RU2801942C9 - COMPOUNDS BASED ON ISOTHIAZOLO[5,4-d]PYRIMIDINE AS IRAK4 INHIBITOR - Google Patents

Abstract

FIELD: pharmaceuticals.

SUBSTANCE: invention relates to a compound represented by formula (III), where R₁-R₃, L₁, L₂, ring A are defined in the claims, and a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing it, and the use of the compound.

EFFECT: compound can be used for the preparation of a medicinal product for the treatment of diseases associated with IRAK4.

20 cl, 2 dwg, 7 tbl, 60 ex

Description

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit and priority of Chinese Patent Application No. 201810662580.9, filed June 25, 2018, and Chinese Patent Application No. 201910463156.6, filed May 30, 2019, with the National Intellectual Property Administration, PR _c , which are incorporated herein by reference in its entirety.

TECHNICAL FIELD

The present invention relates to an IRAK4 inhibitor and its use for the preparation of a medicament for the treatment of IRAK4-related diseases, in particular to a compound of formula (I) and a pharmaceutically acceptable salt thereof.

PREREQUISITES FOR CREATION OF THE INVENTION

Interleukin-1 receptor-associated kinase 4 (IRAK4) is a serine/threonine-specific protein kinase, a member of the tyrosine-like kinase (TLK) family and a key element of the innate immune response, which involves interleukin-1, 18 and 33, as well as toll-like receptors. Upon binding of extracellular signaling molecules to interleukin receptors or toll-like receptors, proteins are recruited to form the MyD88:IRAK4:IRAK1/2 complex, leading to phosphorylation of IRAK1/2, which mediates signal transduction to downstream components of the signaling pathway. Thus, the p38, JNK and NF-κB signaling pathways are activated, ultimately leading to increased expression of proinflammatory cytokines. Clinical pathology studies have shown that individuals with IRAK4 mutations are resistant to chronic lung disease and inflammatory bowel disease. IRAK4 deficiency per se is not lethal, and individuals can survive into adulthood with a reduced risk of infection throughout life. Consequently, IRAK4 is emerging as an important therapeutic target attracting extensive research and development interest.

SHORT DESCRIPTION

The present invention provides a compound of formula (III), its optical isomer or pharmaceutically acceptable salt,

Where

R ₁ is selected from the group consisting of CN, C _1-6 alkyl and 3-6 membered heterocycloalkyl, wherein C _1-6 alkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _a ;

R ₂ is selected from the group consisting of C _1-6 alkyl, C _3-8 cycloalkyl and 3-6 membered heterocycloalkyl, wherein C _1-6 alkyl, C _3-8 cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1. 2 or 3 _Rb ;

R ₃ is selected from the group consisting of H, F, Cl, Br, I, OH, NH ₂ , CN, COOH, C _1-6 alkyl, C _1-6 alkylamino, -C(=O)-O-C _{1 -6} alkyl, -C(=O)-C _1-6 alkyl and C _3-6 cycloalkyl, where C _1-6 alkyl, C _1-6 alkylamino, -C(=O)-O-C _1-6 alkyl , -C(=O)-C _1-6 alkyl and C _3-6 cycloalkyl are optionally substituted with 1, 2 or 3 R _c ;

ring A is selected from the group consisting of 3-10 membered heterocycloalkyl, and ring A contains at least one nitrogen atom, wherein the 3-10 membered heterocycloalkyl is optionally substituted with 1, 2 or 3 R _d ;

L ₁ is selected from the group consisting of O and N (R ₄ );

L ₂ is selected from the group consisting of a single bond, CH ₂ and CH ₂ CH ₂ ;

R ₄ is selected from the group consisting of H and Me;

each R _a is independently selected from the group consisting of H, F, Cl, Br, I, OH, NH ₂ , CN and COOH;

each R _b is independently selected from the group consisting of H, F, Cl, Br, I, OH, NH ₂ , COOH and Me;

each R _c is independently selected from the group consisting of H, F, Cl, Br, I, OH, NH ₂ and CN;

each R _d is independently selected from the group consisting of H, F, Cl, Br, I, OH, NH ₂ and CN;

3-6 membered heterocycloalkyl contains 1, 2 or 3 heteroatoms or heteroatomic groups independently selected from the group consisting of -O-, -S-, -NH- and N; And

3-10 membered heterocycloalkyl contains 1, 2 or 3 heteroatoms or heteroatom groups independently selected from the group consisting of -O-, -S-, -NH-, N and -C(=O)NH-.

The present invention further provides a compound of formula (II), an optical isomer or a pharmaceutically acceptable salt thereof,

Where

R ₁ is selected from the group consisting of C _1-6 alkyl and 3-6 membered heterocycloalkyl, wherein C _1-6 alkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _a ;

R ₂ is selected from the group consisting of C _1-6 alkyl and 3-6 membered heterocycloalkyl, wherein the C _1-6 alkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _b ;

R ₃ is selected from the group consisting of H, F, Cl, Br, I, OH, NH ₂ , CN, C _1-6 alkyl, -C(=O)-O-C _1-6 alkyl, -C(= O)-C _1-6 alkyl and C _3-6 cycloalkyl, where C _1-6 alkyl, -C(=O)-O-C _1-6 alkyl, -C(=O)-C _1-6 alkyl and The C _3-6 cycloalkyl is optionally substituted with 1, 2 or 3 R _c ;

ring A is selected from the group consisting of 3-10 membered heterocycloalkyl, and ring A contains at least one nitrogen atom, wherein the 3-10 membered heterocycloalkyl is optionally substituted with 1, 2 or 3 R _d ;

L ₁ is selected from the group consisting of O and N (R ₄ );

L ₂ is selected from the group consisting of a single bond, CH ₂ and CH ₂ CH ₂ ;

R ₄ is selected from the group consisting of H and Me;

each R _a is independently selected from the group consisting of H, F, Cl, Br, I, OH, NH ₂ , CN and COOH;

each R _b is independently selected from the group consisting of H, F, Cl, Br, I, OH, NH ₂ , COOH and Me;

each R _c is independently selected from the group consisting of H, F, Cl, Br, I, OH, NH ₂ and CN;

each R _d is independently selected from the group consisting of H, F, Cl, Br, I, OH, NH ₂ and CN;

3-6 membered heterocycloalkyl contains 1, 2 or 3 heteroatoms or heteroatomic groups independently selected from the group consisting of -O-, -S-, -NH- and N; And

3-10 membered heterocycloalkyl contains 1, 2 or 3 heteroatoms or heteroatom groups independently selected from the group consisting of -O-, -S-, -NH-, N and -C(=O)NH-.

The present invention further provides a compound of formula (I), an optical isomer or a pharmaceutically acceptable salt thereof,

Where

R ₁ is selected from the group consisting of C _1-6 alkyl and 3-6 membered heterocycloalkyl, wherein C _1-6 alkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _a ;

R ₂ is selected from the group consisting of C _1-6 alkyl and 3-6 membered heterocycloalkyl, wherein C _1-6 alkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _b ;

each R _a is independently selected from the group consisting of F, Cl, Br, I, OH and NH ₂ ;

each R _b is independently selected from the group consisting of F, Cl, Br, I, OH and NH ₂ ;

L ₁ is selected from the group consisting of O, S and NH;

L ₂ is selected from the group consisting of a single bond, CH ₂ and CH ₂ CH ₂ ; And

A 3-6 membered heterocycloalkyl contains 1, 2 or 3 heteroatoms or heteroatom groups independently selected from the group consisting of -O-, -S-, -NH- and N.

In some embodiments of the present invention, the L ₁ described above is selected from the group consisting of O and N(R ₄ ), other variables being as defined herein.

R ₁ is selected from the group consisting of CN, C _1-6 alkyl, 3-membered heterocycloalkyl, 4-membered heterocycloalkyl, 5-membered heterocycloalkyl and 6-membered heterocycloalkyl, wherein C _1-6 alkyl, 3-membered heterocycloalkyl, 4- member heterocycloalkyl, 5 member heterocycloalkyl and 6 member heterocycloalkyl are optionally substituted with 1, 2 or 3 R _a , other variables being as defined herein.

In some embodiments of the present invention, R ₁ above is selected from the group consisting of C _1-3 alkyl and 6-membered heterocycloalkyl, wherein the C _1-3 alkyl and 6-membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _a , while others the variables are as defined in this document.

In some embodiments of the present invention, R ₁ above is selected from the group consisting of C _1-3 alkyl and tetrahydropyranyl, wherein the C _1-3 alkyl and tetrahydropyranyl are optionally substituted with 1, 2 or 3 R _a , other variables being as defined in this document.

In some embodiments of the present invention, R ₁ above is selected from the group consisting of methyl, ethyl, isopropyl and tetrahydropyranyl, wherein methyl, ethyl, isopropyl and tetrahydropyranyl are optionally substituted with 1, 2 or 3 R _a , other variables being as defined in this document.

In some embodiments of the present invention, R ₁ above is selected from the group consisting of Me, Et, , where Me, Et, optionally substituted with 1, 2 or 3 R _a , other variables being as defined herein.

In some embodiments of the present invention, each R _a is independently selected from the group consisting of F, Cl, Br, I, OH, NH ₂ , CN and COOH, other variables being as defined herein.

In some embodiments of the present invention, each R _a is independently selected from the group consisting of OH, CN and COOH, with other variables being as defined herein.

In some embodiments of the present invention, R ₁ above is selected from the group consisting of Me, Et, provided that other variables are as defined herein. In some embodiments of the present invention, R ₁ above is selected from the group consisting of Me, Et, provided that other variables are as defined herein.

In some embodiments of the present invention, R ₂ above is selected from the group consisting of C _1-3 alkyl, C _4-6 cycloalkyl, tetrahydropyranyl, oxetanyl, tetrahydrofuranyl and 1,4-dioxanyl, wherein C _1-3 alkyl, C _4-6 cycloalkyl, tetrahydropyranyl, oxetanyl, tetrahydrofuranyl and 1,4-dioxanyl are optionally substituted with 1, 2 or 3 R _b , other variables being as defined herein.

In some embodiments of the present invention, R ₂ above is selected from the group consisting of C _1-6 alkyl, 3-membered heterocycloalkyl, 4-membered heterocycloalkyl, 5-membered heterocycloalkyl, and 6-membered heterocycloalkyl, wherein C _1-6 alkyl, 3- membered heterocycloalkyl, 4-membered heterocycloalkyl, 5-membered heterocycloalkyl and 6-membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _b , other variables being as defined herein.

In some embodiments of the present invention, R ₂ above is selected from the group consisting of C _1-3 alkyl, 4-membered heterocycloalkyl, 5-membered heterocycloalkyl, and 6-membered heterocycloalkyl, wherein C _1-3 alkyl, 4-membered heterocycloalkyl, 5- member heterocycloalkyl and 6 member heterocycloalkyl are optionally substituted with 1, 2 or 3 R _b , other variables being as defined herein. In some embodiments of the present invention, R ₂ above is selected from the group consisting of C _1-3 alkyl, tetrahydropyranyl, oxetanyl, tetrahydrofuranyl and 1,4-dioxanyl, wherein C _1-3 alkyl, tetrahydropyranyl, oxetanyl, tetrahydrofuranyl and 1,4- dioxanyl is optionally substituted with 1, 2 or 3 R _b , other variables being as defined herein.

In some embodiments of the present invention, R ₂ above is selected from the group consisting of C _1-3 alkyl and tetrahydro-2H-pyranyl, wherein the C _1-3 alkyl and tetrahydropyranyl are optionally substituted with 1, 2 or 3 R _b , other variables being as defined in this document. In some embodiments of the present invention, R ₂ above is selected from the group consisting of Me, Et, and cyclohexyl, where Me, Et, and cyclohexyl are optionally substituted with 1, 2 or 3 R _b , other variables being as defined herein.

In some embodiments of the present invention, R ₂ above is selected from the group consisting of Me, Et, where Me, Et, optionally substituted with 1, 2 or 3 R _b , other variables being as defined herein.

In some embodiments of the present invention, R ₂ above is selected from the group consisting of Me, Et, where Me, Et, optionally substituted with 1, 2 or 3 R _b , other variables being as defined herein.

In some embodiments of the present invention, each R _b is independently selected from the group consisting of F, Cl, Br, I, OH, NH ₂ , COOH and Me.

In some embodiments of the present invention, each R _b is independently selected from the group consisting of F, OH, NH ₂ , COOH and Me.

In some embodiments of the present invention, R ₂ is selected from the group consisting of Me, -CH ₂ OH, Et, provided that other variables are as defined herein.

In some embodiments of the present invention, R ₂ above is selected from the group consisting of Me, -CH ₂ OH, Et, provided that other variables are as defined herein.

In some embodiments of the present invention, R ₂ above is selected from the group consisting of Me, Et, provided that other variables are as defined herein.

In some embodiments of the present invention, L ₂ above is a single bond.

In some embodiments of the present invention, the structural unit above selected from the group consisting of Me, Et, provided that other variables are as defined herein.

In some embodiments of the present invention, the structural unit above selected from the group consisting of Me, Et, provided that other variables are as defined herein.

In some embodiments of the present invention, the structural unit above selected from the group consisting of Me, Et, provided that other variables are as defined herein.

In some embodiments of the present invention, R ₃ above is selected from the group consisting of H, F, Cl, Br, I, OH, NH ₂ , CN, COOH, C _1-3 alkyl, -N(C _1-3 alkyl) ₂ , -C(=O)-O-C _1-3 alkyl, -C(=O)-C _1-3 alkyl and C _3-6 cycloalkyl, where C _1-3 alkyl, -N(C _1-3 alkyl ) ₂ , -C(=O)-O-C _1-3 alkyl, -C(=O)-C _1-3 alkyl and C _3-6 cycloalkyl are optionally substituted with 1, 2 or 3 R _c , other variables are as defined herein.

In some embodiments of the present invention, R ₃ above is selected from the group consisting of H, F, Cl, Br, I, OH, NH ₂ , CN, C _1-3 alkyl, -C(=O)-O-C _{1- 3} alkyl, -C(=O)-C _1-3 alkyl and C _3-6 cycloalkyl, where C _1-3 alkyl, -C(=O)-O-C _1-3 alkyl, -C(=O) -C _1-3 alkyl and C ₃ -bcycloalkyl are optionally substituted with 1, 2 or 3 R _c , other variables being as defined herein.

In some embodiments of the present invention, each R _c is independently selected from the group consisting of F, Cl, Br, I, OH, NH ₂ and CN. In some embodiments of the present invention, R ₃ above is selected from the group consisting of H, F, Cl, Br, I, OH, NH ₂ , CN, COOH, Me, Et, -C(=O)-O-Me, -C(=O)-O-Et and -C(=O)-Me, other variables being as defined herein.

In some embodiments of the present invention, R ₃ above is selected from the group consisting of H, F, Cl, Br, I, OH, NH ₂ , CN, Me, Et, -C(=O)-O-Me, -C(=O)-O-Et and -C(=O)-Me, other variables being as defined herein.

In some embodiments of the present invention, ring A above is selected from the group consisting of 3, 4, 5, 6, 7, 8, 9 or 10 membered heterocycloalkyl, wherein 3, 4, 5, 6, 7, 8, 9 or 10 -membered heterocycloalkyl is optionally substituted with 1, 2 or 3 R _d , other variables being as defined herein.

In some embodiments of the present invention, ring A above is selected from the group consisting of 4, 5, 6, 7 or 8 membered heterocycloalkyl, wherein the 4, 5, 6, 7 or 8 membered heterocycloalkyl is optionally substituted with 1, 2 or 3 R _d , other variables being as defined herein.

In some embodiments of the present invention, ring A above is selected from the group consisting of 4, 6, 7 or 8 membered heterocycloalkyl, wherein the 4, 6, 7 or 8 membered heterocycloalkyl is optionally substituted with 1, 2 or 3 R _d , while others the variables are as defined in this document.

In some embodiments of the present invention, ring A above is selected from the group consisting of 6, 7 or 8 membered heterocycloalkyl, wherein the 6, 7 or 8 membered heterocycloalkyl is optionally substituted with 1, 2 or 3 _Rd , other variables being such as as defined herein.

In some embodiments of the present invention, ring A above is selected from the group consisting of 6 or 7 membered heterocycloalkyl, wherein the 6 or 7 membered heterocycloalkyl is optionally substituted with 1, 2 or 3 _Rd , other variables being as defined herein document.

In some embodiments of the present invention, ring A above is selected from the group consisting of morpholinyl, piperazinyl, 3-morpholinonyl, 2-piperazinonyl, homopiperazinyl, 4,7-diazaspiro[2,5]octyl, 3,6-diazabicyclo[3,1 ,1]heptyl, 2-azacyclohexanonyl, 2,5-diazabicyclo[2.2.1]heptyl and azetidinyl, where morpholinyl, piperazinyl, 3-morpholinonyl, 2-piperazinonyl, homopiperazinyl, 4,7-diazaspiro[2,5]octyl, 3,6-diazabicyclo[3,1,1]heptyl, 2-azacyclohexanonyl, 2,5-diazabicyclo[2.2.1]heptyl and azetidinyl are optionally substituted with 1, 2 or 3 R _d , other variables being as defined in this document.

In some embodiments of the present invention, ring A above is selected from the group consisting of morpholinyl, piperazinyl, 3-morpholinonyl, 2-piperazinonyl, homopiperazinyl, 4,7-diazaspiro[2,5]octyl, 3,6-diazabicyclo[3,1 ,1]heptyl, 2-azacyclohexanonyl and 2,5-diazabicpo[2.2.1]heptyl, where morpholinyl, piperazinyl, 3-morpholinonyl, 2-piperazinonyl, homopiperazinyl, 4,7-diazaspiro[2,5]octyl, 3, 6-diazabicyclo[3,1,1]heptyl, 2-azacyclohexanonyl and 2,5-diazabicyclo[2.2.1]heptyl are optionally substituted with 1, 2 or 3 R _d , other variables being as defined herein.

In some embodiments of the present invention, each R _d is independently selected from the group consisting of F, Cl, Br, I, OH, NH ₂ and CN.

In some embodiments of the present invention, Ring A above is selected from the group consisting of morpholinyl, piperazinyl, 3-morpholinonyl, 2-piperazinonyl, homopiperazinyl, 4,7-diazaspiro[2,5]octyl, 3,6-diazabicpo[3,1 ,1]heptyl, 2-azacyclohexanonyl and 2,5-diazabicyclo[2.2.1]heptyl, other variables being as defined herein.

In some embodiments of the present invention, ring A above is selected from the group consisting of provided that other variables are as defined herein.

In some embodiments of the present invention, ring A above is selected from the group consisting of provided that other variables are as defined herein.

In some embodiments of the present invention, ring A above is selected from the group consisting of provided that other variables are as defined herein.

In some embodiments of the present invention, the structural unit selected from a group consisting of

provided that other variables are as defined herein.

In some embodiments of the present invention, the structural unit selected from a group consisting of

provided that other variables are as defined herein.

In some embodiments of the present invention, the structural unit selected from a group consisting of

provided that other variables are as defined herein.

In some embodiments of the present invention, the compound, its optical isomer, or pharmaceutically acceptable salt is selected from

wherein R ₁ , R ₂ , R ₃ , L ₁ and L ₂ are as defined herein.

Some further embodiments of the present invention are derived from any combination of variables as described above.

The present invention also provides a compound of the following formula, an optical isomer thereof, or a pharmaceutically acceptable salt which is selected from:

In some: embodiments of the present invention, the compound above, an optical isomer thereof, or a pharmaceutically acceptable salt thereof is selected from the group consisting of

The present invention also provides a pharmaceutical composition containing a therapeutically effective amount of the compound above, an isomer thereof, or a pharmaceutically acceptable salt thereof as the active ingredient, and pharmaceutically acceptable carriers.

The present invention also provides a method of treating diseases associated with IRAK4 in a mammal, comprising administering to a mammal, preferably a human, in need of such treatment, a therapeutically effective amount of a compound of the above, an isomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

The present invention also provides the use of a compound of the above, an isomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition based thereon, for the preparation of a medicament for the treatment of diseases associated with IRAK4.

The present invention also provides the use of a compound of the above, an isomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition based thereon, for the treatment of diseases associated with IRAK4. The present invention also provides a compound of the above, an isomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition based thereon, for the treatment of diseases associated with IRAK4.

The present invention also provides for the use of a compound above, an isomer thereof, or a pharmaceutically acceptable salt thereof for the preparation of an IRAK4 inhibitor.

The present invention also provides for the use of the composition above for the production of an IRAK4 inhibitor.

DEFINITIONS

Unless otherwise specified, the following terms and phrases as used herein have the following meanings. A particular term or phrase, unless otherwise specifically defined, shall not be considered vague or unclear, but shall be construed in accordance with its generally accepted meaning. When a trade name is referred to, it refers to its corresponding commercial product or its active ingredient.

The term "pharmaceutically acceptable" is used herein for those compounds, materials, compositions and/or dosage forms that, within the scope of careful medical evaluation, are suitable for use in contact with human and animal tissues without undue toxicity, irritation, or allergic reaction. or other problems or complications, and are commensurate with an acceptable benefit/risk ratio.

The term “pharmaceutically acceptable salt” refers to a salt of a compound disclosed herein that is derived from a compound having the specific substituents disclosed herein and a relatively non-toxic acid or base. If a compound disclosed herein contains a relatively acidic functional group, a base addition salt can be prepared by contacting the neutral form of such compound with a sufficient amount of base in a pure solution or suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salts, or the like. If a compound disclosed herein contains a relatively basic functional group, an acid addition salt can be prepared by contacting the neutral form of such compound with a sufficient amount of acid in a pure solution or suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate radical, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, phosphorous acid; and salts derived from organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p -toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid. Also included are salts of amino acids (eg, arginine, etc.) and salts of organic acids such as glucuronic acid. Certain specific compounds disclosed herein contain both basic and acid functional groups, which allow the compounds to be converted into either base or acid addition salts.

The pharmaceutically acceptable salts disclosed herein can be synthesized from a parent compound having an acidic or basic group using conventional chemical methods. In general, such salts are prepared by the following process: reacting the compound in free acid or base form with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture thereof.

The compounds disclosed herein may be in the form of a geometric isomer or a stereoisomer. All such compounds are provided herein, including cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (5)-enantiomers, diastereomers, (D)-isomers, (L)- isomers and racemic mixtures thereof and other mixtures such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Substituents such as alkyl may have an additional asymmetric carbon atom. All such isomers and mixtures thereof are within the scope of the present invention.

Unless otherwise noted, the term "enantiomer" or "optical isomer" refers to stereoisomers that are mirror images of each other.

Unless otherwise noted, the term "cis-trans isomer" or "geometric isomer" is due to the inability of the double bond or single bond of a carbon atom in the ring to rotate freely.

Unless otherwise specified, the term "diastereomer" or "diastereoisomer" refers to stereoisomers whose molecule has two or more chiral centers and which are not mirror images of each other.

Unless otherwise noted, "(D)" or "(+)" means right rotation, "(L)" or "(-)" means left rotation, and "(DL)" or "(±)" means racemization.

Unless otherwise noted, the absolute configuration of the stereogenic center is represented by a solid wedge-shaped bond and dotted wedge-shaped connection and the relative configuration of the stereogenic center is represented by a direct continuous connection and direct dotted connection Wavy line is a continuous wedge-shaped connection or solid dotted connection or wavy line represents a direct continuous connection and direct dotted connection

Optically active (R)- and (S)-isomers and D and L isomers can be prepared by chiral synthesis, or chiral reagents, or other conventional techniques. The enantiomer of a particular compound disclosed herein can be obtained by asymmetric synthesis or derivatization using a chiral auxiliary, where the resulting diastereomeric mixture is separated and the auxiliary group is cleaved so as to provide the desired pure enantiomer. Alternatively, if the molecule contains a basic functional group (such as amino) or an acidic functional group (such as carboxyl), the compound is reacted with a suitable optically active acid or base to form a diastereoisomer salt, which is then subjected to diastereomeric resolution by means of those conventional in the art methods to obtain the pure enantiomer. In addition, the enantiomer and diastereomer as a whole are isolated by chromatography using a chiral stationary phase, optionally in combination with chemical derivatization (eg, carbamate formation from amines). A compound disclosed herein may contain a non-naturally occurring proportion of atomic isotopes on one or more of the atoms that form the compound. For example, the compound may be labeled with a radioactive isotope such as tritium ( ^3H ), iodine-125 ( ^125I ), or C-14 ( ^14C ). As another example, hydrogen can be replaced by deuterium to form a deuterated drug, and the bond formed between deuterium and carbon is more rigid than the bond that is formed between ordinary hydrogen and carbon. Compared with a non-deuterated drug, a deuterated drug has advantages in terms of reduced toxic side effect, increased stability, increased efficacy, increased biological half-life, and the like. All isotopic variants of the compound described herein, whether radioactive or not, are included within the scope of the present invention. “Optional” or “optional” means that the event or circumstance described may, but does not necessarily, occur in sequence, and the description includes cases in which the event or circumstance occurs and cases in which it does not.

The term "substituted" means that one or more hydrogen atoms on a particular atom are replaced by substituents, which may include deuterium and hydrogen variants, provided that the valence of the particular atom is normal and the compound is stable after substitution. If the substituent is oxygen (i.e. =O), this means that two hydrogen atoms are replaced. Substitution with oxygen does not occur in aromatic groups. The term "optionally substituted" means that the atom may or may not be replaced by a substituent. Unless otherwise indicated, the type and number of substituents may be arbitrary as long as chemically achievable.

If any variable (for example, R) occurs more than once in the composition or structure of a compound, the definition of the variable in each case is independent. Thus, for example, if a group is substituted with 0-2 R's, the group may optionally be substituted with no more than two R's, and the definition of R in each case is independent. In addition, a combination of substituents and/or variants thereof is permitted only if the combination results in a stable compound.

If the variable is a single bond, it means that the two groups are directly connected, for example in A-L-Z, if L is a single bond, it means that the structure is actually A-Z.

For listed related groups, the direction for binding that is not specified is arbitrary. For example, when binding, the group L contained in is -MW-, -MW- can bind either ring A or ring B in a direction similar to the left-to-right reading order to form , or link ring A and ring B in the opposite direction to form . A combination of a linking group, a substituent and/or a variant thereof is permitted only if the combination results in a stable compound.

Unless otherwise stated, the number of atoms in a ring as a whole is defined as the number of ring members. For example, "3-6 membered ring" refers to a "ring" in which 3-6 atoms are arranged in a ring.

Unless otherwise specified, the term "C _1-6 alkyl" refers to a straight or branched saturated hydrocarbon group of 1-6 carbon atoms. C _1-6 alkyl includes C _1-5 , C _1-4 , C _1-3 , C _1-2 , C _2-6 , C _2-4 , C ₆ and C ₅ alkyl, etc., and may be monovalent (eg methyl), divalent (eg methylene) or polyvalent (eg methenyl). Examples of C _1-6 alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, sec-butyl and tert-butyl), pentyl (including n-pentyl, isopentyl, and neopentyl), hexyl, etc.

Unless otherwise specified, the term "C _1-3 alkyl" refers to a linear or branched saturated hydrocarbon group consisting of 1-3 carbon atoms. C _1-3 alkyl includes C _1-2 and C _2-3 alkyl, etc., and may be monovalent (eg methyl), divalent (eg methylene) or polyvalent (eg methenyl). Examples of C _1-3 alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), and the like. Unless otherwise specified, "C _2-8 alkenyl" is used to refer to a straight or branched hydrocarbon group containing 2-8 carbon atoms and at least one carbon-carbon double bond, which can be located anywhere in the group. C _2-8 alkenyl includes C _2-6 , C _2-4 , C _2-3 , C ₄ , C ₃ and C ₂ alkenyl, etc., and may be monovalent, divalent or polyvalent. Examples of C _2-8 alkenyl include, but are not limited to, ethenyl, propenyl, butenyl, pentenyl, hexenyl, butadienyl, 1,3-pentadienyl, 1,3-hexadienyl and the like.

Unless otherwise specified, the term "C _1-6 alkylamino" refers to an alkyl group containing 1-6 carbon atoms that is attached to the rest of the molecule via an amino group. C _1-6 alkylamino includes C _1-4 , C 1-3, C _1-2 , C _2-6 , C _2-4 , C ₆ , C ₅ , _{C 4 ,} C ₃ , C ₂ alkylamino, etc. . Examples of C _1-6 alkylamino include, but are not limited to -NHCH ₃ , -N(CH ₃ )2, -NHCH ₂ CH ₃ , -N(CH ₃ )CH ₂ CH ₃ , -N(CH ₂ CH ₃ )(CH ₂ CH ₃ ), -NHCH ₂ CH ₂ CH ₃ , -NHCH ₂ (CH ₃ ) ₂ , -NHCH ₂ CH ₂ CH ₂ CH ₃ , etc.

Unless otherwise specified, the term "C _1-3 alkylamino" refers to an alkyl group containing 1-3 carbon atoms that is attached to the rest of the molecule via an amino group. The _C1-3 alkylamino group includes _C1-2 , _C3 , _C2 alkylamino and the like. Examples of C _1-3 alkylamino include, but are not limited to -NHCH ₃ , -M(CH ₃ ) ₂ , -NHCH ₂ CH ₃ , -N(CH ₃ )CH ₂ CH ₃ , -NHCH ₂ CH ₂ CH ₃ , -NHCH ₂ ( CH ₃ ) ₂ etc.

Unless otherwise specified, the term "3-10 membered heterocycloalkyl", used alone or in combination with other terms, denotes a saturated cyclic group consisting of 3-10 ring atoms, 1, 2, 3 or 4 of which are heteroatoms, regardless selected from the group consisting of -O-, -S-, -NH-, N and -C(=O)NH-, the remainder being carbon atoms. This includes monocyclic, bicyclic and tricyclic systems, where bicyclic and tricyclic systems include spirocyclic, fused and bridged rings. Additionally, in the case of a "3-10 membered heterocycloalkyl", the heteroatom may occupy a position where the heterocycloalkyl is attached to the rest of the molecule. 3-10 membered heterocycloalkyl includes 3-9 membered, 3-8 membered, 3-7 membered, 3-6 membered, 3-5 membered, 4-6 membered, 5-6 membered, 4 -membered, 5-membered, 6-membered heterocycloalkyl groups, etc. Examples of 3 to 10 membered heterocycloalkyl include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothienyl (including tetrahydrothien-2-yl, tetrahydrothien-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2- yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl, 2-piperazinyl, etc.), morpholinyl ( including 3-morpholinyl, 4-morpholinyl, etc.), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl, homopiperazinyl, homopiperidinyl or dioxepanyl, etc.

Unless otherwise specified, the term "3-6 membered heterocycloalkyl", used alone or in combination with other terms, denotes a saturated cyclic group consisting of 3-6 ring atoms, 1,2,3 or 4 of which are heteroatoms, regardless selected from the group consisting of -O-, -S-, -NH- and N, the remainder being carbon atoms. It includes monocyclic and bicyclic systems, where the bicyclic system includes spirocyclic, fused and bridged rings. Additionally, in the case of a "3-6 membered heterocycloalkyl", the heteroatom may occupy a position where the heterocycloalkyl is attached to the rest of the molecule. 3-6 membered heterocycloalkyl includes 4-6 membered, 5-6 membered, 4 membered, 5 membered, 6 membered heterocycloalkyl and the like. Examples of 3-6 membered heterocycloalkyl include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothienyl (including tetrahydrothien-2-yl, tetrahydrothien-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2- yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl, 2-piperazinyl, etc.), morpholinyl ( including 3-morpholinyl, 4-morpholinyl, etc.), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl, homopiperazinyl, homopiperidinyl, etc.

Unless otherwise specified, "C _3-8 cycloalkyl" refers to a saturated cyclic hydrocarbon group of 3-8 carbon atoms. It includes monocyclic and bicyclic systems, where the bicyclic system includes spirocyclic, fused and bridged rings. _C3-8 cycloalkyl includes _C3-6 , _C3-5 , _C4-8 , _C4-6 , _C4-5 , _C5-8 , _C5-6 cycloalkyl or the like, and may be monovalent, bivalent or polyvalent. Examples of C _3-8 cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl and the like.

Unless otherwise specified, "C _4-6 cycloalkyl" refers to a saturated cyclic hydrocarbon group consisting of 4-6 carbon atoms, including monocyclic and bicyclic systems. _C4-6 cycloalkyl includes _C4-5 , _C5-6 cycloalkyl and the like, and may be monovalent, divalent or polyvalent. Examples of C _4-6 cycloalkyl include, but are not limited to, cyclobutyl, cyclopentyl, cyclohexyl and the like.

Unless otherwise stated, C _n-n+m or C _n -C _n+m includes all of the specific occurrences of carbon atoms n through n+m, for example, C _1-6 includes C ₁ , C ₂ , C ₃ , C ₄ , C ₅ and C ₆ , and also includes any of the ranges from n to n+m, for example, C _1-6 includes C _1-2 , C _1-3 , C _1-4 , C _2-3 , C _2-4 and C _3-5 , etc.; similarly, n-membered to n+m-membered means n to n+m atoms in the ring, for example, 3-6 membered, other rings include 3-membered rings, 4-membered rings, 5-membered rings, 6 -membered rings, and include any range from n to n+m, for example, 3-6 membered rings include 3-5 membered rings, 3-6 membered rings, 4-6 membered rings, 4-5 -membered rings and 5-6-membered rings, etc.

The compounds disclosed herein can be prepared by a variety of synthetic routes well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combinations thereof with other chemical synthetic routes, and equivalents thereof known to those skilled in the art. in this field of technology. Preferred embodiments include, without limitation, the examples disclosed herein.

The solvent used in the present invention may be commercially available. The following abbreviations are used in the present invention: aq. denotes aquatic; HATU stands for O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; EDC stands for N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride; m-CPBA stands for 3-chloroperoxybenzoic acid; eq. denotes equivalent; CDI stands for carbonyldiimidazole; DCM stands for dichloromethane; PE stands for petroleum ether; DIAD stands for diisopropyl azodicarboxylate; DMF stands for N,N-dimethylformamide; DMSO stands for dimethyl sulfoxide; EtOAc stands for ethyl acetate; EtOH means ethanol; MeOH stands for methanol; CBz denotes benzyloxycarbonyl, an amino protecting group; BOC stands for tert-butoxy carbonyl, an amino protecting group; HOAc stands for acetic acid; NaCNBH ₃ denotes sodium cyanoborohydride; k.t. indicates room temperature; O/N stands for overnight; THF stands for tetrahydrofuran; Boc ₂ O denotes di-tert-butyl dicarbonate; TFA stands for trifluoroacetic acid; DIPEA stands for diisopropylethylamine; SOCl ₂ denotes thionyl chloride; CS ₂ denotes carbon disulfide; TsOH stands for p-toluenesulfonic acid; NFSI stands for N-fluoro-N-(phenylsulfonyl)benzenesulfonamide; NCS stands for 1-chloropyrrolidine-2,5-dione; n-Bu ₄ NF means tetrabutylammonium fluoride; iPrOH stands for 2-propanol; m.p. indicates melting point; and LDA stands for lithium diisopropylamide.

Compounds are named according to traditional nomenclature rules from the prior art or using ChemDraw® software, and commercially available compounds are named according to the manufacturer's catalogue.

TECHNICAL EFFECTS

The present invention provides a series of fused ring compounds with increased activity, improved metabolic stability, improved pharmaceutical properties and advantageous pharmacokinetic properties.

The compounds disclosed herein generally demonstrate improved IRAK4 inhibitory activity. The exemplary compounds disclosed herein have impressive advantages over the parent compound (WXR1) for stability in liver microsomes in multiple species, and in particular have up to 20-fold superiority in clearance in some species (eg, mouse). The compounds disclosed herein generally demonstrate improved inhibitory activity against THP-1 cell proliferation. The total systemic exposure, peak concentration, and bioavailability of the various compounds in this project administered orally were equivalent to or superior to those of WXR ₂ (ND-2110) at the same dose, demonstrating superior pharmacokinetic properties. At the same doses, WX001 administered orally in SD rats demonstrated a significant inhibitory effect on lipopolycollagen (LPS)-induced TNF-α secretion, which was significantly superior to that of the parent compounds WXR2(ND-2110), WXR3(BAY -1830839) and WXR4 (BAY-1834845). In this experiment, the efficacy of WX001 was equivalent to that of dexamethasone DEX. At the same doses, orally administered WX001 compounds WX026 and WX044 in SD rats showed significant inhibitory effects on lipopolycollagen (LPS)-induced TNF-α secretion. In this experiment, the efficacy of WX026 was equivalent to that of dexamethasone DEX.

BRIEF DESCRIPTION OF GRAPHIC MATERIALS

Figure 1 shows the results of a pharmacodynamic study evaluating lipopolycollagen (LPS)-induced TNF-α secretion in SD rats at a compound dose of 30 mg/kg; And

Figure 2 shows the results of a pharmacodynamic study evaluating TNF-α secretion induced by lipopolycollagen (LPS) in SD rats at a compound dose of 20 mg/kg.

DETAILED DESCRIPTION

The present invention is described in detail below by way of examples. However, this does not mean that the scope of the present invention is limited to any unfavorable effect. Although the present invention has been described in detail herein and specific examples have also been disclosed, those skilled in the art will appreciate that various changes and modifications can be made with respect to the specific examples without departing from the spirit and scope of the present invention.

Intermediate A1

Synthesis route:

Step 1: Synthesis of compound A1-1

Malononitrile (52.70 g) was dissolved in acetonitrile (1 L) in a 3 L one-neck flask and triethylamine (161.45 g) was added. The mixture was mixed well, isobutyryl chloride (85 g) was added slowly and incubated at 50°C for 2 hours. The acyl chloride was then added and the system was exothermic and changed from a colorless clear solution to a yellow-green suspension. After completion of the reaction, the solvent was removed using a rotary evaporator. The residue was dissolved in water (100 ml) and ethyl acetate (30 ml) was added for three extractions. The organic phases were combined and dried using a rotary evaporator to obtain product A1-1. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 1.23 (d, J=10.4 Hz, 6H), 3.16 (septet, J=10.4 Hz, 1H).

Step 2: Synthesis of compound A1-2

A1-1 (1.1 g) was dissolved in dichloromethane (15 ml) in a 100 ml reaction flask, phosphorus oxychloride (2.26 g) was added dropwise at 0°C and incubated at 20°C for 16 hours. After completion of the reaction, the mixture was directly added to water and the organic phase was separated. The aqueous phase was then extracted twice with dichloromethane (10 ml) and the organic phases were combined and dried using a rotary evaporator to obtain compound A1-2. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 1.7 (d, J=10.4 Hz, 6H), 3.09 (septet, J=10.4 Hz, 1H).

Step 3: Synthesis of compound A1-3

Al-2 (10 g) and aqueous ammonia (7.56 g) were added to a 250 ml reaction flask and incubated at 20°C for 1 hour. Precipitation was observed and water (10 mL) with ethyl acetate (10 mL) was added directly to the reaction system for three extractions. The organic phases were combined, dried using a rotary evaporator to obtain compound A1-3. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 1.24 (d, J=10.4 Hz, 6H), 3.15 (septet, J=10.4 Hz, 1H).

Step 4: Synthesis of compound A1-4

Al-3 (1 g) and triethylamine (2.08 g) were dissolved in pyridine (10 ml) in a 100 ml three-neck flask and then hydrogen sulfide was added and incubated at 30°C for 0.5 hour. After completion of the reaction, the mixture was dried using a rotary evaporator and the residue was added with water and then dichloromethane (10 ml) for two extractions. The solvent was removed using a rotary evaporator to obtain a yellow oily substance, which was then purified using column chromatography (eluent: dichloromethane) to obtain product A1-4. ^1H NMR (400 MHz, CDCl ₃ ) δ = 1.20 (d, J=10.4 Hz, 6H), 3.15 (s, J=10.4 Hz, 1H), 6.04 (br, 1H), 6.55 (br, 1H), 6.87 (br, 1H), 12.18 (br, 1H).

Step 5: Synthesis of compound A1

Compound A1-4 and MeOH (80 ml) were added to a 250 ml one-neck flask with a magnetic stirrer, and H ₂ O ₂ (8.87 g) was added dropwise to the mixture. The reaction system was then stirred at 10-20°C for 16 hours. After completion of the reaction, most of the solvent was removed by concentration under reduced pressure to obtain a solid residue. Ethyl acetate (40 ml) was added to dissolve the solid, followed by the addition of saturated aqueous sodium sulfite (30 ml). The mixture was stirred at room temperature for 30 minutes and the phases were separated. The aqueous phase was extracted with ethyl acetate (20 ml x 3). The combined organic phases were dehydrated using anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give product A1. ^1H NMR (400 MHz, CDCl ₃ ) δ = 5.38 (br s, 2H), 3.13 (td, J=6.8, 13.8 Hz, 1H), 1.33 (d, J= 6.8 Hz, 6H).

Intermediate A2

Synthesis route:

Step 1: Synthesis of compound A2-1

5-amino-3-methylisothiazol-4-carbonitrile (1.00 g) and absolute ethanol (25 mL) were added to a 100 mL round bottom one neck flask with a magnetic stirrer. Concentrated sulfuric acid (7.05 g) was slowly added to the reaction system at 20°C. The reaction mixture in the flask was stirred and incubated in an oil bath at 100°C for 16 hours. After completion of the reaction, the reaction system was cooled to room temperature and slowly added to 100 ml sodium bicarbonate solution. The mixture was adjusted to pH 8, ethyl acetate (30 ml) was added for three extractions, dehydrated using anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give a yellow crude product. The crude product was separated and purified by column chromatography (SiO ₂ , petroleum ether:ethyl acetate = 10:1-3:1, v/v) to give compound A2-1. ^1H NMR (400 MHz, CDCl ₃ ) δ = 6.49 (br s, 2H), 4.35 (q, J=7.2 Hz, 2H), 2.53 (s, 3H), 1.40 (t, J=7.0 Hz, 3H).

Step 2: Synthesis of compound A2-2

A solution of A2-1 (2.5 g) in dry dichloromethane (25 ml) was added to a 50 ml one-neck flask with a magnetic stirrer under a nitrogen atmosphere, cooled to -60°C and chlorosulfonic acid isocyanate (1.79 g) was added dropwise in reaction. The reaction solution was then slowly heated to 25°C and stirred for 30 minutes until the solution became clear. The reaction mixture was concentrated under reduced pressure to obtain a yellow solid, which was suspended in water (10 ml), and the suspension was stirred at 75°C for 30 minutes, filtered and dried to obtain compound A2-2. Stage 3:

Synthesis of compound A2-3

To a solution of A2-2 (1.7 g) in n-butanol (20 ml) in a 50 ml one-neck flask with a magnetic stirrer, potassium carbonate (3.07 g) was added under N ₂ atmosphere and the mixture was incubated at 130°C for 16 hours. The solvent was removed using a rotary evaporator. The residue was suspended with 10 ml of water, filtered and dried to obtain compound A2-3. LCMS (ESI) mass/charge: 183.8 [M+H]+, ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 9.43 (br s, 2H), 2.39-2.34 ( m, 3H).

Step 4: Synthesis of compound A2-4

Phosphorus oxychloride (2.93 g) was added to a solution of dimethylaniline (330.75 mg) and A2-3 (0.5 g) in toluene (5 ml) in a closed 100 ml tube and the mixture was incubated at 120°C for 16 hours. The reaction system was added slowly to 30 ml of water and extracted with dichloromethane (20 ml x 3). The organic phases were combined, dehydrated using anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was subjected to chromatography to obtain compound A2-4. LCMS (ESI) mass/charge: 219.8 [M+H] ⁺ .

Step 5: Synthesis of compound A2

A2-4 (0.2 g), intermediate B1 (303.85 mg) and sodium carbonate (385.28 mg) were dissolved in acetonitrile (4 ml) in a 50 ml one-neck flask with a magnetic stirrer. The reaction system was incubated and stirred at 80°C for 16 hours. The reaction system was cooled to room temperature and concentrated under reduced pressure to obtain a residue. Dichloromethane (10 ml) and water (10 ml) were added to the residue to redissolve. The aqueous phase was extracted with dichloromethane (5 ml). The combined organic phases were washed with brine, dehydrated using anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain a pale yellow solid. The solid was suspended using PE/EtOAc = 10:1 (10 ml), stirred for 1 hour and filtered to obtain compound A2. LCMS (ESI) mass/charge: 368.1 [M+H] ⁺ .

Intermediate connection A3

The synthesis of intermediate A3 is similar to the synthesis of intermediate A1, except that the starting material isobutyryl chloride in step 1 was replaced by tetrahydropyran-4-carbonyl chloride.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 8.16 - 7.89 (m, 2H), 3.99 - 3.80 (m, 2H), 3.40 (dt, J = 3.9, 10.9 Hz, 2H), 2.99 - 2.84 (m, 1H), 1.82 - 1.60 (m, 4H).

Intermediate A4

The synthesis of intermediate A4, starting from A1, is similar to the synthesis of A2, except that the starting material, 5-amino-3-methylisothiazol-4-carbonitrile, in step 1 was replaced by A1.

LCMS (ESI) mass/charge: 396.2,398.2 [M+H] ⁺ .

^1H NMR (400 MHz, CHLOROFORM-d) δ = 5.52 (br d, J=7.6 Hz, 1H), 4.31-4.19 (m, 1H), 4.16 -4.09 (m, 1H), 4.16-4.09 (m, 1H), 3.79-3.70 (m, 4H), 3.22-3.13 (m, 1H), 2.65-2 .56 (m, 4H), 2.41 - 2.27 (m, 3H), 2.06 - 1.96 (m, 2H), 1.60 - 1.48 (m, 2H), 1.46 (d, J=6.8 Hz, 4H), 1.39-1.29 (m, 2H).

Intermediate connection A5

The synthesis of intermediate A5, starting with A3, is similar to the synthesis of A2, except that the starting material, 5-amino-3-methylisothiazol-4-carbonitrile, was replaced in step one with A3.

LCMS (ESI) mass/charge: 438.2, 440.2 [M+H] ⁺

Intermediate connection A6

Synthesis route:

Step 1: Synthesis of compound A6-1

A2-4 (10 g) was dissolved in methanol (200 ml) and a solution of sodium methoxide in methanol (5 M, 36.35 ml) was added. The mixture was incubated at 68°C for 12 hours and then added to saturated ammonium chloride solution (500 ml) and extracted with ethyl acetate (150 ml). The organic phase was dehydrated, filtered and concentrated under reduced pressure. The crude product was purified by column chromatography (ISCO®; 80 g column; eluent: 0-50% ethyl acetate/petroleum ether; flow rate: 40 ml/min) to obtain compound A6-1.

Step 2: Synthesis of compound A6-2

Compound A6-1 (8.79 g) was mixed with 1,2-dichloroethane (100 ml), L-bromosuccinimide (26.66 g) and benzoyl peroxide (453.58 mg) in a flask. After three nitrogen purges, the mixture was incubated at 80°C for 12 hours and added to saturated aqueous sodium thiosulfate (200 ml) and the phases were separated. The aqueous phase was extracted with dichloromethane (50 ml x 2) and the combined organic phases were dried, filtered and concentrated under reduced pressure. The crude product was purified by column chromatography (ISCO® for crude products; 80 g column; eluent: 0-60% ethyl acetate/petroleum ether) to obtain compound A6-2.

Step 3: Synthesis of compound A6-3

A6-2 (4 g) was dissolved in acetonitrile (20 ml), diethyl phosphite (1.95 g) and N,N-diisopropylethylamine (2.80 g) were added and the mixture was incubated at 25°C for 3 hours. The reaction solution was concentrated under reduced pressure and the crude product was purified by column chromatography (ISCO® for crude products; 80 g column; eluent: 0-60% ethyl acetate/petroleum ether)) to obtain compound A6-3.

Step 4: Synthesis of compound A6-4

A6-3 (1.5 g) was dissolved in acetonitrile (40 ml) in a flask and trimethylsilyl cyanide (564.20 mg) and potassium carbonate (786.00 mg) were added successively. The mixture was stirred at 10°C for 12 hours. The reaction system was added to 200 ml of saturated aqueous ammonium chloride and extracted with ethyl acetate (200 ml x 2). The organic phases were combined, dehydrated using anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The resulting crude product was purified by a silica gel column (eluent: dichloromethane:ethyl acetate = 100:0-10:1) to obtain compound A6-4. LCMS (ESI) mass/charge: 237.1 [M+H]. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 4.22 (s, 3H), 4.21 (s, 2H), 4.13 (s, 3H).

Step 5: Synthesis of compound A6-5

A6-4 (0.5 g) was mixed with N,N-dimethylformamide (10 ml) and then pyridine hydrochloride (1.22 g) was added to the flask and the mixture was purged with nitrogen three times. The mixture was stirred at 100°C for 12 hours. The solvent was completely removed under reduced pressure. The residue was suspended with 10 ml of dichloromethane and the mixture was filtered. The filter cake was dried under reduced pressure to obtain compound A6-5. LCMS (ESI) mass/charge: 209.1 [M+H].

Step 6: Synthesis of compound A6-6

The synthesis of intermediate A6-6 starting from A6-5 is similar to step four of the A2 synthesis, except that the starting material A2-3 in step 4 was replaced by A6-5.

Step 7: Synthesis of compound A6

The synthesis of intermediate A6 starting from A6-6 is similar to step four of the A2 synthesis, except that the starting material A2-4 in step 5 was replaced by A6-6.

Intermediate connection A7

Synthesis path

Compound A2 (0.3 g) was added to N,N-dimethylformamide (3 ml) and the mixture was cooled to 0°C before adding sodium hydride (48.93 mg, purity: 60%). The mixture was stirred for 0.5 hour before adding iodomethane (150.47 mg). The mixture was then heated to 15°C followed by stirring for 2.5 hours. The resulting solution was added to water (80 ml) and extracted with ethyl acetate (90 ml x 2). The organic phases were combined and washed with brine (40 ml x 3). The organic phase was then dehydrated using anhydrous sodium sulfate and filtered to remove the drying agent. The filtrate was evaporated under vacuum to obtain compound A7. LCMS (ESI) mass/charge: 382 [M+H]+.

The intermediates in the table below are commercially available reagents.

Intermediate B20

Synthesis route:

Step 1: Synthesis of compound B20-1

2-Bromoethanol (40.52 g) and boron trifluoride diethyl etherate (1.53 g) were added to toluene (80 ml). The mixture was heated to 70°C, epichlorohydrin (20 g) was added dropwise and incubated at 70°C for 1 hour. The reaction system was cooled to 10°C. Aqueous sodium hydroxide (21.61 g, 100 ml) was slowly added dropwise before incubating the mixture at 25°C for 12 hours. The phases were separated. The aqueous phase was extracted with 2-methyltetrahydrofuran (20 ml x 3) and the organic phases were combined. The combined organic phases were washed with water (100 ml), dehydrated, filtered and concentrated under reduced pressure to give compound B20-1.

Step 2: Synthesis of compound B20-2

Aqueous sodium hydroxide (21.61 g, 150 ml) was added with compound B20-1 (29.52 g) and incubated at 90°C for 1 hour before cooling to 15°C. A solution of p-toluenesulfonyl chloride (41.21 g) in anhydrous tetrahydrofuran (150 ml) was then added dropwise and the mixture was incubated at 25°C for 12 hours. The phases were separated. The aqueous phase was extracted with 2-methyltetrahydrofuran (100 ml x 2) and the resulting organic phases were combined and 4 g of dimethylaminopyridine and 30 ml of triethylamine were added before stirring for 10 minutes. Saturated aqueous ammonium chloride (200 ml) was added and the organic phase was separated and concentrated to give the crude product, which was then subjected to column chromatography (ISCO®; 220 g column; eluent: 0-80% ethyl acetate/petroleum ether; flow rate: 60 ml/ min) to obtain compound B20-2.

Step 3: Synthesis of compound B20-3

Compound B20-2 (2 g, 7.34 mmol, 1 eq), 4-nitropyrrole (1.25 g) and cesium carbonate (4.79 g) were added to N,N-dimethylformamide (20 ml) and the mixture was incubated at 70°C for 2 hours. The resulting solution was added to water (100 ml) and extracted with ethyl acetate (180 ml). The resulting organic phase was dehydrated using anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by a silica gel column (petroleum ether:ethyl acetate = 100:0 to 4:1) to give compound B20-3. LCMS (ESI) mass/charge: 214.0 [M+H] ⁺

Step 4: Synthesis of compound B20

Wet palladium on carbon (1 g, 10%) with methanol (5 ml) and B20-3 (1.5 g) were sequentially added to the hydrogenation flask under argon. After three purges of hydrogen (50 psi), the mixture was incubated at 30°C for 3 hours. The resulting mixture was filtered through celite and the filtrate was concentrated under reduced pressure to obtain compound B20. LCMS (ESI) mass/charge: 184.1 [M+H]+

Intermediate B21

Synthesis path

Step 1: Synthesis of compound B21-1

4-Nitropyrrole (10 g) and tert-butyl bromoacetate (17.25 g) were added to a reactor containing acetonitrile (100 ml), and then potassium carbonate (14.67 g) was added. The mixture was stirred at 80°C for 5 hours. The reaction system was added to 100 ml of saturated aqueous ammonium chloride and extracted with ethyl acetate (200 ml x 2). The organic phases were combined, dehydrated using anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by column chromatography (ISCO®; 220 g column; eluent: 0-60% ethyl acetate/petroleum ether; flow rate: 80 ml/min) to obtain compound B21-1.

Step 2: Synthesis of compound B21

Under argon, wet palladium-carbon (3 g, 10%) with methanol (50 ml) was added to the hydrogenation flask and then mixed with B21-1 (10 g), followed by three purges of hydrogen (50 psi). The mixture was stirred at 25°C for 3 hours. The resulting solution was filtered through celite. The solvent in the filtrate was completely removed under reduced pressure and the crude product was purified by column chromatography (ISCO®; 12 g column; eluent: 0-50% DCM/MeOH; flow rate: 40 ml/min) to obtain compound B21. LCMS (ESI) mass/charge: 198.2[M+1]+

Intermediate B22

Synthesis path

Step 1: Synthesis of compound B22-1

Methanol (80 ml) was added with 4-N-benzyloxycarbonylaminocyclohexanone (5 g) and N-Boc-piperazine (3.77 g) and then sodium triacetoxyborohydride (6.43 g) was added slowly. The mixture was stirred at 25°C for 12 hours. The solvent was completely removed under reduced pressure and the residue was added with dichloromethane (100 ml) and water (100 ml) for extraction. The organic phase was dehydrated, filtered and concentrated under reduced pressure to give the crude product. The crude product was subjected to partition chromatography (column: Agela Durashell 10 μm 250 × 50 mm; mobile phase: [water (10 μM NH ₄ HCO ₃ )-ACN]; V%: 35% - 55%, 20 min.) to separate cis -trans isomers. (HPLC Shimadzu 20AD, X-bridge Shield RP18 2.1 × 50 mm, 5 µm, mobile phase [water + 10 mmol/L-ammonium bicarbonate-acetonitrile]; V%: 10% - 80%, 4.5 min. , Rt = 2.615 min) the target fraction was concentrated to remove acetonitrile and dichloromethane (500 ml) was added for extraction. The organic phase was dehydrated, filtered and concentrated under reduced pressure to obtain compound B22-1. The product was used in the next step without purification.

Step 2: Synthesis of compound B22

Under an argon atmosphere, wet palladium on carbon (0.2 g, 10%) was added to the hydrogenation flask and methanol (5 ml) and compound B22-1 (0.2 g) were sequentially added to the reaction system, which was stirred under a hydrogen atmosphere ( 50 psi) at 30°C for 2 hours. The reaction system was filtered through celite, the filter cake was washed with methanol (50 ml x 3) and the solvent was removed from the filtrate under reduced pressure to obtain product B22. LCMS (ESI) mass/charge: 284.1 [M+1]+

Intermediate B23

Synthesis route:

Step 1: Synthesis of compound B23-1

4-Nitropyrrole (1 g) and diethyl 2-bromo-2-methyl maleate (2.69 g) were dissolved in N,N-dimethylformamide (10 ml) and potassium carbonate (2.44 g) was added to carry out the reaction. The mixture was stirred at 100°C for 15 hours. The reaction system was added to ethyl acetate (20 ml) and washed with half-saturated saline (20 ml). The organic phase was dehydrated using anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by column chromatography (ISCO®; 20 g SepaFlash® Silica Flash column, eluent 0-60%, DCM/MeOH gradient @ 40 ml/min) to give compound B23-1. LCMS (ESI) mass/charge: 286.1 [M+1]+

Step 2: Synthesis of compound B23-2

Compound B23-1 (1 g) was dissolved in methanol (10 ml) and sodium borohydride (265.26 mg) was added. The reaction system was stirred at 25°C for 2 hours. 2 ml of saturated ammonium chloride was added to the reaction system, left until bubbles were removed and concentrated under reduced pressure. The crude product was purified by column chromatography (ISCO®; 20 g columns; eluent: 0-50% DCM/MeOH, 40 ml/min.) to obtain compound B23-2. LCMS (ESI) mass/charge: 172.1 [M+1]+

Step 3: Synthesis of compound B23-3

Compound B23-2 (500 mg), di-tert-butyl dicarbonate (956.12 mg) and triethylamine (886.61 mg) were dissolved in anhydrous tetrahydrofuran (10 ml) and the mixture was stirred at 40°C for 3 hours. The reaction system was concentrated under reduced pressure and the crude product was purified by column chromatography (ISCO®; 20 g; eluent: 0-50% DCM/MeOH @ 30 ml/min.) to obtain compound B23-3. LCMS (ESI) mass/charge: 272.1 [M-100+1]+

Step 4: Synthesis of compound B23-4

Compound B23-3 (340 mg) was dissolved in a mixture of pyridine (10 ml) and dichloromethane (20 ml). The solution was cooled to 0°C and then p-toluenesulfonyl chloride (238.91 mg, 1.25 mmol, 1 eq) was added. The mixture was stirred at 0°C for 3 hours. The resulting mixture was concentrated and purified by chromatography (column: Welch Xtimate C18 150×25 mm × 5 μm; mobile phase: [water (10 μM NH ₄ HCO ₃ )-ACN]; V%: 38% - 68%, 10. 5 min) to obtain compound B23-4. LCMS (ESI) mass/charge: 426.2[M+1]+

Step 5: Synthesis of compound B23-5

Compound B23-4 (75 mg) was dissolved in anhydrous tetrahydrofuran (3 ml) and sodium hydride (21.15 mg, 60% purity) was added at 0° C. The reaction system was heated to 67° C. and stirred for 2 hours. The reaction was quenched by adding water (0.1 ml) and mixed directly with a silica gel stirrer. The crude product was purified by column chromatography (ISCO®; 12 g column; eluent: 0-80% ethyl acetate/petroleum ether; 20 ml/min) to give compound B23-5. LCMS (ESI) mass/charge: 254.2[M+1]+

Step 6: Synthesis of compound B23

Compound B23-5 (20 mg) was dissolved in dichloromethane (1.5 ml) and trifluoroacetic acid (1.5 ml) was added. The mixture was stirred at 25°C for 1 hour. The solvent from the resulting mixture was removed under reduced pressure to obtain compound B23. LCMS (ESI) mass/charge: 154.2[M+1]+

The synthesis of the intermediates in the table below is similar to that for intermediate B22, except that N-Boc-piperazine has been replaced by starting material one.

Intermediate B35

Synthesis route:

Step 1: Synthesis of compound B35-1

Diisopropyl azodicarboxylate (854.05 mg) was added to a solution of 4,4-difluorocyclohexanol (0.5 g), 4-nitropyrazole (415.29 mg) and triphenylphosphine (1.06 g) in tetrahydrofuran (20 ml) and the mixture was stirred at 20 °C for 12 hours. The solvent was completely removed under reduced pressure and the crude product was purified by column chromatography (petroleum ether: petroleum ether: ethyl acetate = 3:1) to obtain compound B35-1.

Step 2: Synthesis of compound B35

Compound B35-1 (2 g) was added to methanol (70 ml) and wet palladium on carbon (1 g, purity: 10%) was added. After three hydrogen purges, the mixture was stirred at 30°C for 2 hours under a hydrogen atmosphere (30 psi) and filtered through celite. The solvent from the filtrate was completely removed under reduced pressure to obtain compound B35. LCMS (ESI) mass/charge: 202.2[M+H].

Reference example 1: WXR1

Compound WXR1 was synthesized according to the route specified in patent no. WO 2017205762.

Reference Example 2: WXR2

Compound WXR2 (ND-2110) was synthesized according to the route specified in patent no. WO 2012097013.

Reference Example 3: WXR3

Compound WXR3 ₃ (BAY-1830839) was synthesized according to the route specified in patent no. WO 2017186700.

LCMS (ESI) mass/charge: 451.2[M+H] ⁺ . ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 12.35 (s, 1H), 8.71 (s, 1H), 8.45 (d, J=7.6 Hz, 2H), 8, 37 (t, J=6.0 Hz, 1H), 8.16 (d, J=6.8 Hz, 1H), 7.57 (s, 1H), 5.93 (d, J=8.0 Hz, 1H), 4.51 (s, 1H), 4.49 - 4.45 (m, 2H), 2.05 - 2.01 (m, 2H), 1.62 (s, 6H), 1 .15 (s.6H)

Reference Example 4: WXR4

Compound WXR4 (BAY-1834845) was synthesized according to the route specified in patent no. WO 2017186689.

LCMS (ESI) mass/charge: 471.2[M+H]+. ^1H NMR (400 MHz, DMSO- _d6 ) δ = 12.37 (s, 1H), 8.73 (s, 1H), 8.48-8.42 (m, 2H), 8.40 - 8 .32 (m, 1H), 8.16 (d, J=7.8 Hz, 1H), 7.59 (s, 1H),5.99(s, 1H), 4.86 (t, J= 6.6 Hz, 2H), 3.85 (t, J=6.6 Hz, 2H), 2.90 (s, 3H), 1.62 (s, 6H).

Example 1: Synthesis of compound WX001

Synthesis route:

Example 1: Synthesis of compound WX001

Intermediate A2 (0.15 g) was dissolved in n-butanol (1 ml) followed by the addition of hydrochloric acid/ethyl acetate (4 M, 1.02 ml) and intermediate B2 (81.81 mg). The mixture was stirred at 120°C for 16 hours. The resulting mixture was filtered. The filter cake was washed with ethyl acetate (2 ml x 3) and the filter cake was dried to obtain the crude product. The crude product was purified by preparative chromatography (dichloromethane:methanol = 10:1) to obtain crude compound WX001, which was suspended with 2 ml of methanol to obtain compound WX001.

LCMS (ESI) mass/charge: 499.2 [M+H]+. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 9.05 (s, 1H), 7.90 (s, 1H), 7.55 (s, 1H), 6.10 (d, J=7 ,2 Hz, 1H), 4.40-4.31 (m, 1H), 4.16-4.05 (m, 1H), 4.01-3.94 (m, 2H), 3.83- 3.77 (m, 1H), 3.63-3.57 (m, 4H), 3.48 (dt, J=3.2, 11.2 Hz, 2H), 2.68 (s, 3H) , 2.34 - 2.24 (m, 2H), 2.18-2.10 (m, 2H), 2.03 - 1.91 (m, 7H), 1.60 - 1.48 (m, 2H), 1.44 -1.32 (m, 2H).

Example 2: Synthesis of compound WX002

Synthesis route:

The synthesis of compound WX002, starting with intermediates A2-4, B5 and B2, is similar to step 5 of the synthesis of A2 and the steps of the synthesis of compound WX001.

LCMS (ESI) mass/charge: 500.2 [M+H] ⁺ . ^1H NMR (400 MHz, CHLOROFORM-d) δ = 8.03 (s, 1H), 7.64-7.37 (m, 1H), 7.09-6.83 (m, 1H), 4, 27 (td, J=8.0, 15.7 Hz, 1H), 4.15-4.12 (m, 2H), 4.06 (br d, J=11.8 Hz, 2H), 3, 74 - 3.72(m, 2H), 3.54 - 3.40 (m, 2H), 2.60 (s, 3H), 2.52 (br s, 3H), 2.56-2.47 (m, 1H), 2.24 (br, J=13.1 Hz, 2H), 2.06 (br s, 4H), 1.97 (br d, J=10.8 Hz, 2H), 1 .58 - 1.49 (m, 2H), 1.46 - 1.38 (m, 2H), 0.84 - 0.74 (m, 2H).

Example 3: Synthesis of compound WX003

Synthesis route:

The synthesis of compound WX003, starting with intermediates A4 and B2, is similar to the steps in the synthesis of compound WX001.

LCMS (ESI) mass/charge: 527.2 [M+H]+. ^1H NMR (400 MHz, DMSO- _d6 ) δ = 9.04 (br s, 1H), 7.88 (s, 1H), 7.52 (s, 1H), 6.06 (br s, 1H), 4.38 - 4.27 (m, 1H), 4.20 - 4.08 (m, 1H), 4.00 - 3.89 (m, 2H), 3.60-3.40 (m, 11 H), 2.33-2.22 (m, 1H), 2.15-2.05 (m, 2H), 2.03-1.87 (m, 6H), 1.60 - 1.46 ( m, 2H), 1.42 - 1.25 (m, 8H).

Example 4: Synthesis of compound WX004

Synthesis route:

The synthesis of compound WX004, starting with intermediates A4 and B4, is similar to the steps in the synthesis of compound WX001.

LCMS (ESI) mass/charge: 513.3 [M+H]+. ^1H NMR (400 MHz, METHANOL- _d4 ) δ = 8.60 (s, 2H), 4.89 - 4.79 (m, 2H), 4.73 - 4.60 (m, 2H), 4 .45 - 4.33 (m, 1H), 4.14 - 4.05 (m, 2H), 4.02 - 3.91 t, 2H), 3.82 (d, J=5.2 Hz, 2H), 3.72 - 3.52 (m, 5H), 3.28 - 3.18 (m, 2H), 2.44 - 2.25 (m, 4H), 2.25 - 2.25 ( m, 1H), 1.94 - 1.73 (m, 4H), 1.39 (d, J=6.8 Hz, 6H)

Example 5: Synthesis of compound WX005

Synthesis route:

The synthesis of compound WX005, starting with intermediates A5 and B3, is similar to the steps in the synthesis of compound WX001.

LCMS (ESI) mass/charge: 499.3 [M+H]+. ^1H NMR (400 MHz, DMSO- _d6 ) δ = 9.54 - 9.22 (m, 1H), 8.04 - 7.70 (m, 1H), 7.50 (br s, 1H), 6.51 - 6.09 (m, 1H), 4.12 - 4.00 (br s, 1H), 3.96 -3.86 (m, 3H), 3.80 (s, 3H), 3.72 - 3.49 (m, 7H), 2.67 (s, 3H), 2.32 - 2.22 (m, 1H), 2.17 - 2.07 (m, 2H), 1, 99 - 1.82 (m, 4H), 1.81 - 1.65 (m, 2H), 1.60 - 1.45 (m, 2H), 1.37 (br s, 2H)

Referring to the synthesis procedures in Example 1, the compounds in the following examples were synthesized starting with fragment 1 and fragment 2 in the following table.

Example 25: Synthesis of WX025

Synthesis route:

Stage 1:

Referring to the synthesis of intermediate A2, compound WX025-1 was synthesized starting from intermediates A2-4 and B22.

Stage 2:

Referring to the synthesis of WX001, compound WX025-2 was synthesized starting from intermediates WX025-1 and B2.

Step 3: Synthesis of compound WX025

Compound WX025-2 (0.18 g) was mixed with a solution of hydrochloric acid in methanol (15 ml, 4 M) in a flask. The mixture was stirred at 15°C for 12 hours and the reaction solution was concentrated under reduced pressure. The crude product was purified by chromatography (column: Boston PR ₁ me C18 150×30 mm × 5 µm; mobile phase: [water (0.05% HCl)-ACN]; B%: 1%-22%, 10 min) with receiving connection WX025. LCMS (ESI) mass/charge: 498.1 [M+H]. ^1H NMR (400 MHz, MeOD- _d4 ) δ = 7.99 (s, 1H), 7.80-7.65 (m, 1H), 4.49 - 4.23 (m, 2H), 4.10 -4.08 (d, J=8.0 Hz, 2H), 3.72-3.50 (m, 11H), 2.80 (s, 3H), 2.40-2.36 (m, 4H ), 2.10 (s, 4H), 1.83-1.75 (m, 4H).

Example 26: Synthesis of WX026

Synthesis route:

Step 1: Synthesis of compound WX026

Compound WX025 (0.03 g, HCl salt) and dichloromethane (2 ml) were added to the reaction flask, followed by triethylamine (14.21 mg). The mixture was stirred and methyl chloroformate (6.37 mg, 5.22 μl) was added dropwise. The resulting mixture was stirred at 15°C for 2 hours. The reaction was quenched with 30 mL of water and ethyl acetate (30 mL x 2) was added for extraction. The combined organic phases were dehydrated using anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The resulting crude product was purified using a chromatograph (column: Xtimate C18 150x25 mmx5 μm; mobile phase: [water (0.04% NH ₃ H ₂ O + 10 mM NH ₄ HCO ₃ )-ACN]; B%: 30-55% , 10.5 min) to obtain compound WX026. LCMS (ESI) mass/charge: 556.4 [M+H], ^1H NMR (400 MHz, CDCl ₃ ) δ = 8.00 (s, 1H), 7.55 (s, 1H), 6.81 (s, 1H), 5.19 (s, 1H), 4.34 (m, 1H), 4.14 - 4.11 (m, 3H), 3.72 (s, 3H), 3.55 - 3.50 (m, 6H), 2.70 (s, 3H), 2.56 (s, 4H), 2.41 - 2.30 (m, 3H), 2.13 (s, 4H), 2 .01 - 1.98 (m, 2H), 1.50-1.44 (m, 2H), 1.33-1.30 (m, 2H).

Referring to the synthesis procedures in Example 25 and Example 26, the compounds in the following examples were synthesized starting with intermediate A2-4, fragment 1 and fragment 2 in the following table.

Example 60: Synthesis of WX060

Synthesis route:

Step 1: Synthesis of compound WX060-1

1,3-dibromo-2-propanol (5 g), N-Boc-1,4-cyclohexanediamine (4.47 g) and sodium carbonate (19.90 g) were mixed in a flask containing ethanol (300 ml), and the mixture was stirred at 80°C for 12 hours. The solvent in the mixture was completely removed under reduced pressure. The crude product was suspended with ethyl acetate (100 ml) and filtered. The solvent in the filtrate was completely removed under reduced pressure to obtain compound WX060-1.

Step 2: Synthesis of compound WX060-2

Compound WX060-1 (8 g) was dissolved in N,N-dimethylformamide (100 ml) at 0° C. and then tert-butyldiphenylchlorosilane (6.31 g) and imidazole (3.44 g) were added. The mixture was incubated at 25°C for 4 hours. The resulting mixture was filtered and 200 ml of water was added to the filtrate, followed by extraction with ethyl acetate (150 ml x 4). The organic phases were combined and dehydrated using an appropriate amount of anhydrous sodium sulfate. The dehydrated organic phase was filtered to remove the drying agent and the solvent in the filtrate was completely removed under reduced pressure to obtain the crude product. The crude product was separated using a COMBI-FLASH automated chromatography system (SepaFlash® Silica Flash column, mobile phase: 0-80% petroleum ether/ethyl acetate, flow rate: 80 ml/min) and purified to give compound WX060-2.

Step 3: Synthesis of compound WX060-3

Compound WX060-2 (1 g) was dissolved in dichloromethane (10 ml) at 0°C and trifluoroacetic acid (2 ml) was added. After 1 hour of incubation, additional trifluoroacetic acid (2 ml) was added and the reaction system was incubated for another 2 hours. Thereafter, 30 ml of water was added to the reaction system and ethyl acetate (4 × 20 ml) was added to the mixture for extraction. The organic phases were combined and dehydrated using an appropriate amount of anhydrous sodium sulfate. The dehydrated organic phase was filtered to remove the drying agent and the solvent in the filtrate was completely removed under reduced pressure to obtain the crude product. The crude product was separated using a COMBI-FLASH automated chromatography system (SepaFlash® silicon Flash column, mobile phase: 0-7% dichloromethane/methanol-petroleum ether/ethyl acetate, flow rate: 80 ml/min) to obtain compound WX060-3. LCMS (ESI) mass/charge: 409.1 [M+H] ⁺

Step 4: Synthesis of compound WX060-4

Referring to the synthesis of intermediate A2, compound WX060-4 was synthesized starting from intermediate A2-4 and compound WX060-3.

Step 5: Synthesis of compound WX060-5

Referring to the synthesis of WX001, compound WX060-5 was synthesized starting from compound WX060-4 and intermediate B2.

Step 6: Synthesis of compound WX060

Compound WX060-5 (0.03 g) was dissolved in anhydrous tetrahydrofuran (2 ml) and a solution of tetrabutylammonium fluoride in tetrahydrofuran (1 M, 82.99 μl) was added. The reaction system was incubated at 15°C for 12 hours under a nitrogen atmosphere. The solvent was completely removed under reduced pressure and the crude product was separated by HPLC (column: Welch XTimate C18 150×25 mm × 5 μm; mobile phase: A: 10 mM aqueous NH ₄ HCO ₃ , B: acetonitrile; gradient: B%: 20 %-50%, 10.5 min) to obtain compound WX060. LCMS (ESI) mass/charge: 485.3 [M+1]+. ^1H NMR (400 MHz, DMSO- _d6 ) δ = 9.48-9.36 (m, 1H), 8.10 - 7.81 (m, 1H), 7.63 (s, 1H), 6 .48 - 6.43 (m, 1H), 5.29 (d, J=6.8 Hz, 1H), 4.41 - 4.40 (m, 1H), 4.20 - 4.13 (m , 2H), 4.04 - 4.01 (m, 2H), 3.56 -3.47 (m, 4H), 3.45 - 3.42 (m, 3H), 2.74 (s, 5H ), 2.08 - 1.97 (m, 6H), 1.88-1.86 (m, 2H), 1.58-1.55(m, 1H), 1.11 - 1.10 (m , 1H).

Experimental Example 1: In Vitro Enzyme Activity Evaluation

The inhibitory activity of test compounds against human IRAK4 was assessed by measuring IC50 values in a ³³ P-labeled kinase activity assay (Reaction Biology Corp).

Buffer composition: 20 mM Hepes (pH 7.5), 10 mM MgCl ₂ , 1 mM EGTA, 0.02% Brij35, 0.02 mg/ml BSA, 0.1 mM Na ₃ VO ₄ , 2 mM DTT, 1 %DMSO.

Procedures: The test compound was dissolved in DMSO at room temperature to obtain a 10 mM solution for subsequent use. The substrate was dissolved in fresh buffer, to which the kinase was added and mixed well. A DMSO solution containing the test compound was added to the above stirred reaction system using an acoustic technique (Echo 550). The concentrations of the compounds in the reaction system were as follows: 10 μM, 3.33 μM, 1.11 μM, 0.370 μM, 0.123 μM, 41.2 nM, 13.7 nM, 4.57 nM, 1.52 nM, and 0.508 nM. After 15 minutes of incubation, the reaction was started by adding ³³ P-ATP (activity: 0.01 μCi/μl; the corresponding concentration is indicated in Table 1). Information regarding supplier part number, lot number and concentration in the reaction system for IRAK4 and its substrate is listed in Table 1. After 120 minutes of reaction at room temperature, the resulting solution was loaded onto a P81 ion exchange chromatography paper sheet (Whatman #3698-915). After repeated washing with 0.75% phosphoric acid solution, the radioactivity of the phosphorylated substrate residue on a paper sheet was measured. Kinase activity data are shown as a comparison of the kinase activity of the test compound and the kinase activity of the blank solution (DM SO only), and curve fitting was performed using Prism4 software (GraphPad) to obtain IC ₅₀ values, with the experimental results shown in Table 2.

Conclusion: The compounds disclosed herein generally demonstrate acceptable levels of IRAK4 inhibitory activity.

Experimental Example 2: In vitro stability assay in liver microsomes

Metabolic stability in liver microsomes

Test compounds at 1 μM were incubated together with liver microsomes containing protein at a concentration of 0.5 mg/ml under the action of a reducing coenzyme II regeneration system in a water bath at 37°C.

1) Positive controls included: testosterone (substrate 3A4), propylamine-propiophenone (substrate 2D6) and diclofenac (substrate 2C9). Positive controls were incubated under the same conditions as the test compounds.

2) The reaction was stopped using a stop reagent containing internal standard at time points 0, 5, 10, 20, 30 and 60 minutes. The compounds were incubated with microsomes for 60 minutes in the absence of the reducing coenzyme II regeneration system and served as negative controls.

3) There was no repetition for each time point (n = 1).

4) Samples were analyzed by LC/MS/MS and concentrations are shown as the ratio of the peak area of the compound to the peak area of the internal standard (non-standard curve).

5) In the summary, half-life and clearance were calculated.

6) To calculate clearance, the following formula was used:

Liver to body weight ratio: 88 g/kg for mice

Whole liver clearance was calculated using

internal microsomal clearance;

Microsomal protein during incubation: microsomal protein during incubation;

Microsomes: microsomes;

Liver: liver;

Body weight: body weight

The results are summarized in Table 3.

Conclusion: The exemplary compounds disclosed herein have impressive advantages over the parent compound (WXR1) for stability in liver microsomes in multiple species, and in particular have up to 20-fold superiority in clearance in some species (eg, mouse).

Experimental Example 3: In Vitro Cell Activity Assay

ELISA analysis of TNF-α in THP-1 cells

1. Materials:

THP-1 cells obtained from a person with acute monocytic leukemia were purchased from ATCC (cat. no. TIB-202) and incubated in an incubator at 37°C, 5% CO ₂ .

The medium was RPMI1640 (Gibco, cat. no. 22400-105) supplemented with 10% FBS (Gibco, cat. no. 10091148); 1% PenStrep (Gibco, Cat. No. 15140); 0.05 mM 2-mercaptoethanol (Sigma, cat. no. M6250).

2. Method:

The content of TNF-a in the cell culture supernatant was measured using an Elisa TNF-a kit. TNF-α was produced by stimulating THP-1 cells with 150 ng/ml LPS (Sigma, cat. no. L6529).

Normal THP-1 cells in logarithmic growth phase were added to 96-well plates (Corning No. 3599) at a certain concentration (1 × 10 ⁵ /100 μl) and then incubated in an incubator. After two hours, 16.7 μl of test compound was added at different concentrations (8 × final concentrations) and the mixture was incubated. After one hour, 16.7 μl of LPS at a concentration of 1200 ng/ml was added and the mixture was incubated. After 18 hours, the cell culture was centrifuged and the supernatant was collected. TNF-α content was measured using the TNF-α Elisa kit. Finally, the OD signals (OD450-OD570) were read on an Envision plate reader.

3. Data analysis:

OD450-CD570 signals were converted to percent inhibition.

% Inhibition = (ZPE - sample)/(ZPE - HPE) × 100.

"HPE" represents the CD450-OD570 signal value for the control without LPS-stimulated cells, and "ZPE" represents the OD450-CD570 signal value for the control with LPS-stimulated cells. IC ₅₀ values for compounds were calculated using XLFit in Excel.

Equation: Y = minimum + (maximum - minimum)/(1 + (IC ₅₀ /X) ^∧ slope).

The results are summarized in Table 4.

Conclusion: The compounds disclosed herein generally demonstrate acceptable levels of inhibitory activity against THP-1 cell proliferation.

Experimental Example 4: Pharmacokinetic Study

Pharmacokinetic study of test compounds administered orally and intravenously in SD rats

The test compound was mixed with a solution containing 10% methylpyrrolidone, 10% polyethylene glycol stearate and 80% water. The mixture was vortex-mixed and sonicated to obtain a 1.5 mg/mL clarified solution, which was then filtered through a microporous membrane for subsequent use. Male SD rats, 7-10 weeks old, were selected and intravenously dosed with a candidate compound of 3 mg/kg. The test compound was mixed with a 0.5% aqueous solution of methylcellulose. The mixture was vortex-mixed and sonicated to obtain a 2 mg/mL suspension for subsequent use. Male SD rats, 7-10 weeks old, were selected and orally dosed at 10 mg/kg of the candidate compound. Whole blood was collected at specific time points and plasma was separated. Drug concentration was measured using LC-MS/MS and pharmacokinetic parameters were calculated using Phoenix WinNonlin software (Pharsight, USA). The experimental results are shown in Table 7.

The results demonstrate that the total systemic exposure, peak concentration and bioavailability of the various compounds in this project administered orally were equivalent or superior to the parent compound WXR2 (ND-2110) at the same dose, demonstrating superior pharmacokinetic properties.

Experimental Example 5: Pharmacodynamic Study

Pharmacodynamic study to evaluate lipopolycollagen (LPS)-induced TNF-α secretion in KpbicSD

1. Simulation and introduction

SD rats were orally administered vehicle, dexamethasone (DEX, 0.5 mg/kg) as a positive control, and test compound, and 0.5 hours after administration, LPS was administered intraperitoneally by injection (1 mg/kg). Animals were sacrificed with CO ₂ 2 hours after LPS administration by injection. Blood from the heart was collected into vacuum containers with EDTA-K2, and part of the anticoagulated blood was centrifuged and the blood plasma was frozen at -80°C.

2. Analysis of TNF-α and II-1b

Frozen blood plasma was thawed at room temperature and plasma TNF-α concentration was measured using an ELISA kit.

3. Statistics

Experimental data were presented using mean ± SEM and TNF-α levels were analyzed using one-way ANOVA. Differences were considered significant at p < 0.05. The results of a pharmacodynamic study to evaluate LPS-induced TNF-α secretion in SD rats are shown in Figures 1 and 2.

Figure 1 shows that at the same doses, WX001, administered orally, showed a significant inhibitory effect on lipopolycollagen (LPS)-induced TNF-α secretion, which was significantly superior to that of the parent compounds WXR2(ND-2110). , WXR3(BAY-1830839) and WXR4 (BAY-1834845). In this experiment, the efficacy of WX001 was equivalent to that of dexamethasone DEX.

Figure 2 shows that at the same doses, WX001, WX026 and WX044, administered orally, showed a significant inhibitory effect on lipopolycollagen (LPS)-induced TNF-α secretion. In this experiment, the efficacy of WX026 was equivalent to that of dexamethasone DEX.

Claims (56)

1. A compound of formula (II), its optical isomer or pharmaceutically acceptable salt, Where R ₁ is selected from the group consisting of CN, C _1-6 alkyl and 3-6 membered heterocycloalkyl, wherein C _1-6 alkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _a ; R ₂ is selected from the group consisting of C _1-6 alkyl and 3-6 membered heterocycloalkyl, wherein C _1-6 alkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _b ; R ₃ is selected from the group consisting of H, OH, C _1-6 alkyl, -C(=O)-OC _1-6 alkyl, -C(=O)-C _1-6 alkyl and C _3-6 cycloalkyl; ring A is selected from the group consisting of 3-10 membered heterocycloalkyl, and the 3-10 membered heterocycloalkyl is optionally substituted with 1, 2 or 3 R _d ; L ₁ is selected from the group consisting of O and N(R ₄ ); L ₂ is selected from the group consisting of a single bond, CH ₂ and CH ₂ CH ₂ ; R ₄ is selected from the group consisting of H and Me; each R _a is independently selected from the group consisting of CN and COOH; each R _b is independently selected from the group consisting of F, OH, COOH and Me; each R _d is independently OH; 3-6 membered heterocycloalkyl contains 1, 2 or 3 heteroatoms or heteroatomic groups independently selected from the group consisting of -O-, -NH- and N; And 3-10 membered heterocycloalkyl contains 1, 2 or 3 heteroatoms or heteroatomic groups independently selected from the group consisting of -O-, -NH-, N and -C(=O)NH-. 2. A compound of formula (III), its optical isomer or pharmaceutically acceptable salt, Where R ₁ is selected from the group consisting of CN, C _1-6 alkyl and 3-6 membered heterocycloalkyl, wherein C _1-6 alkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _a ; R ₂ is selected from the group consisting of C _1-6 alkyl, C _3-8 cycloalkyl and 3-6 membered heterocycloalkyl, wherein C _1-6 alkyl, C _3-8 cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1. 2 or 3 _Rb ; R ₃ is selected from the group consisting of H, OH, COOH, C _1-6 alkyl, C _1-6 alkylamino, -C(=O)-OC _1-6 alkyl, -C(=O)-C _1-6 alkyl and C _3-6 cycloalkyl; ring A is selected from the group consisting of 3-10 membered heterocycloalkyl, and the 3-10 membered heterocycloalkyl is optionally substituted with 1, 2 or 3 R _d ; L ₁ is selected from the group consisting of O and N(R ₄ ); L ₂ is selected from the group consisting of a single bond, CH ₂ and CH ₂ CH ₂ ; R ₄ is selected from the group consisting of H and Me; each R _a is independently selected from the group consisting of CN and COOH; each R _b is independently selected from the group consisting of F, OH, COOH and Me; each R _d is independently OH; 3-6 membered heterocycloalkyl contains 1, 2 or 3 heteroatoms or heteroatomic groups independently selected from the group consisting of -O-, -NH- and N; And 3-10 membered heterocycloalkyl contains 1, 2 or 3 heteroatoms or heteroatomic groups independently selected from the group consisting of -O-, -NH-, N and -C(=O)NH-. 3. A compound, its optical isomer or a pharmaceutically acceptable salt according to any one of paragraphs. 1-2, wherein R1 is selected from the group consisting of C _1-3 alkyl and tetrahydropyranyl, wherein C _1-3 alkyl and tetrahydropyranyl are optionally substituted with 1, 2 or 3 R _a . 4. The compound, its optical isomer or pharmaceutically acceptable salt according to claim 3, where R ₁ is selected from the group consisting of Me, Et, And , where Me, Et, And optionally substituted with 1, 2 or 3 R _a . 5. The compound, its optical isomer or pharmaceutically acceptable salt according to claim 4, where R ₁ is selected from the group consisting of Me, Et, 6. The compound, its optical isomer or pharmaceutically acceptable salt according to claim 2, where R ₂ is selected from the group consisting of C _1-3 alkyl, C _4-6 cycloalkyl, tetrahydropyranyl, oxetanyl, tetrahydrofuranyl and 1,4-dioxanyl, where C _1-3 alkyl, C _4-6 cycloalkyl, tetrahydropyranyl, oxetanyl, tetrahydrofuranyl and 1,4-dioxanyl are optionally substituted with 1, 2 or 3 R _b . 7. The compound, its optical isomer or pharmaceutically acceptable salt according to claim 6, where R ₂ is selected from the group consisting of Me, Et, and cyclohexyl, where Me, Et, and cyclohexyl are optionally substituted with 1, 2 or 3 R _b . 8. The compound, its optical isomer or pharmaceutically acceptable salt according to claim 7, where R ₂ is selected from the group consisting of Me, -CH ₂ OH, Et, 9. The compound, its optical isomer or pharmaceutically acceptable salt according to claim 2, where the structural unit selected from the group consisting of Me, Et, 10. The compound, its optical isomer or pharmaceutically acceptable salt according to claim 2, where R ₃ is selected from the group consisting of H, OH, COOH, C _1-3 alkyl, -N(C _1-3 alkyl) ₂ , -C (=O)-OC _1-3 alkyl, -C(=O)-C _1-3 alkyl and C _3-6 cycloalkyl. 11. The compound, its optical isomer or pharmaceutically acceptable salt according to claim 10, where R ₃ is selected from the group consisting of H, OH, COOH, Me, Et, -C(=O)-O-Me, -C(=O)-O-Et and -C(=O)-Me. 12. A compound, its optical isomer or a pharmaceutically acceptable salt according to any one of paragraphs. 1-2, wherein ring A is selected from the group consisting of morpholinyl, piperazinyl, 3-morpholinonyl, 2-piperazinonyl, homopiperazinyl, 4,7-diazaspiro[2,5]octyl, 3,6-diazabicyclo[3,1,1 ]heptyl, 2-azacyclohexanonyl, 2,5-diazabicyclo[2.2.1]heptyl and azetidinyl, where morpholinyl, piperazinyl, 3-morpholinonyl, 2-piperazinonyl, homopiperazinyl, 4,7-diazaspiro[2,5]octyl, 3, 6-diazabicyclo[3,1,1]heptyl, 2-azacyclohexanonyl, 2,5-diazabicyclo[2.2.1]heptyl and azetidinyl are optionally substituted with 1, 2 or 3 R _d . 13. The compound, its optical isomer or pharmaceutically acceptable salt according to claim 12, wherein ring A is selected from the group consisting of , , , , , , , , , , , And . 14. The compound, its optical isomer or pharmaceutically acceptable salt according to claim 13, where the structural unit selected from a group consisting of , , , , , , , , , , , , , , , , , , , , , , , , , , , , And . 15. A compound, its optical isomer or a pharmaceutically acceptable salt according to any one of paragraphs. 2-11, which are selected from: R ₁ , R ₂ , R ₃ , L ₁ and L ₂ are defined in any of paragraphs. 2-11. 16. A compound of the following formula, its optical isomer or a pharmaceutically acceptable salt, which is selected from: 17. The compound, its optical isomer or pharmaceutically acceptable salt according to claim 16, which is selected from: 18. A pharmaceutical composition for producing an IRAK4 inhibitor, containing a therapeutically effective amount of a compound, its isomer or a pharmaceutically acceptable salt according to any one of paragraphs. 1-17 as an active ingredient and a pharmaceutically acceptable carrier. 19. Use of a compound, its isomer or a pharmaceutically acceptable salt thereof according to any one of paragraphs. 1-17 to obtain an IRAK4 inhibitor. 20. Use of the composition according to claim 18 to obtain an IRAK4 inhibitor.