intermediate 1-8 (150 mg, 0.36 mmol) and intermediate 12-1 (111 mg, 0.43 mmol) were dissolved in anhydrous dioxane solution (2 mL), to which was added water (0.4 mL), cesium carbonate ( 234 mg, 0.72 mmol) and 1,1-bis(tert-butylphosphorus)ferrocene palladium chloride (Pd(dtbpf)Cl ₂ ) (23 mg, 0.03 mmol), the reaction solution was stirred at 100° C. for 2 h under nitrogen protection. LC-MS detected the completion of the reaction. After the reaction was cooled to room temperature, the reaction solution was filtered, concentrated to dryness under reduced pressure, and purified by preparative high performance liquid chromatography [YMC-Actus Triart C18 column 5 μm silica, 30 mm diameter, 150 mm length; water (containing 0.05% NH ₄ HCO) ₃ ) and acetonitrile with decreasing polarity mixture as eluent; acetonitrile gradient ratio 55%-80%, elution time 14 minutes] to obtain the title compound (12 mg).

MS m/z (ESI): 469.1 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ )δ8.36(s,1H), 8.27(s,1H), 8.05(s,1H), 7.59-7.19(m,7H), 4.34-4.29(m,2H) ), 4.17(s, 3H), 1.33–1.30(m, 3H).

Example 13, 4-(6-cyclopropylpyridin-3-yl)-2-ethoxy-6-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl) Thiazolo[4,5-b]pyridin-5(4H)-one (Compound 13)

Step 1: 6-Bromo-4-(6-cyclopropylpyridin-3-yl)-2-ethoxythiazolo[4,5-b]pyridin-5(4H)-one (Intermediate 13- 2) Synthesis

Under an oxygen atmosphere, intermediate 1-7 (80.0 mg) was dissolved in dichloroethane (DCE) solution (2 mL), and intermediate 13-1 (56.87 mg), copper acetate (105.6 mg) were added to the reaction solution. ), and pyridine (69.0 mg), the temperature was raised to 45 °C and stirred for 24 h after the addition. Subsequently, the reaction solution was concentrated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (PE/EA=5/1-2/1). The title compound (40 mg) was obtained.

MS m/z (ESI): 392 [M+H] ⁺ .

Step 2: 4-(6-Cyclopropylpyridin-3-yl)-2-ethoxy-6-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)thiazole Synthesis of [4,5-b]pyridin-5(4H)-one (Compound 13)

Under nitrogen atmosphere, intermediate 13-2 (40.0 mg) was dissolved in dioxane solution (0.4 mL), and intermediate 3-1 (28.8 mg), potassium phosphate (43.3 mg) were added to the reaction solution, Water (0.08 mL) and Pd(dtbpf)Cl ₂ (6.7 mg) were added and the temperature was raised to 100 °C and stirred for 2 h. Subsequently, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by high pressure preparation [YMC-Actus Triart C18 column 5 μm silica, 30 mm diameter, 150 mm length; water (containing 0.05% NH ₄ HCO ₃ ) and acetonitrile The mixture of decreasing polarity was used as the eluent; the acetonitrile gradient ratio was 55%-80%, the elution time was 13 minutes] to obtain the title compound (2 mg).

MS m/z (ESI): 421 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ )δ8.32(s,1H),8.22(d,J=2.6Hz,1H),8.20(s,1H),7.71-7.69(m,1H),7.66- 7.63 (m, 1H), 7.41 (d, J=8.3Hz, 1H), 6.37 (d, J=9.5Hz, 1H), 4.28–4.23 (m, 2H), 3.39 (s, 3H), 2.20–2.10 (m,1H),1.24(t,J=7.0Hz,3H),0.99–0.86(m,4H).

Example 14, 2-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-6-(1-methyl-6-oxo-1,6 -Dihydropyridin-3-yl)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 14)

Step 1: Synthesis of 6-bromo-2-(2,2-difluoroethoxy)thiazolo[4,5-b]pyridin-5(4H)-one (Intermediate 14-2)

Intermediate 14-1 (150 mg) was dissolved in tetrahydrofuran (THF) (2 mL), and NaH (77 mg, 60% wt) was added under ice-water bath. After the addition was completed, the mixture was stirred at room temperature for 0.5 hours. 2,2-Difluoroethanol (119 mg) was added under an ice-water bath. After the addition was completed, the mixture was stirred at room temperature for 12 hours. Subsequently, the reaction solution was poured into saturated aqueous ammonium chloride solution (5 mL), extracted with ethyl acetate (5 mL*3), the organic phases were combined, washed with saturated brine (10 mL), and the organic phase was dried over anhydrous sodium sulfate, Concentration to dryness under reduced pressure gave the title compound (110 mg).

MS m/z (ESI): 311.0 [M+H] ⁺ .

Step 2: 6-Bromo-2-(2,2-difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)thiazolo[4,5-b]pyridine-5(4H )-ketone (intermediate 14-4) synthesis

Intermediate 14-2 (120 mg) was dissolved in dichloroethane (2 mL), and copper acetate (140 mg), pyridine (92 mg), and intermediate 14-3 (824 mg) were added to the reaction solution. After the addition was completed, air was introduced into the reaction solution, and the reaction solution was heated to 60° C. and stirred for 12 h. Subsequently, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by column chromatography (DCM/MeOH=10/1) to obtain the title compound (90 mg).

MS m/z (ESI): 453.0 [M+H] ⁺ .

Step 3: 2-(2,2-Difluoroethoxy)-4-(4-(difluoromethoxy)phenyl)-6-(1-methyl-6-oxo-1,6- Synthesis of Dihydropyridin-3-yl)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 14)

Under nitrogen atmosphere, intermediate 14-4 (90 mg) was dissolved in dioxane solution (1 mL), intermediate 3-1 (56 mg), cesium carbonate (129 mg), water (0.2 mL) and Pd ( dtbpf)Cl ₂ (13 mg), the temperature was raised to 100° C. and stirred for 2 hours after the addition was completed. Subsequently, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by high pressure preparative liquid chromatography [YMC-Actus Triart C18 column 5 μm silica, 30 mm diameter, 150 mm length; water (containing 0.05% NH ₄ HCO _{3 )} ) and acetonitrile with decreasing polarity mixture as eluent; acetonitrile gradient ratio 55%-80%, elution time 13 minutes] to obtain the title compound (25 mg).

MS m/z (ESI): 482.1 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ )δ8.24-8.21(m,2H),7.72-7.69(m,1H),7.49-7.12(m,5H),6.43-6.15(m,2H),4.57 –4.49(m, 2H), 3.40(s, 3H).

Example 15, 4-(4-(difluoromethoxy)phenyl)-2-ethoxy-6-(6-methoxypyridazin-3-yl)thiazolo[4,5-b] Pyridin-5(4H)-one (Compound 15)

Step 1: (4-(4-(Difluoromethoxy)phenyl)-2-ethoxy-5-oxo-4,5-dihydrothiazolo[4,5-b]pyridine-6- Synthesis of boronic acid (intermediate 15-1)

Intermediate 1-8 (4.2 g) and double pinacol boronate (3.8 g) were dissolved in anhydrous dioxane (42 mL), to which was added potassium acetate (1.9 g) and Pd(dppf)Cl ₂ (1.1g), the reaction solution was stirred at 100°C for 15h under nitrogen protection. LC-MS detected the completion of the reaction. After the reaction was cooled to room temperature, the solvent was removed by rotary evaporation, water (40 mL) and ethyl acetate (50 mL) were added successively, the organic phase was washed with water (15 mL*2), and the washed organic phase was dried with an appropriate amount of anhydrous sodium sulfate, filtered, The filtrate was concentrated to dryness under reduced pressure. The residue was purified by preparative high performance liquid chromatography (chromatographic column: Gemini NX C18 5μm*10*150mm; mobile phase: A: water (0.225% formic acid v/v), B: acetonitrile; B%: 30%-50%) The title compound (3.2 g) was obtained.

MS m/z (ESI): 383.1 [M+H] ⁺ .

Step 2: 4-(4-(Difluoromethoxy)phenyl)-2-ethoxy-6-(6-methoxypyridazin-3-yl)thiazolo[4,5-b]pyridine Synthesis of -5(4H)-one (Compound 15)

Intermediate 15-1 (36.6 mg) and Intermediate 15-2 (36.24 mg) were dissolved in dioxane (1 mL) and water (0.25 mL), to which was added cesium carbonate (62.48 mg) and Pd (dtbpf )Cl ₂ (6.25mg), the reaction solution was stirred at 90°C for 16h under nitrogen protection. Subsequently, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (chromatographic column: Gemini NX C18 5 μm*10*150 mm; mobile phase: A: water (0.225% formic acid v/v), B: acetonitrile; B%: 30%-50%, 11 min) to obtain the title compound (9.0 mg).

MS m/z (ESI): 447.1 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ )δ8.86(s,1H),8.32(d,J=9.3Hz,1H),7.56-7.19(m,6H),4.37-4.31(m,2H), 4.06(s, 3H), 1.32(t, J=6.0Hz, 3H).

Example 16, 4-(4-(difluoromethoxy)phenyl)-2-ethoxy-6-(2-methoxypyrimidin-5-yl)thiazolo[4,5-b]pyridine -5(4H)-one (Compound 16)

Intermediate 1-8 (40 mg) and intermediate 16-1 (45 mg) were dissolved in dioxane (1 mL) and water (0.2 mL), cesium carbonate (62.47 mg) was added, Pd(dtbpf)Cl ₂ (6.25 mg), nitrogen was passed through, and the reaction was stirred at 90 °C for 1 h. The reaction solution was added with mercapto silica gel (50mg), stirred for 30min, filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (column: Phenomenex Luna C18 100*30mm*5 μm; mobile phase: [water (0.05 % _CF3COOH , v/v)-acetonitrile]; B%: 43%-63%, 9 min) to give the title compound (10 mg).

MS m/z (ESI): 447.1 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d6)δ8.93(s,2H), 8.42(s,1H), 7.56-7.19(m,5H), 4.37-4.31(m,2H), 3.96(s,3H) ,1.32(t,J=6.0Hz,3H).

Example 17, 4-(4-(difluoromethoxy)phenyl)-2-ethoxy-6-(2-(2-hydroxypropan-2-yl)-1-methyl-1H-benzene [d]imidazol-6-yl)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 17)

Step 1: Synthesis of 2-(6-Bromo-1-methyl-1H-benzo[d]imidazol-2-yl)propan-2-ol (Intermediate 17-2)

Intermediate 17-1 (500 mg) was dissolved in dilute hydrochloric acid (4 mL, 2 N), 2-hydroxy-2-methylpropionic acid (388 mg) was added, and the reaction solution was stirred at 100° C. for 8 h. After the reaction was completed, after the reaction was cooled to room temperature, water (15 mL) and ethyl acetate (20 mL) were added successively, the organic phase was washed with water (10 mL*2), and the washed organic phase was dried with an appropriate amount of anhydrous sodium sulfate, and after filtration, It was concentrated to dryness under reduced pressure, and the residue was purified by column chromatography (dichloromethane:methanol=10:1) to obtain the title compound (610 mg).

MS m/z(ESI): 269.0[M+H] ⁺

Step 2: 4-(4-(Difluoromethoxy)phenyl)-2-ethoxy-6-(2-(2-hydroxypropan-2-yl)-1-methyl-1H-benzo Synthesis of [d]imidazol-6-yl)thiazolo[4,5-b]pyridin-5(4H)-one (compound 17)

Intermediate 6-1 (150 mg) and Intermediate 17-2 (104 mg) were dissolved in 1,4-dioxane (1.2 mL) and water (0.3 mL), and Pd(dtbpf)Cl ₂ (32 mg) was added , Cs ₂ CO ₃ (211 mg), and the reaction solution was stirred at 100 °C for 4 h under nitrogen protection. After the reaction was completed, after the reaction was cooled to room temperature, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by column chromatography (dichloromethane:methanol=10:1) to obtain a crude product, which was then prepared by high performance liquid phase Chromatographic purification [YMC-Actus Triart C18 column 5μm silica, 30mm diameter, 150mm length; mixture of water (containing 0.225% formic acid) and acetonitrile with decreasing polarity as eluent; acetonitrile gradient ratio 30%-60%, wash Detox time 15 minutes] to give the title compound (3.1 mg).

MS m/z (ESI): 527.1 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ )δ8.17(s,1H),7.86(s,1H),7.65-7.63(m,1H),7.53-7.51(m,1H),7.46-7.44(m ,2H),7.34–7.32(m,2H),7.14–6.77(m,1H),4.42–4.36(m,2H),4.10(s,3H),1.74(s,6H),1.36(t,J =7.0Hz,3H).

Example 18, 4-(4-(difluoromethoxy)phenyl)-6-(2,3-dihydrobenzofuran-5-yl)-2-ethoxythiazolo[4,5- b] Pyridin-5(4H)-one (Compound 18)

Intermediate 6-1 (80 mg) and Intermediate 18-1 (51 mg) were dissolved in 1,4-dioxane (1.2 mL) and water (0.3 mL), to which was added Pd(dtbpf)Cl ₂ ( 17 mg), Cs ₂ CO ₃ (112 mg), the reaction solution was stirred at 100° C. for 10 h under nitrogen protection. After completion of the reaction, after the reaction was cooled to room temperature, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by column chromatography (dichloromethane:methanol=20:1) to obtain a crude product, which was then prepared by high-performance liquid phase Chromatographic purification [YMC-Actus Triart C18 column 5μm silica, 30mm diameter, 150mm length; mixture of water (containing 0.225% formic acid) and acetonitrile of decreasing polarity as eluent; acetonitrile gradient ratio 40%-70%, wash Detoxification time 15 minutes] to give the title compound (9 mg).

MS m/z (ESI): 457.1 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.16(s, 1H), 7.57(s, 1H), 7.56-7.17(m, 6H), 6.77(d, J=8.3Hz, 1H), 4.55( t, J=8.7Hz, 2H), 4.33–4.28 (m, 2H), 3.19 (t, J=8.7Hz, 2H), 1.31 (t, J=7.0Hz, 3H).

Example 19, 4-(4-(difluoromethoxy)phenyl)-6-(1-methyl-1H-benzo[d]imidazol-6-yl)-2-(oxetine- 3-yloxy)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 19)

Step 1: Synthesis of 2,6-dibromo-5-methoxythiazolo[4,5-b]pyridine (Intermediate 19-1)

Copper bromide (103 g) and isoamyl nitrite (86 g) were dissolved in acetonitrile (1000 mL), cooled to 0 °C, intermediates 1-4 (60 g) were added to the reaction flask in batches, and then the reaction solution was heated at 25 °C The reaction was stirred for 16h. The reaction solution was poured into dilute HCl solution and stirred for 30min, adjusted to pH=8 with sodium bicarbonate, extracted with mixed solvent dichloromethane:methanol=10:1 (200mL*3), and the combined organic phase was dried with anhydrous sodium sulfate . Preparative thin layer chromatography (silica, dichloromethane) gave the title compound (12 g).

MS m/z (ESI): 323.1 [M+H] ⁺ .

Step 2: Synthesis of 2,6-dibromothiazolo[4,5-b]pyridin-5(4H)-one (Intermediate 19-2)

Intermediate 19-1 (8 g) was dissolved in acetic acid (50 mL), 48% hydrobromic acid (52 mL) was added, and the reaction solution was stirred at 80° C. for 5 h. After completion of the reaction, the solvent was removed by concentration under reduced pressure, saturated aqueous sodium bicarbonate solution (60 mL) was added thereto, filtered, and the filter cake was washed with water (40 mL*2) to obtain the title compound (9.5 g).

MS m/z (ESI): 309.1 [M+H] ⁺ .

Step 3: Synthesis of 6-bromo-2-(oxetan-3-yloxy)thiazolo[4,5-b]pyridin-5(4H)-one (Intermediate 19-3)

3-oxetanol (358.5 mg, 4.84 mmol) was dissolved in tetrahydrofuran (10 mL), NaH (193.6 mg, 4.84 mmol) was added at 0°C, and the stirring was completed for 10 minutes, and then Intermediate 19-2 (500 mg) was added. , 1.61mmol), stirred at room temperature for 3h. Water (10 mL) was added to the reaction solution to quench the reaction, extracted with ethyl acetate (20 mL*3), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness under reduced pressure. The residue was purified by column chromatography (DCM:CH3OH ₌ 10:1) to give the title compound (52.0 mg).

MS m/z (ESI): 303.1 [M+H] ⁺ .

Step 4: 6-Bromo-4-(4-(difluoromethoxy)phenyl)-2-(oxetan-3-yloxy)thiazolo[4,5-b]pyridine-5 Synthesis of (4H)-ketone (Intermediate 19-4)

Intermediate 19-3 (50 mg) and 4-(difluoromethoxy)phenylboronic acid (62 mg) were dissolved in dichloroethane (2 ml), followed by addition of Cu(OAc) ₂ (44.9 mg, ), pyridine ( 39.14mg), reacted at 45°C for 12h. The reaction solution was concentrated to dryness under reduced pressure. The residue was purified by column chromatography (DCM:CH3OH ₌ 10:1) to give the title compound (57.9 mg).

MS m/z(ESI): 445.1[M+H] ⁺

Step 5: 4-(4-(Difluoromethoxy)phenyl)-6-(1-methyl-1H-benzo[d]imidazol-6-yl)-2-(oxetane-3 Synthesis of -yloxy)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 19)

Intermediate 5-1 (57.97 mg) and Intermediate 19-1 (50 mg) were dissolved in dioxane (1 mL) and water (0.2 mL), to which was added cesium carbonate (73.17 mg) and Pd(dtbpf) Cl ₂ (7.32 mg), the reaction solution was stirred at 90° C. for 16 h under nitrogen protection. Then the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (chromatographic column: Gemini NX C18 5 μm*10*150 mm; mobile phase: A: water (0.225% formic acid v/v), B : acetonitrile; B%: 30%-50%, 11 min) to obtain the title compound (9.8 mg).

MS m/z (ESI): 497.1 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ )δ8.27(s,1H), 8.13(s,1H), 7.88(s,1H), 7.59-7.49(m,1H), 7.45-7.12(m,6H) ),5.43-5.40(m,1H)4.63-4.60(m,2H)4.55-4.52(m,2H),3.76(s,3H).

Example 20, 4-(4-(difluoromethoxy)phenyl)-2-ethoxy-6-(imidazo[1,2-a]pyridin-6-yl)thiazolo[4,5 -b]pyridin-5(4H)-one (compound 20)

Intermediate 15-1 (40 mg) and Intermediate 20-1 (24.75 mg) were dissolved in dioxane (1 mL) and water (0.25 mL), to which was added cesium carbonate (68.2 mg) and Pd(dtbpf) Cl ₂ (6.82 mg), the reaction solution was stirred at 90° C. for 16 h under nitrogen protection. Subsequently, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (chromatographic column: Gemini NX C185 μm*10*150 mm; mobile phase: A: water (0.225% formic acid v/v), B : acetonitrile; B%: 30%-50%, 11 min) to obtain the title compound (18.0 mg).

MS m/z (ESI): 455.1 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d6)δ9.14(s,1H), 8.45(s,1H), 7.98(s,1H), 7.60-7.19(m,8H), 4.36-4.31(m,2H) ,1.31(t,J=7.1Hz,3H).

Example 21: 4-(4-(Difluoromethoxy)phenyl)-2-ethoxy-6-(1-methyl-2,3-dihydro-1H-benzo[d]imidazole[ 1,2-a]imidazol-6-yl)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 21)

Compound 28 (57 mg) was dissolved in anhydrous tetrahydrofuran (0.8 mL), bistrimethylsilyl amide lithium solution (1 mol/L THF solution, 0.15 mL) was added to it at 0°C, and iodine was added after 30 min Methane (33 mg), the reaction solution was stirred at 30 °C for 4 h. After the reaction was completed, the solvent was removed by concentration under reduced pressure, and the residue was purified by preparative high performance liquid chromatography [YMC-Actus Triart C18 column 5 μm silica, 30 mm diameter, 150 mm length; water (containing 0.225% formic acid) and acetonitrile with decreasing polarity The mixture was used as the eluent; the acetonitrile gradient ratio was 40%-70%, the elution time was 14 minutes] to obtain the title compound (16 mg).

MS m/z (ESI): 510.1 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ )δ8.20-8.16(m,1H),7.59-7.09(m,8H),4.33-4.28(m,2H),4.14-4.08(m,2H),3.94 –3.85(m, 2H), 2.93(s, 3H), 1.31(t, J=7.0Hz, 3H).

Example 22, 6-(benzo[4,5]imidazo[1,2-a]pyrimidin-7-yl)-4-(4-(difluoromethoxy)phenyl)-2-ethoxy Thiazolo[4,5-b]pyridin-5(4H)-one (Compound 22)

Intermediate 22-1 (48.69 mg) and Intermediate 15-1 (50 mg) were dissolved in dioxane (2 mL) and water (0.5 mL), to which was added cesium carbonate (85.26 mg) and Pd (dtbpf) Cl ₂ (8.53 mg), the reaction solution was stirred at 90° C. for 16 h under nitrogen protection. Subsequently, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (chromatographic column: Gemini NX C18 5 μm*10*150 mm; mobile phase: A: water (0.225% formic acid v/v), B: acetonitrile; B%: 30%-50%, 11 min) to obtain the title compound (4.5 mg).

MS m/z (ESI): 506.0 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ )δ9.56-9.54(m,1H), 8.84-8.82(m,1H), 8.74(s,1H), 8.38(s,1H), 7.88(s,2H) ), 7.56–7.14 (m, 6H), 4.37–4.31 (m, 2H), 1.32 (t, J=8.0Hz, 3H).

Example 23, 4-(4-(difluoromethoxy)phenyl)-6-(1-(methyl-d ₃ )-1H-benzo[d]imidazol-6-yl)-2-( Trifluoromethyl)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 23)

Step 1: 1-(Methyl- _d3 )-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo Synthesis of [d]imidazole (intermediate 23-1)

Under nitrogen atmosphere, intermediate 8-2 (1.0 g) was dissolved in dioxane solution (10 mL), potassium acetate (917 mg), bispinacol borate (1.42 g) and Pd (dppf) were added )Cl2 (342 _mg ). The reaction solution was heated to 100 °C and stirred for 2 h. Subsequently, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by column chromatography (PE/EA=5/1-3/1) to obtain the title compound (500 mg).

MS m/z (ESI): 262.0 [M+H] ⁺ .

Step 2: Analysis of 2,6-dibromo-4-(4-(difluoromethoxy)phenyl)thiazolo[4,5-b]pyridin-5(4H)-one (Intermediate 23-3) synthesis

Intermediate 23-2 (450 mg) was dissolved in 1,2-dichloroethane (5 mL), copper acetate (318 mg), pyridine (0.4 mL), intermediate 14-3 (409 mg) were added, and the reaction was completed. The air was bubbling in the middle, and the reaction solution was heated to 60 °C and stirred for 48 h. Subsequently, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by column chromatography (DCM:MeOH=20:1) to obtain the title compound (450 mg).

MS m/z (ESI): 451.0 [M+H] ⁺ .

Step 3: 6-Bromo-4-(4-(difluoromethoxy)phenyl)-2-(trifluoromethyl)thiazolo[4,5-b]pyridin-5(4H)-one (intermediate Synthesis of body 23-4)

Under nitrogen atmosphere, intermediate 23-3 (400 mg) was dissolved in DMF (5 mL), cuprous iodide (202 mg) and methyl 2,2-difluoro-2-(fluorosulfonyl)acetate (204 mg) were added ). The reaction solution was heated to 100 °C and stirred for 12 h. Then, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by column chromatography (DCM:MeOH=20/1-10/1) to obtain the title compound (150 mg).

MS m/z (ESI): 441.0 [M+H] ⁺ .

Step 4: 4-(4-(Difluoromethoxy)phenyl)-6-(1-(methyl- _d3 )-1H-benzo[d]imidazol-6-yl)-2-(tris Synthesis of Fluoromethyl)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 23)

Under nitrogen atmosphere, intermediate 23-4 (120.0 mg) was dissolved in dioxane solution (2 mL), intermediate 23-1 (71 mg), cesium carbonate (177 mg), water (0.2 mL) and Pd were added (dtbpf)Cl2 ( ₁₈ mg). After the addition was completed, the temperature was raised to 100 °C and stirred for 2 h. Subsequently, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by high pressure preparative liquid chromatography [YMC-Actus Triart C18 column 5 μm silica, 30 mm diameter, 150 mm length; water (containing 0.05% NH ₄ HCO _{3 )} ) and acetonitrile with decreasing polarity mixture as eluent; acetonitrile gradient ratio 55%-80%, elution time 13 minutes] to obtain the title compound (25 mg).

MS m/z (ESI): 496.1 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.66(s, 1H), 8.27(s, 1H), 8.03(s, 1H), 7.73(d, J=8.4Hz, 1H), 7.60-7.24( m,6H).

Example 24, 4-(4-(difluoromethoxy)phenyl)-2-methoxy-6-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl )thiazolo[4,5-b]pyridin-5(4H)-one (Compound 24)

Step 1: 6-Bromo-4-(4-(difluoromethoxy)phenyl)-2-methoxythiazolo[4,5-b]pyridin-5(4H)-one (Intermediate 24- 1) Synthesis

Intermediate 23-3 (250 mg) was dissolved in methanol (5 mL), sodium methoxide (120 mg) was added thereto, and the reaction was carried out at 25° C. for 5 h. Subsequently, the reaction solution was concentrated under reduced pressure to remove the solvent, and the residue was washed with water (10 mL). Ethyl acetate (10 mL*2) was extracted, the organic phase was dried with an appropriate amount of anhydrous sodium sulfate, and the title compound (172 mg) was obtained by preparative thin layer chromatography (petroleum ether:ethyl acetate=2:1).

MS m/z (ESI): 402.9 [M+H] ⁺ .

Step 2: 4-(4-(Difluoromethoxy)phenyl)-2-methoxy-6-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl) Synthesis of Thiazolo[4,5-b]pyridin-5(4H)-one (Compound 24)

Intermediate 24-1 (80 mg) and Intermediate 3-1 (70 mg) were dissolved in 1,4-dioxane (1.2 mL) and water (0.3 mL), to which was added Pd(dtbpf)Cl ₂ ( 19mg) and cesium carbonate (196mg), reacted at 100°C for 4h under nitrogen atmosphere, then, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (chromatographic column: Gemini NX C185μm*10* 150mm; mobile phase: A: water (0.225% formic acid v/v), B: acetonitrile; B%: 30%-50%, 17 min) to obtain the title compound (13 mg).

MS m/z (ESI): 432.0 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ )δ8.30(d, J=2.5Hz, 1H), 8.26(s, 1H), 7.79-7.76(m, 1H), 7.62-7.14(m, 5H), 6.44(d, J=9.5Hz, 1H), 3.97(s, 3H), 3.46(s, 3H).

Example 25, 2-ethoxy-4-(4-fluorophenyl)-6-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)thiazolo[4, 5-b]pyridin-5(4H)-one (Compound 25)

Step 1: Synthesis of 6-bromo-2-ethoxy-4-(4-fluorophenyl)thiazolo[4,5-b]pyridin-5(4H)-one (Intermediate 25-2)

Intermediate 1-7 (100 mg) and intermediate 25-1 (156 mg) were dissolved in anhydrous 1,2-dichloroethane (5 mL), pyridine (86 mg) and copper acetate (79 mg) were added, and the addition was completed, The reaction solution was stirred at 40°C for 4 hours. Subsequently, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The residue was purified by residual column chromatography (dichloromethane:methanol=20:1) to obtain the title compound (100 mg).

MS m/z (ESI): 368.0 [M+H] ⁺ .

Step 2: 2-Ethoxy-4-(4-fluorophenyl)-6-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)thiazolo[4,5 Synthesis of -b]pyridin-5(4H)-one (compound 25)

Intermediate 25-2 (55 mg) was dissolved in 1,4-dioxane solution (5 mL) and water (0.5 mL), to which was added intermediate 3-1 (73 mg), Pd(dppf)Cl ₂ ( 11 mg) and cesium carbonate (151 mg), and the reaction solution was stirred at 100° C. for 12 hours under nitrogen protection. After the reaction was cooled to room temperature, water (10 mL) and ethyl acetate (10 mL) were added successively, the organic phase was washed with water (5 mL*2), and the washed organic phase was dried with an appropriate amount of anhydrous sodium sulfate. After filtration, the organic phase was concentrated under reduced pressure to remove the solvent, and the residue was purified by preparative high performance liquid chromatography (column: Waters Xbridge BEH C18 100*25mm*5μm; mobile phase: [A: water (0.225% formic acid), B: acetonitrile]; B%: 23%-50%, 12 minutes) to give the title compound (38 mg).

MS m/z (ESI): 398.0 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.32 (d, J=2.6Hz, 1H), 8.27 (s, 1H), 7.80 (dd, J=9.5, 2.7Hz, 1H), 7.48-7.44 ( m, 2H), 7.42–7.34 (m, 2H), 6.46 (d, J=9.5Hz, 1H), 4.36–4.30 (m, 2H), 1.32 (t, J=7.0Hz, 3H).

Example 26, 4-(3-chlorophenyl)-2-ethoxy-6-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)thiazolo[4, 5-b]pyridin-5(4H)-one (Compound 26)

Step 1: Synthesis of 6-bromo-4-(3-chlorophenyl)-2-ethoxythiazolo[4,5-b]pyridin-5(4H)-one (Intermediate 26-2)

Intermediate 1-7 (100 mg) and intermediate 26-1 (156 mg) were dissolved in anhydrous 1,2-dichloroethane (5 mL), pyridine (86 mg) and copper acetate (79 mg) were added, and the addition was completed, The reaction solution was stirred at 40°C for 4 hours. Subsequently, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The residue was purified by column chromatography (dichloromethane:methanol=20:1) to obtain the title compound (170 mg).

MS m/z (ESI): 385.0 [M+H] ⁺ .

Step 2: 4-(3-Chlorophenyl)-2-ethoxy-6-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)thiazolo[4,5 Synthesis of -b]pyridin-5(4H)-one (compound 26)

Intermediate 26-2 (60 mg) was dissolved in 1,4-dioxane solution (5 mL) and water (0.5 mL), to which was added intermediate 3-1 (73 mg), Pd(dppf)Cl ₂ ( 11 mg) and cesium carbonate (151 mg), and the reaction solution was stirred at 100° C. for 12 hours under nitrogen protection. After the reaction was cooled to room temperature, water (10 mL) and ethyl acetate (10 mL) were added successively, the organic phase was washed with water (5 mL*2), and the washed organic phase was dried with an appropriate amount of anhydrous sodium sulfate. After filtration, the organic phase was concentrated under reduced pressure to remove the solvent, and the residue was purified by preparative high performance liquid chromatography (column: Waters Xbridge BEH C18 100*25mm*5μm; mobile phase: [A: water (0.225% formic acid), B: acetonitrile]; B%: 23%-50%, 12 minutes) to give the title compound (38 mg).

MS m/z (ESI): 414.0 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.31 (d, J=2.6Hz, 1H), 8.27 (s, 1H), 7.80 (dd, J=9.5, 2.6Hz, 1H), 7.62-7.51 ( m, 3H), 7.42–7.38 (m, 1H), 6.45 (d, J=9.5Hz, 1H), 4.44–4.18 (m, 2H), 3.48 (s, 3H), 1.32 (t, J=7.1Hz) , 3H).

Example 27, 4-(4-(difluoromethoxy)phenyl)-6-(3,4-dihydro-2H-benzo[4,5]imidazo[2,1-b][1 ,3]oxazin-7-yl)-2-ethoxythiazolo[4,5-b]pyridin-5(4H)-one (Compound 27)

Step 1: Synthesis of 1-(3-hydroxypropyl)-3-(4-iodophenyl)thiourea (Intermediate 27-3)

Intermediate 27-1 (1.0 g) and Intermediate 27-2 (261.5 mg) were dissolved in anhydrous tetrahydrofuran (10 mL) and stirred at room temperature for 2 h. The reaction solution was concentrated under reduced pressure to obtain the title compound (1.1 g).

MS m/z (ESI): 337.1 [M+H] ⁺ .

Step 2: Synthesis of N-(4-iodophenyl)-5,6-dihydro-4H-1,3-oxazin-2-amine (Intermediate 27-4)

Intermediate 27-3 (1.1 g) was dissolved in tetrahydrofuran (20 mL), a solution of sodium hydroxide (327.19 mg) in water (5 mL) was added, and the mixture was stirred at room temperature for 10 min. Then, p-toluenesulfonyl chloride (TsCl, 748.56 mg) was added thereto, and the mixture was stirred at room temperature for 3 h. Subsequently, the reaction solution was concentrated under reduced pressure, extracted with ethyl acetate (40 ml), the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness under reduced pressure. The residue was purified by column chromatography (EA: _Et3N =100:1) to give the title compound (1.1 g).

MS m/z (ESI): 303.1 [M+H] ⁺ .

Step 3: Synthesis of 7-iodo-3,4-dihydro-2H-benzo[4,5]imidazo[2,1-b][1,3]oxazine (Intermediate 27-6)

Intermediate 27-4 (200 mg) was dissolved in acetonitrile (10 mL), intermediate 27-5 was added at 0°C, and stirred at room temperature for 2 h. Subsequently, the reaction solution was washed with saturated sodium bicarbonate solution (10 mL), extracted with dichloromethane (20 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by column chromatography (EA: _Et3N =100:1) to give the title compound (110 mg).

MS m/z (ESI): 301.1 [M+H] ⁺ .

Step 4: 4-(4-(Difluoromethoxy)phenyl)-6-(3,4-dihydro-2H-benzo[4,5]imidazo[2,1-b][1, 3] Synthesis of oxazin-7-yl)-2-ethoxythiazolo[4,5-b]pyridin-5(4H)-one (Compound 27)

Intermediate 27-6 (94.23 mg) and Intermediate 15-1 (60 mg) were dissolved in dioxane (2 mL) and water (0.5 mL), to which was added cesium carbonate (102.31 mg) and Pd (dtbpf) Cl ₂ (10.22 mg), the reaction solution was stirred at 90° C. for 16 h under nitrogen protection. Subsequently, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (chromatographic column: Gemini NX C18 5 μm*10*150 mm; mobile phase: A: water (0.225% formic acid v/v), B: acetonitrile; B%: 30%-50%, 11 min) to obtain the title compound (14 mg).

MS m/z (ESI): 511.1 [M+H] ⁺ .

¹ H NMR(400MHz,Methanol-d ₄ )δ8.26(s,1H),7.73(s,1H),7.56-7.19(m,7H),4.53-4.50(m,2H),4.34-4.29(m ,2H),4.11-4.08(m,2H),2.26-2.23(m,2H),1.31(t,J＝7.0Hz,3H)

Example 28, 4-(4-(difluoromethoxy)phenyl)-6-(2,3-dihydro-1H-benzo[d]imidazo[1,2-a]imidazole-6- yl)-2-ethoxythiazolo[4,5-b]pyridin-5(4H)-one (Compound 28)

Step 1: Synthesis of 2-((6-Bromo-1H-benzo[d]imidazol-2-yl)amino)ethan-1-ol (Intermediate 28-2)

Intermediate 28-1 (4.6 g) was dissolved in ethanolamine (3.6 mL), and the reaction solution was stirred at 140° C. for 14 h. After the reaction was completed, after the reaction was cooled to room temperature, saturated sodium bicarbonate solution (25 mL) was added to it, a solid was precipitated, the reaction solution was filtered, and the filter cake was washed with water (40 mL) and dried to obtain the crude title compound (4.1 g).

MS m/z(ESI): 255.9[M+H] ⁺

Step 2: Synthesis of 6-bromo-2,3-dihydro-1H-benzo[d]imidazo[1,2-a]imidazole (Intermediate 28-3)

Intermediate 28-2 (2.6 g) was dissolved in 1,2-dichloroethane (20 mL), to which was added thionyl chloride (0.9 mL). The reaction solution was stirred at 80 °C for 50 min. After the reaction was completed, the reaction solution was cooled to room temperature, concentrated under reduced pressure to remove the solvent, added saturated aqueous sodium bicarbonate solution (30 mL) to the residue, extracted with a co-solvent of dichloromethane/methanol (25 mL*2), and the organic phase used An appropriate amount of anhydrous sodium sulfate was dried, filtered, concentrated under reduced pressure to remove the solvent, xylene (20 mL) was added to the residue, and the reaction solution was stirred at 110° C. for 20 h. After the reaction solution was cooled to room temperature, the resulting precipitate was collected by filtration, washed with petroleum ether and dried to obtain the title compound (2.2 g).

MS m/z(ESI): 238.0[M+H] ⁺

Step 3: 4-(4-(Difluoromethoxy)phenyl)-6-(2,3-dihydro-1H-benzo[d]imidazo[1,2-a]imidazol-6-yl )-2-ethoxythiazolo[4,5-b]pyridin-5(4H)-one (Compound 28)

Intermediate 6-1 (180 mg) and Intermediate 28-3 (138 mg) were dissolved in dioxane (1.2 mL) and water (0.3 mL), to which was added cesium carbonate (253 mg) and Pd(dtbpf)Cl ₂ (38 mg), the reaction solution was stirred at 100 °C for 4 h under nitrogen protection. After completion of the reaction, after the reaction solution was cooled to room temperature, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by preparative high performance liquid chromatography [YMC-Actus Triart C18 column 5 μm silica, 30 mm in diameter, 150 mm in length; A mixture of decreasing polarity of water (containing 0.225% formic acid) and acetonitrile as eluent; acetonitrile gradient ratio 40%-70%, elution time 12 minutes] gave the title compound (88.4 mg).

MS m/z (ESI): 496.0 [M+H] ⁺ .

¹ H NMR (400MHz, Chloroform-d)δ8.21-8.18(m,1H),7.55(s,1H),7.52-7.08(m,7H),6.96(d,J=17.1Hz,1H),4.33 –4.28(m,2H),4.04-4.08(m,2H),3.98-3.95(m,2H),1.31(t,J=7.0Hz,3H).

Example 29, 4-(4-(difluoromethoxy)phenyl)-2-ethoxy-6-(1-oxo-2,3-dihydro-1H-benzo[d]pyrrole[ 1,2-a]imidazol-7-yl)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 29)

Step 1: Synthesis of 1-(5-Bromo-2-nitrophenyl)pyrrolidine-2,5-dione (Intermediate 29-1)

4-Bromonitrobenzene (1 g) and succinimide (588 mg) were dissolved in anhydrous N,N-dimethylformamide (20 mL), potassium carbonate (2.38 g) was added, and the mixture was stirred at 25°C for 12 hours . After the reaction, water (50mL) and ethyl acetate (50mL*3) were added in turn, the organic phase was washed with saturated brine (25mL*2), and the washed organic phase was dried with an appropriate amount of anhydrous sodium sulfate. The phase was concentrated under reduced pressure to remove the solvent, and the residue was purified by column chromatography (petroleum ether:ethyl acetate=1:1) to obtain the title compound (1 g).

MS m/z(ESI): 299.0[M+H] ⁺

Step 2: Synthesis of 1-(2-Amino-5-bromophenyl)pyrrolidine-2,5-dione (Intermediate 29-2)

Intermediate 29-1 (1 g) was dissolved in acetic acid (10 mL), iron powder (1.87 g) was added to the reaction solution, and the reaction solution was stirred at 25° C. for 12 hours. After the reaction, water (100 mL) and ethyl acetate (100 mL) were added in sequence, the organic phase was washed with water (50 mL*2), and the washed organic phase was dried with an appropriate amount of anhydrous sodium sulfate. After filtration, the organic phase was concentrated under reduced pressure to remove the solvent, and the residue was purified by column chromatography (petroleum ether:ethyl acetate=1:1) to obtain the title compound (0.6 g).

MS m/z(ESI): 269.0[M+H] ⁺

Step 3: Synthesis of 7-bromo-2,3-dihydro-1H-benzo[d]pyrro[1,2-a]imidazol-1-one (Intermediate 29-3)

Intermediate 29-2 (1 g) was dissolved in acetic acid (10 mL), and the reaction solution was stirred at 40°C for 12 hours. After the reaction, water (100 mL) and ethyl acetate (100 mL) were added in sequence, the organic phase was washed with water (50 mL*2), and the washed organic phase was dried with an appropriate amount of anhydrous sodium sulfate. After filtration, the organic phase was concentrated under reduced pressure to remove the solvent, and the residue was purified by column chromatography (petroleum ether:ethyl acetate=1:1) to obtain the title compound (0.2 g).

MS m/z(ESI): 251.0[M+H] ⁺

Step 4: 4-(4-(Difluoromethoxy)phenyl)-2-ethoxy-6-(trimethyltinyl)thiazolo[4,5-b]pyridine-5(4H)- Synthesis of Ketones (Intermediate 29-4)

Under nitrogen protection, intermediate 1-8 (50 mg) was added to anhydrous dioxane (6 mL), and tetrakis(triphenylphosphine) palladium (13,8 mg) and hexamethyl dioxane were added to the reaction solution. Tin (58 mg). Subsequently, the reaction solution was stirred at 120° C. for 16 hours under nitrogen protection. After completion of the reaction, the reactant was filtered and concentrated to dryness under reduced pressure. The residue was subjected to preparative thin layer chromatography (petroleum ether:ethyl acetate=10:1) to obtain the title compound (40 mg).

MS m/z(ESI): 503.0[M+H] ⁺

Step 5: 4-(4-(Difluoromethoxy)phenyl)-2-ethoxy-6-(1-oxo-2,3-dihydro-1H-benzo[d]pyrrole[1 Synthesis of ,2-a]imidazol-7-yl)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 29)

Under nitrogen protection, intermediate 29-4 (40 mg) and intermediate 29-3 (10 mg) were added to anhydrous dioxane (6 mL), and tetrakis(triphenylphosphine) palladium (9.2 mL) was added to the reaction solution. mg). Subsequently, the reaction solution was stirred at 120° C. for 16 hours under nitrogen protection. After the reaction, the reactant was filtered and concentrated under reduced pressure to remove the solvent. The residue was purified by preparative high performance liquid chromatography (column: Waters Xbridge BEH C18 100*25mm*5μm; mobile phase: [A: water (0.225% formic acid), B: acetonitrile]; B%: 27%-47%, 14 min) to give the title compound (1.5 mg).

MS m/z(ESI): 509.0[M+H] ⁺

¹ H NMR (400MHz, Chloroform-d)δ8.01(d,J=1.6Hz,1H),7.93(d,J=8.4Hz,1H),7.85(s,1H),7.79-7.77(m,1H) ), 7.42(s, 1H), 7.40(s, 1H), 7.28(s, 2H), 6.58(t, J=73.6Hz, 1H), 4.41–4.36(m, 2H), 3.35–3.32(m, 2H), 3.26–3.23 (m, 2H), 1.39 (t, J=7.1Hz, 3H).

Example 30, 4-(4-(difluoromethoxy)phenyl)-6-(1-(methyl-d ₃ )-1H-benzo[d]imidazol-6-yl)-2-propane Oxythiazolo[4,5-b]pyridin-5(4H)-one (Compound 30)

Step 1: 6-Bromo-4-(4-(difluoromethoxy)phenyl)-2-propoxythiazolo[4,5-b]pyridin-5(4H)-one (Intermediate 30- 1) Synthesis

Sodium hydride (133 mg) was dissolved in anhydrous tetrahydrofuran (2 mL), n-propanol (198 mg) was added to it at 0°C, intermediate 23-3 (150 mg) was added after 30 min, and the reaction solution was stirred at 25°C 4 hours. After the reaction was completed, the pH was adjusted to 7 with dilute hydrochloric acid (2N), the solvent was removed by concentration under reduced pressure, water (10 mL) was added, ethyl acetate (10 mL*2) was added for extraction, and the organic phase was dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by column chromatography (petroleum ether:ethyl acetate=2:1) to obtain the title compound (71 mg).

MS m/z(ESI): 431.8[M+H] ⁺

Step 2: 4-(4-(Difluoromethoxy)phenyl)-6-(1-(methyl- _d3 )-1H-benzo[d]imidazol-6-yl)-2-propoxy Synthesis of Thiazolo[4,5-b]pyridin-5(4H)-one (Compound 30)

Intermediate 30-1 (68 mg) and Intermediate 23-1 (62 mg) were dissolved in dioxane (1.2 mL) and water (0.3 mL), to which was added cesium carbonate (103 mg) and Pd(dtbpf)Cl ₂ (15mg), the reaction solution was stirred at 90°C for 10h under nitrogen protection. After the completion of the reaction, after the reaction was cooled to room temperature, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and purified by preparative high performance liquid chromatography [YMC-Actus Triart C18 column 5 μm silica, 30 mm in diameter, 150 mm in length; water (containing A mixture of decreasing polarity of 0.225% formic acid) and acetonitrile as eluent; acetonitrile gradient ratio 40%-70%, elution time 14 minutes] to give the title compound (17 mg).

MS m/z (ESI): 486.1 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ )δ8.32-8.27(m,1H),8.19-8.18(m,1H),7.96(s,1H),7.70-7.36(m,5H),7.36-7.31 (m, 2H), 4.25–4.22 (m, 2H), 1.77–1.68 (m, 2H), 0.91 (t, J=7.4Hz, 3H).

Example 31, 4-(4-(difluoromethoxy)phenyl)-2-ethoxy-6-(2-(methyl- _d3 )-2H-indazol-5-yl)thiazolo [4,5-b]pyridin-5(4H)-one (Compound 31)

Under nitrogen atmosphere, intermediate 6-1 (100 mg) was dissolved in dioxane solution (1 mL), intermediate 31-1 (55 mg), cesium carbonate (140 mg), water (0.2 mL) and Pd ( dtbpf)Cl2 ( ₁₄ mg). The reaction solution was heated to 100 °C and stirred for 2 h. Subsequently, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by high pressure preparative liquid chromatography [YMC-Actus Triart C18 column 5 μm silica, 30 mm diameter, 150 mm length; water (containing 0.05% NH ₄ HCO _{3 )} ) and acetonitrile with decreasing polarity mixture as eluent; acetonitrile gradient ratio 55%-80%, elution time 13 minutes] to obtain the title compound (15 mg).

MS m/z(ESI): 472.1[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ8.35(s,1H), 8.28(s,1H), 8.05(s,1H), 7.59-7.19(m,7H), 4.35-4.29(m,2H) ),1.32(t,J=7.0Hz,3H).

Example 32, 4-(4-(difluoromethoxy)phenyl)-2-ethoxy-6-(2-(ethoxymethyl)-1-methyl-1H-benzo[d ]imidazol-6-yl)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 32)

Step 1: Synthesis of 6-bromo-2-(ethoxymethyl)-1-methyl-1H-benzo[d]imidazole (Intermediate 32-2)

Intermediate 32-1 (200 mg) was dissolved in THF (2 mL), NaH (66 mg, 60% wt) was added under ice-water bath, and the mixture was stirred at room temperature for 0.5 hour. Under an ice-water bath, iodoethane (388 mg) was added, followed by stirring at room temperature for 12 h. TLC detection showed that the reaction was over, the reaction solution was poured into saturated aqueous ammonium chloride solution (5 mL), extracted with ethyl acetate (5 mL*3), the organic phases were combined, washed with saturated brine (10 mL), and the organic phase was washed with anhydrous Dry over sodium sulfate and spin to remove the solvent under reduced pressure to give the title compound (140 mg).

MS m/z(ESI): 269.0[M+H] ⁺

Step 2: 4-(4-(Difluoromethoxy)phenyl)-2-ethoxy-6-(2-(ethoxymethyl)-1-methyl-1H-benzo[d] Synthesis of Imidazol-6-yl)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 32)

Under nitrogen atmosphere, intermediate 32-2 (75 mg) was dissolved in dioxane (1 mL), intermediate 6-1 (129 mg), cesium carbonate (182 mg), water (0.2 mL) and Pd (dtbpf) were added Cl2 ( ₁₈ mg). The reaction solution was heated to 100 °C and stirred for 2 h. Subsequently, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by high pressure preparative liquid chromatography [YMC-Actus Triart C18 column 5 μm silica, 30 mm diameter, 150 mm length; water (containing 0.05% NH ₄ HCO _{3 )} ) and acetonitrile with decreasing polarity mixture as eluent; acetonitrile gradient ratio 55%-80%, elution time 13 minutes] to obtain the title compound (25 mg).

MS m/z(ESI): 527.1[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.34(s, 1H), 7.95(s, 1H), 7.63-7.20(m, 7H), 4.75(s, 2H), 4.33(t, J=7.0 Hz, 2H), 3.82(s, 3H), 3.53(t, J＝7.0Hz, 2H), 1.33(t, J＝7.0Hz, 3H), 1.16(t, J＝7.0Hz, 3H).

Example 33, 6-(2-(aminomethyl)-1-methyl-1H-benzo[d]imidazol-6-yl)-4-(4-(difluoromethoxy)phenyl)- 2-Ethoxythiazolo[4,5-b]pyridin-5(4H)-one (Compound 33)

Step 1: Synthesis of 2-(azidomethyl)-6-bromo-1-methyl-1H-benzo[d]imidazole (Intermediate 33-1)

Intermediate 32-1 (500 mg) was dissolved in THF (5 mL), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) (474 mg), diphenylphosphonium azide were added ester (DPPA) (685 mg), stirred at room temperature for 12 h. Subsequently, the reaction solution was poured into water (5 mL), extracted with ethyl acetate (5 mL*3), the organic phases were combined, washed with saturated brine (10 mL), the organic phase was dried over anhydrous sodium sulfate, and removed under reduced pressure solvent, and the residue was purified by column chromatography (PE/EA=5/1-2/1) to give the title compound (450 mg).

MS m/z(ESI): 266.0[M+H] ⁺

Step 2: Synthesis of (6-bromo-1-methyl-1H-benzo[d]imidazol-2-yl)methanamine (Intermediate 33-2)

Intermediate 33-1 (500 mg) was dissolved in THF (5 mL), water (1 mL), triphenylphosphine (591 mg) were added. After the addition was completed, the mixture was stirred at room temperature for 12 h. Subsequently, the reaction solution was poured into water (5 mL), extracted with ethyl acetate (5 mL*3), the organic phases were combined, washed with saturated brine (10 mL), the organic phase was dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. The concentrated residue was purified by column chromatography (DCM/MeOH=20/1) to obtain the title compound (160 mg).

MS m/z(ESI): 240.0[M+H] ⁺

Step 3: 6-(2-(Aminomethyl)-1-methyl-1H-benzo[d]imidazol-6-yl)-4-(4-(difluoromethoxy)phenyl)-2 - Synthesis of Ethoxythiazolo[4,5-b]pyridin-5(4H)-one (Compound 33)

Under nitrogen atmosphere, intermediate 6-1 (100 mg) was dissolved in dioxane solution (1 mL), intermediate 33-2 (52 mg), cesium carbonate (140 mg), water (0.2 mL) and Pd ( dtbpf)Cl ₂ (14 mg),. The reaction solution was heated to 100 °C and stirred for 2 h. LCMS monitoring showed that the reaction was over, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by high pressure preparative liquid chromatography [YMC-Actus Triart C18 column 5 μm silica, 30 mm diameter, 150 mm length; water (containing 0.05% A mixture of decreasing polarity of _NH4HCO3 ₎ and acetonitrile as eluent; acetonitrile gradient ratio 55%-80%, elution time 13 minutes] to give the title compound (13 mg).

MS m/z(ESI): 498.1[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ8.31(s,1H), 7.89(s,1H), 7.57-7.55(m,1H), 7.50-7.19(m,6H), 4.32(d,J =7.1Hz,2H),3.98(s,2H),3.77(s,3H),1.32(t,J=7.0Hz,3H).

Example 34, 2-ethoxy-4-(4-ethynylphenyl)-6-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)thiazolo[4 ,5-b]pyridin-5(4H)-one (Compound 34)

Step 1: Synthesis of 6-bromo-2-ethoxy-4-(4-iodophenyl)thiazolo[4,5-b]pyridin-5(4H)-one (Intermediate 34-2)

Intermediate 1-7 (100 mg) and Intermediate 34-1 (135 mg) were dissolved in anhydrous 1,2-dichloroethane (5 mL), and pyridine (86 mg) and copper acetate (79 mg) were added dropwise thereto, After the addition was completed, the mixture was stirred at 40°C for 4 hours. After the reaction was completed, the solvent was removed by concentration under reduced pressure. The residue was purified by preparative high performance liquid chromatography (column: Waters Xbridge BEH C18 100*25mm*5μm; mobile phase: [A: water (0.225% formic acid), B: acetonitrile]; B%: 23%-50%, 12 min) to give the title compound (170 mg).

MS m/z(ESI): 477.0[M+H] ⁺

Step 2: 6-Bromo-2-ethoxy-4-(4-((trimethylsilyl)ethynyl)phenyl)thiazolo[4,5-b]pyridin-5(4H)-one ( Synthesis of intermediate 34-3)

Intermediate 34-2 (100 mg) and 1-(trimethylsilyl)acetylene (25 mg) were dissolved in anhydrous tetrahydrofuran solution (5 mL), cuprous iodide (12 mg), triethylamine (21 mg) and Tetrakistriphenylphosphine palladium (12.10 mg). The reaction solution was stirred at 25°C for 12 hours under nitrogen protection. After the reaction was cooled to room temperature, water (10 mL) and ethyl acetate (10 mL) were added successively, the organic phase was washed with water (50 mL*2), and the washed organic phase was dried with an appropriate amount of anhydrous sodium sulfate. After filtration, the organic phase was concentrated under reduced pressure to remove the solvent, and the residue was purified by column chromatography (dichloromethane:methanol=20:1) to give the title compound (70 mg).

MS m/z(ESI): 447.0[M+H] ⁺

Step 3: 2-Ethoxy-6-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-4-(4-((trimethylsilyl)ethynyl) )Phenyl)thiazolo[4,5-b]pyridin-5(4H)-one (Intermediate 34-4) Synthesis

Intermediate 34-3 (70 mg) was dissolved in 1,4-dioxane (5 mL) and water (0.5 mL), to which was added intermediate 3-1 (73.56 mg), Pd(dppf)Cl ₂ ( 11 mg) and potassium phosphate (99 mg), and the reaction solution was stirred at 100° C. for 12 hours under nitrogen protection. After the reaction was cooled to room temperature, water (10 mL) and ethyl acetate (10 mL) were added successively, the organic phase was washed with water (5 mL*2), and the washed organic phase was dried with an appropriate amount of anhydrous sodium sulfate. After filtration, the organic phase was concentrated under reduced pressure to remove the solvent, and the residue was purified by preparative high performance liquid chromatography (column: Waters Xbridge BEH C18 100*25mm*5μm; mobile phase: [A: water (0.225% formic acid), B: acetonitrile) ]; B%: 23%-50%, 12 minutes) to obtain the title compound (40 mg).

MS m/z(ESI): 476.0[M+H] ⁺

Step 4: 2-Ethoxy-4-(4-ethynylphenyl)-6-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)thiazolo[4, Synthesis of 5-b]pyridin-5(4H)-one (Compound 34)

Intermediate 34-4 (40 mg) was dissolved in anhydrous ethanol solution (2 mL), potassium carbonate (35 mg) was added thereto, and the reaction solution was stirred at 25° C. for 2 hours. After the reaction was completed, after filtration, the organic phase was concentrated under reduced pressure to remove the solvent, and the residue was purified by preparative high performance liquid chromatography (column: Waters Xbridge BEH C18 100*25mm*5 μm; mobile phase: [A: water (0.225% formic acid) ), B: acetonitrile]; B%: 23%-50%, 12 minutes) to give the title compound (15 mg).

MS m/z(ESI): 404.0[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.30 (d, J=2.6Hz, 1H), 8.25 (s, 1H), 7.78 (dd, J=9.5, 2.7Hz, 1H), 7.66-7.62 ( m, 2H), 7.45–7.35 (m, 2H), 6.44 (d, J=9.5Hz, 1H), 4.31 (d, J=6.8Hz, 3H), 3.46 (s, 3H), 1.30 (t, J =7.0Hz,3H).

Example 35, 2-ethoxy-6-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-4-(4-(prop-1-yn-1- yl)phenyl)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 35)

Step 1: 6-Bromo-2-ethoxy-4-(4-(prop-1-yn-1-yl)phenyl)thiazolo[4,5-b]pyridin-5(4H)-one ( Synthesis of Intermediate 35-1)

Intermediate 34-2 (100mg) and 1-(trimethylsilyl)propyne (25mg) were dissolved in anhydrous tetrahydrofuran solution (5mL), cuprous iodide (12mg), triethylamine (21mg) were added , tetrabutylammonium fluoride (58mg) and tetrakistriphenylphosphine palladium (12.10mg), the reaction solution was stirred at 25°C for 12 hours under nitrogen protection. After the reaction was cooled to room temperature, water (10 mL) and ethyl acetate (10 mL) were added successively, the organic phase was washed with water (50 mL*2), and the washed organic phase was dried with an appropriate amount of anhydrous sodium sulfate. After filtration, the organic phase was concentrated under reduced pressure to remove the solvent, and the residue was purified by column chromatography (dichloromethane:methanol=20:1) to obtain the title compound (60 mg).

MS m/z(ESI): 389.0[M+H] ⁺

Step 2: 2-Ethoxy-6-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-4-(4-(prop-1-yn-1-yl) )Phenyl)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 35) Synthesis

Intermediate 35-1 (60 mg) was dissolved in 1,4-dioxane (5 mL) and water (0.5 mL), to which was added Intermediate 3-1 (73 mg), Pd(dppf)Cl ₂ (11 mg) ) and potassium phosphate (99 mg). The reaction solution was stirred at 100°C for 12 hours under nitrogen protection. After the reaction was cooled to room temperature, water (10 mL) and ethyl acetate (10 mL) were added successively, the organic phase was washed with water (5 mL*2), and the washed organic phase was dried with an appropriate amount of anhydrous sodium sulfate. After filtration, the organic phase was concentrated under reduced pressure to remove the solvent, and the residue was purified by preparative high performance liquid chromatography (column: Waters Xbridge BEH C18 100*25mm*5μm; mobile phase: [A: water (0.225% formic acid), B: acetonitrile) ]; B%: 23%-50%, 12 minutes) to obtain the title compound (40 mg).

MS m/z(ESI): 418.0[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.31 (d, J=2.6Hz, 1H), 8.26 (s, 1H), 7.79 (dd, J=9.5, 2.7Hz, 1H), 7.58-7.51 ( m, 2H), 7.41-7.32(m, 2H), 6.45(d, J=9.6Hz, 1H), 4.32(q, J=7.0Hz, 2H), 3.47(s, 3H), 2.10(s, 3H ),1.31(t,J=7.0Hz,3H).

Example 36, 4-(4-(difluoromethoxy)phenyl)-2-ethoxy-6-(1,2,3,4-tetrahydrobenzo[4,5]imidazo[1 ,2-a]pyridin-8-yl)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 36)

Intermediate 6-1 (80 mg) and Intermediate 36-1 (65 mg) were dissolved in 1,4-dioxane (1.2 mL) and water (0.3 mL), to which was added Pd(dtbpf)Cl ₂ ( 21 mg), Cs ₂ CO ₃ (143 mg), the reaction solution was stirred at 100° C. for 12 h under nitrogen protection. After completion of the reaction, after the reaction was cooled to room temperature, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by column chromatography (dichloromethane:methanol=20:1) to obtain a crude product, which was then prepared by high-performance liquid phase Chromatographic purification [YMC-Actus Triart C18 column 5μm silica, 30mm diameter, 150mm length; mixture of water (containing 0.225% formic acid) and acetonitrile with decreasing polarity as eluent; acetonitrile gradient ratio 40%-70%, wash Detox time 15 min] to give the title compound (25 mg).

MS m/z (ESI): 509.1 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ )δ8.21(s,1H),7.77(s,1H),7.51-7.08(m,7H),4.29-4.20(m,2H),4.00(t,J = 5.8Hz, 2H), 2.89 (t, J = 6.1Hz, 2H), 2.02–1.94 (m, 2H), 1.89–1.85 (m, 2H), 1.25 (t, J = 7.0Hz, 3H).

Example 37, 4-(4-(difluoromethoxy)phenyl)-2-ethoxy-6-(3-methyl-4-oxo-3,4-dihydroquinazoline-6 -yl)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 37)

Step 1: Synthesis of 6-bromo-3-methylquinazolin-4(3H)-one (Intermediate 37-2)

Intermediate 37-1 (600 mg) was dissolved in acetonitrile (6 mL), potassium carbonate (1.1 g) and methyl iodide (3.8 g) were added thereto, and the reaction solution was stirred at 30° C. for 10 h. After completion of the reaction, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by column chromatography (petroleum ether:ethyl acetate=3:1) to obtain the title compound (305 mg).

MS m/z(ESI): 238.9[M+H] ⁺

Step 2: 3-Methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)quinazolin-4(3H)-one Synthesis of (Intermediate 37-3)

Intermediate 37-2 (100 mg) and bispinacol boronate (170 mg) were dissolved in anhydrous dioxane (2 mL), to which were added potassium acetate (82 mg) and Pd(dppf)Cl ₂ ( 46 mg), the reaction solution was stirred at 100 °C for 4 h under nitrogen protection. After the completion of the reaction, after the reaction was cooled to room temperature, the solvent was concentrated under reduced pressure to remove the solvent, water (8 mL) and ethyl acetate (10 mL*2) were added in turn, the organic phase was washed with water (10 mL*2), and the washed organic phase was washed with an appropriate amount of Dry over aqueous sodium sulfate, filter, and concentrate under reduced pressure. The residue was purified by column chromatography (silica, petroleum ether:ethyl acetate=4:1) to give the title compound (105 mg).

MS m/z(ESI): 287.1[M+H] ⁺

Step 3: 4-(4-(Difluoromethoxy)phenyl)-2-ethoxy-6-(3-methyl-4-oxo-3,4-dihydroquinazoline-6- Synthesis of yl)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 37)

Intermediate 37-3 (98 mg) and Intermediate 1-8 (110 mg) were dissolved in dioxane (1.2 mL) and water (0.3 mL), to which was added cesium carbonate (172 mg) and Pd(dtbpf)Cl ₂ (26 mg), the reaction solution was stirred at 80 °C for 10 h under nitrogen protection. After the completion of the reaction, after the reaction was cooled to room temperature, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and purified by preparative high performance liquid chromatography [YMC-Actus Triart C18 column 5 μm silica, 30 mm in diameter, 150 mm in length; water (containing 0.225% formic acid) and acetonitrile of decreasing polarity mixture as eluent; acetonitrile gradient ratio 40%-70%, elution time 15 minutes] to give the title compound (21 mg).

MS m/z (ESI): 497.0 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.55(s, 1H), 8.44(s, 1H), 8.37(s, 1H), 8.15(d, J=8.6Hz, 1H), 7.68(d, J=8.6Hz, 1H), 7.58–7.15 (m, 5H), 4.36–4.30 (m, 2H), 3.51 (s, 3H), 1.32 (t, J=7.0Hz, 3H).

Example 38, 4-(4-(difluoromethoxy)phenyl)-2-ethoxy-6-(4-oxo-4H-pyrido[1,2-a]pyrimidin-7-yl )thiazolo[4,5-b]pyridin-5(4H)-one (Compound 38)

Intermediate 15-1 (79 mg) and Intermediate 38-1 (70 mg) were dissolved in dioxane (1.2 mL) and water (0.3 mL), to which was added cesium carbonate (136 mg) and Pd(dtbpf)Cl ₂ (25mg), the reaction solution was stirred at 90°C for 7h under nitrogen protection. After the reaction was completed, after the reaction was cooled to room temperature, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by preparative high performance liquid chromatography [YMC-Actus Triart C18 column 5 μm silica, 30 mm diameter, 150 mm length; water A mixture of decreasing polarity (containing 0.225% formic acid) and acetonitrile as eluent; acetonitrile gradient ratio 40%-70%, elution time 18 min] gave the title compound (28 mg).

MS m/z (ESI): 483.0 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ )δ9.66(d,J=1.8Hz,1H),8.65(s,1H),8.40-8.26(m,2H),7.75(d,J=9.3Hz, 1H), 7.60–7.11 (m, 5H), 6.40 (d, J=6.3Hz, 1H), 4.38–4.32 (m, 2H), 1.32 (t, J=7.0Hz, 3H).

Example 39, 4-(4-(difluoromethoxy)phenyl)-2-ethoxy-6-(4-methyl-3-oxy-3,4-dihydroquinoxaline-6 -yl)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 39)

Step 1: Synthesis of 7-bromo-1-methylquinoxalin-2(1H)-one (Intermediate 39-2)

Intermediate 39-1 (400 mg) was dissolved in acetonitrile (4 mL), potassium carbonate (736 mg) and methyl iodide (2.5 g) were added thereto, and the reaction solution was stirred at 30° C. for 10 h. After completion of the reaction, the reaction solution was filtered, the filtrate was concentrated under reduced pressure to remove the solvent, and the residue was purified by column chromatography (petroleum ether:ethyl acetate=2:1) to obtain the title compound (160 mg).

MS m/z(ESI): 238.9[M+H] ⁺

Step 2: 1-Methyl-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)quinoxalin-2(1H)-one Synthesis of (Intermediate 39-3)

Intermediate 39-2 (100 mg) and bispinacol boronate (170 mg) were dissolved in anhydrous dioxane (2 mL), to which were added potassium acetate (82 mg) and Pd(dppf)Cl ₂ ( 46 mg), the reaction solution was stirred at 100 °C for 4 h under nitrogen protection. After the completion of the reaction, after the reaction was cooled to room temperature, the solvent was concentrated under reduced pressure to remove the solvent, water (8 mL) and ethyl acetate (10 mL*2) were added in turn, the organic phase was washed with water (10 mL*2), and the washed organic phase was washed with an appropriate amount of Dry over aqueous sodium sulfate, filter, and concentrate under reduced pressure. The residue was purified by column chromatography (silica, petroleum ether:ethyl acetate=4:1) to give the title compound (98 mg).

MS m/z(ESI): 287.1[M+H] ⁺

Step 3: 4-(4-(Difluoromethoxy)phenyl)-2-ethoxy-6-(4-methyl-3-oxo-3,4-dihydroquinoxaline-6- Synthesis of yl)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 39)

Intermediate 39-3 (98 mg) and Intermediate 1-8 (110 mg) were dissolved in dioxane (1.2 mL) and water (0.3 mL), to which was added cesium carbonate (172 mg) and Pd(dtbpf)Cl ₂ (26 mg), the reaction solution was stirred at 80 °C for 10 h under nitrogen protection. After the completion of the reaction, after the reaction was cooled to room temperature, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and purified by preparative high performance liquid chromatography [YMC-Actus Triart C18 column 5 μm silica, 30 mm in diameter, 150 mm in length; water (containing A mixture of decreasing polarity of 0.225% formic acid) and acetonitrile as eluent; acetonitrile gradient ratio 40%-70%, elution time 15 minutes] to give the title compound (23 mg).

MS m/z (ESI): 497.0 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ )δ8.53(s,1H),8.21(s,1H),7.98(d,J=1.4Hz,1H),7.84-7.82(m,1H),7.77- 7.75 (m, 1H), 7.57–7.18 (m, 5H), 4.37–4.32 (m, 2H), 3.62 (s, 3H), 1.32 (t, J=7.0Hz, 3H).

Example 40, 4-(4-(difluoromethoxy)-3-fluorophenyl)-2-ethoxy-6-(1-methyl-6-oxo-1,6-dihydropyridine Synthesis of -3-yl)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 40)

Step 1: Synthesis of 6-bromo-2-ethoxy-5-oxothiazolo[4,5-b]pyridine-4(5H)-carboxylate tert-butyl ester (Intermediate 40-1)

Intermediate 1-7 (2 g) was dissolved in anhydrous tetrahydrofuran (20 mL), tert-butyl carbonate (2.38 g), triethylamine (2.21 g) and 4-dimethylaminopyridine (266.43 mg) were added, and the addition was completed. , and stirred at 25°C for 12 hours. After the reaction, the organic phase was concentrated under reduced pressure to remove the solvent. The residue was purified by flash (

20g

Silica Flash chromatography column, gradient 0-5% methanol/dichloromethane, 80 mL/min), the collected fractions were concentrated to give the title compound (1.5 g).

MS m/z(ESI): 375.0[M+H] ⁺

Step 2: 2-Ethoxy-6-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-5-oxothiazolo[4,5-b]pyridine-4 Synthesis of (5H)-tert-Butyl Carboxylic Acid (Intermediate 40-2)

Intermediate 40-1 (1.5 g) was dissolved in anhydrous 1,4-dioxane solution (20 mL) and water (5 mL), intermediate 3-1 (1.13 g) was added, Pd(dppf)Cl ₂ (260 mg) and potassium phosphate (2.54 g, 11.99 mmol), the reaction solution was stirred at 100° C. for 12 hours under nitrogen protection. After the reaction was completed, the reaction was cooled to room temperature, water (100 mL) and ethyl acetate (100 mL) were added in sequence, the organic phase was washed with water (50 mL*2), and the washed organic phase was dried with an appropriate amount of anhydrous sodium sulfate. After filtration, the organic phase was concentrated under reduced pressure to remove the solvent, and the residue was purified by column chromatography (petroleum ether:ethyl acetate=1:1) to give the title compound (1 g).

MS m/z(ESI): 404.0[M+H] ⁺

Step 3: 2-Ethoxy-6-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)thiazolo[4,5-b]pyridine-5(4H)- Synthesis of Ketones (Intermediate 40-3)

Intermediate 40-2 (1 g) was dissolved in anhydrous dichloromethane (10 mL), hydrochloric acid/dioxane (6 mL, 4N) was added dropwise, the addition was completed, and the mixture was stirred at 25° C. for 4 hours. After the reaction, the reaction solution was concentrated under reduced pressure to remove the solvent, then saturated sodium bicarbonate (50mL) and ethyl acetate (50mL*3) were added in turn, the organic phase was washed with water (20mL*2), and the washed organic phase was washed with an appropriate amount of Dry over anhydrous sodium sulfate, filter, and concentrate to give the title compound (450 mg).

MS m/z(ESI): 304.0[M+H] ⁺

Step 4: Synthesis of (4-(difluoromethoxy)-3-fluorophenyl)boronic acid (Intermediate 40-4)

4-Bromo-1-(difluoromethoxy)-2-fluorobenzene (300 mg) and triisopropyl borate (375 mg) were dissolved in anhydrous tetrahydrofuran solution (3 mL), to which was added n- A solution of BuLi (1.6M, 0.895mL) in tetrahydrofuran, the reaction solution was stirred at 0°C for 4h under nitrogen protection. Subsequently, the reaction solution was concentrated to dryness under reduced pressure, and the residue was purified by column chromatography (PE/EA=10/1-3/1) to obtain the title compound (120 mg).

Step 5: 4-(4-(Difluoromethoxy)-3-fluorophenyl)-2-ethoxy-6-(1-methyl-6-oxo-1,6-dihydropyridine- Synthesis of 3-yl)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 40)

Intermediate 40-4 (36 mg) and intermediate 40-3 (35 mg) were dissolved in 1,2-dichloroethane (1 mL), and anhydrous copper acetate (25 mg) and pyridine (27 mg) were added thereto to react The liquid was stirred at 50°C for 12h under nitrogen protection. Subsequently, after the reaction was cooled to room temperature, the reaction solution was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by high pressure preparation [YMC-Actus Triart C18 column 5 μm silica, 30 mm diameter, 150 mm length; water (containing 0.05% NH) A mixture of decreasing polarity of ₄ HCO ₃ ) and acetonitrile as eluent; acetonitrile gradient ratio 55%-75%, elution time 12 minutes] to give the title compound (12 mg).

MS m/z (ESI): 464.0 [M+H] ⁺ .

¹ H NMR(400MHz, DMSO-d ₆ )δ8.30(d,J=2.7Hz,1H),8.27(s,1H),7.80-7.76(m,1H),7.65-7.61(m,1H), 7.56–7.20 (m, 3H), 6.44 (d, J=9.5Hz, 1H), 4.36–4.31 (m, 2H), 3.46 (s, 3H), 1.32 (t, J=7.0Hz, 3H).

Example 41, 4-(6-(difluoromethoxy)pyridin-3-yl)-2-ethoxy-6-(1-methyl-6-oxo-1,6-dihydropyridine- 3-yl)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 41)

Step 1: Synthesis of 2-(difluoromethoxy)-5-iodopyridine (Intermediate 41-2)

Intermediate 41-1 (1.11 g) and sodium difluorochloroacetate (1.54 g) were dissolved in anhydrous N,N-dimethylformamide (20 mL), to which was added cesium carbonate (4.92 g), the addition was complete Stir at 100°C for 12 hours. After the reaction, saturated ammonium chloride (100mL) and ethyl acetate (100mL*3) were added in sequence, the organic phase was washed with water (50mL*2), and the washed organic phase was dried with an appropriate amount of anhydrous sodium sulfate. After filtration, the organic phase The solvent was removed by concentration under reduced pressure. The residue was purified by column chromatography (dichloromethane:methanol=20:1) to obtain the title compound (600 mg).

MS m/z(ESI): 272.0[M+H] ⁺

Step 2: Synthesis of (6-(difluoromethoxy)pyridin-3-yl)boronic acid (Intermediate 41-3)

Intermediate 41-2 (550 mg) and trimethyl borate (314 mg) were dissolved in anhydrous tetrahydrofuran (10 mL), and n-butyllithium (2.5 mL, 1.6 M) was added dropwise to it at -78°C. Stir at -78°C for 4 hours. After the reaction, hydrochloric acid (3 mL, 1N) was added successively, and the tetrahydrofuran was removed by concentration under reduced pressure. The residue was purified by preparative high performance liquid chromatography (column: Waters Xbridge BEH C18 100*25mm*5μm; mobile phase: [A: water (0.225% formic acid), B: acetonitrile]; B%: 23%-50%, 12 min) to give the title compound (80.0 mg).

MS m/z(ESI): 189.0[M+H] ⁺

Step 3: 6-Bromo-4-(6-(difluoromethoxy)pyridin-3-yl)-2-ethoxythiazolo[4,5-b]pyridin-5(4H)-one (middle Synthesis of body 41-4)

Intermediate 41-3 (53 mg) and intermediate 1-7 (51 mg) were dissolved in anhydrous 1,2-dichloroethane (10 mL), pyridine (66 mg) and copper acetate (51 mg) were added, and 40 was added. Stir in air for 4 hours. After the reaction was completed, the solvent was removed by concentration under reduced pressure. The residue was purified by preparative high performance liquid chromatography (column: Waters Xbridge BEH C18 100*25mm*5μm; mobile phase: [A: water (0.225% formic acid), B: acetonitrile]; B%: 23%-50%, 12 minutes) to give the title compound (40 mg).

MS m/z(ESI): 418.1[M+H] ⁺

Step 4: 4-(6-(Difluoromethoxy)pyridin-3-yl)-2-ethoxy-6-(1-methyl-6-oxo-1,6-dihydropyridine-3 Synthesis of -yl)thiazolo[4,5-b]pyridin-5(4H)-one (Compound 41)

Intermediate 41-4 (40 mg) was dissolved in 1,4-dioxane solution (2 mL) and water (0.5 mL), to which was added intermediate 3-1 (45 mg), Pd(dppf)Cl ₂ ( 7 mg) and cesium carbonate (93 mg), and the reaction solution was stirred at 100° C. for 12 hours under nitrogen protection. After the reaction was cooled to room temperature, water (10 mL) and ethyl acetate (10 mL) were added successively, the organic phase was washed with water (50 mL*2), and the washed organic phase was dried with an appropriate amount of anhydrous sodium sulfate. After filtration, the organic phase was concentrated under reduced pressure to remove the solvent, and the residue was purified by preparative high performance liquid chromatography (column: Waters Xbridge BEH C18 100*25mm*5μm; mobile phase: [A: water (0.225% formic acid), B: acetonitrile) ]; B%: 23%-50%, 12 minutes) to obtain the title compound (10 mg).

MS m/z(ESI): 447.0[M+H] ⁺

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.37 (d, J=2.6 Hz, 1H), 8.29 (d, J=3.7 Hz, 2H), 8.06 (dd, J=8.7, 2.6 Hz, 1H) ,7.99–7.63(m,2H),7.30(d,J=8.7Hz,1H),6.46(d,J=9.5Hz,1H),4.37–4.32(m,2H),3.48(s,3H), 1.33(t,J=7.0Hz,3H).

Example 42, 4-(4-(difluoromethoxy)-3-methylphenyl)-2-ethoxy-6-(1-methyl-6-oxo-1,6-dihydro Pyridin-3-yl)thiazolo[4,5-b]pyridin-5-(4H)-one (Compound 42)

Step 1: Synthesis of (4-(difluoromethoxy)-3-methylphenyl)boronic anhydride (Intermediate 42-2)

Intermediate 42-1 (500 mg) and triisopropyl borate (800 mg) were dissolved in tetrahydrofuran (5 mL), n-butyllithium (4.26 mL) was added dropwise to it at -78 °C, and the mixture was stirred at 0 °C for 2 h. Subsequently, H ₂ O (20 mL) and ethyl acetate (20 mL*3) were sequentially added to the reaction solution, the organic phase was washed with saturated brine (30 mL*2), and the washed organic phase was dried with an appropriate amount of anhydrous sodium sulfate, Concentrate to give the title compound (350 mg).

MS m/z (ESI): 247.2 [M+H] ⁺ .

Step 2: 4-(4-(Difluoromethoxy)-3-methylphenyl)-2-ethoxy-6-(1-methyl-6-oxo-1,6-dihydropyridine Synthesis of -3-yl)thiazolo[4,5-b]pyridin-5-(4H)-one (Compound 42)

Intermediate 42-2 (48 mg) and Intermediate 40-2 (25 mg) were dissolved in dichloromethane (2 mL), pyridine (10 mg) and copper acetate (4 mg) were added thereto, and the reaction solution was placed under nitrogen protection at 45° C. Stir for 16 hours. After the reaction was cooled to room temperature, water (30 mL) and dichloromethane (40 mL) were added successively, the organic phase was washed with saturated brine (30 mL*2), and the washed organic phase was dried with an appropriate amount of anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by preparative high performance liquid chromatography (chromatographic column: Gemini NX C18 5μm*10*150mm; mobile phase: A: 0.05% TFA v/v, B: acetonitrile; B%: 40%-45%, 12min) to obtain The title compound (26 mg).

MS m/z (ESI): 460.1 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ )δ8.31(d, J=2.6Hz, 1H), 8.26(s, 1H), 7.84-7.75(m, 1H), 7.54-7.08(m, 4H), 6.45 (d, J=9.5Hz, 1H), 4.38–4.28 (m, 2H), 3.48 (s, 3H), 2.29 (s, 3H), 1.37–1.28 (m, 3H).

Example 43, 2-(difluoromethoxy)-5-(2-ethoxy-6-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-5 -Oxothiazolo[4,5-b]pyridin-4(5H)-yl)benzonitrile (Compound 43)

Step 1: Synthesis of 5-bromo-2-(difluoromethoxy)benzonitrile (Intermediate 43-2)

Intermediate 43-1 (1 g) and sodium difluorochloroacetate (1.54 g) were dissolved in anhydrous N,N-dimethylformamide (20 mL), to which was added cesium carbonate (4.92 g), after adding 100 °C was stirred for 12 hours. After the reaction, saturated ammonium chloride (100mL) and ethyl acetate (100mL*3) were added in sequence, the organic phase was washed with water (50mL*2), and the washed organic phase was dried with an appropriate amount of anhydrous sodium sulfate. After filtration, the organic phase The solvent was removed by concentration under reduced pressure. The residue was purified by column chromatography (

20g

Silica Flash column, gradient 0-5% methanol/dichloromethane, 80 mL/min) to give the title compound (500 mg).

MS m/z(ESI): 248.0[M+H] ⁺

Step 2: Synthesis of (3-cyano-4-(difluoromethoxy)phenyl)boronic acid (Intermediate 43-3)

Intermediate 43-2 (500 mg) and trimethyl borate (314 mg) were dissolved in anhydrous tetrahydrofuran (10 mL), n-butyllithium (2.5 mL, 1.6 M) was added dropwise to it at -78 °C, and the addition of - Stir at 78°C for 4 hours. After the reaction, hydrochloric acid (3mL, 1N) was added successively, the tetrahydrofuran was removed by concentration under reduced pressure, and the residue was purified by high performance liquid chromatography (column: Waters Xbridge BEH C18 100*25mm*5 μm; mobile phase: [A: water (0.225 μm) % formic acid), B: acetonitrile]; B%: 23%-50%, 12 minutes) to give the title compound (80.0 mg).

Step 3: 2-(Difluoromethoxy)-5-(2-ethoxy-6-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-5- Synthesis of oxothiazolo[4,5-b]pyridin-4(5H)-yl)benzonitrile (Compound 43)

Intermediate 43-3 (60 mg) and intermediate 40-2 (51.28 mg) were dissolved in anhydrous 1,2-dichloroethane (10 mL), pyridine (66.86 mg) and copper acetate (51 mg) were added, After stirring at 40°C in air for 4 hours. After the reaction was completed, the solvent was removed by concentration under reduced pressure. The residue was purified by preparative high performance liquid chromatography (column: Waters Xbridge BEH C18 100*25mm*5μm; mobile phase: [A: water (0.225% formic acid), B: acetonitrile]; B%: 23%-50%, 12 min) to give the title compound (10.4 mg).

MS m/z(ESI): 471.0[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.34-8.27(m, 2H), 8.17(d, J=2.5Hz, 1H), 7.93-7.90(m, 1H), 7.82-7.40(m, 2H) ), 6.47(d, J=9.5Hz, 1H), 4.38–4.33(m, 2H), 3.49(s, 3H), 1.34(t, J=7.0Hz, 3H).

Example 44, 3-(2-ethoxy-6-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-5-oxothiazolo[4,5- b] Pyridin-4(5H)-yl)benzonitrile (Compound 44)

Intermediate 40-2 (20 mg) and 3-cyanophenylboronic acid (14.53 mg) were dissolved in dry 1,2-dichloroethane (5 mL), pyridine (15.65 mg) and copper acetate (11.93 mg) were added , the addition was completed, and the mixture was stirred at 40 °C for 4 hours. After the reaction was completed, the solvent was removed by concentration under reduced pressure. The residue was purified by preparative high performance liquid chromatography (column: Waters Xbridge BEH C18 100*25mm*5μm; mobile phase: [A: water (0.225% formic acid), B: acetonitrile]; B%: 23%-50%, 12 minutes) to give the title compound (2.8 mg).

MS m/z(ESI): 405.0[M+H] ⁺

¹ H NMR (400MHz, Chloroform-d) δ8.34 (d, J=2.7Hz, 1H), 7.84-7.74 (m, 2H), 7.71-7.58 (m, 4H), 6.64 (d, J=9.6Hz) ,1H),4.38–4.33(m,2H),3.58(s,3H),1.39(t,J=7.1Hz,3H).

Example 45, 4-(3-(difluoromethoxy)phenyl)-2-ethoxy-6-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl )thiazolo[4,5-b]pyridin-5(4H)-one (Compound 45)

Intermediate 40-3 (15 mg) and intermediate 45-1 (13.94 mg) were dissolved in anhydrous 1,2-dichloroethane (5 mL), pyridine (15.65 mg) and copper acetate (11.93 mg) were added, After the addition was completed, the mixture was stirred at 40°C for 4 hours. After the reaction was completed, the solvent was removed by concentration under reduced pressure. The residue was purified by preparative high performance liquid chromatography (column: Waters Xbridge BEH C18100*25mm*5μm; mobile phase: [A: water (0.225% formic acid), B: acetonitrile]; B%: 23%-50%, 12 minutes ) to give the title compound (5.4 mg).

MS m/z(ESI): 446.0[M+H] ⁺

¹ H NMR(400MHz, DMSO-d ₆ )δ8.30(d,J=2.6Hz,1H),8.26(s,1H),7.80-7.77(m,1H),7.68-7.53(m,1H), 7.50–7.10 (m, 4H), 6.44 (d, J=9.5Hz, 1H), 4.34–4.29 (m, 2H), 3.46 (s, 3H), 1.30 (t, J=7.0Hz, 3H).

Biological Activity and Related Properties Test Examples

Test example 1. Biochemical test

Introduction to the experimental principle: L-methionine and ATP can be converted into SAM, inorganic phosphate and inorganic diphosphate under the catalysis conditions of MAT2A enzyme. By adding a chromogenic reagent, such as ammonium molybdate, etc., to the enzymatic reaction mixture, the content of inorganic phosphate in the sample can be quantitatively detected, and then the enzymatic activity of MAT2A can be reflected.

Materials: MAT2A screening kit was purchased from BPS bioscience (USA); 384-well plate was purchased from Corning (USA).

1. MAT2a protein (Kanglong Chemical (Beijing) New Drug Technology Co., Ltd.);

2. L-Methionine (Sigma#M9625-5G)

3. ATP (Sigma#A7699-1G)

4. KCl (Sigma#60142-500ML-F)

5.Tris(Sigma#T2663-1L)

6. MgCl ₂ (Sigma#M1028)

7. EDTA (Invitrogen #AM9260G)

8. BSA (Sangon Biotech #A500023-0100)

9. PiColorLock (abcam #ab270004)

Detection method:

Compounds were dissolved in DMSO, diluted to a final concentration of 10 μM using Echo, diluted 3-fold, and 80 nL were transferred to a 384-well plate.

Prepare assay buffer (50 mM Tris, 50 mM KCl, 15 mM _MgCl2 , 100 [mu]M EDTA, 0.005% BSA). MAT2a protein was diluted with assay buffer (final concentration 4 μg/mL). Add 40 μL of 2X MAT2a solution to a 384-well plate, centrifuge at 1000 rpm for 1 minute, and re-incubate for 120 minutes.

Dilute L-methionine and ATP with assay buffer (final L-methionine concentration 200 μM, ATP final concentration 400 μM). Add 40 μL of 2X L-methionine and ATP solution to start the reaction, centrifuge at 1000 rpm for 1 minute, and incubate at room temperature for 90 minutes.

Mix PiColorLock ^TM reaction catalyst with PiColorLock ^TM buffer at 1:100 according to the instructions, add 20 μL to each well, and shake for 30 seconds. Add 8 μL of stabilization reagent and shake for 30 seconds. Signal values were detected after incubation at room temperature for 30 minutes.

data analysis:

Calculate the % Compound inhibition and fit the _IC50 of the compound.

Compound inhibition=(100-100*(Signal-Bottom)/(Top-Bottom))%

Experimental results:

Under the experimental conditions, the inhibitory effect of the test compound on MAT2A can be expressed by the _IC50 value of the inhibition of the phosphate production level during the enzymatic reaction. The MAT2A inhibitory activities of the compounds of the present invention are shown in Table 1.

Table 1

Test example 2. Detection of intracellular SAM levels

Introduction to the test principle: SAM, as the catalytic product of MAT2A, can reflect the inhibitory effect of the test compound on MAT2A by measuring the level of SAM generated in cells. After co-incubating the test MAT2A inhibitor with the cancer cells for a period of time, the cells were lysed with a stop reagent to quench the MAT2A enzymatic activity. Quantitative determination of MAT2A catalytic product SAM in cell lysate was performed by LC-MS/MS method to reflect the activity of MAT2A in cells.

Materials and cells: HCT116 MTAP ^-/- cells were purchased from Kangyuan Borchuang; fetal bovine serum, McCoy's 5a medium and penicillin-streptomycin were purchased from Gibco (USA), 96-well plates were purchased from Corning (USA), PBS was purchased from Cytiva (USA).

Cell culture: HCT116 MTAP ^-/- cells were cultured in McCoy's 5a medium containing 10% fetal bovine serum + 1% penicillin-streptomycin at 37°C, 5% CO ₂ . Cells in logarithmic growth phase can be used for experiments.

LC-MS/MS detection: LC-MS/MS was used to detect the effect of compounds on SAM production level in HCT116 MTAP ^-/- cell line. The cell concentration was adjusted to 50,000 cells per well, seeded in a 96-well plate, and cultured overnight at 37°C and 5% CO ₂ . Compounds were solubilized in DMSO, diluted in DMSO followed by medium and transferred to cell plates to a final concentration of 10 μM in 3-fold dilutions. Incubate for 6 hours at 37°C and 5% CO ₂ . The supernatant was aspirated, and after washing with PBS, glacial acetic acid was added to lyse the cells. After the lysate was processed, the SAM concentration was determined by LC-MS/MS injection analysis.

data analysis:

Calculate the % Compound inhibition and fit the _IC50 of the compound.

%Compound inhibition=1-100%*(Signal-Bottom)/(Top-Bottom)

Experimental results:

The experimental results in Table 2 show that the compounds of the present invention can inhibit the production of intracellular SAM to a nanomolar level, and the compounds of the present invention exhibit strong MAT2A inhibitory activity.

Table 2

Test example 3. Human colon cancer HCT116 cell proliferation inhibition test

Introduction to the test principle: After incubating the test MAT2A inhibitor with cancer cells for a period of time, the cell proliferation counting method based on ATP content was used to measure the effect of the test compound on cell proliferation.

Materials and cells: HCT116 WT cells and HCT116 MTAP ^-/- cells were purchased from Kangyuan Borchuang; fetal bovine serum, McCoy's5a medium and penicillin-streptomycin were purchased from Gibco (USA), and 96-well plates were purchased from Corning Company (USA), Cell-Titer Glo reagent was purchased from Promega (USA).

Cell culture: HCT116 WT cells and HCT116 MTAP ^-/- cells were cultured in McCoy's 5a medium containing 10% fetal bovine serum + 1% penicillin-streptomycin at 37°C and 5% CO ₂ . Cells in logarithmic growth phase can be used for experiments.

Detection of cell proliferation activity: Cell-Titer Glo reagent was used to detect the inhibitory activity of compounds on the proliferation of HCT116 WT and HCT116 MTAP ^-/- cell lines. The cell concentration was adjusted to 400 cells per well, seeded in a 96-well plate, and cultured overnight at 37°C and 5% CO ₂ .

Compounds were solubilized in DMSO, diluted in DMSO followed by medium and transferred to cell plates to a final concentration of 10 μM in 3-fold dilutions. Culture was continued for 6 days at 37°C and 5% CO ₂ . Cell-Titer Glo reagent was added to detect cell viability.

data analysis:

Calculate the % Compound inhibition and fit the _IC50 of the compound.

%Compound inhibition=1-100%*(Signal-Bottom)/(Top-Bottom)

HCT116 MTAP ^-/- cells are cells that do not express MTAP protein obtained by targeted knockout of wild-type HCT116 (HCT116 WT) cells by gene knockout. Inhibition of MAT2A and loss of MTAP can produce synthetic lethal effects, resulting in tumor cell death. The activity and selectivity of MAT2A inhibitors can be assessed by testing the antiproliferative activity of MAT2A inhibitors on HCT116 MTAP ^-/- cells and HCT116 WT.

Experimental results:

Under the conditions of this experiment, the compounds to be tested exhibited strong proliferation inhibitory activity on HCT116 MTAP ^-/- cells, and showed better selectivity compared with HCT116 MTAP ^+/+ cells. The specific anti-cell proliferation activities of the compounds to be tested are shown in Table 3.

table 3

Test Example 4. Measurement of metabolic stability of the compounds of the present invention in liver microsomes

The metabolic stability of the compounds of the present invention in liver microsomes was determined by the following test method.

1. Test materials and instruments

1. Human liver microsomes (Corning 452117), Beagle dog liver microsomes (XENOTECH D1000), SD rat liver microsomes (XENOTECH R1000) and CD-1 mouse liver microsomes (XENOTECH M1000)

2.Na ₂ HPO ₄ (Tianjin Guangfu Institute of Fine Chemicals 20180130)

3. KH ₂ PO ₄ (Tianjin Guangfu Institute of Fine Chemicals 20180920)

4.MgCl ₂ (Tianjin Guangfu Institute of Fine Chemicals 20191216)

5. NADPH (Solarbio 1216C022)

6. Positive control compound verapamil (Sigma MKBV4993V)

7. AB Sciex API4000 LC/MS

2. Test steps

1. Preparation of 100 mM Phosphate Buffered Saline (PBS): Weigh 7.098 g of Na ₂ HPO ₄ , add 500 mL of pure water to dissolve by ultrasonic wave, and use as solution A. Weigh 3.400 g of KH ₂ PO ₄ , add 250 mL of pure water for ultrasonic dissolution, and use it as solution B. Place solution A on a stirrer and slowly add solution B until the pH value reaches 7.4 to prepare a 100 mM PBS buffer.

2. Preparation of the reaction system

Prepare the reaction system according to the following table:

Table 4 Reaction system preparation information

3. Pre-incubate the reaction system in a 37°C water bath for 10 minutes. 40 μL of 10 mM NADPH solution (NADPH was dissolved in 100 mM phosphate buffer) was added to the reaction system, and the final concentration of NADPH was 1 mM. The NADPH solution was replaced with 40 μL of phosphate buffer as a negative control. The role of the negative control is to exclude the influence of the chemical stability of the compound itself.

4. Add 4 μL of 100 μM compounds of the present invention and positive control compound verapamil to the reaction system to initiate the reaction, and the final concentration of the compounds is 1 μM.

5. After fully mixing on a vortex shaker, 50 μL of the incubation samples were taken out at 0.5, 15, 30, 45 and 60 minutes, respectively, and the reaction was terminated with 200 μL of ice acetonitrile containing the internal standard. The samples were centrifuged at 3220g for 45 minutes. After centrifugation, 90 μL of supernatant was transferred to the sample injection plate, and 90 μL of ultrapure water was added to mix well for LC-MS/MS analysis.

All data were calculated by Microsoft Excel software. The peak area was detected by extracting the ion spectrum, and the in vitro half-life (t _1/2 ) of the compound was determined by linear fitting of the natural logarithm of the elimination percentage of the compound with time.

The in vitro half-life (t _1/2 ) was calculated from the slope:

in vitro t _1/2 = 0.693/k

In vitro intrinsic clearance (unit: μL/min/mg protein) was calculated with the following formula:

in vitro CL _int =k×volume of incubation(μL)/amount of proteins(mg)

Table 4

The experimental results in Table 4 show that the compounds of the present invention all exhibit good metabolic stability of liver microsomes.

Test Example 5. Determination of membrane permeability and transport properties of the compounds of the present invention

The membrane permeability and transport properties of the compounds of the present invention were determined using the following test methods.

1. Test materials and instruments

1. Caco-2 cells (ATCC)

2. HEPES (Solarbio 804D049), penicillin/streptomycin (Solarbio 20200109), PBS (Solarbio 20200620) and kanamycin (MP Biomedicals 194531)

3. Fetal Bovine Serum (FBS) (Sigma WXBD0055V), Lucifer Yellow (Sigma MKCJ3738) and NaHCO ₃ (Sigma SLBZ4647)

4. Hank's Balanced Salt Solution (HBSS) (Gibco 2085528), Non-Essential Amino Acids (NEAA) (Gibco 2211548) and Trypsin/EDTA (Gibco 2120732)

5. High glucose DMEM (Corning 20319014)

6. HTS Transwell-96 Well Permeable (Corning, 3391)

7. Resistance detector (Millipore,

ERS-2)

8.

Vision (Nexcelom Bioscience)

9. Infinite 200 PRO microplate reader (Tecan, Infinite M200PRO)

10. Positive control compounds Metoprolol (Sinopharm 100084-201403), Erythromycin (MCE 84550) and Cimetidine (Sinopharm 100158-201406)

11. ABI QTrap 5500 LC/MS

2. Test steps

1. Caco-2 cell culture

1) Preparation of transport buffer (HBSS containing 25mM HEPES, pH 7.4): Accurately weigh 5.958g HEPES and 0.35g NaHCO ₃ , add 900 mL of pure water to dissolve it, then add 100 mL of 10×HBSS, stir evenly, and adjust the pH to 7.4, Filtering.

2) Preparation of Caco-2 cell culture medium: FBS, penicillin/streptomycin, kanamycin and NEAA were added to high glucose DMEM (containing L-glutamine) medium to prepare 10% FBS and 100 units of penicillin /0.1 mg/mL streptomycin, 0.6 μg/mL kanamycin, and 1×NEAA in cell culture medium.

3) Incubate the cells in a T-75 flask in a 37°C, 5% _CO2 incubator, discard the medium when the cells grow to 80-90% density. Rinse cells with 5 mL of PBS, add 1.5 mL of Trypsin/EDTA, then incubate in a 37°C incubator for 5-10 minutes until cells fall off as quicksand, and finally neutralize Trypsin/EDTA with FBS-containing medium.

4) The cell suspension was centrifuged at 120 g for 10 minutes, and the supernatant was discarded.

5) Add cell culture medium to resuspend the cells and adjust to a cell suspension with a density of 6.86×10 ⁵ cells/mL.

2. Caco-2 Cell Seeding

1) Add 50 μL of medium to each well of the Transwell chamber, add 25 mL of medium to the lower layer, and place it in a 37°C, 5% CO ₂ incubator to preheat for 1 hour.

2) 50 μL of cell suspension was added to each well of the preheated Transwell chamber, and the final seeding density was 2.4×10 ⁵ cells/cm ² .

3) Culture for 14-18 days, change the medium every other day, and change the medium within 48 hours after the initial seeding. The medium must be changed the day before the experiment.

3. Assessing Monolayer Membrane Integrity

1) Cells were fused and differentiated after 14 days of culture, ready for transport experiments.

2) Measure the resistance of the single-layer film with a resistance meter, and record the resistance of each hole.

3) After the measurement, re-incubate the Transwell plate.

4) Calculate the TEER value:

TEER value=TEER measurement value (Ω)×film area (cm ² )

The resistance of the monolayer cell membrane is less than 230Ω·cm ² , indicating that the monolayer cell membrane has poor compactness and cannot be used for the test.

4. Transport Experiment

1) Dilute a 10 mM stock solution of the present compound or positive control compound with DMSO to obtain a 2 mM stock solution, and then dilute the 2 mM stock solution with transport buffer to obtain a 10 μM working solution of the present compound or positive control compound.

2) Remove the Caco-2 cell plate from the incubator, then wash the Transwell plate twice with pre-warmed transport buffer, and then place it in a 37°C incubator for 30 minutes.

3) To determine the transport rate of compounds from apical to basolateral (A→B), add 108 μL of compound working solution to the Transwell chamber (top), and immediately remove 8 μL of sample from the top to 72 μL of transport buffer, add 240 μL of The target stop solution terminates the transport to serve as the initial apical sample. At the same time, 300 μL of transport buffer was added to the receiving end (basal end). The experiment has two samples.

4) To determine the transport rate of the compound from the basolateral to the apical (B→A), add 308 μL of compound working solution to the basal end, and immediately remove 8 μL of the sample from the basal end to 72 μL of transport buffer, and add 240 μL of stopper containing internal standard Liquid terminated transport as the initial basolateral sample. At the same time, 100 μL of transport buffer was added to the Transwell chamber (top). The experiment has two samples.

5) Incubate the cell culture plate in a 37°C _CO2 incubator for 2 hours.

6) After the transport experiment, take 8 μL of sample from the administration end (ie, the apical end in the A→B direction and the basal end in the B→A direction) into 72 μL of transport buffer, and then add 240 μL of stop solution containing internal standard to terminate the transport. Take 80 μL of sample from the receiving end (ie, the basal end in the A→B direction and the apical end in the B→A direction) into 240 μL of stop solution containing internal standard, vortex at 1000 rpm for 10 minutes, and centrifuge at 3220 g for 30 minutes. Take 100 μL of supernatant to the injection plate, add 100 μL of ultrapure water and mix well for LC-MS/MS analysis.

7) Measure the fluorescence value after the transport experiment, prepare a 10 mM fluorescent yellow stock solution with water, and then dilute to 100 μM with transport buffer solution. Add 100 μL of fluorescent yellow solution to the Transwell chamber (top), add 300 μL of transport buffer solution to the basal end, and incubate in a CO ₂ incubator at 37°C for 30 minutes. 80 μL of the solution was taken from the apical and basal ends into a 96-well plate, and the fluorescence value of the cells was measured with a microplate reader (detecting membrane integrity) at an excitation wavelength of 485 nm and an emission wavelength of 530 nm.

Calculate the fluorescence value of the Caco-2 cell monolayer with the following formula:

LY Leakage={I _acceptor ×0.3/(I _acceptor ×0.3+I _donor ×0.1)}×100%

I _acceptor refers to the fluorescence density on the receiving side (0.3 mL), and I _donor refers to the fluorescence density on the dosing side (0.1 mL). LY > 1.0% indicates poor monolayer membrane compaction and corresponding results will be excluded from the evaluation.

Determine the peak area of the compound on the dosing side and the receiving side, and calculate the apparent permeability coefficient (P _app , unit: cm/s) and efflux ratio (Efflux ratio) of the compound:

P _app = {V _A ×[drug] _acceptor /(Area×incubation time×[drug] _{initial donor} }

V _A is the volume of the receiving end solution (A→B is 0.3 mL, B→A is 0.1 mL), Area is the membrane area of the Transwell-96-well plate (0.143 cm ² ); incubation time is the incubation time (unit: s).

P _app(BA) is the apparent permeability coefficient from the basal end to the apical end; P _app(AB) is the apparent permeability coefficient from the apical end to the basal end.

table 5

化合物compound	P _app(A-B)(10 ^-6cm/s) P _app (AB)(10 ^-6 cm/s)	P _app(B-A)(10 ^-6cm/s) P _app (BA)(10 ^-6 cm/s)	Efflux RatioEfflux Ratio
化合物5Compound 5	3.443.44	4.704.70	1.371.37
化合物8Compound 8	2.662.66	4.294.29	1.361.36
化合物9Compound 9	3.653.65	3.953.95	1.081.08
化合物10Compound 10	1.781.78	6.326.32	3.543.54
化合物11Compound 11	2.292.29	3.873.87	1.701.70

Caco-2 is a human cloned colon adenocarcinoma cell, similar in structure and function to differentiated small intestinal epithelial cells, and can be used to perform experiments that mimic intestinal transit in vivo. The higher the Papp(A-B), the better the permeability of the compound, indicating that the bioavailability of the compound through oral absorption may be higher. The Efflux Ratio indicated that the compound is affected by the transporter, and the greater the possibility of being excreted by the transporter, it indicates that the membrane permeability of the compound is more affected by the transporter, which is not conducive to the oral absorption of the compound and the penetration of the target cell membrane to exert its curative effect. The results in Table 5 show that the compound of the present invention has good membrane permeability, indicating that the compound of the present invention has a strong ability to be absorbed orally.

Test Example 6. Inhibitory effect of the compounds of the present invention on the enzymatic activities of CYP2C9, CYP2D6 and CYP3A4

The inhibition of CYP2C9, CYP2D6 and CYP3A4 enzymatic activities by the compounds of the present invention was determined by the following test method.

1. Test materials and instruments

1. Human liver microsomes (Corning 452117)

2.Na ₂ HPO ₄ (Sigma SLBZ6180)

3. KH ₂ PO ₄ (Sigma SLBT6559)

4.NADPH (Solarbio 705Y021)

5. Positive substrates Diclofenac (Sigma SLBV3438), Dextromethorphan (TRC 3-EDO-175-1) and Midazolam (Cerilliant FE01161704)

6. Positive inhibitors sulfaphenazole (D.Ehrenstorfer GmbH 109012), quinidine (TCI WEODL-RE) and ketoconazole (Sigma 100M1091V)

7. AB Sciex Triple Quad 5500 LC/MS

2. Test steps

2. Prepare a 10 mM NADPH solution with 100 mM PBS buffer. 10 mM stock solutions of compounds of the invention were diluted with DMSO to give 200X concentrations of compound working solutions (6000, 2000, 600, 200, 60, 20, 0 [mu]M). Dilute the positive inhibitor stock solution with DMSO to obtain a 200× concentration of the positive inhibitor working solution (sulfaphenazole, 1000, 300, 100, 30, 10, 3, 0 μM; quinidine/ketoconazole, 100, 30, 10, 3, 1, 0.3, 0 μM). Substrate working solutions (120 μM diclofenac, 400 μM dextromethorphan, and 200 μM midazolam) were prepared in water, acetonitrile, or acetonitrile/methanol at a concentration of 200×.

3. Take 2 μL of 20 mg/ml liver microsome solution, 1 μL of substrate working solution, 1 μL of compound working solution and 176 μL of PBS buffer, mix well, and pre-incubate in a 37°C water bath for 15 minutes. In the positive control group, 1 μL of sulfaphenazole, quinidine or ketoconazole working solution was added to replace the compound working solution. Simultaneously, 10 mM NADPH solution was pre-incubated in a 37°C water bath for 15 minutes. After 15 minutes, 20 μL of NADPH was added to each well to initiate the reaction and incubated at 37°C for 5 minutes (CYP2C9), 20 minutes (CYP2D6) or 5 minutes (CYP3A4). All incubation samples were double-sampled. The reaction was terminated by adding 400 μL of ice methanol containing the internal standard to all samples after the corresponding incubation time. Vortex to mix and centrifuge at 3220g, 4°C for 40 minutes. After centrifugation, 100 μL of the supernatant was transferred to the injection plate, and 100 μL of ultrapure water was added to mix well for LC-MS/MS analysis.

Table 6

Drug-drug interaction (DDI) refers to the physical or chemical changes produced by two or more drugs, as well as the changes in drug efficacy caused by these changes. Understanding drug interactions can provide better pharmaceutical services for patients, promote rational drug use, and maximize the avoidance of adverse reactions. Drug interactions are mainly metabolic interactions, which are mainly related to CYP450 enzymes involved in drug metabolism. The experimental results in Table 6 show that the compounds of the present invention have weak inhibitory ability to CYP450, indicating that the compounds of the present invention have less potential risk of DDI.

Claims

A compound represented by general formula (A) or a pharmaceutically acceptable salt thereof:

Wherein, ring Q is a 5-6-membered heterocyclic group, a C 6 -C 10 -membered aryl group or a 5-10-membered heteroaryl group, and the 5-6-membered heterocyclic group, a C 6 -C 10 -membered aryl group or a 5-10 A membered heteroaryl group is optionally substituted with R a selected from F, Cl, Br, I, OH, CN, C 2 -C 3 alkynyl, C 1 -C 10 alkyl, C 3 -C 10 cycloalkyl or C 1 -C 10 alkoxy, said C 1 -C 10 alkyl, C 3 -C 10 cycloalkyl or C 1 -C 10 alkoxy optionally substituted by R b ;

R b is selected from F, Cl, Br, I, OH or CN;

Ring W is phenyl, pyridyl, pyrimidinyl, pyridazinyl, pyridinonyl, pyridazinonyl or
wherein Ring M is a 5-10 membered heteroaryl group or a 4-10 membered heterocyclic group, and said phenyl, pyridyl, pyrimidinyl, pyridazinyl, pyridinyl, pyridazinonyl or ring M is optionally replaced by R c is substituted, said R c is selected from OH, =O or the following groups optionally substituted by R c1 : C 1 -C 10 alkyl, C 1 -C 10 alkoxy, C 3 -C 10 cycloalkyl , P(O)(C 1 -C 3 alkyl) 2 , 4-6 membered heterocycloalkyl, NHC(O)(C 1 -C 6 alkyl); X 1 is selected from C or N, when X 1 When selected from C,
Represents a double bond, when X 1 is selected from N,
represents a single key;

R c1 is selected from deuterium, F, Cl, Br, I, CN, OH, NH 2 or the following groups optionally substituted by R c2 : C 1 -C 3 alkyl, C 1 -C 3 alkoxy, NHC (O)O(C 1 -C 6 alkyl), (C 1 -C 6 alkyl) NHC(O)O, NH(C 1 -C 3 alkyl), S(O) 2 (C 1 -C 3 alkyl), 4-10 membered heterocyclyl, 4-10 membered heterocyclyloxy, (C 3 -C 10 cycloalkyl) CH 2 O, 5-6 membered heteroaryl, 5-6 membered heterocyclyl Aryloxy;

R c2 is selected from 4-6 membered heterocycloalkyl, =O, C 1 -C 3 alkyl, halogenated C 1 -C 3 alkyl, F, Cl, Br, I, CN, OH, CH 2 OH or NH 2 ;

X is selected from O, S or NR 2 ;

L is selected from O, NH or a chemical bond;

R 1 and R 2 are independently selected from H, C 1 -C 10 alkyl, C 3 -C 10 cycloalkyl or 4-10 membered heterocyclic group, the C 1 -C 10 alkyl, C 3 -C The 10 -cycloalkyl or 4-10 membered heterocyclyl is optionally substituted with Rd selected from F, Cl, Br, I, OH, CN or C1 - C3 alkyl.
The compound represented by the general formula (A) according to claim 1 or a pharmaceutically acceptable salt thereof, wherein ring Q is a phenyl group or a 5-6 membered heteroaryl group, and the phenyl group or a 5-6 membered heteroaryl group A membered heteroaryl group is optionally substituted with Ra .
The compound represented by the general formula (A) or a pharmaceutically acceptable salt thereof according to claim 1, wherein ring W is phenyl, pyridyl, pyrimidinyl, pyridazinyl, 2-pyridone,
wherein ring M is a 5-10-membered heteroaryl group or a 5-10-membered heterocyclic group, the phenyl, pyridyl, pyrimidinyl, pyridazinyl, 2-pyridone,
or Ring M is optionally substituted with Rc .
The compound represented by general formula (A) according to claim 1 or a pharmaceutically acceptable salt thereof, is characterized in that, ring W is the following group optionally substituted by R c :
phenyl,
The compound represented by the general formula (A) according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R a is F, Cl, Br, I, CN, C 2 -C 3 alkynyl, C 1 - C6alkyl , C3 - C6cycloalkyl or C1 - C6alkoxy optionally substituted by F.
The compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to claim 1, wherein R c is selected from =O or the following groups optionally substituted by R c1 : C 1 -C 6 alkyl, C 1 -C 6 alkoxy, NHC(O) (C 1 -C 6 alkyl), the R c1 is selected from deuterium atom, F, Cl, Br, I, CN, OH, NH 2 or ethoxy.
The compound represented by general formula (A) according to claim 1 or a pharmaceutically acceptable salt thereof, wherein ring W is selected from
The compound represented by the general formula (A) according to claim 1 or a pharmaceutically acceptable salt thereof, wherein ring Q is selected from
The compound represented by the general formula (A) according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the R 1 is selected from H, a 4-6 membered heterocyclic group, optionally by C 1 - C3 - C6 cycloalkyl substituted with C3 alkyl or C1 - C6 alkyl optionally substituted with F.
The compound represented by the general formula (A) or a pharmaceutically acceptable salt thereof according to claim 1, wherein the compound represented by the general formula (A) or a pharmaceutically acceptable salt thereof is selected from the group consisting of the formula ( B) the compound shown or a pharmaceutically acceptable salt:

wherein rings W, R a , R 1 are as defined in claim 1 and n is selected from 0, 1, 2, 3, 4 or 5.
The compound of general formula (A) or a pharmaceutically acceptable salt thereof according to claim 1, wherein the compound of general formula (A) or a pharmaceutically acceptable salt thereof is selected from the group consisting of formula (C) Compound or pharmaceutically acceptable salt:

wherein the rings W, R a , R 1 are as defined in claim 1 .
The compound represented by the general formula (A) according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from one of the following structures:
A pharmaceutical composition comprising the compound of any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.
Use of the compound of any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 13 in the preparation of a medicament for preventing or treating tumors with reduced or absent MTAP activity.