WO2015135461A1

WO2015135461A1 - Substituted dihydrobenzofuran-piperidine-ketone derivative, preparation and use thereof

Info

Publication number: WO2015135461A1
Application number: PCT/CN2015/073932
Authority: WO
Inventors: 魏用刚; 邱关鹏; 郑苏欣; 刘建余; 钱枚林
Original assignee: 四川海思科制药有限公司
Priority date: 2014-03-10
Filing date: 2015-03-10
Publication date: 2015-09-17
Also published as: CN105793264A; CN105793264B

Abstract

The present invention relates to a substituted dihydrobenzofuran-piperidine-ketone derivative, and a preparation and use thereof. In particular, the present invention relates to a substituted dihydrobenzofuran-piperidine-ketone derivative as shown by general formula (A), a stereoisomer or pharmaceutically acceptable salt thereof, the preparation method thereof, pharmaceutical compositions comprising them, and a use in the preparation of drugs for treating cancer-related diseases.

Description

Substituted dihydrobenzofuran-piperidine-methanone derivatives, preparation and use thereof

Technical field

The present invention relates to a substituted dihydrobenzofuran-piperidine-methanone derivative represented by the formula (A), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, a process for producing the same, and a medicament containing the same Compositions and uses in the manufacture of a medicament for the treatment of cancer-related diseases.

Background technique

The invention provides a series of inhibitors of the Wnt signaling pathway. These inhibitors are capable of inhibiting tumor cell proliferation and preventing tumor metastasis.

The Wnt signaling pathway is a key regulator in various cellular processes, including maintenance of stem cells, determination of stem cell fate, and control of cell cycle (Huang, Xie et al, 2006, J Immunol, 176, 4880-4887). It plays an important role in the development and maintenance of adult homeostasis, and also exhibits an abnormal activation state in many types of tumors. The classical Wnt signaling pathway is regulated by the stability of its major participant, catenin β-catenin. The stability of β-catenin is controlled by the destruction of the complex. --catenin has many important biological functions. On the cell membrane, β-catenin is linked to cadherin E-cadherin and participates in the formation of adhesion junctions. In cytoplasm, β-catenin is capable of forming downstream β-catenin destruction complexes with APC, AXIN, GSK3 and CK1. When the Wnt signaling pathway is not activated, the β-catenin in the cytosol is degraded by the phosphorylation of the complex. When Wnt activates the Wnt signaling pathway, the β-catenin destruction complex degrades, resulting in the accumulation of stable β-catenin after entering the nucleus, causing Wnt pathway target genes (such as c-myc, cyclin D, Axin2, and Nkd1). Transcriptional activation.

In many tumors, overexpression of Wnt proteins or mutations in individual constituent proteins in the β-catenin destruction complex stabilizes β-catenin, resulting in abnormal activation of the Wnt pathway. In particular, the truncation mutation of the tumor suppressor APC is the most prevalent genetic alteration in colon cancer (Miyaki, Konishi et al, 1994, Cancer Res, 54, 3011-3020). Mutations in Axin1 and Axin2 were also found in human hepatocellular carcinoma and colon cancer, respectively. These mutations result in non-Wnt-dependent β-catenin stabilization and β-catenin-mediated transcriptional activation (Taniguchi, Roberts et al., 2002, Oncogene, 21, 4863-4871). In addition, mutations in β-catenin cause a reversal of β-catenin degradation and an increase in β-catenin signaling. Unregulated activation of the Wnt pathway is also involved in many other types of tumors, including colorectal cancer, melanoma, breast cancer, liver cancer, lung cancer, and gastric cancer (Barker and Clevers, 2006, Nat Rev Drug Discov, 5, 997- 1014). In addition to this tumor, there are also some disorders that are also associated with abnormal Wnt pathways, including osteoporosis. Arthritis, polycystic kidney disease, pulmonary fibrosis, diabetes, schizophrenia, cardiovascular disease, non-carcinogenic proliferative diseases, and neurodegenerative diseases such as Alzheimer's disease. The efficient assembly of the polyprotein β-catenin destructive complex depends on the level of steady state of its various essential components. Axin regulates the function of the β-catenin destruction complex in a concentration-dependent manner (Salic, Lee et al., 2000, Mol Cell, 5, 523-532). Increased expression of Axin in cell lines expressing APC truncation mutations enhances the degradation of β-catenin (Behrens, Jerchow et al., 1998, Science, 280, 596-599). Therefore, the level of Axin protein is likely to be tightly regulated to ensure that a suitable Wnt pathway signal is produced.

The Tankyrase protein is a multifunctional poly(ADP-ribose) polymerase (PRAP) that utilizes NAD+ as a substrate to transfer ADP-ribose polymers to target proteins for post-translational modification. Tankyrase is able to directly bind to Axin protein to regulate its stability. Studies have shown that inhibition of Tankyrase can stabilize Axin to degrade β-catenin and inhibit Wnt signaling pathway (Huang, Mishina et al, 2009, Nature, 461, 614-620). The tankyrase subtype can react with a highly conserved sequence of Axin to promote the degradation of Axin via the ubiquitin-protease pathway. Axin has important regulatory roles in a wide range of physiological processes, including differentiation of glioma progenitor cells during remyelination (Fancy, Harrington et al., 2011, Nat Neurosci, 14, 1009-1016) and pulmonary fibrosis Transformation of the epithelium into the stroma (Ulsamer, Wei et al, 2012, J Biol Chem, 287, 5164-5172). Inhibition of Tankyrase can stabilize Axin and inhibit Wnt pathway signaling, and this process can be used to develop a treatment for disorders associated with Wnt signaling. Tankyrase has multiple chaperones, including TRF1, a double-stranded telomere repeat binding protein; NuMA, a protein that plays an important role in mitotic spindle assembly; IRAP, a membrane protein associated with insulin-responsive glucose uptake And Mcl-1, a pro-apoptotic protein. In addition to regulating the Wnt signaling pathway, Tankyrase proteins have a variety of biological functions by interacting with various proteins. Tankyrase releases TRF1 from telomeres, causing telomeres to contact telomerase. Thus, Tankyrase is able to positively regulate telomerase elongation of telomeres (Cook, Dynek et al, 2002, Mol Cell Biol, 22, 332-342). In normal cells, telomerase expression is usually inhibited, and most tumor cells express telomerase, leading to telomere elongation (Hahn, Stewart et al., 1999, Nat Med, 5, 1164-1170). ). This suggests that Tankyrase can be used as a target for tumor therapy by inhibiting telomerase access to telomeres and inhibiting tumors. Inhibitors of Tankyrase can be used as effective tumor treatments to inhibit a variety of tumors, including leukemia, lymphoma, multiple myeloma, lung cancer and breast cancer. Tankyrase also plays an important role in cell mitosis: (1) regulates the regulation of NuMA in the mitosis on the spindle pole (Chang, Dynek et al, 2005, Biochem J, 391, 177-184); (2) regulates the spindle Assembly and structure of bodies (Chang, Jacobson et al, 2004, Nature, 432, 645-649); (3) maintenance of dissociation of sister chromatids (Dynek and Smith, 2004, Science, 304, 97-100). Inhibition of tankyrase causes cell mitosis arrest or cell aging, so this strategy can be developed to treat diseases associated with mitotic abnormalities, such as tumors (breast, lung, ovarian, leukemia, lymphoma, and melanin). Tumor, etc.). In addition, Tankyrase1 is an important gene required for centrosome aggregation, and tumor cells have redundant centrosomes to inhibit multipolar mitosis and undergo bipolar mitosis (Kwon, Godinho et al., 2008, Genes Dev, 22, 2189-2203). ). Therefore, inhibition of Tankyrase can be developed into tumors that inhibit centrosome expansion, including various solid tumors and hematological cancers such as breast cancer, bladder cancer, lung cancer, colon cancer, and leukemia.

Some synthetic inhibitors of Tankyrase have been disclosed in existing studies. Some of these studies have found that inhibitors of Tankyrase are capable of blocking Wnt signaling in APC-mutated colon cancer cells (Chen, Dodge et al, 2009, Nat Chem Biol, 5, 100-107; Huang, Mishina et al, 2009, Nature, 461, 614-620) and Wnt signaling in breast cancer (Bao, Christova et al, 2012, PLoS One, 7, e48670). Although the Wnt signaling pathway is a highly potential target for anti-tumor therapy, there are currently fewer effective tankyrase inhibitors.

WO2013012723 describes novel 2-piperidin-2-yl-acetamide derivatives and their use as Tankyrase inhibitors for the treatment of diseases associated with Wnt, Tankyrase1 and Tankyrase2, such as cancer. Wherein X is NH or O, Y is CH or N, and R ^1a and R ^1b are alkyl groups or together form a 5- to 7-membered ring which may be substituted, R ² is H or an alkyl group, and R ³ is H, an alkyl group, An alkynyl group or the like, and R ⁵ is a benzene, a 2,3-dihydrobenzofuran, a quinoline or the like which may be substituted. The specific description in this patent is not considered to be part of the present invention and its structure is as follows:

WO2013008217 describes novel 4-piperidine derivatives and their use as Tankyrase inhibitors for the treatment of diseases associated with Wnt, Tankyrase 1 and Tankyrase 2, such as cancer. Wherein R ¹ is a substitutable phenyl group, a 5-membered heterocyclic ring, an 8- to 10-membered heterocyclic ring or the like, and R ⁴ is H or a substitutable phenyl group. The specific description in this patent is not considered to be part of the present invention and its structure is as follows:

WO2013010092 describes a novel 4-oxo-3,5,7,8-tetrahydro-4H-pyrano[4,3-d]pyrimidine derivative and its use as a Tankyrase inhibitor for treatment with Wnt , Tankyrase1 and Tankyrase2 related diseases, such as cancer. Wherein R ¹ is H or a substitutable alkyl group, and R ² is R ³ -alkenyl-C(O)-, R ³ -alkenyl-S(O) ₂ - or R ³ -alkenyl-C(O) O-, R ³ is a phenyl group, a 6-membered heteroaryl ring or an anthracene which may be substituted. The detailed description in this patent is not considered to be part of the present invention and is structured as follows:

The object of the present invention is to provide a novel and effective Tankyrase inhibitor, and a use for treating a cancer-related disease, wherein the cancer-related diseases include bladder cancer, colon cancer, ovarian cancer, melanoma, leukemia, lymphoma , multiple myeloma, lung cancer and breast cancer.

Summary of the invention

The present invention provides a compound represented by the formula (A), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein:

R ^{1 is} selected from a C _6-10 carbocyclic ring or a 5- to 10-membered heterocyclic ring containing at least 1 to 4 hetero atoms selected from N, O or S, optionally further 0 to 4 are selected from the group consisting of F, Cl, Br, I, (=O), cyano, hydroxy, carboxy, amino, nitro, R ^1a , NR ^1a R ^1b , COR ^1a , CONR ^1a R ^1b or NR ^1a COR Substituted by a substituent of ^1b ;

R ^1a and R ^1b are each independently selected from H, C _1-6 alkyl or C _1-6 alkoxy, and the alkyl and alkoxy are optionally further from 0 to 4 selected from F, Cl, Br. Substituted by a substituent of I, cyano, hydroxy, carboxy, amino or nitro;

R ² , R ³ and R ⁴ are each independently selected from the group consisting of H, F, Cl, Br, I, cyano, hydroxy, carboxy, amino, nitro, C _1-6 alkyl or C _1-6 alkoxy, The alkyl group and the alkoxy group are optionally further substituted with from 0 to 4 substituents selected from the group consisting of F, Cl, Br, I, cyano, hydroxy, carboxy, amino or nitro;

R ⁵ , R ⁶ , R ⁷ and R ⁸ are each independently selected from H or C _1-6 alkyl.

A preferred embodiment of the invention, a compound of the formula (A), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of a compound represented by the formula (I), a stereoisomer thereof Or pharmaceutically acceptable salts:

A preferred embodiment of the invention, a compound of the formula (A) or the formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein R ^{1 is} selected from a 6-membered carbocyclic ring and a 5-membered heterocyclic ring. a ring, a 6-membered heterocyclic ring, a 7-membered heterocyclic ring, an 8-membered heterocyclic ring, a 9-membered heterocyclic ring or a 10-membered heterocyclic ring, the heterocyclic ring having at least 1 to 4 hetero atoms selected from N, O or S, Carbocyclic or heterocyclic ring optionally further selected from 0 to 4 selected from the group consisting of F, Cl, Br, I, (=O), cyano, hydroxy, carboxy, amino, nitro, R ^1a , NR ^1a R ^1b or COR Substituted by a substituent of ^1a ;

R ^1a and R ^1b are each independently selected from H, C _1-4 alkyl or C _1-4 alkoxy, and the alkyl and alkoxy groups are further further selected from 0 to 4 selected from F, Cl, Br. Substituted by a substituent of I, cyano, hydroxy, carboxy, amino or nitro.

A preferred embodiment of the invention, a compound of the formula (A) or the formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein R ^{1 is} selected from the following structures which are substituted or unsubstituted One:

When substituted, optionally further 1 to 4 are selected from the group consisting of F, Cl, Br, I, (=O), cyano, hydroxy, carboxy, amino, nitro, R ^1a , NR ^1a R ^1b or COR ^1a Substituted by a substituent;

R ^1a and R ^1b are each independently selected from H, C _1-4 alkyl or C _1-4 alkoxy, and the alkyl and alkoxy groups are further further selected from 0 to 4 selected from F, Cl, Br. Substituted with a cyano or hydroxy substituent;

Preferred substituted or unsubstituted

When substituted, optionally further 1 to 4 are selected from the group consisting of F, Cl, Br, cyano, hydroxy, carboxy, amino, nitro, methyl, ethyl, trifluoromethyl, methoxy or ethoxy Substituted by a substituent of the group, it is preferably substituted with from 0 to 4 substituents selected from the group consisting of F, Cl, Br, cyano, methyl, ethyl or trifluoromethyl.

A preferred embodiment of the invention, a compound of the formula (A) or the formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein: R ^{1 is} selected from the following structures which are substituted or unsubstituted One:

When substituted, optionally further substituted with from 1 to 4 substituents selected from the group consisting of F, Cl, cyano, methyl, trifluoromethyl or methoxy;

R ² , R ³ and R ⁴ are each independently selected from H, F, Cl or Br;

R ⁵ , R ⁶ , R ⁷ and R ⁸ are each independently H.

A preferred embodiment of the invention, a compound of the formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of a compound represented by the formula (II), a stereoisomer thereof Or pharmaceutically acceptable salts:

A preferred embodiment of the invention, a compound of the formula (II), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein:

R ² , R ³ and R ⁴ are each independently selected from H, F, Cl, Br, trifluoromethyl, methyl, ethyl, methoxy or ethoxy, preferably H, F, Cl or Br, more Preferred H or F

R ⁵ , R ⁶ , R ⁷ and R ⁸ are each independently selected from H, methyl or ethyl, preferably H.

A preferred embodiment of the invention, a compound of the formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein the compound is selected from, but not limited to, one of the following structures:

The present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of the present invention, and a stereoisomer or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

Further, the present invention provides a compound of the present invention, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, Or the use of the pharmaceutical composition of the present invention for the preparation of a medicament for treating cancer.

Still further, the present invention provides a method of treating cancer comprising administering a compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, as hereinbefore described, or as described above in the present invention Pharmaceutical composition.

Terms used in the specification and claims have the following meanings unless stated to the contrary.

The present invention relates to the substitution of a plurality of substituents, which may be the same or different.

The present invention relates to the inclusion of a plurality of heteroatoms, each of which may be the same or different.

The elements carbon, hydrogen, oxygen, sulfur, nitrogen or halogen involved in the groups and derivatives of the present invention include their isotopic conditions, and the elements carbon and hydrogen involved in the groups and derivatives of the present invention. , oxygen, sulfur or nitrogen are optionally further replaced by one or more of their corresponding isotopes, wherein the carbon isotopes include ¹² C, ¹³ C and ¹⁴ C, and the hydrogen isotopes include strontium (H), strontium (D, also known as Heavy hydrogen), strontium (T, also known as super heavy hydrogen), oxygen isotopes include ¹⁶ O, ¹⁷ O and ¹⁸ O, sulfur isotopes include ³² S, ³³ S, ³⁴ S and ³⁶ S, nitrogen isotopes include ¹⁴ N and ¹⁵ N, the fluorine isotope ¹⁹ F, the chlorine isotope includes ³⁵ Cl and ³⁷ Cl, and the bromine isotopes include ⁷⁹ Br and ⁸¹ Br.

The term "alkyl" refers to a saturated aliphatic hydrocarbon group, including straight chain and branched chain groups of 1 to 20 carbon atoms. Preference is given to alkyl groups having 1 to 10 carbon atoms, non-limiting examples including methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, positive a mercapto group, and various branched isomers thereof; more preferred are lower alkyl groups having 1 to 4 carbon atoms, and non-limiting examples include methyl, ethyl, propyl, isopropyl, and Butyl, isobutyl or tert-butyl. The alkyl group may be substituted or unsubstituted, and when substituted, the substituent is preferably from 1 to 5, independently selected from the group consisting of H, F, Cl, Br, I, =O, alkyl, alkenyl, alkynyl. , alkoxy, alkylthio, alkylamino, fluorenyl, hydroxy, nitro, cyano, amino, alkyl acylamino, cycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy A cycloalkylalkyl group, a hydroxyalkyl group, a carboxylic acid, a carboxylic acid ester or a heterocycloalkyl group.

"Alkoxy" means an -O-alkyl group wherein alkyl is as defined above. The alkoxy group may be substituted or unsubstituted, and non-limiting examples thereof include methoxy, ethoxy, propoxy, isopropoxy, butoxy, tert-butoxy, pentyloxy or The hexyloxy group preferably has a 1 to 12 member alkoxy group. When substituted, the substituent is preferably from 1 to 5, independently selected from the group consisting of H, F, Cl, Br, I, =O, alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkyl Amino, mercapto, hydroxy, nitro, cyano, amino, alkyl acylamino, cycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylindolyl, hydroxyalkyl, A carboxylic acid, a carboxylic acid ester or a heterocycloalkyl fluorenyl group.

"Carbocycle" means a saturated or unsaturated aromatic ring or a non-aromatic ring. The aromatic ring or non-aromatic may be a single ring of 3 to 8 members, a 4 to 12 membered double ring or a 10 to 15 membered three ring system. Linked to a bridged or spiro ring, non-limiting examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclopentene, ring Hexadiene, cycloheptatriene, phenyl, naphthyl, benzocyclopentyl, bicyclo[3.2.1]octyl, bicyclo[5.2.0]decyl, tricyclo[5.3.1.1] Dodecyl, adamantyl or spiro[3.3]heptyl and the like. The carbocyclic ring may be substituted, and when substituted, the substituent is preferably from 1 to 5, independently selected from the group consisting of H, F, Cl, Br, I, =O, alkyl, alkenyl, alkynyl, alkoxy, Alkyl, alkylamino, sulfhydryl, hydroxy, nitro, cyano, amino, alkyl acylamino, cycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkyl fluorenyl A hydroxyalkyl group, a carboxylic acid, a carboxylic acid ester or a heterocycloalkyl fluorenyl group.

"Heterocycle" means a substituted or unsubstituted saturated or unsaturated aromatic or non-aromatic ring, and the aromatic and non-aromatic rings may be a 3 to 8 membered monocyclic ring, a 4 to 12 membered bicyclic ring or a 10 to 15 membered member. A tricyclic system consisting of at least one hetero atom selected from N, O or S, preferably a 3 to 10 membered heterocyclic ring, and optionally substituted N, S in the heterocyclic ring can be oxidized to various oxidation states. The heterocyclic ring can be attached to a hetero atom or a carbon atom. The heterocyclic ring may be attached to a bridged or spiro ring, and non-limiting examples include, ethylene oxide, aziridine, oxetanyl, azetidinyl, 1,3-dioxolane, 1,4-dioxolane, 1,3-dioxane, azepanyl, pyridyl, furyl, thienyl, pyranyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl , pyridazinyl, imidazolyl, piperidinyl, piperidinyl, morpholinyl, thiomorpholinyl, 1,3-dithiane, dihydrofuran, dihydropyran, dithiapentane, tetrahydrofuran, Tetrahydropyrrolyl, tetrahydroimidazole, tetrahydrothiazole, tetrahydropyran, benzimidazole, benzopyridine, pyrrolopyridine, benzodihydrofuran, azabicyclo[3.2.1]octyl, nitrogen Heterobicyclo[5.2.0]nonane, oxatricyclo[5.3.1.1]dodecyl, azaadamantyl, oxaspiro[3.3]heptyl, etc.; when substituted, substituent Preferably from 1 to 5, the substituents are independently selected from the group consisting of H, F, Cl, Br, I, =O, alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, fluorenyl, hydroxy , nitro, cyano, amino, alkyl acylamino, heterocycloalkyl, ring Alkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylfluorenyl, hydroxyalkyl, carboxylic acid, carboxylic acid ester, heterocycloalkyl fluorenyl, -(CH ₂ ) _n S (=O) _p R ⁶ , -(CH ₂ ) _n -alkenyl-R ⁶ or -(CH ₂ ) _n -alkynyl-R ⁶ .

"Amino" means -NH _2, may be substituted or unsubstituted. When substituted, the substituent is preferably 1 to 3, independently selected from alkyl, alkenyl, alkynyl, alkoxy, Thio, hydroxy, amino, alkylamino, alkyl acylamino, heterocycloalkyl, cycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, hydroxyalkyl, carboxylic acid or Carboxylic acid ester.

"Aryl" means a substituted or unsubstituted 6 to 14 membered all carbon monocyclic or fused polycyclic group having a polycyclic group of a conjugated π-electron system, preferably a 6 to 10 membered aromatic ring, Non-limiting examples include phenyl or naphthyl; the aryl group may be fused to a heteroaryl, heterocyclyl or cycloalkyl group, and the moiety attached to the parent structure is an aryl group, non-limiting examples of which include benzo Furan, benzocyclopentyl or benzothiazole. When substituted, the substituent is preferably from 1 to 5, and the substituent is independently selected from the group consisting of H, F, Cl, Br, I, =O, alkyl, alkenyl, alkynyl, alkoxy, alkylthio, Alkyl Amino, mercapto, hydroxy, nitro, cyano, amino, alkyl acylamino, cycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylindolyl, hydroxyalkyl, A carboxylic acid, a carboxylic acid ester or a heterocycloalkyl fluorenyl group.

"Heteroaryl" means a substituted or unsubstituted 5 to 15 membered aromatic ring and contains 1 to 3 heteroatoms selected from N, O or S heteroatoms, preferably 5 to 10 members, and non-limiting heteroaryl groups. Examples include pyridyl, furyl, thienyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, benzofuran, benzimidazole, benzopyridine or pyrrolopyridine, and the like. When substituted, the substituent is preferably from 1 to 5, and the substituent is independently selected from the group consisting of H, F, Cl, Br, I, =O, alkyl, alkenyl, alkynyl, alkoxy, alkylthio, Alkylamino, fluorenyl, hydroxy, nitro, cyano, amino, alkyl acylamino, cycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylindolyl, hydroxyalkane a carboxylic acid, a carboxylic acid ester or a heterocycloalkyl fluorenyl group.

"Natural or pharmaceutically acceptable amino acids": The basic skeleton of a protein molecule is an amino acid sequence, and there are 20 basic amino acids constituting a protein. These 20 basic amino acids are the basis for biological late modification of proteins, and in addition, based on these basic amino acids. The organism also synthesizes amino acid types derived from hydroxyproline, hydroxylysine, etc. These biosynthesized amino acids are collectively referred to as "natural amino acids"; artificially synthesized are "unnatural amino acids". "Pharmaceutically acceptable amino acid" refers to a pharmaceutically acceptable natural or unnatural amino acid.

The "=O" of the present invention is generally used in the art and refers to an oxygen atom bonded by a double bond, such as a double bond oxygen atom to a carbon atom in a carbonyl group.

"Pharmaceutically acceptable salt" means a pharmaceutically acceptable salt of a non-toxic acid or base, including salts of inorganic acids and bases, organic acids and bases.

"Crystal" refers to a combination of an active pharmaceutical ingredient (API) and a cocrystal former (CCF) under the action of hydrogen bonds or other non-covalent bonds, of which API and CCF The pure state is solid at room temperature and there is a fixed stoichiometric ratio between the components. Eutectic is a multi-component crystal that contains both a binary eutectic formed between two neutral solids and a multi-component eutectic formed by a neutral solid with a salt or solvate. The "eutectic former" includes, but is not limited to, various pharmaceutically acceptable acid, base, nonionic compounds.

"Stereoisomer" refers to isomers resulting from the spatial arrangement of atoms in a molecule, including cis and trans isomers, enantiomers and conformational isomers.

"Pharmaceutical composition" means a derivative of one or more of the above-mentioned derivatives or physiological/pharmaceutically acceptable salts or prodrugs thereof and other chemical components, other components such as physiological/pharmaceutically acceptable carriers And excipients. The purpose of the pharmaceutical composition is to facilitate the administration of the compound to the organism.

"Prodrug" means a derivative of the invention which can be converted to biological activity under physiological conditions or by solvolysis. Things. Prodrugs of the invention are prepared by modifying functional groups in the derivative which can be removed by conventional procedures or in vivo to provide the parent compound. A prodrug includes a compound formed by attaching a hydroxyl group, an amino group or a thiol group to any group in the derivative of the present invention. When a prodrug of the derivative of the present invention is administered to a mammalian individual, the prodrug is cleaved to form a free form. Hydroxyl group, free amino group or free sulfhydryl group. Examples of prodrugs include, but are not limited to, compounds formed by the hydroxyl or amino functional groups of the derivatives of the invention with formic acid, acetic acid or benzoic acid.

"Optional," "optional," or "optionally" means that the event or environment described subsequently can, but need not, occur, including where the event or environment occurs or does not occur. For example, "aryl substituted with an alkyl group" means that an alkyl group may, but need not, be present, and the description includes the case where the aryl group is substituted with an alkyl group and the case where the aryl group is not substituted with an alkyl group.

"Substituted or unsubstituted" refers to a situation in which a group may be substituted or unsubstituted, and if it is not indicated in the present invention that a group may be substituted, it means that the group is unsubstituted.

“As a choice” means that the scheme after “as a choice” is a side-by-side relationship with the scheme before “as a choice” rather than a further selection in the previous scheme.

"Substitution" refers to the case where one or more hydrogen atoms in a group are substituted by another group, and if the group is substituted by a hydrogen atom, the group formed is the same as the group substituted by a hydrogen atom. Where the group is substituted, for example, amino, C _1-4 alkyl, C _1-4 alkoxy, C _3-6 carbocyclic, 3 to 6 membered heterocyclic ring, optionally further from 0 to 4 selected from H, Substituted by a substituent of F, Cl, Br, I, hydroxy, cyano, amino, C _1-4 alkyl or C _1-4 alkoxy, the groups formed include, but are not limited to, methyl, chloromethyl, Trichloromethyl, hydroxymethyl, -CH ₂ OCH ₃ , -CH ₂ SH, -CH ₂ CH ₂ CN, -CH ₂ NH ₂ , -NHOH, -NHCH ₃ , -OCH ₂ Cl, -OCH ₂ OCH ₂ CH ₃ , -OCH ₂ CH ₂ NH ₂ , -OCH ₂ CH ₂ SH, -OCH ₂ CH ₂ OH, 1-hydroxycyclopropyl, 2-hydroxycyclopropyl, 2-aminocyclopropyl, 4-methyl Furanyl, 2-hydroxyphenyl, 4-aminophenyl or phenyl.

detailed description

The technical solutions of the present invention are described in detail below with reference to the accompanying drawings and embodiments, but the scope of protection of the present invention includes but is not limited thereto.

The structure of the compound is determined by nuclear magnetic resonance (NMR) or (and) mass spectrometry (MS). The NMR shift (δ) is given in units of 10 ^-6 (ppm). The NMR was measured using a (Bruker Avance III 400 and Bruker Avance 300) nuclear magnetic apparatus, and the solvent was deuterated dimethyl sulfoxide (DMSO-d ₆ ), deuterated chloroform (CDCl ₃ ), deuterated methanol (CD ₃ OD). ), the internal standard is tetramethylsilane (TMS).

The measurement of MS was carried out (Agilent 6120B (ESI) and Agilent 6120B (APCI)).

The HPLC was measured using an Agilent 1260 DAD high pressure liquid chromatograph (Zorbax SB-C18 100 x 4.6 mm).

Thin layer chromatography silica gel plate uses Yantai Yellow Sea HSGF254 or Qingdao GF254 silica gel plate. The specification of silica gel plate used for thin layer chromatography (TLC) is 0.15mm~0.20mm. The specification for thin layer chromatography separation and purification is 0.4mm. ~0.5mm.

Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as a carrier.

The known starting materials of the present invention may be synthesized by or according to methods known in the art, or may be purchased from Titan Technology, Anike Chemical, Shanghai Demo, Chengdu Kelon Chemical, Suiyuan Chemical Technology, and Belling Technology. And other companies.

The nitrogen atmosphere means that the reaction flask is connected to a nitrogen balloon having a volume of about 1 L.

The hydrogen atmosphere means that the reaction flask is connected to a hydrogen balloon of about 1 L volume.

The hydrogenation reaction is usually evacuated, charged with hydrogen, and operated three times.

Unless otherwise stated in the examples, the reaction was carried out under a nitrogen atmosphere.

Unless otherwise stated in the examples, the solution means an aqueous solution.

There is no particular description in the examples, and the reaction temperature is room temperature.

The room temperature is an optimum reaction temperature of 20 ° C to 30 ° C.

Example 1

2-[[3-[4-(2,3-dihydrobenzofuran-5-carbonyl)-1-piperidinyl]-2-oxo-pyrrolidin-1-yl]methyl]-3, 5,7,8-tetrahydropyrano[4,3-d]pyrimidin-4-one (Compound 1)

2-[[3-[4-(2,3-dihydrobenzofuran-5-carbonyl)-1-piperidyl]-2-oxo-pyrrolidin-1-yl]methyl]-3,5,7,8-tetrahydropyrano[4 ,3-d]pyrimidin-4-one

First step: 4-methoxy(methyl)carbamoyl)piperidine-1-carboxylic acid tert-butyl ester (1B)

Tert-butyl 4-(methoxy(methyl)carbamoyl)piperidine-1-carboxylate

N,N'-carbonyldiimidazole (21.0 g, 0.13 mol) was dissolved in anhydrous dichloromethane (200 mL), and 1-t-butoxycarbonylpiperidine-4-carboxylic acid (1A) (23.0 g, A 0.10 mmol) solution of anhydrous dichloromethane (100 mL) was stirred for 30 minutes, and dimethylhydroxylamine hydrochloride (12.7 g, 0.13 mol) was added portionwise, and reacted at room temperature for 3 hours. A citric acid solution (wt = 10%, 250 mL) was added to the reaction mixture, and the organic layer was washed with brine (300 mL) Formyl) piperidine-1-carboxylic acid tert-butyl ester (1B), pale yellow liquid (25.0 g, yield 92%).

^{_{1 H NMR (400MHz, CDCl 3}} ) δ4.39-4.24 (m, 3H), 3.71 (t, 2H), 3.65-3.63 (m, 2H), 3.55-3.53 (m, 2H), 3.38 (s, 3H ), 1.44 (s, 9H), 1.39 (m, 3H).

LCMS m/z = 217.0 [M-55].

Second step: 4-(2,3-dihydrobenzofuran-5-carbonyl)piperidine-1-carboxylic acid tert-butyl ester (1C)

Tert-butyl 4-(2,3-dihydrobenzofuran-5-carbonyl)piperidine-1-carboxylate

5-Bromo-2,3-dihydrobenzofuran (10.0 g, 50.2 mmol) was dissolved in anhydrous tetrahydrofuran (80 mL), cooled to -78 ° C under nitrogen, and n-butyl lithium (31.3 mL) was added dropwise. , 50.2 mmol), stirring for 30 minutes, adding a solution of tert-butyl 4-methoxy(methyl)carbamoyl)piperidine-1-carboxylate (1B) (3.1 g, 33.5 mmol) in tetrahydrofuran (200 mL) , liter at room temperature for 1 hour. A solution of saturated ammonium chloride (150 mL) was added, and the mixture was evaporated. Tert-butyl ester of acyl)piperidine-1-carboxylate (1C), pale yellow liquid (10.0 g, yield 91%).

^{_{1 H NMR (400MHz, CDCl 3}} ) δ7.84 (d, 1H), 7.79 (dd, 1H), 6.81 (d, 1H), 4.66 (t, 2H), 4.16 (m, 2H), 3.34 (m, 1H), 3.26 (t, 2H), 2.88 (m, 2H), 1.81 (m, 2H), 1.76-1.64 (m, 2H), 1.47 (s, 9H).

The third step: (2,3-dihydrobenzofuran-5-yl)(piperidin-4-yl)methanone (1D)

(2,3-dihydrobenzofuran-5-yl)(piperidin-4-yl)methanone

tert-Butyl 4-methoxy(methyl)carbamoyl)piperidine-1-carboxylate (1C) (4.0 g, 0.012 mol) was dissolved in anhydrous dichloromethane (50 mL) Trifluoroacetic acid (13.7 g, 0.12 mol) was added, and the mixture was reacted at room temperature for 4 hours. Water (50 mL) was added to the reaction mixture, and the mixture was evaporated. -Dihydrobenzofuran-5-yl)(piperidin-4-yl)methanone (1D), pale yellow solid (1.2 g, yield 43%).

^{_{1 H NMR (400MHz, CDCl 3}} ) δ7.84 (d, 1H), 7.79 (dd, 1H), 6.81 (d, 1H), 4.66 (t, 2H), 3.34 (m, 1H), 3.25 (t, 2H), 3.18 (m, 2H), 2.77 (td, 2H), 2.18 (s, 1H), 1.83 (m, 2H), 1.75-1.64 (m, 2H).

LCMS m/z = 232.0 [M + 1].

The fourth step: (2-oxopyrrolidin-3-yl)methanesulfonate (1F)

2-oxopyrrolidin-3-yl methanesulfonate

3-Hydroxy-2-pyrrolidone (1E) (5.0 g, 0.05 mol) was dissolved in anhydrous dichloromethane (50 mL), triethylamine (10.1 g, 0.10 mol) was added, cooled to 0 ° C, and added dropwise Sulfonyl chloride (6.8 g, 0.06 mol) was allowed to react to room temperature for 2 hours. Saturated brine (50 mL) was added to the mixture, and the mixture was evaporated. -yl)methanesulfonate (1F), white solid (4.0 g, yield 45%).

^{_{1 H NMR (400MHz, CDCl 3}} ) δ6.64 (s, 1H), 5.15 (t, 1H), 3.54-3.48 (m, 1H), 3.43-3.37 (m, 1H), 3.26 (s, 3H), 2.69-2.61 (m, 1H), 2.41-2.31 (m, 1H).

LCMS m/z = 179.9 [M + 1].

Step 5: 3-(4-(2,3-Dihydrobenzofuran-5-carbonyl)piperidin-1-yl)pyrrolidin-2-one (1G)

3-(4-(2,3-dihydrobenzofuran-5-carbonyl)piperidin-1-yl)pyrrolidin-2-one

(2-Oxopyrrolidin-3-yl)methanesulfonate (1F) (1.0 g, 5.58 mmol) was dissolved in acetonitrile (30 mL) and (2,3-dihydrobenzofuran-5-yl was added. (piperidin-4-yl)methanone (1D) (1.3 g, 5.58 mmol) and N,N-diisopropylethylamine (1.3 g, 5.58 mmol), and the mixture was heated to 85 ° C for 4 hours. The reaction mixture was concentrated to give purified crystalljjjjjjjjjjjjjjjjjjjj And furan-5-carbonyl)piperidin-1-yl)pyrrolidin-2-one (1G) as a pale yellow solid (0.73 g, yield 42%).

1H NMR (400MHz, CDCl3) δ 7.83 (d, 1H), 7.78 (dd, 1H), 6.80 (d, 1H), 5.99 (s, 1H), 4.65 (t, 2H), 3.50 (t, 1H) , 3.40-3.31 (m, 2H), 3.25 (m, 3H), 3.13 (m, 1H), 2.98 (m, 2H), 2.53 (m, 1H), 2.32 - 2.15 (m, 2H), 1.90 (m) , 4H).

LCMS m/z = 315.0 [M + 1].

Step 6: 2-[[3-[4-(2,3-Dihydrobenzofuran-5-carbonyl)-1-piperidinyl]-2-oxo-pyrrolidin-1-yl]methyl ]-3,5,7,8-tetrahydropyrano[4,3-d]pyrimidin-4-one (Compound 1)

3-(2,3-Dihydrobenzofuran-5-carbonyl)piperidin-1-yl)pyrrolidin-2-one (1G) (0.73 g, 2.32 mmol) was dissolved in tetrahydrofuran (20 mL) , 2-(Chloromethyl)-7,8-dihydro-3H-pyrano[4,3-d]pyrimidin-4(5H)-one (512 mg, 2.55 mol, synthesized according to WO2013008217 Intermediate 3) The method was prepared), cooled to 0 ° C, and under a nitrogen atmosphere, sodium hydride (278 mg, 6.96 mol) was added, and the mixture was heated at 70 ° C for 1 hour. The reaction solution was cooled to 0 ° C, and excess sodium hydride was evaporated, and the residue was purified by silica gel column chromatography (methanol: methylene chloride (v/v) = 1:1 to 5:95). Compound 2-[[3-[4-(2,3-dihydrobenzofuran-5-carbonyl)-1-piperidinyl]-2-oxo-pyrrolidin-1-yl]methyl]-3 , 5,7,8-tetrahydropyrano[4,3-d]pyrimidin-4-one (Compound 1), white solid (0.60 g, yield 55%).

^{_{1 H NMR (400MHz, CDCl 3}} ) δ7.83 (d, 1H), 7.78 (dd, 1H), 6.80 (d, 1H), 4.66 (t, 2H), 4.55 (s, 2H), 4.39 (s, 2H), 3.93 (t, 2H), 3.66 (t, 1H), 3.55-3.43 (m, 2H), 3.25 (m, 3H), 3.15 (m, 1H), 3.05 (m, 2H), 2.66 (t) , 2H), 2.54 (m, 1H), 2.34 (m, 1H), 2.20 (m, 1H), 1.91 (m, 4H).

LCMS m/z = 478.9 [M + 1].

Example 2

2-[[3-[4-(2,3-dihydrobenzofuran-5-carbonyl)-1-piperidinyl]-2-oxo-pyrrolidin-1-yl]methyl]-3H- Cyclohepta[d]imidazol-4-one (Compound 2)

2-[[3-[4-(2,3-dihydrobenzofuran-5-carbonyl)-1-piperidyl]-2-oxo-pyrrolidin-1-yl]methyl]- 3H-cyclohepta[d]imidazol-4-one

first step:

2-[3-[4-(2,3-dihydrobenzofuran-5-carbonyl)-1-piperidinyl]-2-oxo-pyrrolidin-1-yl]acetonitrile (2B)

2-[3-[4-(2,3-dihydrobenzofuran-5-carbonyl)-1-piperidyl]-2-oxo-pyrrolidin-1-yl]acetonitrile

3-3-4(2,3-Dihydrobenzofuran-5-carbonyl)piperidin-1-yl)pyrrolidin-2-one (1G) (0.53 g, 1.68 mmol) was dissolved in dry tetrahydrofuran (10 mL) To the reaction mixture, 2-bromoacetonitrile (0.61 g, 5.04 mmol) was added, and the mixture was cooled to 0 ° C under nitrogen atmosphere, and sodium hydride (0.12 g, 5.04 mol) was added to the reaction mixture, and the mixture was reacted at 0 ° C for 30 minutes. The reaction was quenched with EtOAc (EtOAc)EtOAc. Dichloromethane:methanol (v/v) = 49:1 to 1:0) to give the title compound 2-[3-[4-(2,3-dihydrobenzofuran-5-carbonyl)-1- Piperidinyl]-2-oxo-pyrrolidin-1-yl]acetonitrile (2B), yellow solid (0.30 g, yield 50%).

^{_{1 H NMR (400MHz, CDCl 3}} ) δ7.82 (s, 1H), 7.77 (dd, 1H), 6.80 (d, 1H), 4.65 (t, 2H), 4.27 (q, 2H), 3.60-3.46 ( m, 2H), 3.46-3.38 (m, 1H), 3.22 (m, 3H), 3.06 (dt, 1H), 2.92 (dd, 2H), 2.47-2.38 (m, 1H), 2.36-2.26 (m, 1H), 2.19-2.09 (m, 1H), 1.91-1.78 (m, 4H).

LCMS m/z = 354.1 [M + 1].

The second step:

2-[3-[4-(2,3-dihydrobenzofuran-5-carbonyl)-1-piperidinyl]-2-oxo-pyrrolidin-1-yl]acetamidine hydrochloride (2C )

2-[3-[4-(2,3-dihydrobenzofuran-5-carbonyl)-1-piperidyl]-2-oxo-pyrrolidin-1-yl]acetamidi ne Hydrochloride salt

2-[3-[4-(2,3-Dihydrobenzofuran-5-carbonyl)-1-piperidinyl]-2-oxo-pyrrolidin-1-yl]acetonitrile (2B) (0.3 g, 0.8m mol) dissolved in anhydrous methanol (6mL), added sodium methoxide (0.005g, 0.08mmol), reacted at 25 ° C for 2 hours, then added ammonium chloride (0.05g, 1.0mmol), after the addition, continue The reaction was carried out at 25 ° C for 2 hours. Concentrate to dryness under reduced pressure. EtOAc (EtOAc/EtOAc/EtOAc/EtOAc (v/v = 2/3, 20 mL) was washed, and the filtered cake was dried under reduced pressure to give the title compound 2-[3-[4-(2,3-dihydrobenzofuran-5-carbonyl)-1-piperidine. 2-Oxo-pyrrolidin-1-yl]acetamidine hydrochloride (2C), yellow solid (0.2 g, yield 58.1%).

^{1 H NMR (400MHz, DMSO)} δ9.04 (b, 2H), 7.88 (s, 1H), 7.81 (d, 1H), 7.36 (b, 2H), 6.86 (d, 1H), 4.63 (t, 2H ), 4.29 (d, 1H), 4.11 (m, 1H), 3.40-3.28 (m, 5H), 3.23 (t, 2H), 3.17 (d, 1H), 2.84-2.74 (m, 1H), 2.40- 2.30 (m, 1H), 2.23 - 2.13 (m, 1H), 2.13-1.99 (m, 1H), 1.82-1.66 (m, 2H), 1.66-1.46 (m, 2H).

LCMS m/z = 371.1 [M + 1].

third step:

2-[[3-[4-(2,3-dihydrobenzofuran-5-carbonyl)-1-piperidyl]-2-oxo-pyrrolidin-1-yl]methyl]-3H-cyclohepta[d]imidazol-4-one

2-[3-[4-(2,3-Dihydrobenzofuran-5-carbonyl)-1-piperidinyl]-2-oxo-pyrrolidin-1-yl]acetamidine hydrochloride ( 2C) (0.2 g, 0.49 mmol) was suspended in toluene (20 mL), 2-p-toluenesulfonyloxycycloheptenone (0.135 g, 0.49 mmol) was added, and tetrabutylammonium bromide (0.063 g, 0.19 mmol), an additional 30% aqueous solution of sodium hydroxide (0.078 g, 1.9 mmol) was added. After the addition, the mixture was heated to 30 ° C overnight. The liquid layer was separated, the toluene was collected, the methylene chloride (20 mL) was added, and the mixture was separated. The aqueous layer was extracted with a mixture of methanol/dichloromethane (v/v = 1/9, 20 mL). The residue was washed with EtOAc (EtOAc)EtOAc. -[4-(2,3-dihydrobenzofuran-5-carbonyl)-1-piperidinyl]-2-oxo-pyrrolidin-1-yl]methyl]-3H-cyclohepta[d Imidazol-4-one (Compound 2), pale yellow solid (0.01 g, yield 4%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.83 (s, 1H), 7.78-7.72 (t, 2H), 7.40-7.36 (t, 1H), 7.29 (d, 1H), 7.07-6.96 (m, 1H) ), 6.80 (d, 1H), 4.76 (s, 2H), 4.65 (t, 2H), 3.66-3.56 (m, 1H), 3.55-3.40 (m, 2H), 3.24 (t, 3H), 3.18- 3.08 (m, 1H), 3.05-2.90 (m, 2H), 2.55-2.45 (m, 1H), 2.35-2.25 (m, 1H), 2.20-2.09 (m, 2H), 1.92-1.80 (m, 4H) ).

LCMS m/z = 473.3 [M + 1].

Example 3

2-[[3-[4-(6-fluoro-2,3-dihydrobenzofuran-7-carbonyl)-1-piperidinyl]-2-oxo-pyrrolidin-1-yl]methyl ]-3,5,7,8-tetrahydropyrano[4,3-d]pyrimidin-4-one (Compound 3)

2-[[3-[4-(6-fluoro-2,3-dihydrobenzofuran-7-carbonyl)-1-piperidyl]-2-oxo-pyrrolidin-1-yl]methyl]-3,5,7,8 -tetrahydropyrano[4,3-d]pyrimidin-4-one

First step: 5-bromo-6-fluoro-2,3-dihydrobenzofuran (3B)

5-bromo-6-fluoro-2,3-dihydrobenzofuran

A solution of 48% hydrobromic acid (160 mL) was added to 6-fluoro-2,3-dihydrobenzofuran-5-amine (3A) (10.0 g, 65.30 mmol) at room temperature and stirred to dissolve. After cooling to 0 ° C, an aqueous solution (160 mL) of sodium nitrite (9.79 g, 142 mmol) was slowly added dropwise, and the mixture was dropped over 1 hour. The mixture was stirred at room temperature for 30 minutes, cooled again to 0 ° C, and cuprous bromide (14.20 g, 98.99 mmol) was added portionwise. After the addition, the mixture was reacted at room temperature for 40 minutes, and heated to 140 ° C for 1.5 hours. After cooling to room temperature, it was extracted with dichloromethane (100 mL×3), and the organic phase was combined, and the organic phase was washed with water (200 mL×2), dried over anhydrous sodium sulfate and evaporated. Ethyl ester (v/v) = 100:0 to 20:1) The title compound 5-bromo-6-fluoro-2,3-dihydrobenzofuran (Compound 3B) was obtained as a white solid (4.50 g, yield 31.8%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.32 - 7.23 (m, 1H), 6.57 (d, 1H), 4.62 (t, 2H), 3.22-3.11 (m, 2H).

Step 2: 4-(6-Fluoro-2,3-dihydrobenzofuran-7-carbonyl)piperidine-1-carboxylic acid tert-butyl ester (3C)

Tert-butyl 4-(6-fluoro-2,3-dihydrobenzofuran-7-carbonyl)piperidine-1-carboxylate

Slowly add n-butyllithium to a solution of 5-bromo-6-fluoro-2,3-dihydrobenzofuran (3B) (2.17 g, 10.0 mmol) in tetrahydrofuran (30 mL) at -78 °C. Alkane solution (5.00 mL, 12.5 mmol), stirred for 30 min, added 4-[methoxy(methyl)carbamoyl]piperidine-1-carboxylic acid tert-butyl ester (2.50 g, 9.18 mmol). 1 hour. After adding 20 mL of a saturated aqueous solution of ammonium chloride and ethyl acetate (100 mL × 2), the organic phase was combined, dried over anhydrous sodium sulfate and evaporated to dryness. The title compound 4-(6-fluoro-2,3-dihydrobenzofuran-7-carbonyl)piperidine-1-carboxylic acid tert-butyl ester (3C) was obtained as pale yellow solid. (1.90 g, yield 54.4%).

^{_{1 H NMR (400MHz, CDCl 3}} ) δ7.68 (d, 1H), 6.50 (d, 1H), 4.69 (t, 2H), 4.15-4.09 (m, 2H), 3.27-3.15 (m, 3H), 2.90-2.85 (m, 2H), 1.87 (d, 2H), 1.66-1.60 (m, 2H), 1.46 (s, 9H).

LCMS m/z = 372.1 [M+23].

The third step: (6-fluoro-2,3-dihydrobenzofuran-7-yl)(piperidin-4-yl)methanone (3D)

(6-fluoro-2,3-dihydrobenzofuran-7-yl)(piperidin-4-yl)methanone

tert-Butyl 4-(6-fluoro-2,3-dihydrobenzofuran-7-carbonyl)piperidine-1-carboxylate (3C) (1.90 g, 5.4 mmol) in dichloromethane (8 mL) Trifluoroacetic acid (4 mL) was added to the solution, and the mixture was reacted for 1 hour. The aqueous solution was extracted with aq. Yield 81.4%).

LCMS m/z = 250.1 [M + 1].

Fourth step: 3-[4-(6-fluoro-2,3-dihydrobenzofuran-7-carbonyl)-1-piperidinyl]pyrrolidin-2-one (3E)

3-[4-(6-fluoro-2,3-dihydrobenzofuran-7-carbonyl)-1-piperidyl]pyrrolidin-2-one

Add (6-fluoro-2,3-dihydrobenzofuran-7-yl)(piperidin-4-yl)methanone (3D) (0.5 g, 2.0 mmol) in 20 mL of acetonitrile at room temperature (2) -Oxopyrrol-3-yl)methanesulfonate (0.7 g, 4.0 mmol) and diisopropylethylamine (0.5 g, 4.0 mmol). After the addition, the mixture was heated to 80 ° C for 5 hours. After adding 30 mL of water and ethyl acetate (100 mL × 2), the organic phase was combined, and the organic phase was washed with water (100 mL × 2), dried over anhydrous sodium sulfate The residue was purified by silica gel column chromatography (dichloromethanol:methanol (v/v) = 100:1 to 30:1) to give the title compound 3-[4-(6-fluoro-2,3-dihydrobenzofuran) -7-carbonyl)-1-piperidinyl]pyrrolidin-2-one (3E), pale yellow solid (0.35 g, yield 52.2%).

^{_{1 H NMR (400MHz, CDCl 3}} ) δ7.16-7.13 (m, 1H), 6.56 (dd, 1H), 6.05 (s, 1H), 4.714.64 (m, 2H), 3.47-3.43 (t, 1H ), 3.38-3.24 (m, 2H), 3.19-3.14 (q, 2H), 3.05-2.99 (m, 2H), 2.92-2.85 (m, 1H), 2.80-2.74 (m, 1H), 2.45-2.37 (m, 1H), 2.29-2.09 (m, 2H), 1.99-1.88 (m, 2H), 1.86-1.71 (m, 2H).

LCMS m/z = 333.3 [M + 1].

Step 5: 2-[[3-[4-(6-Fluoro-2,3-dihydrobenzofuran-7-carbonyl)-1-piperidinyl]-2-oxo-pyrrolidine-1- Methyl]-3,5,7,8-tetrahydropyrano[4,3-d]pyrimidin-4-one (Compound 3)

In the compound 3-[4-(6-fluoro-2,3-dihydrobenzofuran-7-carbonyl)-1-piperidinyl]pyrrolidin-2-one (3D) (0.35 g, 1.05 mmol) To the tetrahydrofuran (15 mL) was added 2-(chloromethyl)-7,8-dihydro-3H-pyrano[4,3-d]pyrimidin-4(5H)-one (0.25 g, 1.26 mmol, Referring to the synthesis method of Intermediate 3 of WO2013008217, after the addition, sodium hydride (0.05 g, 1.26 mmol) was added, and the reaction was heated to 80 ° C for 2 hours. The reaction was quenched by the addition of 5 mL of EtOAc (EtOAc) (EtOAc (EtOAc) [4-(6-Fluoro-2,3-dihydrobenzofuran-7-carbonyl)-1-piperidinyl]-2-oxo-pyrrolidin-1-yl]methyl]-3,5, 7,8-Tetrahydropyrano[4,3-d]pyrimidin-4-one (Compound 3), white solid (0.03 g, yield 5.7%).

^{_{1 H NMR (400MHz, CDCl 3}} ) δ7.18-7.12 (m, 1H), 6.56 (dd, 1H), 4.70-4.66 (t, 2H), 4.54 (s, 2H), 4.40-4.38 (m, 2H ), 3.94-3.91(t,2H), 3.63-3.59(t,1H), 3.50-3.46(m,2H), 3.19-3.15(t,2H),3.07-2.98(m,2H),2.92-2.87 (m, 1H), 2.82-2.77 (m, 1H), 2.66-2.64 (t, 2H), 2.45-2.37 (m, 1H), 2.31-2.20 (m, 1H), 2.17-2.07 (m, 1H) , 1.94-1.89 (m, 2H), 1.83-1.69 (m, 2H).

LCMS m/z = 497.1 [M + 1].

Example 4

2-Chloro-5-[[3-[4-(2,3-dihydrobenzofuran-5-carbonyl)-1-piperidinyl]-2-oxopyrrolidin-1-yl]methyl] Benzoonitrile (Compound 4)

2-chloro-5-[[3-[4-(2,3-dihydrobenzofuran-5-carbonyl)-1-piperidyl]-2-oxo-pyrrolidin-1-yl]methyl]benzonitrile

3-(4-(2,3-Dihydrobenzofuran-5-carbonyl)piperidin-1-yl)pyrrolidin-2-one (1G) (0.314 g, 1.0 mmol) was dissolved in THF (10 mL) To the mixture, 5-bromomethyl-2-chlorobenzonitrile (276 mg, 1.2 mol) was added, and the mixture was cooled to 0 ° C, and then filtered, and then evaporated, and then, then, sodium hydride (120 mg, 3.0 mol) was added, and the mixture was heated to 70 ° C for 1 hour. The reaction solution was cooled to 0 ° C, and excess sodium hydride was evaporated, and the residue was evaporated. The title compound 2-chloro-5-[[3-[4-(2,3-dihydrobenzofuran-5-carbonyl)-1-piperidinyl]-2-oxopyrrolidin-1-yl ]Methyl]benzonitrile (Compound 4), pale yellow solid (0.60 g, yield 54.5%).

^{_{1 H NMR (400MHz, CDCl 3}} ) δ7.83 (s, 1H), 7.78 (dd, 1H), 7.55 (d, 1H), 7.47 (q, 2H), 6.81 (d, 1H), 4.66 (t, 2H), 4.51 (d, 1H), 4.40 (d, 1H), 3.57 (t, 1H), 3.21 (ddd, 5H), 3.11 (s, 1H), 3.00 (s, 2H), 2.48 (s, 1H) ), 2.24 (s, 1H), 2.07 (s, 1H), 1.90 (s, 4H).

LCMS m/z = 464.3 [M + 1].

Example 5

2-((3-(4-(2,3-dihydrobenzofuran-5-carbonyl)piperidin-1-yl)-2-oxopyrrolidin-1-yl)methyl)-6,7 -dihydro-3H-cyclopenta[d]pyrimidin-4(5H)-one (Compound 5)

2-((3-(4-(2,3-dihydrobenzofuran-5-carbonyl)piperidin-1-yl)-2-oxopyrrolidin-1-yl)methyl)-6,7-dihydro-3H-cyclopenta[d] Pyrimidin-4(5H)-one

First step: 2-(chloromethyl)-6,7-dihydro-3H-cyclopenta[d]pyrimidin-4(5H)-one (5B)

2-(chloromethyl)-6,7-dihydro-3H-cyclopenta[d]pyrimidin-4(5H)-one

Triethylamine (4.20 g, 41.5 mmol) and 2-chloroethyl hydrazine (3.00) were added to a solution of methyl 2-oxocyclopentanecarboxylate (5A) (7.10 g, 49.9 mmol) in methanol (150 mL). g, 0.649 mmol), and reacted at room temperature for 5 hours after the addition. After completion of the reaction, water (200 mL) was added, and the mixture was evaporated. v/v)=100:1 to 30:1) to give the title compound 2-(chloromethyl)-6,7-dihydro-3H-cyclopenta[d]pyrimidin-4(5H)-one (5B) , pale yellow solid (1.00 g, yield 10.8%).

¹ H NMR (400 MHz, DMSO) δ 12.58 (s, 1H), 4.46 (s, 2H), 2.76 (t, 2H), 2.63 (t, 2H), 2.04-1.90 (m, 2H).

LCMS m/z = 185.1 [M + 1].

Second step: 2-((3-(4-(2,3-dihydrobenzofuran-5-carbonyl)piperidin-1-yl)-2-oxopyrrolidin-1-yl)methyl) -6,7-dihydro-3H-cyclopenta[d]pyrimidin-4(5H)-one (compound 5)

Tetrahydrofuran in the compound 3-(4-(2,3-dihydrobenzofuran-5-carbonyl)piperidin-1-yl)pyrrolidin-2-one (1G) (0.314 g, 1.00 mmol) (10 mL) was added 2-(chloromethyl)-6,7-dihydro-3H-cyclopenta[d]pyrimidin-4(5H)-one (5B) (0.400 g, 2.17 mmol). Thereafter, sodium hydride (0.100 g, 4.17 mmol) was added in portions, and the mixture was heated to 80 ° C for 1 hour. After cooling to zero, methanol (5 mL) was added to quench the reaction, and the residue was concentrated with silica gel. Purification by spectral separation (dichloromethane:methanol (v/v) = 100:1 to 30:1) to give the title compound 2-((3-(4-(2,3-dihydrobenzofuran-5-carbonyl)) Piperidin-1-yl)-2-oxopyrrolidin-1-yl)methyl)-6,7-dihydro-3H-cyclopenta[d]pyrimidin-4(5H)-one (Compound 5) White solid (80 mg, yield 17.0%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 11.17 (s, 1H), 7.83 (s, 1H), 7.79 (d, 1H), 6.80 (d, 1H), 4.66 (t, 2H), 4.40 (dd, 2H) ), 3.64 (t, 1H), 3.45 (t, 2H), 3.25 (t, 3H), 3.05 (d, 3H), 2.89-2.74 (m, 4H), 2.48 (s, 1H), 2.32 (s, 1H), 2.07 (m, 3H), 1.87 (s, 4H).

LCMS m/z = 463.3 [M + 1].

Example 6

2-(3-(4-(2,3-dihydrobenzofuran-5-carbonyl)piperidin-1-yl)-2-oxopyrrolidin-1-yl)-N-(5,6- Dihydrothiazole [2,3-c][1,2,4]triazol-3-yl)acetamide (Compound 6)

2-(3-(4-(2,3-dihydrobenzofuran-5-carbonyl)piperidin-1-yl)-2-oxopyrrolidin-1-yl)-N-(5,6-dihydrothiazolo[2,3-c] [1,2,4]triazol-3-yl)acetamide

First step: methyl 2-(3-(4-(2,3-dihydrobenzofuran-5-carbonyl)piperidin-1-yl)-2-oxopyrrolidin-1-yl)acetate ( 6B)

Methyl 2-(3-(4-(2,3-dihydrobenzofuran-5-carbonyl)piperidin-1-yl)-2-oxopyrrolidin-1-yl)acetate

3-(4-(2,3-Dihydrobenzofuran-5-carbonyl)piperidin-1-yl)pyrrolidin-2-one (1G) (1.0 g, 3.18 mmol) was dissolved in tetrahydrofuran (10 mL) In addition, ethyl bromoacetate (6A) (1.06g, 6.36mol) was added and cooled to 0 ° C. The sodium hydride (153 m g, 6.36 mol) was added and the mixture was reacted at 0 ° C for 0.5 hour. The reaction solution was cooled to 0 ° C, and excess sodium hydride was evaporated to dryness, and the residue was evaporated to dryness (yield: methylene chloride (v/v) (v/v) = 1:1 to 5:95). The title compound 2-(3-(4-(2,3-dihydrobenzofuran-5-carbonyl)piperidin-1-yl)-2-oxopyrrolidin-1-yl)acetic acid methyl ester (6B) ), yellow oil (0.370 g, yield 30.1%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.82 (s, 1H), 7.77 (dd, 1H), 6.80 (d, 1H), 4.65 (t, 2H), 4.13 (d, 1H), 4.02 (d, 1H) ), 3.74 (s, 3H), 3.65-3.55 (m, 1H), 3.48-3.34 (m, 2H), 3.31-3.16 (m, 3H), 3.11 (d, 1H), 3.00 (d, 2H), 2.58-2.43 (m, 1H), 2.28 (ddd, 1H), 2.21-2.07 (m, 1H), 1.87 (t, 4H).

LCMS m/z = 387.1 [M + 1].

Second step: 2-(3-(4-(2,3-dihydrobenzofuran-5-carbonyl)piperidin-1-yl)-2-oxopyrrolidin-1-yl)acetic acid (6C)

2-(3-(4-(2,3-dihydrobenzofuran-5-carbonyl)piperidin-1-yl)-2-oxopyrrolidin-1-yl)acetic acid

Methyl 2-(3-(4-(2,3-dihydrobenzofuran-5-carbonyl)piperidin-1-yl)-2-oxopyrrolidin-1-yl)acetate (6B) ( 0.370 g, 0.958 mmol) was dissolved in methanol (8 mL), and aqueous solution (2 mL) of sodium hydroxide (0.0153 g, 0.383 mmol) was added and allowed to react at room temperature for 1 hour. The reaction solution was adjusted to pH 4 with hydrochloric acid (1 mol/L) and concentrated under reduced pressure. Dichloromethane (10 mL) and a saturated aqueous solution (3 mL) were added to the mixture, and the mixture was evaporated. 2-(3-(4-(2,3-dihydrobenzofuran-5-carbonyl)piperidin-1-yl)-2-oxopyrrolidin-1-yl)acetic acid (6C), white solid ( 0.310 g, yield 86.9%).

¹ H NMR (400 MHz, DMSO) δ 13.03 (s, 1H), 7.93 (s, 1H), 7.86 (d, 1H), 6.88 (d, 1H), 4.65 (t, 2H), 4.36 (s, 1H) , 4.05 (s, 2H), 3.77 (d, 1H), 3.68 (s, 1H), 3.46 (dd, 3H), 3.24 (t, 2H), 3.15 (d, 1H), 2.49-2.28 (m, 3H) ), 1.93 (d, 4H).

LCMS m/z = 373.3 [M + 1].

The third step: 5-((2-bromoethyl)thio)-4H-1,2,4-triazole-3-ammonia (6E)

5-((2-bromoethyl)thio)-4H-1,2,4-triazol-3-amine

The starting material 3-amino-5-mercapto-1,2,4-triazole (6D) (2.320 g, 19.98 mmol) was dissolved in methanol (25 mL) Sodium methoxide (1.079 g, 19.98 mmol) was added, and 1,2-dibromoethane (30.02 g, 159.8 mmol) was added, and the mixture was reacted at room temperature for 3.5 hours. After the reaction was completed, the title compound (5-(2-bromoethyl)) was obtained. Thio)-4H-1,2,4-triazole-3-ammonia (6E), white solid (2.1 g, yield 47.1%).

¹ H NMR (400 MHz, DMSO) δ 12.03 (s, 1H), 6.11 (s, 2H), 3.73 - 3.65 (m, 2H), 3.37 (m, 2H).

LCMS m/z = 224.9 [M + 1].

Step 4: 3-Amino-5,6-dihydrothiazolo[2,3-C][1,2,4]triazole hydrobromide (6F)

5,6-dihydrothiazolo[2,3-c][1,2,4]triazol-3-aminehydrobromide

The starting material 5-((2-bromoethyl)thio)-4H-1,2,4-triazole-3-ammonia (6E) (2.1 g, 9.41 mmol) was dissolved in N,N-dimethyl In formamide (10 mL), the mixture was heated to 50 ° C for 3.5 hours. After the reaction, a white solid precipitated and the solid was filtered to give the title compound 3-amino-5,6-dihydrothiazolo[2,3-C][1,2,4]triazole hydrobromide (6F ), white solid (1.2 g, yield 57%).

¹ H NMR (400 MHz, DMSO) δ 13.47 (s, 1H), 8.49 (s, 2H), 4.20 - 4.13 (m, 2H), 4.09 - 4.02 (m, 2H).

LCMS m/z = 143.2 [M + 1].

The fifth step: 2-(3-(4-(2,3-dihydrobenzofuran-5-carbonyl)piperidin-1-yl)-2-oxopyrrolidin-1-yl)-N-( 5,6-dihydrothiazole [2,3-c][1,2,4]triazol-3-yl)acetamide (compound 6)

2-(3-(4-(2,3-Dihydrobenzofuran-5-carbonyl)piperidin-1-yl)-2-oxopyrrolidin-1-yl)acetic acid (6C) (0.300 g) , 0.806 mmol) dissolved in anhydrous dichloromethane (10 mL), added 3-amino-5,6-dihydrothiazolo[2,3-C][1,2,4]triazole hydrobromide (6F) (0.198 g, 0.886 mmol) and diisopropylethylamine (0.416 g, 3.22 mmol), with O-(7-nitrobenzotriazole)-N,N,N',N '-Tetramethylurea hexafluorophosphate (HATU) (0.337g, 0.886 Methyl) was reacted at room temperature for 48 hours. The reaction mixture was concentrated, and the residue was purified (jjjjjjjjjjj Hydrobenzofuran-5-carbonyl)piperidin-1-yl)-2-oxopyrrolidin-1-yl)-N-(5,6-dihydrothiazole[2,3-c][1,2 , 4] Triazol-3-yl)acetamide (Compound 6), white solid (0.070 g, yield 17%).

¹ H NMR (400 MHz, DMSO) δ 9.21 (s, 1H), 7.88 (s, 1H), 7.84 - 7.76 (m, 1H), 6.85 (d, 1H), 4.63 (t, 2H), 4.17 - 3.95 ( m, 6H), 3.57 (dd, 2H), 3.41-3.33 (m, 2H), 3.23 (t, 2H), 3.11 (dd, 2H), 2.81 (s, 2H), 2.16 (d, 1H), 1.99 (s, 1H), 1.73 (s, 2H), 1.57 (s, 2H).

LCMS m/z = 497.1 [M + 1].

Example 7

2-((3-(4-(6-fluoro-2,3-dihydrobenzofuran-5-carbonyl)piperidin-1-yl)-2-oxopyrrolidin-1-yl)methyl) -7,8-dihydro-3H-pyrano[4,3-d]pyrimidin-4(5H)-one (compound 7)

2-((3-(4-(6-fluoro-2,3-dihydrobenzofuran-5-carbonyl)piperidin-1-yl)-2-oxopyrrolidin-1-yl)methyl)-7,8-dihydro-3H- Pyrano[4,3-d]pyrimidin-4(5H)-one

First step: 4-(6-fluoro-2,3-dihydrobenzofuran-5-carbonyl)piperidine-1-carboxylic acid tert-butyl ester (7C)

Tert-butyl4-(6-fluoro-2,3-dihydrobenzofuran-5-carbonyl)piperidine-1-carboxylate

Slowly add n-butyllithium to a solution of 5-bromo-6-fluoro-2,3-dihydrobenzofuran (3B) (2.17 g, 10.0 mmol) in tetrahydrofuran (30 mL) at -78 °C. Alkane solution (5.00 mL, 12.5 mmol), stirred for 30 min, added 4-[Methoxy(methyl)carbamoyl]piperidine-1-carboxylic acid tert-butyl ester (2.50 g, 9.18 mmol, CAS: 139290-70 -3), continue to react for 1 hour after the addition. A saturated aqueous solution of ammonium chloride (20 mL) was added and ethyl acetate (100 mL×2) The organic phases were combined, dried over anhydrous sodium sulfate and dried. The residue was purified by silica gel column chromatography (EtOAc (EtOAc:EtOAc) tert-Butyl carbonyl)piperidine-l-carboxylate (7C), pale yellow solid (1.10 g, yield 31.5%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.68 (d, 1H), 6.50 (d, 1H), 4.69 (t, 2H), 4.12 (m, 2H), 3.21 (m, 3H), 2.96 - 2.80 (m) , 2H), 1.87 (d, 2H), 1.61 (m, 2H), 1.46 (s, 9H).

LCMS m/z = 372.1 [M+23].

Second step: (6-fluoro-2,3-dihydrobenzofuran-5-yl)(piperidin-4-yl)methanone (7D)

(6-fluoro-2,3-dihydrobenzofuran-5-yl)(piperidin-4-yl)methanone

tert-Butyl 4-(6-fluoro-2,3-dihydrobenzofuran-5-carbonyl)piperidine-1-carboxylate (7C) (1.10 g, 3.15 mmol) in dichloromethane (8 mL) Trifluoroacetic acid (4 mL) was added to the solution, and the mixture was reacted for 1 hour. The organic layer was extracted with EtOAc (3 mL) (EtOAc) Benzofuran-5-yl)(piperidin-4-yl)methanone (7D), white solid (0.750 g, yield 95.6%).

¹ H NMR (400 MHz, DMSO) δ 7.68 (d, 1H), 6.78 (d, 1H), 4.69 (t, 2H), 3.28 (dd, 1H), 3.18 (dd, 4H), 2.80 (m, 2H) , 1.83 (d, 2H), 1.55 (m, 2H).

LCMS m/z = 250.1 [M + 1].

Third step: 3-(4-(6-fluoro-2,3-dihydrobenzofuran-5-carbonyl)piperidin-1-yl)pyrrolidin-2-one (7E)

3-(4-(6-fluoro-2,3-dihydrobenzofuran-5-carbonyl)piperidin-1-yl)pyrrolidin-2-one

Add (6-fluoro-2,3-dihydrobenzofuran-5-yl)(piperidin-4-yl)methanone (7D) (0.400, 1.60 mmol) in EtOAc (20 mL) 2-oxopyrrol-3-yl)methanesulfonate (1F) (1.20 g, 6.70 mmol) and diisopropylethylamine (0.850 g, 6.58 mmol) were added, and the mixture was warmed to 80 ° C for 5 hours. Water (30 mL), EtOAc (EtOAc) The residue was purified by silica gel column chromatography (dichloromethanol:methanol (v/v) = 100:1 to 30:1) to give the title compound 3-(4-(6-fluoro-2,3-dihydrobenzofuran) 5-5-Carbonylpiperidin-1-yl)pyrrolidin-2-one (7E), pale yellow solid (0.400 g, yield 75.0%).

¹ H NMR (400 MHz, DMSO) δ 8.17 (d, 1H), 7.66 (d, 1H), 6.76 (d, 1H), 4.68 (t, 2H), 3.37-3.22 (m, 3H), 3.23-3.08 ( m,3H), 3.02 (m, 2H), 2.74 (s, 1H), 2.29 (d, 1H), 2.17-2.01 (m, 2H), 1.76 (m, 2H), 1.59-1.38 (m, 2H) .

LCMS m/z = 333.3 [M + 1].

Fourth step: 2-((3-(4-(6-fluoro-2,3-dihydrobenzofuran-5-carbonyl)piperidin-1-yl)-2-oxopyrrolidin-1-yl )methyl)-7,8-dihydro-3H-pyrano[4,3-d]pyrimidin-4(5H)-one (compound 7)

2-((3-(4-(6-fluoro-2,3-dihydrobenzofuran-5-carbonyl)piperidin-1-yl)-2-oxopyrrolidin-1-y l)methyl)-7,8-dihydro-3H -pyrano[4,3-d]pyrimidin-4(5H)-one

Compound 0-(4-(6-fluoro-2,3-dihydrobenzofuran-5-carbonyl)piperidin-1-yl)pyrrolidin-2-one (7E) (0.334 g, 1.00) To the tetrahydrofuran (15 mL) was added 2-(chloromethyl)-7,8-dihydro-3H-pyrano[4,3-d]pyrimidin-4(5H)-one (0.500 g, 2.49 mmol) Refer to WO2013008217 Intermediate 3 synthesis method to prepare), after the addition, sodium hydride (0.100 g, 4.17 mmol) was added in portions, and the reaction was heated to 80 ° C for 1 hour. After cooling to 0 ° C, methanol (5 mL) was added and the residue was evaporated. mjjjjjjjjjjjj 3-(4-(6-Fluoro-2,3-dihydrobenzofuran-5-carbonyl)piperidin-1-yl)-2-oxopyrrolidin-1-yl)methyl)-7,8 -Dihydro-3H-pyrano[4,3-d]pyrimidin-4(5H)-one (Compound 7), white solid (110 mg, yield 22.0%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.67 (d, 1H), 6.48 (d, 1H), 4.68 (t, 2H), 4.55 (s, 2H), 4.39 (d, 2H), 3.93 (t, 2H) ), 3.61 (t, 1H), 3.51-3.42 (m, 2H), 3.38 (dd, 1H), 3.19 (t, 2H), 3.09 (d, 2H), 2.94 (d, 1H), 2.66 (t, 2H), 2.38 (t, 1H), 2.32-2.21 (m, 1H), 2.20-2.09 (m, 1H), 1.93 (d, 2H), 1.79 (dd, 2H).

LCMS m/z = 497.2 [M + 1].

Test case

Test Example 1: Inhibition of Wnt signaling pathway by compounds tested by reporter gene assay

Super-TOpFlash (STF) is a luciferase reporter gene system that specifically responds to the wnt signaling pathway. The Super-TOpFlash (STF) plasmid was transferred into HEK293 cells, and the reporter gene detection method was used to reflect the inhibition of the activity of the compound on the intracellular wnt signaling pathway. The continuously cultured HEK293 cells were cultured in a six-well plate and incubated at 37 ° C in a 5% CO ₂ carbon dioxide incubator overnight; when the cells reached 90% confluence, the reporter plasmid was transfected into the cells using Lipofectamine 2000 (Invitrogen) transfection reagent. After 4.5 hours, the cells were plated into 96-well plates at 10,000 cells per well, and cultured overnight at 37 ° C in a 5% CO ₂ carbon dioxide incubator, and the test compound was added the next day. The compound was dissolved in DMSO, the highest concentration was 10 μM, and the cell culture medium was diluted 5 times, 10 concentrations, and 50% wnt3A conditioned medium was added to each well, and cultured in a 5% CO ₂ carbon dioxide incubator for 24 hours at 37 ° C, using fluorescence. The fluorescence intensity was measured by a Luciferase Assay System Freezer Pack (Promega, Cat. #E4530) and a Perkin Elmer Envision plate reader, and the IC ₅₀ value was calculated. The test results are shown in Table 1.

Table 1: Inhibitory activity of compounds on the activity of wnt signaling pathway

化合物编号Compound number	STF IC₅₀(nM)STF IC ₅₀ (nM)
化合物编号Compound number	STF IC₅₀(nM)STF IC ₅₀ (nM)	11	8.278.27
22	1.511.51	11	8.278.27
22	1.511.51	33	66.1766.17
44	24.9924.99	33	66.1766.17
44	24.9924.99	55	5.745.74
66	21.6921.69	55	5.745.74
66	21.6921.69	77	4.624.62

Conclusion: The compounds of the present invention have significant inhibitory activity on the wnt signaling pathway.

Claims

A compound of the formula (A), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein:

R 1 is selected from a C 6-10 carbocyclic ring or a 5- to 10-membered heterocyclic ring containing at least 1 to 4 hetero atoms selected from N, O or S, optionally further a carbocyclic or heterocyclic ring. From 0 to 4 selected from the group consisting of F, Cl, Br, I, (=O), cyano, hydroxy, carboxy, amino, nitro, R 1a , NR 1a R 1b , COR 1a , CONR 1a R 1b or NR 1a Substituted by a substituent of COR 1b ;

R 1a and R 1b are each independently selected from H, C 1-6 alkyl or C 1-6 alkoxy, and the alkyl and alkoxy are optionally further from 0 to 4 selected from F, Cl, Br. Substituted by a substituent of I, cyano, hydroxy, carboxy, amino or nitro;

R 2 , R 3 and R 4 are each independently selected from the group consisting of H, F, Cl, Br, I, cyano, hydroxy, carboxy, amino, nitro, C 1-6 alkyl or C 1-6 alkoxy, The alkyl group and the alkoxy group are optionally further substituted with from 0 to 4 substituents selected from the group consisting of F, Cl, Br, I, cyano, hydroxy, carboxy, amino or nitro;

R 5 , R 6 , R 7 and R 8 are each independently selected from H or C 1-6 alkyl.
The compound, stereoisomer or pharmaceutically acceptable salt thereof according to claim 1, wherein the compound is selected from the group consisting of the compound of the formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof:
The compound according to claim 1 or 2, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein:

R 1 is selected from one of the following structures substituted or unsubstituted:

When substituted, optionally further 1 to 4 are selected from the group consisting of F, Cl, Br, I, (=O), cyano, hydroxy, carboxy, amino, nitro, R 1a , NR 1a R 1b or COR 1a Substituted by a substituent;

R 1a and R 1b are each independently selected from H, C 1-4 alkyl or C 1-4 alkoxy, and the alkyl and alkoxy groups are further further selected from 0 to 4 selected from F, Cl, Br. Substituted by a substituent of a cyano group or a hydroxyl group.
A compound according to claim 3, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein:

R 1 is selected from one of the following structures substituted or unsubstituted:

When substituted, optionally further 1 to 4 are selected from the group consisting of F, Cl, Br, cyano, hydroxy, carboxy, amino, nitro, methyl, ethyl, trifluoromethyl, methoxy or ethoxy Substituted by a substituent.
A compound according to claim 4, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein:

R 1 is selected from one of the following structures substituted or unsubstituted:

When substituted, optionally further substituted with from 1 to 4 substituents selected from the group consisting of F, Cl, cyano, methyl, trifluoromethyl or methoxy;

R 2 , R 3 and R 4 are each independently selected from H, F, Cl or Br;

R 5 , R 6 , R 7 and R 8 are each independently selected from H.
The compound according to claim 4, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of a compound represented by the formula (II), a stereoisomer thereof or a pharmaceutically acceptable salt thereof. among them:
The compound according to claim 6, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein

R 2 , R 3 and R 4 are each independently selected from H, F, Cl, Br, trifluoromethyl, methyl, ethyl, methoxy or ethoxy;

R 5 , R 6 , R 7 and R 8 are each independently selected from H, methyl or ethyl.
A compound according to claim 1, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein the compound Selected from one of the following structures:
A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 1 to 8, a stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable Carrier or excipient.
Use of the compound according to any one of claims 1 to 8, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 9 for the preparation of a medicament for treating cancer.
A method of treating cancer, the method comprising administering the compound of any one of claims 1-8, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 9. Things.