WO2018228369A1 - Heteroarylpyrimidinone derivative, method for preparation thereof, and medicinal use thereof - Google Patents

Abstract

The present invention relates to a heteroarylpyrimidinone derivative represented by formula (I), a method for preparation thereof, and a use thereof as a therapeutic agent, in particular a use as an acetyl-CoA carboxylase (ACC) inhibitor, the definitions of the substituents in formula (I) being the same as the definitions in the description.

Description

Heteroarylpyrimidinone derivative, preparation method thereof and use thereof in medicine

This application claims to be submitted to the Chinese Patent Office on June 15, 2017, the application number is 201710450023.6, and the Chinese name of the invention entitled "heteroarylpyrimidinone derivatives, their preparation methods and their use in medicine" is preferred. The entire contents are hereby incorporated by reference.

Technical field

The present invention relates to a heteroarylpyridinone derivative, a process for the preparation thereof, a pharmaceutical composition containing the same, and its use as a therapeutic agent, particularly as an acetyl-CoA carboxylase (ACC) inhibitor.

Background technique

Acetyl-CoA carboxylase (ACC) is one of the important proteins involved in fatty acid metabolism. It uses biotin as a coenzyme to catalyze the production of malonyl-CoA from acetyl-CoA. This irreversible reaction (malonyl-CoA), in turn, provides a substrate for the synthesis of subsequent fatty acids or modulates the fatty acid oxidation signal, which is the first step of fatty acid metabolism and is a rate limiting step. The catalytic reaction can be divided into two steps, depending on the biotin carboxylase (BC) and carboxyltransferase (CT) activities of ACC.

There are two subtypes of ACC in human body, namely ACC1 and ACC2, which are separately encoded by two genes, ACACA and ACACB. There are differences in tissue distribution and intracellular distribution. ACC1 is a cytosolic enzyme that is mainly expressed in fat synthesis tissues (such as fat and breast tissue); ACC2 is localized in the mitochondrial membrane and is mainly enriched in oxidized tissues (such as the heart). In skeletal muscle, both are expressed at high levels in the liver. Therefore, ACC1 is mainly involved in the regulation of fatty acid synthesis, and ACC2 is mainly responsible for the regulation of the oxidation process of fatty acids. The activity of ACC is regulated by a variety of proteins, cytokines, endocrine hormones and receptors. Among them, AMPK is the main substance regulating ACC activity, which can inhibit the activity by direct phosphorylation of ACC; and protein phosphorylase 2 can dephosphorylate ACC, thereby enhancing the effect of ACC. Under physiological conditions, free fatty acids synthesized in the cytosol are transported to the mitochondria via the mitochondrial membrane on carnitine palmitoyltransferase 1 (CPT1) for oxidative energy supply. Malonyl-CoA in the cytosol allosterically inhibits CPT1, leaving its activity at a lower level, thereby limiting fatty acid oxidation. When the body is under stress or increased energy consumption, the AMPK pathway can be activated immediately, and the downstream ACC is inactivated. The level of malonyl-CoA is rapidly decreased, further inhibiting the inhibition of CPT1, promoting the oxidation of fatty acids, and providing the body with More ATP.

Increased fatty acid synthesis and fatty acid metabolism disorders caused by impaired fatty acid oxidation are common features of many metabolic diseases, including liver steatosis, dyslipidemia, obesity, metabolic syndrome, nonalcoholic fatty Hepatitis (NASH), type 2 diabetes (T2DM) and atherosclerosis. In addition, abnormal fatty acid metabolism is also one of the characteristics of tumor diseases, and participates in the cell proliferation process that regulates abnormal malignant tumors. Since ACC is a key regulatory protein of lipid metabolism, drug inhibition of ACC can stimulate the synthesis of fatty acids in lipid-derived tissues while stimulating the oxidation of fatty acids in oxidized tissues, thus providing treatment for the above-mentioned diseases with abnormal lipid metabolism. A very attractive treatment.

A series of ACC inhibitor patents have been published, including WO2014182943, WO2014182945, WO2014182950, etc. The research and application of ACC inhibitors have made some progress. For example, the current Girard company's firsocostat is in clinical phase II, but existing The compounds disclosed in the technology as well as the test drugs are still unsatisfactory in terms of effectiveness, safety or applicability, and it is still necessary to continue research and development of new ACC inhibitors to meet the growing medical and health needs of people.

Summary of the invention

The inventors have unexpectedly discovered through experimental research that the compound of the following formula (I) can effectively inhibit ACC.

Accordingly, in a first aspect, the present invention provides a class of heteroaryl-pyrimidinone derivatives of the formula (I):

Including stereoisomers, tautomers or pharmaceutically acceptable salts thereof,

among them:

X is selected from -NH-, -O- or -S-; preferably -S-;

Ring A is selected from cycloalkyl, and R ² and N attached to ring A are not attached to the same carbon atom;

R ^{1 is} selected from a hydrogen atom, an alkyl group or a halogen, wherein the alkyl group is further further selected from one or more selected from the group consisting of halogen, hydroxy, cyano, nitro, alkoxy, cycloalkyl, heterocyclic, Aryl, heteroaryl, -NR ⁸ R ⁹ , -C(O)NR ⁸ R ⁹ , -C(O)R ¹⁰ , -OC(O)R ¹⁰ , -S(O) _q NR ⁸ R ⁹ , Substituted by a substituent of -NR ⁸ S(O) ₂ R ⁹ or -NR ⁸ C(O)R ⁹ ;

R ^{2 is} selected from the group consisting of a hydrogen atom, a hydroxyl group, a halogen, a nitro group, a cyano group, an alkyl group, an alkoxy group, a cycloalkyl group, a heterocyclic group, an aryl group, a heteroaryl group, -NR ⁸ R ⁹ , -C(O) NR ⁸ R ⁹ , -C(O)R ¹⁰ , -OC(O)R ¹⁰ , -S(O) _q NR ⁸ R ⁹ , -NR ⁸ S(O) ₂ R ⁹ or -NR ⁸ C(O) R ⁹ wherein said alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl group is further further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl , alkoxy, cycloalkyl, heterocyclic, aryl, heteroaryl, -NR ¹¹ R ¹² , -C(O)NR ¹¹ R ¹² , -C(O)R ¹³ , -C(O)OR Substituted by a substituent of ¹³ or -NR ¹¹ C(O)R ¹² ;

R ^{3 is} selected from aryl or heteroaryl, wherein said aryl or heteroaryl is optionally further substituted with one or more substituents selected from R ⁷ ;

R ⁴ and R ⁵ are each independently selected from a hydrogen atom, an alkyl group, -OR ¹⁰ , -SR ¹⁰ , -NR ⁸ R ⁹ , -C(O)NR ⁸ R ⁹ , -C(O)R ¹⁰ , -OC (O) R ¹⁰ , -S(O) _q NR ⁸ R ⁹ , -NR ⁸ S(O) ₂ R ⁹ or -NR ⁸ C(O)R ⁹ ;

Alternatively, R ⁴ , R ⁵ together with the atom to which they are attached form a 3 to 8 membered saturated or partially unsaturated cycloalkyl group, or form a hetero atom having one or more selected from N, O, S(O) _q a 4 to 8 membered saturated or partially unsaturated heterocyclic group; wherein the cycloalkyl or heterocyclic group is further further selected from one or more selected from the group consisting of a hydroxyl group, a halogen, a nitro group, a cyano group, an alkyl group, an alkoxy group, Cycloalkyl, heterocyclyl, aryl, heteroaryl, -NR ⁸ R ⁹ , -C(O)NR ⁸ R ⁹ , -C(O)R ¹⁰ , -OC(O)R ¹⁰ , -S( _{^{O) q NR 8 R 9,}} -NR 8 S (O) 2 R 9 or -NR ⁸ C (O) R ⁹ is substituted with a substituent;

R ^{6 is} selected from the group consisting of halogen, cyano, cycloalkyl, heterocyclic, aryl, heteroaryl, -NR ⁸ R ⁹ , -C(O)NR ⁸ R ⁹ , -C(O)R ¹⁰ , -OC (O) R ¹⁰ , -S(O) _q NR ⁸ R ⁹ , -NR ⁸ S(O) ₂ R ⁹ or -NR ⁸ C(O)R ⁹ ; preferably a heteroaryl group;

Alternatively, R ¹ , R ⁶ together with the atom to which they are attached form a 3 to 8 membered saturated or partially unsaturated cycloalkyl group, or form a hetero atom having one or more selected from N, O, S(O) _q a 4 to 8 membered saturated or partially unsaturated heterocyclic group, or a 5 to 10 membered aryl or heteroaryl group; wherein the cycloalkyl group, heterocyclic group, aryl group or heteroaryl group is optionally further one or more One selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, -NR ⁸ R ⁹ , -C(O)NR ⁸ R ⁹ , -C(O)R ¹⁰ , -OC(O)R ¹⁰ , -S(O) _q NR ⁸ R ⁹ , -NR ⁸ S(O) ₂ R ⁹ or -NR ⁸ C(O)R ⁹ Substituted by

R ^{7 is} each independently selected from the group consisting of hydroxyl, halogen, cyano, nitro, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, -NR ⁸ R ⁹ , -C(O) NR ⁸ R ⁹ , -C(O)R ¹⁰ , -OC(O)R ¹⁰ , -S(O) _q NR ⁸ R ⁹ , -NR ⁸ S(O) ₂ R ⁹ or -NR ⁸ C(O) R ⁹ wherein said alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl group is further further hydroxy, halo, nitro, cyano, alkyl, alkoxy, cycloalkane Base, heterocyclic group, aryl group, heteroaryl group, -NR ⁸ R ⁹ , -C(O)NR ⁸ R ⁹ , -C(O)R ¹⁰ , -OC(O)R ¹⁰ , -S(O) _{^{q NR 8 R 9, -NR 8}} S (O) 2 R 9 or -NR ⁸ C (O) R ⁹ is substituted with a substituent;

R ⁸ , R ⁹ and R ¹⁰ are each independently selected from a hydrogen atom, an alkyl group, -OR ¹³ , a cyano group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, wherein the alkyl group, cycloalkyl group Or a heterocyclyl, aryl or heteroaryl group optionally further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, hetero Substituted with a substituent of aryl, -NR ¹¹ R ¹² , -C(O)NR ¹¹ R ¹² , -C(O)R ¹³ , -C(O)OR ¹³ or -NR ¹¹ C(O)R ¹² ;

Alternatively, R ⁸ and R ⁹ together with the N atom to which they are attached form a 4 to 8 membered heterocyclic group, wherein the 4 to 8 membered heterocyclic ring contains one or more N, O or S(O) _q atoms, And the 4-8-membered heterocyclic ring is further selected from one or more selected from the group consisting of a hydroxyl group, a halogen, a nitro group, a cyano group, an alkyl group, an alkoxy group, a cycloalkyl group, a heterocyclic group, an aryl group, a heteroaryl group, Substituted by a substituent of =O, -NR ¹¹ R ¹² , -C(O)NR ¹¹ R ¹² , -C(O)R ¹³ , -C(O)OR ¹³ or -NR ¹¹ C(O)R ¹² ;

R ¹¹ , R ¹² and R ¹³ are each independently selected from a hydrogen atom, an alkyl group, an alkenyl group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, wherein the alkyl group, a cycloalkyl group, a heterocyclic group Or an aryl or heteroaryl group optionally further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, carboxy Substituted by a substituent of an acid or a carboxylic acid ester;

q is 0, 1, or 2.

Herein, the compound of the formula (I) (and the compound of the formula (II) to the formula (IV)) also includes, in scope, stereoisomers, tautomers or pharmaceutically acceptable salts thereof.

In some preferred embodiments of the invention, the compound of formula (I) has the structure of formula (II):

among them:

m is 1, 2, 3, 4 or 5;

The definitions of ring A, R ¹ , R ² , R ⁶ , R ⁷ and R ¹⁰ are as described in formula (I).

In some preferred embodiments of the invention, the compound of formula (I) has a specific stereo configuration, ie, has the structure of formula (III):

among them:

m is 1, 2, 3, 4 or 5;

The definitions of ring A, R ¹ , R ² , R ⁶ , R ⁷ and R ¹⁰ are as described in formula (I).

In some preferred embodiments of the invention, the compound of formula (I) has the structure of formula (IV):

among them:

m is 1, 2, 3, 4 or 5;

The definitions of ring A, R ¹ , R ² , R ⁶ , R ⁷ and R ¹⁰ are as described in formula (I).

In some preferred embodiment of the present invention, there is provided a compound of formula (I), (II), (III) or the compound (IV), in which R ¹ is selected from methyl or trifluoromethyl.

In some preferred embodiments of the invention there is provided a compound of formula (I), (II), (III) or (IV), wherein:

R ^{2 is} selected from the group consisting of tetrazolyl, -C(O)OR ¹³ or -C(O)NR ⁸ R ⁹ ;

R ^{8 is} selected from a hydrogen atom or an alkyl group;

R ^{9 is} selected from cyano or -OR ¹³ ;

R ^{13 is} selected from a hydrogen atom, an alkyl group, an alkenyl group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, wherein the alkyl group, cycloalkyl group, heterocyclic group, aryl group or heteroaryl group is optional Further substituted by one or more substituents selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, carboxylic acid or carboxylic acid esters Replace

R ² is preferably -C(O)OH.

In some preferred embodiments of the invention there is provided a compound of formula (I), (II), (III) or (IV), wherein ring A is selected from the group consisting of:

Preferred

In some preferred embodiments of the invention there is provided a compound of formula (I), (II), (III) or (IV), wherein R ^{6 is} selected from a 5-membered heteroaryl group, preferably a thiazolyl group.

In some preferred embodiments of the invention there is provided a compound of formula (I), (II), (III) or (IV), wherein R ^{7 is} selected from halo or alkoxy, preferably methoxy.

In some preferred embodiment of the present invention, there is provided a compound of formula (I), (II), (III) or the compound (IV), wherein R ¹⁰ is tetrahydropyran-4-yl.

Typical compounds of the invention include, but are not limited to:

Typical compounds described above include stereoisomers, tautomers or pharmaceutically acceptable salts thereof.

Further, the present invention provides a process for the preparation of a compound of formula (I), which process comprises:

The compound of formula (IA) is reacted with R ⁶ -substituted tributylstannane such that the resulting compound is optionally further hydrolyzed, and the resulting compound is optionally further resolved to the optically pure isomer to provide a compound of formula (I);

Wherein: X ^{1 is} selected from halogen; and X, ring A, R ¹ to R ⁶ are as defined in formula (I).

The present invention provides a compound of formula (IA):

among them:

X ^{1 is} selected from halogen; and X, ring A, R ¹ to R ⁵ are as defined in formula (I).

Typical compounds of formula (IA) include, but are not limited to:

Typical compounds described above include stereoisomers, tautomers or pharmaceutically acceptable salts thereof.

Further, the present invention provides a process for the preparation of a compound of formula (IA), the process comprising:

The compound of formula (IB) is reacted with a compound of formula (IC) in the presence of triphenylphosphine to provide a compound of formula (IA);

among them:

X ^{1 is} selected from halogen; and X, ring A, R ¹ to R ⁵ are as defined in formula (I).

In another aspect, the invention provides a pharmaceutical composition comprising an effective amount of a compound of formula (I), (II), (III) or (IV), and optionally Pharmaceutically acceptable carriers, excipients or combinations thereof.

In still another aspect, the present invention provides a method of inhibiting ACC comprising contacting ACC with a compound of formula (I), (II), (III) or (IV) of the present invention or a pharmaceutical composition thereof. The invention accordingly also provides a method of preventing or treating a disease or condition associated with ACC comprising administering a compound or pharmaceutical composition according to the invention to a subject in need thereof.

In another aspect, the present invention provides the use of a compound of formula (I), (II), (III) or (IV) or a pharmaceutical composition thereof for the manufacture of a medicament for use as an ACC inhibitor.

The invention also provides the use of a compound of formula (I), (II), (III) or (IV) or a pharmaceutical composition thereof for the manufacture of a medicament for the prevention or treatment of a disease or condition associated with ACC, wherein said The disease or condition is preferably a metabolic disease, a cancer, a fungus, a parasite or a bacterial infection, wherein the metabolic disease is preferably hepatic steatosis, nonalcoholic fatty liver, obesity, dyslipidemia, hyperlipidemia, type II Diabetes or metabolic syndrome, wherein the obesity is preferably Prader-Willi syndrome, Bardet-Biedl syndrome or Cohen syndrome Or MOMO syndrome, wherein the cancer is preferably hepatocellular carcinoma, non-small cell lung cancer, small cell lung cancer, gastric cancer, colorectal cancer, head and neck cancer, melanoma, ovarian cancer or cervical cancer, more preferably hepatocellular carcinoma And non-small cell lung cancer.

Detailed description of the invention

Unless otherwise stated, some of the terms used in the specification and claims of the invention are defined as follows:

"Alkyl" as a group or part of a group means a C ₁ -C ₂₀ straight or branched C ₁ -C ₂₀ aliphatic hydrocarbon group, preferably a C ₁ -C ₁₀ alkyl group More preferably, it is a C ₁ -C ₆ alkyl group, and particularly preferably a C ₁ -C ₄ alkane. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, sec-butyl, n-pentyl, 1, 1-di Methylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1 -ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethyl Butyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl Wait. The alkyl group can be substituted or unsubstituted.

"Alkylene" is a divalent alkyl group. It is preferably a C ₁ -C ₁₀ alkylene group, more preferably a C ₁ -C ₆ alkylene group, and particularly preferably a C ₁ -C ₄ alkylene group. Examples of alkylene groups include, but are not limited to, methylene, ethylene,

Acetylene and so on. The alkylene group may be substituted or unsubstituted.

"Alkenyl" refers to an alkyl radical as defined above consisting of at least two carbon atoms and at least one carbon-carbon double bond, representative examples including, but not limited to, ethenyl, 1-propenyl, 2-propenyl, 1-, 2- or 3-butenyl and the like. A C ₂ -C ₄ alkylene group is preferred. The alkenyl group can be optionally substituted or unsubstituted.

"Alkynyl" as a group or part of a group refers to an aliphatic hydrocarbon group containing a carbon-carbon triple bond which may be straight or branched. Preference is given to C ₂ -C ₁₀ alkynyl, more preferably C ₂ -C ₆ alkynyl, most preferably C ₂ -C ₄ alkynyl. Examples of alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-, 2- or 3-butynyl, and the like. An alkynyl group can be substituted or unsubstituted.

"Cycloalkyl" means a saturated or partially saturated monocyclic, fused, bridged or spiro carbon ring. It is preferably a C ₃ -C ₁₂ cycloalkyl group, more preferably a C ₃ -C ₈ cycloalkyl group, and most preferably a C ₃ -C ₆ cycloalkyl group. Examples of monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatriene The alkenyl group, the cyclooctyl group and the like are preferably a cyclopropyl group or a cyclohexenyl group.

"Cycloalkylene" is a divalent cycloalkyl group. It is preferably a C ₃ -C ₁₂ cycloalkylene group, more preferably a C ₃ -C ₈ cycloalkylene group, and most preferably a C ₃ -C ₆ cycloalkylene group. Examples of alkylene groups include, but are not limited to, cyclopropylene, cyclobutylene, cyclopentylene, and the like. A cycloalkylene group can be substituted or unsubstituted.

"Spirocycloalkyl" means a 5- to 18-membered polycyclic group having two or more cyclic structures and sharing a carbon atom (called a spiro atom) with each other, and the ring may contain 1 One or more double bonds, but none of the rings have a fully conjugated π-electron aromatic system. It is preferably 6 to 14 members, more preferably 7 to 10 members. The spirocycloalkyl group is classified into a monospiro, a spiro- or a spirocycloalkyl group, preferably a mono- and bi-spirocycloalkyl group, preferably 4 yuan/5 yuan, 4, depending on the number of common spiro atoms between the rings. Yuan / 6 yuan, 5 yuan / 5 yuan or 5 yuan / 6 yuan. Non-limiting examples of "spirocycloalkyl" include, but are not limited to, spiro[4.5]decyl, spiro[4.4]decyl, spiro[3.5]decyl, spiro[2.4]heptyl.

"Fused cycloalkyl" refers to a 5 to 18 membered all carbon polycyclic group having two or more cyclic structures that share a pair of carbon atoms with each other, wherein one or more of the rings may contain one or more A double bond, but none of the rings have a fully conjugated π-electron aromatic system, preferably 6 to 12 members, more preferably 7 to 10 members. Depending on the number of constituent rings, it may be classified into a bicyclic ring, a tricyclic ring, a pyridone or a polycyclic fused ring alkyl group, preferably a bicyclic ring or a tricyclic ring, more preferably a 5-membered/5-membered or 5-membered/6-membered bicycloalkyl group. Non-limiting examples of "fused cycloalkyl" include, but are not limited to, bicyclo[3.1.0]hexyl, bicyclo[3.2.0]hept-1-enyl, bicyclo[3.2.0]heptyl, Decalinyl or tetradecafluorophenanyl.

"Bridge cycloalkyl" refers to a 5- to 18-membered, all-carbon polycyclic group containing two or more cyclic structures that share two non-directly bonded carbon atoms, wherein one or more rings may contain One or more double bonds, but none of the rings have a fully conjugated π-electron aromatic system. It is preferably 6 to 14 members, more preferably 7 to 10 members. Depending on the number of constituent rings, it may be classified into a bicyclic ring, a tricyclic ring, a pyridone or a polycyclic bridged cycloalkyl group, preferably a bicyclic ring, a tricyclic ring or a pyridone, and more preferably a bicyclic ring or a tricyclic ring. Non-limiting examples of "bridged cycloalkyl" include, but are not limited to: (1s, 4s)-bicyclo[2.2.1]heptyl, bicyclo[3.2.1]octyl, (1s,5s)-di Ring [3.3.1] fluorenyl, bicyclo [2.2.2] octyl, (1r, 5r)-bicyclo[3.3.2] fluorenyl.

The cycloalkyl ring may be fused to an aryl, heteroaryl or heterocyclyl ring, wherein the ring to which the parent structure is attached is a cycloalkyl group, non-limiting examples include indanyl, tetrahydronaphthalene Base, benzocycloheptyl and the like. The cycloalkyl group can be optionally substituted or unsubstituted.

"Heterocyclyl", "heterocyclic" or "heterocyclic" are used interchangeably herein to refer to a non-aromatic heterocyclic group wherein one or more of the ring-forming atoms are heteroatoms such as oxygen, Nitrogen, sulfur atoms, etc., including monocyclic, fused, bridged, and spiro rings. It preferably has a 5- to 7-membered monocyclic ring or a 7- to 10-membered double- or tricyclic ring which may contain 1, 2 or 3 atoms selected from nitrogen, oxygen and/or sulfur. Examples of "heterocyclyl" include, but are not limited to, morpholinyl, oxetane, thiomorpholinyl, tetrahydropyranyl, 1,1-dioxo-thiomorpholinyl, piperidine , 2-oxo-piperidinyl, pyrrolidinyl, 2-oxo-pyrrolidinyl, piperazin-2-one, 8-oxa-3-aza-bicyclo[3.2.1]octyl and Piperazinyl. The heterocyclic group may be substituted or unsubstituted.

"Spiroheterocyclyl" means a 5- to 18-membered polycyclic group having two or more cyclic structures and sharing one atom between the single rings, and having one or more double bonds in the ring. , but none of the rings have a fully conjugated π-electron aromatic system in which one or more ring atoms are selected from nitrogen, oxygen or S(O) _q (where q is selected from 0, 1 or 2) heteroatoms, the remainder The ring atom is carbon. It is preferably 6 to 14 members, more preferably 7 to 10 members. The spirocycloalkyl group is classified into a monospiroheterocyclic group, a dispiroheterocyclic group or a polyspirocyclic group according to the number of common spiro atoms between the ring and the ring, and is preferably a monospiroheterocyclic group and a dispiroheterocyclic group. More preferably, it is 4 yuan / 4 yuan, 4 yuan / 5 yuan, 4 yuan / 6 yuan, 5 yuan / 5 yuan or 5 yuan / 6-membered monospiroheterocyclic group. Non-limiting examples of "spiroheterocyclyl" include, but are not limited to, 1,7-dioxaspiro[4.5]fluorenyl, 2-oxa-7-azaspiro[4.4]decyl, 7-oxo Heterospiro[3.5]decyl and 5-oxaspiro[2.4]heptyl.

"Fused heterocyclic group" refers to an all-carbon polycyclic group containing two or more cyclic structures that share a pair of atoms with each other, wherein one or more of the rings may contain one or more double bonds, but none of the rings have A fully conjugated π-electron aromatic system in which one or more ring atoms are heteroatoms selected from nitrogen, oxygen or S(O) _q (where q is selected from 0, 1 or 2) and the remaining ring atoms are carbon. It is preferably 6 to 14 members, more preferably 7 to 10 members. Depending on the number of constituent rings, it may be classified into a bicyclic ring, a tricyclic ring, a pyridone or a polycyclic fused heterocyclic group, preferably a bicyclic ring or a tricyclic ring, and more preferably a 5-membered/5-membered or 5-membered/6-membered bicyclic fused heterocyclic group. Non-limiting examples of "fused heterocyclic groups" include, but are not limited to, octahydropyrrolo[3,4-c]pyrrolyl, octahydro-1H-isoindenyl, 3-azabicyclo[3.1. 0] hexyl, octahydrobenzo[b][1,4]dioxine.

"Bridge heterocyclyl" refers to a polycyclic group of 5 to 18 members, preferably 5 to 14 members containing two or more cyclic structures and sharing two atoms which are not directly bonded to each other, wherein one or more rings An aromatic system which may contain one or more double bonds, but none of which has a fully conjugated π-electron, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) _q (where q is selected from 0, 1 Or 2) a hetero atom, the remaining ring atoms being carbon. It is preferably 6 to 14 members, more preferably 7 to 10 members. Depending on the number of constituent rings, it may be classified into a bicyclic ring, a tricyclic ring, a pyridone or a polycyclic bridged heterocyclic group, preferably a bicyclic ring, a tricyclic ring or a pyridone, and more preferably a bicyclic ring or a tricyclic ring. Non-limiting examples of "fused heterocyclic groups" include, but are not limited to, 2-azabicyclo[2.2.1]heptyl, 2-azabicyclo[2.2.2]octyl and 2-aza-di Ring [3.3.2] sulfhydryl. The heterocyclyl ring may be fused to an aryl, heteroaryl or cycloalkyl ring wherein the ring to which the parent structure is attached is a heterocyclic group. The heterocyclic group may be optionally substituted or unsubstituted.

"Heterocyclylene" means a divalent heterocyclic group. It preferably has a 5- to 7-membered monocyclic heterocyclic group or a 7 to 10 membered bicyclic heterocyclic group or a tricyclic heterocyclic group which may contain 1, 2 or 3 atoms selected from nitrogen, oxygen and/or sulfur. . The heterocyclylene group may be substituted or unsubstituted.

"Aryl" means a carbocyclic aromatic system containing one or two rings wherein the rings may be joined together in a fused manner. The term "aryl" includes aryl groups such as phenyl, naphthyl, tetrahydronaphthyl. Preferably, the aryl group is a C ₆ -C ₁₀ aryl group, more preferably the aryl group is a phenyl group and a naphthyl group, and most preferably a phenyl group. The aryl group can be substituted or unsubstituted. The "aryl" may be fused to a heteroaryl, heterocyclyl or cycloalkyl group, wherein the parent structure is attached to an aryl ring, non-limiting examples include, but are not limited to:

"Heteroaryl" means an aromatic 5 to 6 membered monocyclic or 9 to 10 membered bicyclic ring which may contain from 1 to 4 atoms selected from nitrogen, oxygen and/or sulfur. Examples of "heteroaryl" include, but are not limited to, furyl, pyridyl, 2-oxo-1,2-dihydropyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, thienyl, isoxazolyl , oxazolyl, oxadiazolyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, benzo Dioxolyl, benzimidazolyl, fluorenyl, isodecyl, 1,3-dioxo-isoindenyl, quinolyl, oxazolyl, benzisothiazolyl, benzene And oxazolyl and benzoisoxazolyl. Heteroaryl groups can be substituted or unsubstituted. The heteroaryl ring can be fused to an aryl, heterocyclic or cycloalkyl ring wherein the ring to which the parent structure is attached is a heteroaryl ring, non-limiting examples include, but are not limited to:

"Alkoxy" means a group of (alkyl-O-). Among them, the alkyl group is defined in the relevant definition herein. The C ₁ -C ₆ alkoxy group is preferred, and a C ₁ -C ₄ alkoxy group is particularly preferred. Examples thereof include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy and the like.

"Hydroxy" refers to an -OH group.

"Halogen" means fluoro, chloro, bromo and iodo, preferably chloro, bromo and iodo.

"Amino" means -NH ₂ .

"Cyano" means -CN.

"Nitro" means -NO ₂ .

"Benzyl" refers to -CH ₂ - phenyl.

"Carboxy" refers to -C(O)OH.

"Carboxylic acid ester group" means -C(O)O(alkyl) or (cycloalkyl) wherein alkyl, cycloalkyl are as defined above.

"DMSO" refers to dimethyl sulfoxide.

"Et" means ethyl.

"Substituted" refers to one or more hydrogen atoms in the group, preferably up to 5, more preferably 1 to 3, hydrogen atoms are replaced by a corresponding number of substituents independently of one another. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art will be able to determine (by experiment or theory) substitutions that may or may not be possible without undue effort. For example, an amino group or a hydroxyl group having a free hydrogen may be unstable when combined with a carbon atom having an unsaturated bond such as an ethylenic bond.

As used herein, "substituted" or "substituted", unless otherwise indicated, means that the group may be substituted by one or more groups selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy. , alkylthio, alkylamino, halogen, fluorenyl, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkane Base, heterocycloalkylthio, amino, haloalkyl, hydroxyalkyl, carboxyl, carboxylate, =O, -NR ⁸ R ⁹ , -C(O)NR ⁸ R ⁹ , -C(O)R ¹⁰ Substituted by a substituent of -OC(O)R ¹⁰ , -S(O) _q NR ⁸ R ⁹ , -NR ⁸ S(O) ₂ R ⁹ or -NR ⁸ C(O)R ⁹ ;

"Pharmaceutically acceptable salt" refers to certain salts of the above compounds which retain their original biological activity and are suitable for pharmaceutical use. The pharmaceutically acceptable salt of the compound of the formula (I) may be a metal salt, an amine salt formed with a suitable acid, a metal salt preferably an alkali metal or an alkaline earth metal salt, and suitable acids including inorganic acids and organic acids such as acetic acid and benzenesulfonate. Acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, malic acid, maleic acid, mandelic acid , methanesulfonic acid, nitric acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, p-toluenesulfonic acid, and the like. Particularly preferred are hydrochloric acid, hydrobromic acid, phosphoric acid and sulfuric acid, and most preferred is the hydrochloride salt.

"Pharmaceutical composition" means containing one or more of the compounds described herein, including pharmaceutically acceptable salts or stereoisomers, tautomers or prodrugs thereof, and optionally other pharmaceutically active ingredients. A mixture, which may contain other optional ingredients such as pharmaceutically acceptable carriers and/or excipients. The purpose of the pharmaceutical composition is to promote the administration of the organism, which facilitates the absorption of the active ingredient and thereby exerts biological activity.

As used herein, the term "plurality" includes two or more, such as two, three, four, and the like.

Method for synthesizing the compound of the present invention

In order to accomplish the object of the present invention, the present invention adopts the following technical solutions:

The preparation method of the compound of the formula (I) of the invention comprises the following steps:

The compound of formula (IB) is reacted with a compound of formula (IC) in the presence of triphenylphosphine to provide a compound of formula (IA);

The compound of formula (IA) is reacted with R ⁶ -substituted tributylstannane such that the resulting compound is optionally further hydrolyzed, and the resulting compound is optionally further resolved to the optically pure isomer to provide a compound of formula (I);

Wherein: X ^{1 is} selected from halogen; and X, ring A, R ¹ to R ⁶ are as defined in formula (I).

detailed description

The invention is further described in the following examples, but these examples are not intended to limit the scope of the invention.

The examples give the preparation of representative compounds represented by formula (I) and related structural identification data. It is to be understood that the following examples are intended to illustrate the invention and not to limit the invention. ^{The 1} H NMR spectrum was measured using a Bruker instrument (400 MHz) and the chemical shift was expressed in ppm. The internal standard of tetramethylsilane (0.00 ppm) was used. ¹ H NMR representation: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, br = broadened, dd = doublet of doublet, dt = triple The double peak of the peak. If a coupling constant is provided, its unit is Hz.

Mass spectrometry was measured by LC/MS, and the ionization method was ESI or APCI.

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~0.2mm. 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.

In the following examples, all temperatures are in degrees Celsius unless otherwise indicated, and unless otherwise indicated, the various starting materials and reagents are either commercially available or synthesized according to known methods, and the commercially available starting materials and reagents are not further purified. For direct use, unless otherwise indicated, commercial manufacturers include, but are not limited to, Aldrich Chemical Company, ABCR GmbH & Co. KG, Acros Organics, Guangzan Chemical Technology Co., Ltd. and Jingyan Chemical Technology Co., Ltd., and the like.

CD ₃ OD: Deuterated methanol.

CDCl ₃ : deuterated chloroform.

DMSO-d ₆ : deuterated dimethyl sulfoxide.

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

There is no particular description in the examples, and the solution in the reaction means an aqueous solution.

The compound is purified by silica gel column chromatography and silica gel sheet chromatography, wherein the developing solvent or eluent system is selected from the group consisting of: A: petroleum ether and ethyl acetate system; B: dichloromethane and methanol system; C: two Methyl chloride: ethyl acetate; wherein the volume ratio of the solvent varies depending on the polarity of the compound, and it may be adjusted by adding a small amount of an acidic or alkaline agent such as acetic acid or triethylamine.

Example 1

(1R,3R)-3-(1-(2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-5-A -6-(oxazol-2-yl)-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl)cyclobutane-1 -formic acid

first step

2-(2-methoxyphenyl)oxirane

2-Methoxybenzaldehyde 1a (20.0 g, 146.9 mmol) was dissolved in 100 mL of dimethyl sulfoxide, followed by t-butylthiohypoiodate (36.0 g, 173.3 mmol) and sodium hydroxide (24.7 g). , 441.0 mmol), heated to 80 ° C for 1.5 hours. The reaction solution was cooled to room temperature, and then added with EtOAc (EtOAc) (EtOAc (EtOAc) Purification by silica gel column chromatography (eluent: EtOAc)

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.27 (t, J = 1.2 Hz, 1H), 7.17 (d, J = 7.6 Hz, 1H), 6.98 (t, J = 1.2 Hz, 1H), 6.89 (d) , J = 7.6 Hz, 1H), 4.22 (t, J = 0.4 Hz, 1H), 3.87 (s, 3H), 2.71 (dd, J = 5.6, 2.4 Hz, 1H) 3.14 (dd, J = 5.6, 2.4 Hz, 1H).

Second step

2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)ethanol

(2-Methoxyphenyl)oxirane 1b (26.0 g, 173.0 mmol) was added to stirred tetrahydro-2H-pyran-4-ol 1c (53.1 g, 519.7 mmol) and trifluoromethanesulfonate The aluminum acid (4.10 g, 8.65 mmol) was reacted at room temperature for 3 hours. 200 mL of dichloromethane and 200 mL of water were added to the reaction mixture, and the organic layer was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: A system) to give 2-(2-methoxybenzene) 2-((tetrahydro-2H-pyran-4-yl)oxy)ethanol 1d (13.0 g, white solid), yield: 30%.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ7.42 (d, J = 8.0Hz, 1H), 7.26 (t, J = 7.2Hz, 1H), 6.98 (t, J = 7.2Hz, 1H), 6.87 (d , J=8.0 Hz, 1H), 5.07 (dd, J=8.0, 4.0 Hz, 1H), 3.87-4.00 (m, 2H), 3.83 (s, 3H), 3.62-3.72 (m, 1H), 3.46- 3.58 (m, 2H), 3.32-3.43 (m, 2H), 2.35-2.37 (m, 1H), 1.99-2.03 (m, 1H), 1.77-1.80 (m, 1H), 1.60-1.70 (m, 2H) ).

third step

2-(tributylstannyl)oxazole

Oxazole 1e (500 mg, 7.24 mmol) was dissolved in 12 mL of tetrahydrofuran. Under nitrogen, the mixture was cooled to -78 ° C for 5 minutes, and n-butyllithium (4.56 mL, 7.29 mmol) was slowly added. After the addition, the mixture was stirred at -78 ° C for 30 minutes. Then, tributyltin chloride (1.96 mL, 7.24 mmol) was added, and the mixture was stirred at -78 ° C for 10 minutes, and allowed to react to room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure. 15 mL of hexane was evaporated, and then filtered, and the filtrate was concentrated under reduced pressure to give 2-(tributylstannyl) oxazole 1f (1.8 g, pale yellow liquid), yield: 70%.

^{1 H NMR (400MHz, CDCl3)} : 7.84 (1H, s), 7.18 (1H, s), 1.67-1.53 (6H, m), 1.42-1.29 (6H, m), 1.20 (6H, m), 0.89 ( 9H, t, J = 7Hz).

the fourth step

Ethyl 2-amino-4-methylthiophene-3-carboxylate

Under argon protection, 1 g of ethyl 2-cyanoacetate (185.5 g, 1.64 mol), acetone (100 g, 1.72 mol), sulfur (53 g, 1.64 mol) were dissolved in 500 mL of absolute ethanol, slowly added dropwise? The porphyrin 1 h (149.6 g, 1.64 mol) was added over 20 minutes. The reaction was carried out at 45 ° C for 10 hours. The reaction solution was cooled to room temperature, and the remaining sulfur powder was removed by filtration. The filtrate was concentrated under reduced pressure, and then added with 900 mL of 75% ethanol and stirred at room temperature for 30 min. The yellow solid was precipitated, filtered, and the solid was evaporated. The filtrate was concentrated under reduced pressure. <RTI ID=0.0>&&&&&&&&&&&&&&&& Ethyl 3-carboxylate 1i (95 g, green solid), yield: 31.4%.

MS m/z (ESI): 185.9 [M+1]

the fifth step

3-oxocyclobutane-1-carboxylic acid tert-butyl ester

3-oxocyclobutane-1-carboxylic acid 1j (5.7 g, 50 mmol), tert-butanol (9.25 g, 125 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide Hydrochloride (11.5 g, 60 mmol) and 4-dimethylaminopyridine (3.1 g, 25 mmol) were dissolved in dichloromethane (1 mL) and stirred at room temperature for 12 hr. The reaction mixture was diluted with water (100 mL), EtOAc (EtOAc) Tert-butyl butane-1-carboxylic acid tert-butyl ester 1k (8.5 g, reddish brown oil), yield: 100%.

Step 6

(1S,3S)-3-hydroxycyclobutane-1-carboxylic acid tert-butyl ester

3-tert-cyclobutane-1-carboxylic acid tert-butyl ester 1k (6.5 g, 38.2 mmol) was dissolved in 72 mL of tetrahydrofuran/methanol (V:V=8:1), and sodium borohydride was added portionwise at 0 °C. 0.71 g, 19.1 mmol), and reacted at room temperature for one hour. After adding 80 ml of a saturated aqueous solution of potassium carbonate, and ethyl acetate (100 mL × 3), the organic layer was evaporated. , (1S,3S)-3-hydroxycyclobutane-1-carboxylic acid tert-butyl ester 1 l (6.58 g, viscous solid), yield: 100%;

Seventh step

(1R,3R)-3-(1,3-dioxoisoindol-2-yl)cyclobutane-1-carboxylic acid tert-butyl ester

(1S,3S)-3-hydroxycyclobutane-1-carboxylic acid tert-butyl ester 1 l (4.9 g, 28.45 mmol), isoindoline-1,3-dione 1 m (4.6 g, 31.3 mmol) and Triphenylphosphine (11.2 g, 42.1 mmol) was dissolved in 80 mL of tetrahydrofuran, and azodiisopropyldicarboxylic acid (8.6 g, 42.7 mmol) was added with stirring, and the mixture was reacted at room temperature for 2 hours. The organic layer was concentrated under reduced pressure. The residue was dissolved in ethyl acetate (20 mL), and then evaporated to ethyl ether. Purification of system A) gave (1R,3R)-3-(1,3-dioxoisoindolin-2-yl)cyclobutane-1-carboxylic acid tert-butyl ester 1n (7.5 g, white solid). Yield: 87.5%.

Eighth step

(1R,3R)-3-aminocyclobutane-1-carboxylic acid tert-butyl ester

(1R,3R)-3-(1,3-dioxoisoindol-2-yl)cyclobutane-1-carboxylic acid tert-butyl ester 1n (4.5 g, 14.9 mmol) was dissolved in 45 mL of ethanol. Hydrazine hydrate (2.2 g, 44.8 mmol) was added and the reaction was carried out for 12 hours at room temperature. Filtration, washing the filter cake with ethanol (10 mL × 2), the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: A system) to give (1R,3R)-3-aminocyclobutane- 1-tert-butyl formate 1o (1.2 g, colorless oil), yield: 66.4%.

Step 9

Ethyl 2-(3-(1R,3R)-3-(tert-butoxycarbonyl)cyclobutyl)ureido)-4-methylthiophene-3-carboxylate

2-Amino-4-methylthiophene-3-carboxylic acid ethyl ester 1i (1.44 g, 7.8 mmol) was dissolved in 60 mL of dichloromethane, and bis(trichloromethyl) carbonate (810 mg, A solution of 2.7 mmol) in dichloromethane (10 mL) was stirred at room temperature for 30 min. Triethylamine (2.4 g, 23.4 mmol) was added at 0 ° C and stirred at room temperature for 30 min. (1R,3R)-3-aminocyclobutane-1-carboxylic acid tert-butyl ester 1o (1.3 g, 7.8 mmol) was added at 0 ° C and allowed to react at room temperature for 30 min. The reaction mixture was diluted with EtOAc (EtOAc) (EtOAc) :A system) Purification to give ethyl 2-(3-(1R,3R)-3-(tert-butoxycarbonyl)cyclobutyl)ureido)-4-methylthiophene-3-carboxylate 1p (2.5 g , white solid), Yield: 83%.

MS m/z (ESI): 383.0 [M+1]

Step 10

(1R,3R)-3-(5-methyl-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl)cyclobutane- 1-carboxylic acid tert-butyl ester

2-(3-(1R,3R)-3-(tert-Butoxycarbonyl)cyclobutyl)ureido)-4-methylthiophene-3-carboxylate 1p (250 mg, 0.65 mmol) and cesium carbonate (426 mg, 1.31 mmol) was dissolved in 3 mL of N,N-dimethylformamide and reacted at 100 ° C for 2 hours. The reaction mixture was diluted with EtOAc (EtOAc EtOAc (EtOAc) Eluent: A system) purified to give (1R,3R)-3-(5-methyl-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidine-3 tert-Butyl (4H)-yl)cyclobutane-1-carboxylate 1q (200 mg, white solid).

MS m/z (ESI): 280.9 [M-55]

The eleventh step

(1R,3R)-3-(6-bromo-5-methyl-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl) Cyclobutane-1-carboxylic acid tert-butyl ester

(1R,3R)-3-(5-methyl-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl)cyclobutane Tert-butyl 1-carboxylic acid ester 1q (2.2 g, 6.54 mmol) was dissolved in 50 mL of dichloromethane, and N-bromosuccinimide (1.4 g, 7.85 mmol) was added at 0 ° C, and reacted at 0 ° C for 30 min. . The reaction mixture was diluted with EtOAc (EtOAc) (EtOAc) Deprotection: A system) purification, to obtain (1R, 3R)-3-(6-bromo-5-methyl-2,4-dioxo-1,2-dihydrothieno[2,3-d] Pyrimidine-3(4H)-yl)cyclobutane-1-carboxylic acid tert-butyl ester 1r (2.5 g, white solid), yield: 92.0%.

MS m/z (ESI): 358.8 [M-55]

Step 12

(1R,3R)-3-(6-bromo-1-(2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl) -5-Methyl-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl)cyclobutane-1-carboxylic acid tert-butyl ester

(1R,3R)-3-(6-bromo-5-methyl-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidine-3 (under nitrogen) 4H)-yl)cyclobutane-1-carboxylic acid tert-butyl ester 1r (723 mg, 1.74 mmol), 2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl) Ethoxy)ethanol 1d (834 mg, 3.30 mmol) and triphenylphosphine (912 mg, 3.48 mmol) were dissolved in 20 mL of anhydrous tetrahydrofuran. After cooling to 0 ° C for 3 minutes, a solution of diisopropyl azodicarboxylate (0.69 mL, 3.48 mmol) in 4 mL of tetrahydrofuran was added. After the addition was completed, the mixture was allowed to react at room temperature for 18 hours. The reaction mixture was concentrated under reduced pressure. mjjjjjjjjjjjjjjjjjjjjj 2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-5-methyl-2,4-dioxo-1,2-dihydrothieno[2, 3-d]pyrimidine-3(4H)-yl)cyclobutane-1-carboxylic acid tert-butyl ester 1 s (986 mg, white solid).

MS m/z (ESI): 649.8 [M+1]

Step 13

(1R,3R)-3-(1-(2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-5-A -6-(oxazol-2-yl)-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl)cyclobutane-1 - tert-butyl formate

(1R,3R)-3-(6-bromo-1-(2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy) under nitrogen Ethyl)-5-methyl-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl)cyclobutane-1-carboxylic acid tert-Butyl ester 1 s (986 mg, 1.52 mmol), 2-(tributylstannyl)oxazole 1f (817 mg, 2.28 mmol) and tetratriphenylphosphine palladium (245 mg, 0.21 mmol) were dissolved in 12 mL of toluene and heated to The reaction was carried out at 110 ° C for 7 hours. The reaction mixture was cooled to room temperature, EtOAc (EtOAc) (EtOAc) Purification of system A) gives (1R,3R)-3-(1-(2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)) 5-methyl-6-(oxazol-2-yl)-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl tert-Butylbutane-1-carboxylate 1t (756 mg, pale yellow solid), yield: 78%.

MS m/z (ESI): 638.8 [M+1]

Fourteenth step

(1R,3R)-3-(1-(2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-5-A -6-(oxazol-2-yl)-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl)cyclobutane-1 -formic acid

(1R,3R)-3-(1-(2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-5- Methyl-6-(oxazol-2-yl)-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl)cyclobutane- 1-t-carboxylic acid tert-butyl ester 1t (756 mg, 1.18 mmol) was dissolved in 10 mL dichloromethane. 2 mL of trifluoroacetic acid was added dropwise at 0 ° C, and after the addition was completed, the reaction was allowed to proceed to room temperature for 3 hours. 25 mL of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (10 mL×3). :B system) purification to give (1R,3R)-3-(1-(2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)) Ethyl)-5-methyl-6-(oxazol-2-yl)-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)- Cyclobutane-1-carboxylic acid 1 (185 mg, white solid), yield: 27%. MS m/z (ESI): 582.2 [M+1]

^{1 H NMR (400MHz, DMSO)} δ12.31 (s, 1H), 8.23 (s, 1H), 7.49 (d, J = 7.4Hz, 1H), 7.39 (s, 1H), 7.29 (t, J = 7.4 Hz,1H),7.13-6.89(m,2H),5.53(m,1H),5.31(m,1H),4.02(m,2H),3.78(s,3H),3.55(m,2H),3.36 (m, 1H), 3.24 (m, 2H), 3.07 (m, 3H), 2.78 (s, 3H), 2.41 (s, 2H), 1.63 (s, 2H), 1.42-1.12 (m, 2H).

Embodiment 2 and Embodiment 3

(1R,3R)-3-(1-((S)-2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl) 5-5-methyl-6-(oxazol-2-yl)-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl) ring Butane-1-carboxylic acid

(1R,3R)-3-(1-((R)-2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl) 5-5-methyl-6-(oxazol-2-yl)-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl) ring Butane-1-carboxylic acid

first step

(1R,3R)-3-(1-((S)-2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl) 5-5-methyl-6-(oxazol-2-yl)-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl) ring Butane-1-carboxylic acid

(1R,3R)-3-(1-((R)-2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl) 5-5-methyl-6-(oxazol-2-yl)-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl) ring Butane-1-carboxylic acid

(1R,3R)-3-(1-(2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-5- Methyl-6-(oxazol-2-yl)-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl)cyclobutane- 1-carboxylic acid 1 (120 mg, 0.60 mmol) was resolved by preparative chromatography and chiral column chiral isomers using a supercritical fluid chromatography (SFC) method (chiral Pak AS, 250 x 30 mm ID, 5 μm; 70 mL/min; mobile phase is A (for CO ₂ ) and B (for methanol) (0.1% NH ₃ .H ₂ O)) to give (1R,3R)-3-(1-((S)-2- (2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-5-methyl-6-(oxazol-2-yl)-2 , 4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl)cyclobutane-1-carboxylic acid 2 (56.3 mg, white solid), yield: 46.9%, retention time 5.289min, ee value 99.7%; (1R,3R)-3-(1-((R)-2-(2-methoxyphenyl)-2-((tetrahydro-2H-) Pyran-4-yl)oxy)ethyl)-5-methyl-6-(oxazol-2-yl)-2,4-dioxo-1,2-dihydrothieno[2,3 -d]pyrimidin-3(4H)-yl)cyclobutane-1-carboxylic acid 3 (63.3 mg, white solid), yield: 52.28%, retention time 4.505 min, ee value 99.8%.

Compound 2

MS m/z (ESI): 582.2 [M+1]

^{1 H NMR (400MHz, DMSO)} δ12.31 (s, 1H), 8.23 (s, 1H), 7.49 (d, J = 7.4Hz, 1H), 7.39 (s, 1H), 7.29 (t, J = 7.4 Hz,1H),7.13-6.89(m,2H),5.53(m,1H),5.31(m,1H),4.02(m,2H),3.78(s,3H),3.55(m,2H),3.36 (m, 1H), 3.24 (m, 2H), 3.07 (m, 3H), 2.78 (s, 3H), 2.41 (s, 2H), 1.63 (s, 2H), 1.42-1.12 (m, 2H).

Compound 3

MS m/z (ESI): 582.2 [M+1]

^{1 H NMR (400MHz, DMSO)} δ12.31 (s, 1H), 8.23 (s, 1H), 7.49 (d, J = 7.4Hz, 1H), 7.39 (s, 1H), 7.29 (t, J = 7.4 Hz,1H),7.13-6.89(m,2H),5.53(m,1H),5.31(m,1H),4.02(m,2H),3.78(s,3H),3.55(m,2H),3.36 (m, 1H), 3.24 (m, 2H), 3.07 (m, 3H), 2.78 (s, 3H), 2.41 (s, 2H), 1.63 (s, 2H), 1.42-1.12 (m, 2H).

Example 4

(1S,3S)-3-(1-(2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-5-A -6-(oxazol-2-yl)-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl)cyclobutane-1 -formic acid

first step

Ethyl 3-oxocyclobutane-1-carboxylate

3-oxocyclobutane-1-carboxylic acid 1j (4.56 g, 40 mmol), ethanol (2.3 g, 50 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride The salt (9.2 g, 48 mmol) and 4-dimethylaminopyridine (2.4 g, 20 mmol) were dissolved in 100 mL dichloromethane and stirred at room temperature for 12 hr. The reaction mixture was diluted with water (150 mL), EtOAc (EtOAc) Ethyl cyclobutane-1-carboxylate 4a (5.5 g, colorless oil), yield: 96.7%.

Second step

(1S,3S)-3-(dibenzylamino)cyclobutane-1-carboxylic acid ethyl ester

Ethyl 3-oxocyclobutane-1-carboxylate 4a (5.5 g, 40 mmol) and dibenzylamine (8.58 g, 44 mmol) were dissolved in EtOAc EtOAc Sodium triacetoxyborohydride (17.1 g, 80 mmol) and 20 mL of acetic acid were added, and the mixture was reacted at room temperature for 12 hours. The tetrahydrofuran was removed under reduced pressure, and a saturated aqueous solution of sodium hydrogencarbonate was added, and the mixture was adjusted to basic, ethyl acetate (50mL×3), and the organic phase was combined and washed with 50 mL of saturated aqueous sodium chloride. The residue was purified by silica gel chromatography (yield: A) to give ethyl (1S,3S)-3-(dibenzylamino)cyclobutane-1-carboxylate 4b. (6.0 g, colorless oil), yield: 46.5%.

third step

(1S,3S)-3-aminocyclobutane-1-carboxylic acid ethyl ester

Ethyl (1S,3S)-3-(dibenzylamino)cyclobutane-1-carboxylate 4b (5.0 g, 15.5 mmol) was dissolved in 150 mL of ethanol, then 10% Pd-C (200 mg, 4%) The hydrogen was replaced 3 times and reacted at room temperature for 12 hours. Filtration, the residual Pd-C was removed, and the filtrate was concentrated under reduced pressure to give ethyl (1S,3S)-3-aminocyclobutane-1-carboxylate 4c (2.1 g, colorless oil), yield: 95.5%.

the fourth step

Ethyl 2-(3-((1S,3S)-3-(ethoxycarbonyl)cyclobutyl)ureido)-4-methylthiophene-3-carboxylate

2-Amino-4-methylthiophene-3-carboxylic acid ethyl ester 1i (2.78 g, 15.0 mmol) was dissolved in 100 mL of dichloromethane, and bis(trichloromethyl) carbonate (1.56 g) was added dropwise at 0 °C. , 5.25 mmol) in dichloromethane (20 mL), stirred at room temperature for 30 min. Triethylamine (4.5 g, 45 mmol) was added at 0 ° C and stirred at room temperature for 30 min. (1S,3S)ethyl-3-aminocyclobutane-1-carboxylate 4c (2.2 g, 15.5 mmol) was added at 0 ° C and allowed to react at room temperature for 30 min. The reaction mixture was diluted with EtOAc (EtOAc) (EtOAc) :A system) Purification to give ethyl 2-(3-((1S,3S)-3-(ethoxycarbonyl)cyclobutyl)ureido)-4-methylthiophene-3-carboxylate 4d (4.7 g , pale yellow solid), yield: 89%.

MS m/z (ESI): 354.9 [M+1]

the fifth step

(1S,3S)-3-(5-methyl-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl)cyclobutane- Ethyl 1-carboxylate

Ethyl 2-(3-((1S,3S)-3-(ethoxycarbonyl)cyclobutyl)ureido)-4-methylthiophene-3-carboxylate 4d (4.7 g, 13.3 mmol) and carbonic acid The hydrazine (8.67 g, 26.6 mmol) was dissolved in 30 mL of N,N-dimethylformamide and reacted at 100 ° C for 1 hour. The reaction mixture was diluted with ethyl acetate (150 mL), EtOAc (EtOAc) Purification by the method (eluent: A system) to give (1S,3S)-3-(5-methyl-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidine Ethyl 3-(4H)-yl)cyclobutane-1-carboxylate 4e (2.0 g, pale yellow solid), yield: 49.0%.

MS m/z (ESI): 308.9 [M+1]

Step 6

(1S,3S)-3-(6-bromo-5-methyl-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl) Ethyl cyclobutane-1-carboxylate

(1S,3S)-3-(5-methyl-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl)cyclobutane Ethyl 1-ethylcarboxylate 4e (2.0 g, 6.5 mmol) was dissolved in dichloromethane (EtOAc), and then evaporated tolud. The reaction mixture was diluted with EtOAc (EtOAc) (EtOAc) :A system) purified to give (1S,3S)-3-(6-bromo-5-methyl-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidine- 3(4H)-yl)cyclobutane-1-carboxylic acid ethyl ester 4f (2.0 g, white solid), yield: 80.0%.

MS m/z (ESI): 386.8 [M+1]

Seventh step

(1S,3S)-3-(6-bromo-1-(2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl) -5-Methyl-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl)cyclobutane-1-carboxylate

(1S,3S)-3-(6-bromo-5-methyl-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidine-3 (under nitrogen) 4H)-yl)cyclobutane-1-carboxylic acid ethyl ester 4f (2.0 mg, 5.2 mmol), 2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl) Ethoxy)ethanol 1d (1.52 g, 6.0 mmol) and triphenylphosphine (1.58 g, 6.0 mmol) were dissolved in 25 mL of anhydrous tetrahydrofuran. After cooling to 0 ° C for 3 minutes, a solution of diisopropyl azodicarboxylate (1.2 g, 6.0 mmol) in 4 mL of tetrahydrofuran was added. After the addition was completed, the mixture was allowed to react at room temperature for 18 hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified mjjjjjjjjjj Phenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-5-methyl-2,4-dioxo-1,2-dihydrothieno[ 2,3-d]Pyridine-3(4H)-yl)cyclobutane-1-carboxylic acid ethyl ester 4 g (2.9 g, white solid), yield: 90%.

MS m/z (ESI): 621.2 [M+1]

Eighth step

(1S,3S)-3-(1-(2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-5-A -6-(oxazol-2-yl)-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl)cyclobutane-1 -ethyl formate

(1S,3S)-3-(6-bromo-1-(2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy) Ethyl)-5-methyl-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl)cyclobutane-1-carboxylic acid Ethyl ester 4 g (1.24 g, 2.0 mmol), 2-(tributylstannyl)oxazole 1f (1.4 g, 3 mmol) and tetratriphenylphosphine palladium (323.5 mg, 0.28 mmol) were dissolved in 20 mL of toluene. The reaction was heated to 110 ° C for 7 hours. The reaction solution was cooled to room temperature, and then added with ethyl acetate (30 mL×3), and the mixture was evaporated. Deprotection: A system) purification, to obtain (1S,3S)-3-(1-(2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy) Ethyl)-5-methyl-6-(oxazol-2-yl)-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidine-3 (4H Ethyl 4-butane-l-carboxylate 4 h (936 mg, pale yellow solid), yield: 78%.

MS m/z (ESI): 609.9 [M+1]

Step 9

(1S,3S)-3-(1-(2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-5-A -6-(oxazol-2-yl)-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl)cyclobutane-1 -formic acid

(1S,3S)-3-(1-(2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-5- Methyl-6-(oxazol-2-yl)-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl)cyclobutane- Ethyl 1-carboxylate 4h (936mg, 1.5mmol) and lithium hydroxide (108mg, 4.5mmol) were dissolved in 10mL of tetrahydrofuran / methanol (V: V = 1:1). The reaction was carried out for 12 hours at room temperature. The residue was purified by silica gel chromatography (yield: B system) to give (1S,3S)-3-(1-(2-(2-methoxyphenyl)-2- ((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-5-methyl-6-(oxazol-2-yl)-2,4-dioxo-1,2-di Hydrothieno[2,3-d]pyrimidin-3(4H)-yl)cyclobutane-1-carboxylic acid 4 (760 mg, white solid), yield: 87.3%.

MS m/z (ESI): 582.2 [M+1]

¹ H NMR (400 MHz, DMSO) δ 12.17 (s, 1H), 8.22 (s, 1H), 7.49 (d, J = 6.7 Hz, 1H), 7.39 (s, 1H), 7.30 (t, J = 7.2) Hz, 1H), 7.10-6.92 (m, 2H), 5.33-5.26 (m, 1H), 5.04 (m, 1H), 4.16-3.89 (m, 2H), 3.78 (s, 3H), 3.56 (m, 2H), 3.43-3.35 (m, 1H), 3.24 (m, 2H), 3.00-2.81 (m, 3H), 2.77 (s, 3H), 2.46 (m, 2H), 1.63 (s, 2H), 1.34 -1.13(m,2H).

Example 5 and Example 6

(1S,3S)-3-(1-((S)-2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl) 5-5-methyl-6-(oxazol-2-yl)-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl) ring Butane-1-carboxylic acid

(1S,3S)-3-(1-((R)-2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl) 5-5-methyl-6-(oxazol-2-yl)-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl) ring Butane-1-carboxylic acid

first step

(1S,3S)-3-(1-((S)-2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl) 5-5-methyl-6-(oxazol-2-yl)-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl) ring Butane-1-carboxylic acid

(1S,3S)-3-(1-((R)-2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl) 5-5-methyl-6-(oxazol-2-yl)-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl) ring Butane-1-carboxylic acid

(1S,3S)-3-(1-(2-(2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-5- Methyl-6-(oxazol-2-yl)-2,4-dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl)cyclobutane- 1-carboxylic acid 4 (130 mg, 0.23 mmol) was resolved by preparative chromatography using chiral column ChiralCel OJ, 250 x 30 mm ID, 5 μm; 70 mL by supercritical fluid chromatography (SFC). /min; mobile phase is A (for CO ₂ ) and B (for methanol) (0.1% NH ₃ .H ₂ O)), yielding (1S,3S)-3-(1-((S)-2-( 2-methoxyphenyl)-2-((tetrahydro-2H-pyran-4-yl)oxy)ethyl)-5-methyl-6-(oxazol-2-yl)-2, 4-Dioxo-1,2-dihydrothieno[2,3-d]pyrimidin-3(4H)-yl)cyclobutane-1-carboxylic acid 5 (59.12 mg, white solid), yield: 45.48 %, retention time 3.736min, ee value 100%; (1S, 3S)-3-(1-((R)-2-(2-methoxyphenyl)-2-((tetrahydro-2H-py)喃-4-yl)oxy)ethyl)-5-methyl-6-(oxazol-2-yl)-2,4-dioxo-1,2-dihydrothieno[2,3- d] Pyrimidine-3(4H)-yl)cyclobutane-1-carboxylic acid 6 (68.73 mg, white solid), yield: 52.87%, retention time 4.066 min, ee value 99.8%.

Compound 5

MS m/z (ESI): 582.2 [M+1]

¹ H NMR (400 MHz, DMSO) δ 12.17 (s, 1H), 8.22 (s, 1H), 7.49 (d, J = 6.7 Hz, 1H), 7.39 (s, 1H), 7.30 (t, J = 7.2) Hz, 1H), 7.10-6.92 (m, 2H), 5.33-5.26 (m, 1H), 5.04 (m, 1H), 4.16-3.89 (m, 2H), 3.78 (s, 3H), 3.56 (m, 2H), 3.43-3.35 (m, 1H), 3.24 (m, 2H), 3.00-2.81 (m, 3H), 2.77 (s, 3H), 2.46 (m, 2H), 1.63 (s, 2H), 1.34 -1.13(m,2H).

Compound 6

MS m/z (ESI): 582.2 [M+1]

¹ H NMR (400 MHz, DMSO) δ 12.17 (s, 1H), 8.22 (s, 1H), 7.49 (d, J = 6.7 Hz, 1H), 7.39 (s, 1H), 7.30 (t, J = 7.2) Hz, 1H), 7.10-6.92 (m, 2H), 5.33-5.26 (m, 1H), 5.04 (m, 1H), 4.16-3.89 (m, 2H), 3.78 (s, 3H), 3.56 (m, 2H), 3.43-3.35 (m, 1H), 3.24 (m, 2H), 3.00-2.81 (m, 3H), 2.77 (s, 3H), 2.46 (m, 2H), 1.63 (s, 2H), 1.34 -1.13(m,2H).

Biological evaluation

Test Example 1. Determination of IC ₅₀ of inhibition of enzymatic activity of ACC1 and ACC2 by the compound of the present invention

The following method was used to determine the degree of inhibition of the enzymatic activity of recombinant human ACC1, ACC2 protein by the preferred compounds of the invention under in vitro conditions.

The principle of the method is based on the reaction of AMC-catalyzed acetyl-CoA to form malonyl-CoA. ATP is consumed during this reaction and ADP is produced. The resulting reaction can be reconverted to ADP by the kinase using ADP-Glo ^TM kit from Promega (Promega) to ATP, which may be part of the kit in the ATP luciferase - luciferin reaction, and generates a chemical Illuminated signal. Therefore, by measuring the intensity of the chemiluminescent signal, the amount of ADP produced in the catalytic reaction can be reflected, thereby indirectly determining the enzymatic activity of the ACC protein and the effect of the test compound on the enzyme activity. The main reagents used were: ACC1, ACC2 protein (purchased from BPS bioscience, ACC1 Cat. No. 50200, ACC2 Cat. No. 50201), Acetyl CoA (acetyl-CoA, purchased from Sigma, Cat. No. A2056), NaHCO3 (purchased from Sigma, Cat. No. S6014). ), ADP-Glo ^TM Kinase assay kit (purchased from Promega, Cat. No. V9102).

The test procedure is briefly described as follows: First, the 1x buffer required for the reaction was prepared, and its composition was as follows: 50 mM HEPES (pH 7.4 purchased from Invitrogen, Cat. No. 15630), 2 mM magnesium chloride (MgCl ₂ , purchased from Sigma, article number M1028), 2 mM lemon Potassium citrate (purchased from Sigma, Cat. No. 89306), 0.01% Brij-35 detergent (available from Merck, Cat. No. 203728), 2 mM DTT (purchased from Sigma, Cat. No. D0632). The test compound powder was dissolved in DMSO to prepare a stock solution having a concentration of 10 mM, and then subjected to a 3-fold dilution to prepare a concentration required for the test, and each compound was set at 10 concentration points in a concentration range of 10 μM to 0.5 nM. First, add appropriate amount of ACC protein (2nM) to the 384-well microplate, and then add diluted test compound solutions to each well. Each concentration is provided with a duplicate well control and a solvent control (blank group) ), negative control group (DMSO group). The 384-well plates were then shaken on a microplate shaker and incubated for 15 minutes at room temperature. Thereafter, a mixture of substrates containing ATP, acetyl-CoA and NaHCO3 diluted with the aforementioned buffer was added to each well to start the reaction, and the final concentrations of the three components were ATP 20 μM, acetyl-CoA 10 μM, and NaHCO 3 30 mM, respectively. After the reaction at room temperature for 30 minutes in accordance with the ADP-Glo ^TM Kinase assay kit instructions in the kit, the reaction solution was added to each well and the liquid corresponding to the detection (refer to the specific method of operation instructions of the kit), and finally the Envision 2104 Relative light unit (RLU) values for each well were determined on a multi-plate reader (Perkin Elmer). The percent inhibition of a compound's inhibition of ACC enzyme activity at a concentration is calculated by the following formula:

Inhibition rate % = [(negative control well RLU mean - blank well RLU average) - (test well RLU mean - blank well RLU mean)] / (negative control well RLU mean - blank well RLU mean) * 100

Finally, a non-linear regression analysis was performed on the logarithm of the concentration of the compound and the percent inhibition of the corresponding concentration in the GraphPad Prism 5 software to obtain the half-inhibitory concentration value (IC ₅₀ ) of the compound.

Table 1 IC ₅₀ data for inhibition of ACC1 enzyme and ACC2 enzyme activity by the compounds of the present invention

Example number	IC 50 (nM)/ACC1	IC 50 (nM)/ACC2
1	1.3	ND
3	1.5	4.0
4	0.9	ND
5	ND	2.0
6	0.5	ND

Note: ND means not measured.

Conclusion: The compounds of the present invention have a good inhibitory effect on both ACC1 and ACC2 enzymes.

Test Example 2: Inhibitory activity of the compound of the present invention for [ ¹⁴ C]-acetate incorporation into HepG2 cells

The following method was used to determine the degree of inhibition of [ ^14C ]-acetate incorporation into HepG2 cells by the compounds of the invention under in vitro conditions.

1. Reagents and instruments

1.1. Reagents and consumables

Table 2 Reagents and consumables

Table 3 Reagents and consumables

Reagent	Vendor	Reagent
Sodium hydroxide	Tianjin Fuchen Chemical Reagent Factory	Sodium hydroxide
Potassium hydroxide	Beijing Jingqi Chemical Products Co., Ltd.	Potassium hydroxide
acetic acid	Tianjin Guangfu Technology Development Co., Ltd.	acetic acid
Trichloromethane	Beijing Chemical Factory	Trichloromethane
Ether	Tianjin Jindong Tianzheng Fine Chemical Reagent Factory	Ether
Petroleum ether	Tianjin Jindong Tianzheng Fine Chemical Reagent Factory	Petroleum ether
hydrochloric acid	Beijing Xingqinghong Fine Chemical Technology Co., Ltd.	hydrochloric acid
Hexane	Beijing Chemical Factory	Hexane

N-heptane	Beijing Chemical Factory	N-heptane
Absolute ethanol	Beijing Chemical Factory	Absolute ethanol

1.2. Instrument

Table 4 Instruments

instrument	Vendor	Item number
Biological safety cabinet	Thermo Scientific	1300 Series A2
Centrifuge	Eppendorf	5702
Carbon dioxide incubator	Thermo Scientific	1300 SERIES A2
Cell counter	Invitrogen	C10281
Pipette	BIOHIT	Easypet
microscope	Olympus	CKX41
Pipette	BIOHIT	Proline Plus
Vortex oscillator	IKA	MS3 basic
MicroBeta	PerkinElmer	2450

1.3 Preparation of the compounds of the invention

The compounds of the invention to be tested were all dissolved in DMSO at 10 mM and stored at 4 °C prior to use.

2. Experimental steps

2.1. Cell culture

HepG2 cells were purchased and purchased in the American Type Culture Collection (ATCC) resource bank. The cells were incubated in DMEM containing 10% fetal bovine serum, penicillin (100 units/mL) and streptomycin (100 μg/mL) in a 37 ° C incubator containing 5% carbon dioxide, and passaged every 2 to 3 days.

2.2.[2- ¹⁴ C]-Acetate Intake Experiment

(1) On the first day, HepG2 cells were seeded at 2 × 105 cells per well in a 24-well plate, and incubated in a 37 ° C incubator containing 5% carbon dioxide.

(2) On the fourth day, the medium containing the compound was replaced. The initial concentration of the compound of the present invention was 3 μM, 4 fold dilution, 5 concentration gradients, and a final DMSO concentration of 0.5% (v/v), and incubated for 1 hour in a 37 ° C incubator containing 5% carbon dioxide.

(3) 2 μCi of [2- ¹⁴ C]-Acetate was added to each well, and incubation was continued for 5 hours in a 37 ° C incubator containing 5% carbon dioxide.

(4) Transfer the medium to a 15 mL centrifuge tube, add 0.5 mL of 0.1 M NaOH to each well, and transfer the cell lysate to the corresponding 15 mL centrifuge tube.

(5) 1 mL of ethanol and 0.17 mL of 50% KOH were added to each tube and a water bath at 90 ° C for 1 hour.

(6) The sample was taken out, and after cooling to room temperature, 5 mL of petroleum ether was added to each tube, inverted several times, and centrifuged at 1000 rpm for 5 minutes. The upper organic phase is discarded and the aqueous phase is retained for fatty acid extraction.

(7) Add 1 mL of concentrated hydrochloric acid to each tube (ensure that its pH is below 1).

(8) Add 5 mL of petroleum ether per tube, invert several times, 1000 rpm, centrifuge for 5 minutes, transfer 4 mL of petroleum ether layer to a new glass tube (18 x 180 mm).

(9) Repeat step (8).

10) The pooled extracts were placed in a 64 ° C water bath and evaporated overnight.

(11) On the fifth day, fatty acids were dissolved in 240 μL of chloroform/n-hexane (1:1) containing 200 μg of linoleic acid.

(12) 10 μL of the sample was spotted on a silica gel plate and chromatographed for 10 minutes in a mixture of n-heptane:ethyl ether:acetic acid (90:30:1 by volume).

(13) Develop a fatty acid band with iodine vapor and cut into a scintillation vial and incubate with 2 mL of ULTIMA GOLD for 10 minutes at room temperature.

14) Record the scintillation signal with MicroBeta.

Non-linear regression of the concentration and the blinking signal corresponding concentration of compound in GraphPad Prism5 analysis software to give the half-maximum inhibition concentration value (IC _50).

TABLE 5 Compound of the invention ^[14 C] - ₅₀ incorporated inhibition value IC HepG2 cells acetate

Example number	IC 50 (nM)
Firsocostat (control compound)	50
3	6.25
6	19.8

Conclusion: The compounds of the present invention have a significant inhibitory effect on the incorporation of [ ¹⁴ C]-acetate into HepG2 cells, which is superior to Firsocostat as a control compound.

Test Example 3: Oral pharmacokinetic study of ICR mice of the present invention

1. Summary

ICR mice were used as test animals, and the compounds of Example 3 and the control firsocostat were intragastrically administered by LC-MS/MS method. The drug concentrations in plasma and liver were measured at different times. The compounds of the present invention were studied in mice. Pharmacokinetic characteristics in vivo.

2, the experimental program

2.1 Experimental drugs and animals

Example 3 compound and Firsocostat;

Healthy adult ICR male mice were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., and the animal body weight was 29.3-35.4 g.

2.2 drug configuration and drug delivery

Weigh an appropriate amount of the experimental drug, add 0.5% sodium carboxymethyl cellulose (CMC-Na), and grind to prepare a suspension of 1 mg / mL;

Eighteen healthy adult ICR male mice were divided into two groups. After fasting overnight, the rats were intragastrically administered at a dose of 10 mg/kg, a dose of 10 mL/kg, and a dose of 4 hours after administration.

Group A mice were intragastrically administered with a preparation of 10 mg·kg ^-1 of the compound of Example 3, and Group B mice were orally administered with a preparation of 10 mg·kg ^-1 firsocostat.

2.3 sample collection

80 μL of blood was collected through the eyelids before administration, and 0.20 mL of blood was collected through the heart after deep anesthesia with CO _{2 at} each set time point. The whole blood sample was placed in an anticoagulation tube containing EDTA-K2; and the liver tissue was immediately removed. . The sampling time points are as follows:

Plasma: 0 hours before administration, 0.5 hours after administration, 1 hour, 4 hours.

Liver: 1 hour after administration.

Blood samples were collected and placed in an EDTA-K ₂ anticoagulant tube, and plasma was centrifuged (centrifugation conditions: 1500 g, 10 minutes), and the upper plasma sample was collected into a sample tube. A portion of the liver tissue sample was weighed and homogenized by adding 20% methanol water in proportion (tissue: homogenate = 1:5, w/v). The collected biological samples were stored in a refrigerator at -40 to -20 °C before analysis.

The content of the test compound in the plasma and liver of the mice after the intragastric administration of the compound was analyzed by LC-MS/MS.

3, pharmacokinetic parameters results

The pharmacokinetic parameters of the compound of Example 3 of the present invention and Firsocostat are as follows:

Conclusion: Compared with Firsocostat, the compound of the present invention has good pharmacophore absorption and good pharmacokinetic properties; after 1 hour of administration, the drug concentration in the liver is 10740 ng/g, and the compound of Example 3 is There is a good enrichment in the liver.

Remarks: The structure of Firsocostat is as follows, prepared according to WO2013071169

All documents mentioned in the present application are hereby incorporated by reference in their entirety in their entireties in the the the the the the the the In addition, it should be understood that various modifications and changes may be made by those skilled in the art in the form of the appended claims.

Claims (19)

1. a compound of formula (I):

   

   Including stereoisomers, tautomers or pharmaceutically acceptable salts thereof,

   among them:

   X is selected from -NH-, -O- or -S-; preferably -S-;

   Ring A is selected from cycloalkyl, and R ² and N attached to ring A are not attached to the same carbon atom;

   R ^{1 is} selected from a hydrogen atom, an alkyl group or a halogen, wherein the alkyl group is further further selected from one or more selected from the group consisting of halogen, hydroxy, cyano, nitro, alkoxy, cycloalkyl, heterocyclic, Aryl, heteroaryl, -NR ⁸ R ⁹ , -C(O)NR ⁸ R ⁹ , -C(O)R ¹⁰ , -OC(O)R ¹⁰ , -S(O) _q NR ⁸ R ⁹ , Substituted by a substituent of -NR ⁸ S(O) ₂ R ⁹ or -NR ⁸ C(O)R ⁹ ;

   R ^{2 is} selected from the group consisting of a hydrogen atom, a hydroxyl group, a halogen, a nitro group, a cyano group, an alkyl group, an alkoxy group, a cycloalkyl group, a heterocyclic group, an aryl group, a heteroaryl group, -NR ⁸ R ⁹ , -C(O) NR ⁸ R ⁹ , -C(O)R ¹⁰ , -OC(O)R ¹⁰ , -S(O) _q NR ⁸ R ⁹ , -NR ⁸ S(O) ₂ R ⁹ or -NR ⁸ C(O) R ⁹ wherein said alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl group is further further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl , alkoxy, cycloalkyl, heterocyclic, aryl, heteroaryl, -NR ¹¹ R ¹² , -C(O)NR ¹¹ R ¹² , -C(O)R ¹³ , -C(O)OR Substituted by a substituent of ¹³ or -NR ¹¹ C(O)R ¹² ;

   R ^{3 is} selected from aryl or heteroaryl, wherein said aryl or heteroaryl is optionally further substituted with one or more substituents selected from R ⁷ ;

   R ⁴ and R ⁵ are each independently selected from a hydrogen atom, an alkyl group, -OR ¹⁰ , -SR ¹⁰ , -NR ⁸ R ⁹ , -C(O)NR ⁸ R ⁹ , -C(O)R ¹⁰ , -OC (O) R ¹⁰ , -S(O) _q NR ⁸ R ⁹ , -NR ⁸ S(O) ₂ R ⁹ or -NR ⁸ C(O)R ⁹ ;

   Alternatively, R ⁴ , R ⁵ together with the atom to which they are attached form a 3 to 8 membered saturated or partially unsaturated cycloalkyl group, or form a hetero atom having one or more selected from N, O, S(O) _q a 4 to 8 membered saturated or partially unsaturated heterocyclic group, wherein the cycloalkyl or heterocyclic group is further optionally further selected from one or more selected from the group consisting of a hydroxyl group, a halogen, a nitro group, a cyano group, an alkyl group, an alkoxy group, Cycloalkyl, heterocyclyl, aryl, heteroaryl, -NR ⁸ R ⁹ , -C(O)NR ⁸ R ⁹ , -C(O)R ¹⁰ , -OC(O)R ¹⁰ , -S( _{^{O) q NR 8 R 9,}} -NR 8 S (O) 2 R 9 or -NR ⁸ C (O) R ⁹ is substituted with a substituent;

   R ^{6 is} selected from the group consisting of halogen, cyano, cycloalkyl, heterocyclic, aryl, heteroaryl, -NR ⁸ R ⁹ , -C(O)NR ⁸ R ⁹ , -C(O)R ¹⁰ , -OC (O) R ¹⁰ , -S(O) _q NR ⁸ R ⁹ , -NR ⁸ S(O) ₂ R ⁹ or -NR ⁸ C(O)R ⁹ , preferably a heteroaryl group;

   Alternatively, R ¹ , R ⁶ together with the atom to which they are attached form a 3 to 8 membered saturated or partially unsaturated cycloalkyl group, or form a hetero atom having one or more selected from N, O, S(O) _q a 4 to 8 membered saturated or partially unsaturated heterocyclic group, or a 5 to 10 membered aryl or heteroaryl group, wherein the cycloalkyl group, heterocyclic group, aryl group or heteroaryl group is optionally further one or more One selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, -NR ⁸ R ⁹ , -C(O)NR ⁸ R ⁹ , -C(O)R ¹⁰ , -OC(O)R ¹⁰ , -S(O) _q NR ⁸ R ⁹ , -NR ⁸ S(O) ₂ R ⁹ or -NR ⁸ C(O)R ⁹ Substituted by

   R ^{7 is} each independently selected from the group consisting of hydroxyl, halogen, cyano, nitro, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, -NR ⁸ R ⁹ , -C(O) NR ⁸ R ⁹ , -C(O)R ¹⁰ , -OC(O)R ¹⁰ , -S(O) _q NR ⁸ R ⁹ , -NR ⁸ S(O) ₂ R ⁹ or -NR ⁸ C(O) R ⁹ wherein said alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl group is further further hydroxy, halo, nitro, cyano, alkyl, alkoxy, cycloalkane Base, heterocyclic group, aryl group, heteroaryl group, -NR ⁸ R ⁹ , -C(O)NR ⁸ R ⁹ , -C(O)R ¹⁰ , -OC(O)R ¹⁰ , -S(O) _{^{q NR 8 R 9, -NR 8}} S (O) 2 R 9 or -NR ⁸ C (O) R ⁹ is substituted with a substituent;

   R ⁸ , R ⁹ and R ¹⁰ are each independently selected from a hydrogen atom, an alkyl group, -OR ¹³ , a cyano group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, wherein the alkyl group, cycloalkyl group Or a heterocyclyl, aryl or heteroaryl group optionally further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, hetero Substituted with a substituent of aryl, -NR ¹¹ R ¹² , -C(O)NR ¹¹ R ¹² , -C(O)R ¹³ , -C(O)OR ¹³ or -NR ¹¹ C(O)R ¹² ;

   Alternatively, R ⁸ and R ⁹ together with the N atom to which they are attached form a 4 to 8 membered heterocyclic group, wherein the 4 to 8 membered heterocyclic ring contains one or more N, O, S(O) _q atoms, And the 4-8-membered heterocyclic ring is further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclic, aryl, heteroaryl, =0. Substituting a substituent of -NR ¹¹ R ¹² , -C(O)NR ¹¹ R ¹² , -C(O)R ¹³ , -C(O)OR ¹³ or -NR ¹¹ C(O)R ¹² ;

   R ¹¹ , R ¹² and R ¹³ are each independently selected from a hydrogen atom, an alkyl group, an alkenyl group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, wherein the alkyl group, a cycloalkyl group, a heterocyclic group Or an aryl or heteroaryl group optionally further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, carboxy Substituted by a substituent of an acid or a carboxylic acid ester;

   q is 0, 1, or 2.
2. A compound according to claim 1 having the structure of formula (II):

   

   among them:

   m is 1, 2, 3, 4 or 5;

   The definition of ring A, R ¹ , R ² , R ⁶ , R ⁷ and R ¹⁰ is as set forth in claim 1.
3. A compound according to claim 2 having the structure of formula (III):

   

   among them:

   m is 1, 2, 3, 4 or 5;

   The definition of ring A, R ¹ , R ² , R ⁶ , R ⁷ and R ¹⁰ is as set forth in claim 1.
4. A compound according to claim 2 having the structure of formula (IV):

   

   among them:

   m is 1, 2, 3, 4 or 5;

   The definition of ring A, R ¹ , R ² , R ⁶ , R ⁷ and R ¹⁰ is as set forth in claim 1.
5. The compound according to any one of claims 1 to 4, wherein ring A is selected from the group consisting of:

   

   Preferred

   
6. The compound according to any one of claims 1 to 5, wherein R ^{1 is} selected from methyl or trifluoromethyl.
7. The compound according to any one of claims 1 to 6, wherein:

   R ^{2 is} selected from the group consisting of tetrazolyl, -C(O)OR ¹³ or -C(O)NR ⁸ R ⁹ ;

   R ^{8 is} selected from a hydrogen atom or an alkyl group;

   R ^{9 is} selected from cyano or -OR ¹³ ;

   R ^{13 is} selected from a hydrogen atom, an alkyl group, an alkenyl group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, wherein the alkyl group, cycloalkyl group, heterocyclic group, aryl group or heteroaryl group is optional Further substituted by one or more substituents selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, carboxylic acid or carboxylic acid esters Replace

   R ² is -C(O)OH.
8. The compound according to any one of claims 1 to 7, wherein R ^{6 is} selected from a 5-membered heteroaryl group, preferably a thiazolyl group.
9. The compound according to any one of claims 1 to 8, wherein R ^{7 is} selected from halogen or alkoxy, preferably methoxy.
10. The compound according to any one of claims 1 to 9, or a stereoisomer, tautomer thereof or a pharmaceutically acceptable salt thereof, wherein R ¹⁰ is tetrahydropyran-4-yl.
11. A compound according to claim 1 selected from the group consisting of:

    
12. A method of preparing a compound of formula (I) according to claim 1, the method comprising:

    

    The compound of the formula (IA) is reacted with an R ⁶ -substituted tributylstannane such that the resulting compound is optionally further hydrolyzed, and the hydrolyzed compound is optionally further resolved to the optically pure isomer to give the compound of the formula (I);

    Wherein: X ^{1 is} selected from halogen; and X, ring A, R ¹ to R ⁶ are as defined in claim 1.
13. a compound of the formula (IA):

    

    Including stereoisomers, tautomers or pharmaceutically acceptable salts thereof,

    among them:

    X ^{1 is} selected from halogen; and X, ring A, R ¹ to R ⁵ are as defined in claim 1.
14. A compound according to claim 13 selected from the group consisting of:

    
15. A method of preparing a compound of formula (IA) according to claim 13, the method comprising:

    

    The compound of formula (IB) is reacted with a compound of formula (IC) in the presence of triphenylphosphine to provide a compound of formula (IA);

    among them:

    X ^{1 is} selected from halogen; and X, ring A, R ¹ to R ⁵ are as defined in claim 1.
16. A pharmaceutical composition comprising an effective amount of a compound according to any one of claims 1 to 11, and optionally a pharmaceutically acceptable carrier, excipient or combination thereof.
17. Use of a compound according to any one of claims 1 to 11 or a pharmaceutical composition according to claim 16 for the preparation of a medicament for use as an ACC inhibitor.
18. Use of a compound according to any one of claims 1 to 11 or a pharmaceutical composition according to claim 16 for the preparation of a medicament for preventing or treating a disease or condition associated with ACC, wherein the disease or The condition is preferably a metabolic disease, a cancer, a fungus, a parasite or a bacterial infection; wherein the metabolic disease is preferably hepatic steatosis, nonalcoholic fatty liver, obesity, dyslipidemia, hyperlipidemia, type II diabetes or Metabolic syndrome, wherein the obesity is preferably Prader-Willi syndrome, Bardet-Biedl syndrome or Cohen syndrome or MOMO A syndrome wherein the cancer is preferably hepatocellular carcinoma, non-small cell lung cancer, small cell lung cancer, gastric cancer, colorectal cancer, head and neck cancer, melanoma, ovarian cancer or cervical cancer, more preferably hepatocellular carcinoma and non- Small Cell Lung Cancer.
19. A method of preventing or treating a disease or condition associated with ACC, comprising administering a compound according to any one of claims 1 to 11 or a pharmaceutical composition according to claim 16 to a subject in need thereof.