CN114174282A - Pyridazinone derivatives as thyroxine receptor-beta agonists and uses thereof - Google Patents

Abstract

Pyridazinone derivatives as thyroxine receptor-beta agonists and their use in the manufacture of medicaments for treating diseases associated with thyroxine receptor-beta agonists. Specifically disclosed is a compound represented by the formula (I) or a pharmaceutically acceptable salt thereof.

Description

Pyridazinone derivatives as thyroxine receptor-beta agonists and uses thereof

The following priority is claimed in the present application:

cn201910459299.x, application No. 2019-05-29;

cn201910741177.x, application No. 2019-08-12;

CN201911130068.0, application date 2019-11-18;

CN202010076525.9, App. No. 2020-01-23;

CN202010188909.X, App. 2020-03-17;

CN202010409420.0, application No. 2020-05-14.

Technical Field

The invention relates to a series of pyridazinone derivatives serving as thyroxine receptor-beta agonists and application thereof in preparing medicaments for treating thyroxine receptor-beta agonist related diseases. In particular to a compound shown in a formula (I) or pharmaceutically acceptable salt thereof.

Background

Thyroid hormones play very important roles in the human body, including controlling the body's overall metabolism, protein synthesis, fat metabolism, neuronal and skeletal growth, and regulating cardiac and renal functions. It is secreted by the thyroid gland. Hypothalamus and pituitary gland strictly regulate thyroid secretion by the thyroid gland through the classical negative feedback mechanism of secretion of thyroid stimulating hormone releasing hormone and thyroid stimulating hormone. In recent years, the incidence of obesity and its complications, diabetes, metabolic syndrome, atherosclerosis, and nonalcoholic steatohepatitis has been on the rise. Therefore, it is desired to develop thyroid hormone and its analogs for clinical treatment.

Thyroid hormones exert transcriptional regulation by binding to two highly homologous receptors, the thyroid receptor alpha (THR α) and the thyroid receptor beta (THR β). THR alpha is mainly distributed in brain, heart and skeletal muscle and can control heart rate; THR beta is distributed mainly in the liver and brain and can lower cholesterol and increase metabolic rate (Curr Atheroscher Rep (2016)18(3): 14; PNAS (2003),100(17), 10067-. During the development of thyroid hormone and thyroid hormone analogue, side effects of heart and skeleton are found, and further clinical development is hindered. It was found that this side effect could be caused by binding of thyroid hormone analogues to THR α (j.med.chem. (2014),57, 3912-. Therefore, the development of thyroid hormone analogues with high liver tissue distribution specificity and thyroid hormone receptor subtype selectivity has great clinical value.

Based on literature (J.Med.chem.2014,57, 3912-:

disclosure of Invention

The invention provides a compound shown in a formula (I) or a pharmaceutically acceptable salt thereof,

wherein,

selected from single and double bonds;

T ₁selected from NR_a；

T ₂Selected from C, CH and N;

T ₃selected from the group consisting of CR_bAnd N;

T ₄selected from the group consisting of CR_cAnd O;

R _aselected from H and C_1-3An alkyl group;

R _band R_cEach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl and C_1-3Alkoxy radical, said C_1-3Alkyl and C_{1- 3}Alkoxy is optionally substituted by 1,2 or 3R;

and when R is_cSelected from H or CN, R₁、R ₂And the atoms to which they are attached together form C_3-8Cycloalkenyl or 3-8 membered heterocycloalkenyl, said C_3-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

when R is_cSelected from F, Cl, Br, I, OH, NH₂And C optionally substituted by 1,2 or 3R_1-3Alkyl and C_1-3At alkoxy radical, R₁And R₂Each independently selected from H and optionally substituted by 1,2 or 3R_dSubstituted C_1-6Alkyl, or, R₁、R ₂And the atoms to which they are attached together form C_3-8Cycloalkenyl or 3-8 membered heterocycloalkenyl, said C_3-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

R ₃and R₄Each independently selected from H, F, Cl, Br, I, OH, NH₂CN and optionally substituted by 1,2 or 3R_fSubstituted C_1-3An alkyl group;

R _d、R _eand R_fEach independently selected from H, F, Cl, Br, I, OH, NH₂CN and C_1-3An alkyl group;

each R is independently selected from H, F, Cl, Br, I, OH and NH₂CN and C_1-3An alkyl group;

the 3-8 membered heterocycloalkenyl contains 1,2,3, or 4 heteroatoms or groups of heteroatoms independently selected from-NH-, -O-, -S-, and N.

The invention provides a compound shown in a formula (I) or a pharmaceutically acceptable salt thereof,

wherein,

selected from single and double bonds;

T ₁selected from NR_a；

T ₂Selected from C, CH and N;

T ₃selected from the group consisting of CR_bAnd N;

T ₄selected from the group consisting of CR_cAnd O;

R _aselected from H and C_1-3An alkyl group;

R _band R_cEach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl and C_1-3Alkoxy radical, said C_1-3Alkyl and C_{1- 3}Alkoxy is optionally substituted with 1,2 or 3R;

and when R is_cSelected from H or CN, R₁、R ₂And the atoms to which they are attached together form C_3-8Cycloalkenyl or 3-8 membered heterocycloalkenyl, said C_3-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

when R is_cSelected from F, Cl, Br, I, OH, NH₂And C optionally substituted by 1,2 or 3R_1-3Alkyl and C_1-3At alkoxy radical, R₁And R₂Each independently selected from H and optionally substituted by 1,2 or 3R_dSubstituted C_1-6Alkyl, or, R₁、R ₂And the atoms to which they are attached together form thienyl, C_3-8Cycloalkenyl or 3-8 membered heterocycloalkenyl, said C_3-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

R ₃and R₄Each independently selected from H, F, Cl, Br, I, OH, NH₂CN and optionally substituted by 1,2 or 3R_fSubstituted C_1-3An alkyl group;

R _d、R _eand R_fEach independently selected from H, F, Cl, Br, I, OH, NH₂CN and C_1-3An alkyl group;

each R is independently selected from H, F, Cl, Br, I, OH and NH₂CN and C_1-3An alkyl group;

the 3-8 membered heterocycloalkenyl contains 1,2,3, or 4 heteroatoms or groups of heteroatoms independently selected from-NH-, -O-, -S-, and N.

The invention provides a compound shown in a formula (I) or a pharmaceutically acceptable salt thereof,

wherein,

selected from single and double bonds;

T ₁is NR_a；

T ₂Selected from C, CH and N;

T ₃selected from the group consisting of CR_bAnd N;

T ₄selected from the group consisting of CR_cAnd O;

R _aselected from H and C_1-3An alkyl group;

R _band R_cEach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl and C_1-3Alkoxy radical, said C_1-3Alkyl and C_{1- 3}Alkoxy is optionally substituted by 12 or 3R;

and when R is_cSelected from F, Cl, Br, I, OH, NH₂And C optionally substituted by 1,2 or 3R_1-3Alkyl and C_1-3At alkoxy radical, R₁And R₂Each independently selected from H and optionally substituted by 1,2 or 3R_dSubstituted C_1-6Alkyl, or, R₁、R ₂And the atoms to which they are attached together form thienyl, C_3-8Cycloalkenyl or 3-8 membered heterocycloalkenyl, said thienyl, C_3-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

when R is_cSelected from H or CN, R₁、R ₂And the atoms to which they are attached together form C_3-8Cycloalkenyl or 3-8 membered heterocycloalkenyl, said C_3-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

R ₃and R₄Each independently selected from H, F, Cl, Br, I, OH, NH₂CN and optionally substituted by 1,2 or 3R_fSubstituted C_1-3An alkyl group;

R _d、R _eand R_fEach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl and C_1-3An alkoxy group;

each R is independently selected from H, F, Cl, Br, I, OH and NH₂CN and C_1-3An alkyl group;

the 3-8 membered heterocycloalkenyl contains 1,2,3, or 4 heteroatoms or groups of heteroatoms independently selected from-NH-, -O-, -S-, and N.

The invention provides a compound shown in a formula (I) or a pharmaceutically acceptable salt thereof,

wherein,

selected from single and double bonds;

T ₁is NR_a；

T ₂Selected from C, CH and N;

T ₃selected from the group consisting of CR_bAnd N;

T ₄selected from the group consisting of CR_cAnd O;

R _aselected from H and C_1-3An alkyl group;

R _band R_cEach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl and C_1-3Alkoxy radical, said C_1-3Alkyl and C_{1- 3}Alkoxy is optionally substituted with 1,2 or 3R;

and when R is_cSelected from F, Cl, Br, I, OH, NH₂And C optionally substituted by 1,2 or 3R_1-3Alkyl and C_1-3At alkoxy radical, R₁And R₂Each independently selected from H and optionally substituted by 1,2 or 3R_dSubstituted C_1-6Alkyl, or, R₁、R ₂And the atoms to which they are attached together form thienyl, C_3-8Cycloalkenyl or 3-8 membered heterocycloalkenyl, said thienyl, C_3-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

when R is_cSelected from H and CN, R₁、R ₂And the atoms to which they are attached together form C_6-8Cycloalkenyl or 3-8 membered heterocycloalkenyl, said C_6-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

R ₃and R₄Each independently selected from H, F, Cl, Br, I, OH, NH₂CN and optionally substituted by 1,2 or 3R_fSubstituted C_1-3An alkyl group;

R _dand R_eEach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、OCD ₃、C _1-3Alkyl radical, C_1-3Alkoxy and-O-C_3-5A cycloalkyl group;

R _findependently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl and C_1-3An alkoxy group;

each R is independently selected from H, F, Cl, Br, I, OH and NH₂CN and C_1-3An alkyl group;

the 3-8 membered heterocycloalkenyl contains 1,2,3, or 4 heteroatoms or groups of heteroatoms independently selected from-NH-, -O-, -S-, and N.

The invention provides a compound shown in a formula (I) or a pharmaceutically acceptable salt thereof,

wherein,

selected from single and double bonds;

T ₁is NR_a；

T ₂Selected from C, CH and N;

T ₃selected from the group consisting of CR_bAnd N;

T ₄selected from the group consisting of CR_cAnd O;

R _ais selected from H andC _1-3an alkyl group;

R _band R_cEach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl and C_1-3Alkoxy radical, said C_1-3Alkyl and C_{1- 3}Alkoxy is optionally substituted with 1,2 or 3R;

and when R is_cSelected from F, Cl, Br, I, OH, NH₂And C optionally substituted by 1,2 or 3R_1-3Alkyl and C_1-3At alkoxy radical, R₁And R₂Each independently selected from H and optionally substituted by 1,2 or 3R_dSubstituted C_1-6Alkyl, or, R₁、R ₂And the atoms to which they are attached together form thienyl, C_3-8Cycloalkenyl or 3-8 membered heterocycloalkenyl, said thienyl, C_3-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

when R is_cSelected from H and CN, R₁、R ₂And the atoms to which they are attached together form thienyl, C_6-8Cycloalkenyl or 3-8 membered heterocycloalkenyl, said thienyl, C_6-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

R ₃and R₄Each independently selected from H, F, Cl, Br, I, OH, NH₂CN and optionally substituted by 1,2 or 3R_fSubstituted C_1-3An alkyl group;

R _dand R_eEach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、OCD ₃、C _1-3Alkyl, OCH₃、OCH ₂CH ₃、OCH(CH ₃) ₂and-O-C_3-5A cycloalkyl group;

R _findependently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl and C_1-3An alkoxy group;

each R is independently selected from H, F, Cl, Br, I, OH and NH₂CN and C_1-3An alkyl group;

the 3-8 membered heterocycloalkenyl contains 1,2,3, or 4 heteroatoms or groups of heteroatoms independently selected from-NH-, -O-, -S-, and N.

The invention provides a compound shown in a formula (I) or a pharmaceutically acceptable salt thereof,

wherein,

selected from single and double bonds;

T ₁is NR_a；

T ₂Selected from C, CH and N;

T ₃selected from the group consisting of CR_bAnd N;

T ₄selected from the group consisting of CR_cAnd O;

R _aselected from H and C_1-3An alkyl group;

R _band R_cEach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl and C_1-3Alkoxy radical, said C_1-3Alkyl and C_{1- 3}Alkoxy is optionally substituted with 1,2 or 3R;

and when R is_cSelected from F, Cl, Br, I, OH, NH₂And C optionally substituted by 1,2 or 3R_1-3Alkyl and C_1-3At alkoxy radical, R₁And R₂Are each independently selected from H andoptionally substituted by 1,2 or 3R_dSubstituted C_1-6Alkyl, or, R₁、R ₂And the atoms to which they are attached together form thienyl, C_3-8Cycloalkenyl or 3-8 membered heterocycloalkenyl, said thienyl, C_3-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

when R is_cSelected from H and CN, R₁、R ₂And the atoms to which they are attached together form thienyl, C_6-8Cycloalkenyl or 3-8 membered heterocycloalkenyl, said thienyl, C_6-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

R ₃and R₄Each independently selected from H, F, Cl, Br, I, OH, NH₂CN and optionally substituted by 1,2 or 3R_fSubstituted C_1-3An alkyl group;

R _dand R_eEach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl radical, C_1-3Alkoxy and-O-C_3-5Cycloalkyl radical, said C_1-3Alkyl radical, C_1-3Alkoxy and-O-C_3-5Cycloalkyl is optionally substituted with 1,2 or 3R';

R _findependently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl and C_1-3An alkoxy group;

each R is independently selected from H, F, Cl, Br, I, OH and NH₂CN and C_1-3An alkyl group;

each R' is independently selected from D, F, Cl, Br and I;

the 3-8 membered heterocycloalkenyl contains 1,2,3, or 4 heteroatoms or groups of heteroatoms independently selected from-NH-, -O-, -S-, and N.

In some embodiments of the invention, the compound or pharmaceutically acceptable salt thereof is selected from

Wherein,

selected from single and double bonds;

T ₂selected from C and N;

T ₄selected from the group consisting of CR_cAnd O;

R _cselected from F, Cl, Br, I, OH, NH₂、C _1-3Alkyl and C_1-3Alkoxy radical, said C_1-3Alkyl and C_1-3Alkoxy is optionally substituted with 1,2 or 3R;

R ₁and R₂Each independently selected from H and optionally substituted by 1,2 or 3R_dSubstituted C_1-6Alkyl, or, R₁、R ₂And the atoms to which they are attached together form C_3-8Cycloalkenyl or 3-8 membered heterocycloalkenyl, said C_3-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

R ₃and R₄Each independently selected from H, F, Cl, Br, I, OH, NH₂CN and optionally substituted by 1,2 or 3R_fSubstituted C_1-3An alkyl group;

R _d、R _eand R_fEach independently selected from H, F, Cl, Br, I, OH, NH₂CN and C_1-3An alkyl group;

each R is independently selected from H, F, Cl, Br, I, OH and NH₂CN and C_1-3An alkyl group;

the 3-8 membered heterocycloalkenyl contains 1,2,3, or 4 heteroatoms or groups of heteroatoms independently selected from-NH-, -O-, -S-, and N.

In some embodiments of the invention, the compound or pharmaceutically acceptable salt thereof is selected from

Wherein,

selected from single and double bonds;

T ₂selected from C and N;

T ₄selected from the group consisting of CR_cAnd O;

R _cselected from F, Cl, Br, I, OH, NH₂、C _1-3Alkyl and C_1-3Alkoxy radical, said C_1-3Alkyl and C_1-3Alkoxy is optionally substituted with 1,2 or 3R;

R ₁and R₂Each independently selected from H and optionally substituted by 1,2 or 3R_dSubstituted C_1-6Alkyl, or, R₁、R ₂And the atoms to which they are attached together form thienyl, C_3-8Cycloalkenyl or 3-8 membered heterocycloalkenyl, said C_3-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

R ₃and R₄Each independently selected from H, F, Cl, Br, I, OH, NH₂CN and optionally substituted by 1,2 or 3R_fSubstituted C_1-3An alkyl group;

R _d、R _eand R_fEach independently selected from H, F, Cl, Br, I, OH, NH₂CN and C_1-3An alkyl group;

each R is independently selected from H, F, Cl, Br, I, OH and NH₂CN and C_1-3An alkyl group;

the 3-8 membered heterocycloalkenyl contains 1,2,3, or 4 heteroatoms or groups of heteroatoms independently selected from-NH-, -O-, -S-, and N.

In some embodiments of the invention, the compound or pharmaceutically acceptable salt thereof is selected from

Wherein,

selected from single and double bonds;

T ₂selected from C and N;

T ₄selected from the group consisting of CR_cAnd O;

R _cselected from F, Cl, Br, I, OH, NH₂、C _1-3Alkyl and C_1-3Alkoxy radical, said C_1-3Alkyl and C_1-3Alkoxy is optionally substituted with 1,2 or 3R;

R ₁and R₂Each independently selected from H and optionally substituted by 1,2 or 3R_dSubstituted C_1-6Alkyl, or, R₁、R ₂And the atoms to which they are attached together form thienyl, C_3-8Cycloalkenyl or 3-8 membered heterocycloalkenyl, said thienyl, C_3-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

R ₃and R₄Each independently selected from H, F, Cl, Br, I, OH, NH₂CN and optionally substituted by 1,2 or 3R_fSubstituted C_1-3An alkyl group;

R _d、R _eand R_fEach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl and C_1-3An alkoxy group;

each R is independently selected from H, F, Cl, Br, I, OH and NH₂CN and C_1-3An alkyl group;

the 3-8 membered heterocycloalkenyl contains 1,2,3, or 4 heteroatoms or groups of heteroatoms independently selected from-NH-, -O-, -S-, and N.

In some embodiments of the invention, the compound or pharmaceutically acceptable salt thereof is selected from

Wherein,

selected from single and double bonds;

T ₂selected from C and N;

T ₄selected from the group consisting of CR_cAnd O;

R _cselected from F, Cl, Br, I, OH, NH₂、C _1-3Alkyl and C_1-3Alkoxy radical, said C_1-3Alkyl and C_1-3Alkoxy is optionally substituted with 1,2 or 3R;

R ₁and R₂Each independently selected from H and optionally substituted by 1,2 or 3R_dSubstituted C_1-6Alkyl, or, R₁、R ₂And the atoms to which they are attached together form thienyl, C_3-8Cycloalkenyl or 3-8 membered heterocycloalkenyl, said thienyl, C_3-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

R ₃and R₄Each independently selected from H, F, Cl, Br, I, OH, NH₂CN and optionally substituted by 1,2 or 3R_fSubstituted C_1-3An alkyl group;

R _dand R_eEach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、OCD ₃、C _1-3Alkyl radical, C_1-3Alkoxy and-O-C_3-5A cycloalkyl group;

R _findependently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl and C_1-3An alkoxy group;

each R is independently selected from H, F, Cl, Br, I, OH and NH₂CN and C_1-3An alkyl group;

the 3-8 membered heterocycloalkenyl contains 1,2,3, or 4 heteroatoms or groups of heteroatoms independently selected from-NH-, -O-, -S-, and N.

In some embodiments of the invention, the compound or pharmaceutically acceptable salt thereof is selected from

Wherein,

selected from single and double bonds;

T ₂selected from C and N;

T ₄selected from the group consisting of CR_cAnd O;

R _cselected from F, Cl, Br, I, OH, NH₂、C _1-3Alkyl and C_1-3Alkoxy radical, said C_1-3Alkyl and C_1-3Alkoxy is optionally substituted with 1,2 or 3R;

R ₁and R₂Each independently selected from H and optionally substituted by 1,2 or 3R_dSubstituted C_1-6Alkyl, or, R₁、R ₂And the atoms to which they are attached together form thienyl, C_3-8Cycloalkenyl or 3-8 membered heterocycloalkenyl, said thienyl, C_3-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

R ₃and R₄Each independently selected from H, F, Cl, Br, I, OH, NH₂CN and optionally substituted by 1,2 or 3R_fSubstituted C_1-3An alkyl group;

R _dand R_eEach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、OCD ₃、C _1-3Alkyl, OCH₃、OCH ₂CH ₃、OCH(CH ₃) ₂and-O-C_3-5A cycloalkyl group;

R _findependently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl and C_1-3An alkoxy group;

each R is independently selected from H, F, Cl, Br, I, OH and NH₂CN and C_1-3An alkyl group;

the 3-8 membered heterocycloalkenyl contains 1,2,3, or 4 heteroatoms or groups of heteroatoms independently selected from-NH-, -O-, -S-, and N.

In some embodiments of the invention, the compound or pharmaceutically acceptable salt thereof is selected from

Wherein,

selected from single and double bonds;

T ₂selected from C and N;

T ₄selected from the group consisting of CR_cAnd O;

R _cselected from F, Cl, Br, I, OH, NH₂、C _1-3Alkyl and C_1-3Alkoxy radical, said C_1-3Alkyl and C_1-3Alkoxy is optionally substituted with 1,2 or 3R;

R ₁and R₂Each independently selected from H and optionally substituted by 1,2 or 3R_dSubstituted C_1-6Alkyl, or, R₁、R ₂And the atoms to which they are attached together form thienyl, C_3-8Cycloalkenyl or 3-8 membered heterocycloalkenyl, said thienyl, C_3-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

R ₃and R₄Each independently selected from H, F, Cl, Br, I, OH, NH₂CN and optionally substituted by 1,2 or 3R_fSubstituted C_1-3An alkyl group;

R _dand R_eEach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl radical, C_1-3Alkoxy and-O-C_3-5Cycloalkyl radical, said C_1-3Alkyl radical, C_1-3Alkoxy and-O-C_3-5Cycloalkyl is optionally substituted with 1,2 or 3R';

R _findependently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl and C_1-3An alkoxy group;

each R is independently selected from H, F, Cl, Br, I, OH and NH₂CN and C_1-3An alkyl group;

each R' is independently selected from D, F, Cl, Br and I;

the 3-8 membered heterocycloalkenyl contains 1,2,3, or 4 heteroatoms or groups of heteroatoms independently selected from-NH-, -O-, -S-, and N.

In some embodiments of the invention, the compound or pharmaceutically acceptable salt thereof is selected from

Wherein,

R _cselected from H and CN;

R ₁、R ₂and the atoms to which they are attached together form C_3-8Cycloalkenyl and 3-8 membered heterocycloalkenyl, said C_3-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

R ₃and R₄Each independently selected from H, F, Cl, Br, I, OH, NH₂CN and optionally substituted by 1,2 or 3R_fSubstituted C_1-3An alkyl group;

R _eand R_fEach independently selected from H, F, Cl, Br, I, OH, NH₂CN and C_1-3An alkyl group;

the 3-8 membered heterocycloalkenyl contains 1,2,3, or 4 heteroatoms or groups of heteroatoms independently selected from-NH-, -O-, -S-, and N.

In some embodiments of the invention, the compound or pharmaceutically acceptable salt thereof is selected from

Wherein,

R _cselected from H and CN;

R ₁、R ₂and the atoms to which they are attached together form thienyl, C_3-8Cycloalkenyl and 3-8 membered heterocycloalkenyl, said C_3-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

R ₃and R₄Each independently selected from H, F, Cl, Br, I, OH, NH₂CN and optionally substituted by 1,2 or 3R_fSubstituted C_1-3An alkyl group;

R _eand R_fEach independently selected from H, F, Cl, Br, I, OH, NH₂CN and C_1-3An alkyl group;

the 3-8 membered heterocycloalkenyl contains 1,2,3, or 4 heteroatoms or groups of heteroatoms independently selected from-NH-, -O-, -S-, and N.

In some embodiments of the invention, the compound or pharmaceutically acceptable salt thereof is selected from

Wherein,

R _cselected from H and CN;

R ₁、R ₂and the atoms to which they are attached together form thienyl, C_3-8Cycloalkenyl and 3-8 membered heterocycloalkenyl, said thienyl, C_3-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

R ₃and R₄Each independently selected from H, F, Cl, Br, I, OH, NH₂CN and optionally substituted by 1,2 or 3R_fSubstituted C_1-3An alkyl group;

R _eand R_fEach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl and C_1-3An alkoxy group;

the 3-8 membered heterocycloalkenyl contains 1,2,3, or 4 heteroatoms or groups of heteroatoms independently selected from-NH-, -O-, -S-, and N.

In some embodiments of the invention, the compound or pharmaceutically acceptable salt thereof is selected from

Wherein,

R _cselected from H and CN;

R ₁、R ₂and the atoms to which they are attached together form thienyl, C_6-8Cycloalkenyl and 3-8 membered heterocycloalkenyl, said thienyl, C_6-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

R ₃and R₄Each independently selected from H, F, Cl, Br, I, OH, NH₂CN and optionally substituted by 1,2 or 3R_fSubstituted C_1-3An alkyl group;

R _eeach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、OCD ₃、C _1-3Alkyl radical, C_1-3Alkoxy and-O-C_3-5A cycloalkyl group;

R _findependently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl and C_1-3An alkoxy group;

the 3-8 membered heterocycloalkenyl contains 1,2,3, or 4 heteroatoms or groups of heteroatoms independently selected from-NH-, -O-, -S-, and N.

In some embodiments of the invention, the compound or pharmaceutically acceptable salt thereof is selected from

Wherein,

R _cselected from H and CN;

R ₁、R ₂and the atoms to which they are attached together form thienyl, C_6-8Cycloalkenyl and 3-8 membered heterocycloalkenyl, said thienyl, C_6-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

R ₃and R₄Each independently selected from H, F, Cl, Br, I, OH, NH₂CN and optionally substituted by 1,2 or 3R_fSubstituted C_1-3An alkyl group;

R _eeach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、OCD ₃、C _1-3Alkyl, OCH₃、OCH ₂CH ₃、OCH(CH ₃) ₂and-O-C_3-5A cycloalkyl group;

R _findependently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl and C_1-3An alkoxy group;

the 3-8 membered heterocycloalkenyl contains 1,2,3, or 4 heteroatoms or groups of heteroatoms independently selected from-NH-, -O-, -S-, and N.

In some embodiments of the invention, the compound or pharmaceutically acceptable salt thereof is selected from

Wherein,

R _cselected from H and CN;

R ₁、R ₂and the atoms to which they are attached together form thienyl, C_6-8Cycloalkenyl and 3-8 membered heterocycloalkenyl, said thienyl, C _6-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

R ₃and R₄Each independently selected from H, F, Cl, Br, I, OH, NH₂CN and optionally substituted by 1,2 or 3R_fSubstituted C_1-3An alkyl group;

R _eeach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl radical, C_1-3Alkoxy and-O-C_3-5Cycloalkyl radical, said C_1-3Alkyl radical, C_1-3Alkoxy and-O-C_3-5Cycloalkyl is optionally substituted with 1,2 or 3R';

R _findependently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl and C_1-3An alkoxy group;

each R' is independently selected from D, F, Cl, Br and I;

the 3-8 membered heterocycloalkenyl contains 1,2,3, or 4 heteroatoms or groups of heteroatoms independently selected from-NH-, -O-, -S-, and N.

In some embodiments of the invention, T is₂Selected from N, T₃When selected from N, T₂And T₃Connected thereto

Is a single bond; t is₄Selected from the group consisting of CR_cWhen, T₃And T₄Connected thereto

Is a double bond; t is₄Selected from O, T₃And T₄Connected thereto

Is a single bond.

In some embodiments of the invention, R is as defined above_bAnd R_cEach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、CH ₃、CH ₂CH ₃And

the CH₃、CH ₂CH ₃And

optionally substituted with 1,2 or 3R, the other variables being as defined herein.

In some embodiments of the invention, R is as defined above_bAnd R_cEach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、CH ₃、CH ₂F、CHF ₂、CF ₃、CH ₂CH ₃、

Other variables are as defined herein.

In some embodiments of the invention, R is as defined above_bAnd R_cEach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、CH ₃、CH ₂F、CHF ₂、CF ₃、CH ₂CH ₃、CF ₂CH ₃、

Other variables are as defined herein.

In some embodiments of the invention, R is as defined above_bAnd R _cEach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、CH ₃、CH ₂F、CH ₂CN、CHF ₂、CF ₃、CH ₂CH ₃、CF ₂CH ₃、

Other variables are as defined herein.

In some embodiments of the invention, R is as defined above_cSelected from F, Cl, Br, I, OH, NH₂、CH ₃、CH ₂F、CHF ₂、CF ₃、CH ₂CH ₃、

Other variables are as defined herein.

In some embodiments of the invention, R is as defined above_cSelected from F, Cl, Br, I, OH, NH₂、CH ₃、CH ₂F、CHF ₂、CF ₃、CH ₂CH ₃、CF ₂CH ₃、

Other variables are as defined herein.

In some embodiments of the invention, R is as defined above_cSelected from F, Cl, Br, I, OH, NH₂、CH ₃、CH ₂F、CHF ₂、CH ₂CN、CF ₃、CH ₂CH ₃、CF ₂CH ₃、

Other variables are as defined herein.

In some embodiments of the invention, R is as defined above₁And R₂Each independently selected from H, CH₃、CH ₂CH ₃、CH(CH ₃) ₂And C (CH)₃) ₃The other variables are as defined herein.

In some embodiments of the invention, R is as defined above₁、R ₂And the atoms to which they are attached together form a cyclohexenyl, bicyclo [2.2.1 ]]Hept-2-enyl, 1,2,3, 4-tetrahydropyridinyl, bicyclo [4.1.0]Hept-3-enyl, 3, 6-dihydro-2H-pyranyl and 7-oxabicyclo [2.2.1]Hept-2-enyl, said cyclohexenyl, bicyclo [2.2.1]Hept-2-enyl, 1,2,3, 4-tetrahydropyridinyl, bicyclo [4.1.0]Hept-3-enyl, 3, 6-dihydro-2H-pyranyl and 7-oxabicyclo [2.2.1]Hept-2-enyl optionally substituted by 1,2 or 3R_eAnd the other variables are as defined herein.

In some embodiments of the invention, R is as defined above₁、R ₂And the atoms to which they are attached together form a cyclopentenyl, cyclohexenyl, bicyclo [2.2.1 ] group]Hept-2-enyl, 1,2,3, 4-tetrahydropyridinyl, bicyclo [4.1.0]Hept-3-enyl, 3, 6-dihydro-2H-pyranyl, 7-oxabicyclo [2.2.1]Hept-2-enyl and thienyl, said cyclopentenyl, cyclohexenyl, bicyclo [2.2.1]Hept-2-enyl, 1,2,3, 4-tetrahydropyridinyl, bicyclo [4.1.0]Hept-3-enyl, 3, 6-dihydro-2H-pyranyl, 7-oxabicyclo [2.2.1]Hept-2-enyl and thienyl are optionally substituted by 1,2 or 3R_eAnd the other variables are as defined herein.

In some embodiments of the invention, R is as defined above₁、R ₂And the atoms to which they are attached together form a cyclopentenyl, cyclohexenyl, bicyclo [2.2.1 ] group]Hept-2-enyl, 1,2,3, 4-tetrahydropyridinyl, bicyclo [4.1.0]Hept-3-enyl, 3, 6-dihydro-2H-pyranyl, 7-oxabicyclo [2.2.1]Hept-2-enyl, thienyl, 3, 4-dihydro-2H-pyranyl and bicyclo [4.1.0]Hept-2-enyl, cyclopentenyl, cyclohexenyl, bicyclo [2.2.1]Hept-2-enyl, 1,2,3, 4-tetrahydropyridinyl, bicyclo [4.1.0]Hept-3-enyl, 3, 6-dihydro-2H-pyranyl, 7-oxabicyclo [2.2.1]Hept-2-enyl, thienyl, 3, 4-dihydro-2H-pyranyl and bicyclo [4.1.0]Hept-2-enyl optionally substituted by 1,2 or 3R_eAnd the other variables are as defined herein.

In some embodiments of the invention, R is as defined above₁、R ₂And the atoms to which they are attached together form a cyclohexenyl, bicyclo [2.2.1 ]]Hept-2-enyl, 1,2,3, 4-tetrahydropyridinyl, bicyclo [4.1.0]Hept-3-enyl, 3, 6-dihydro-2H-pyranyl, 7-oxabicyclo [2.2.1]Hept-2-enyl, thienyl, 3, 4-dihydro-2H-pyranyl and bicyclo [4.1.0]Hept-2-enyl, said cyclohexenyl, bicyclo [2.2.1]Hept-2-enyl, 1,2,3, 4-tetrahydropyridinyl, bicyclo [4.1.0]Hept-3-enyl, 3, 6-dihydro-2H-pyranyl, 7-oxabicyclo [2.2.1]Hept-2-enyl, thienyl, 3, 4-dihydro-2H-pyranyl and bicyclo [4.1.0]Hept-2-enyl optionally substituted by 1,2 or 3R_eAnd the other variables are as defined herein.

In some embodiments of the invention, the structural unit

Is selected from

Other variables are as defined herein.

In some embodiments of the invention, the structural unit

Is selected from

Other variables are as defined herein.

In some embodiments of the invention, the structural unit

Is selected from

Other variables are as defined herein.

In some embodiments of the invention, the structural unit

Is selected from

Other variables are as defined herein.

In some embodiments of the invention, the structural unit

Is selected from

Other variables are as defined herein.

In some embodiments of the invention, the structural unit

Is selected from

Other variables are as defined herein.

In some embodiments of the invention, the structural unit

Is selected from

Other variables are as defined herein.

In some embodiments of the invention, the structural unit

Is selected from

Other variables are as defined herein.

In some embodiments of the invention, the structural unit

Is selected from

Other variables are as defined herein.

In some embodiments of the invention, the structural unit

Is selected from

Other variantsThe amounts are as defined herein.

In some embodiments of the invention, the structural unit

Is selected from

Other variables are as defined herein.

In some embodiments of the invention, the structural unit

Is selected from

Other variables are as defined herein.

In some embodiments of the invention, the structural unit

Is selected from

Other variables are as defined herein.

In some embodiments of the invention, R is as defined above₃And R₄Each independently selected from H, F, Cl, Br, I, OH, NH₂、CN、CH ₃And CH₂CH ₃Said CH₃And CH₂CH ₃Optionally substituted by 1,2 or 3R_fAnd the other variables are as defined herein.

In some embodiments of the invention, R is as defined above₃And R₄Each independently selected from F, Cl, Br, I, OH and NH₂。

In some embodiments of the invention, R is as defined above₃And R₄Each independently selected from F, Cl, Br, I, OH and NH₂The other variables are as defined herein.

In some embodiments of the invention, R is as defined above₃And R₄Each independently selected from F, Cl, Br, I, OH, NH₂And CF₃The other variables are as defined herein.

In some embodiments of the invention, the structural unit

Is selected from

OthersThe variables are as defined herein.

In some embodiments of the invention, the structural unit

Is selected from

Other variables are as defined herein.

In some embodiments of the invention, the structural unit

Is selected from

Other variables are as defined herein.

In some embodiments of the invention, the structural unit

Is selected from

Other variables are as defined herein.

Still other embodiments of the present invention are derived from any combination of the above variables.

In some embodiments of the invention, the compound or pharmaceutically acceptable salt thereof is selected from

Wherein,

Z ₁and Z₂Are each independently selected from CH (R)_e) O and N (R)_e)；

Z ₃Is selected from CH₂And O;

Z ₄is selected from O;

Z ₅is selected from C (R)_e) And N;

Z ₆is selected from N (R)_e) And O;

R _cselected from H and CN;

R ₃、R ₄and R_eAs defined herein.

In some embodiments of the invention, the compound or pharmaceutically acceptable salt thereof is selected from

Wherein,

R ₁、R ₂、R ₃、R ₄and R_cAs defined herein.

The invention also provides a compound shown as the following formula or a pharmaceutically acceptable salt thereof,

in some embodiments of the invention, the compound, or a pharmaceutically acceptable salt thereof, is selected from the group consisting of:

the invention also provides a pharmaceutical composition comprising as an active ingredient a therapeutically effective amount of a compound according to the above or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

In some embodiments of the present invention, the use of the above compound or a pharmaceutically acceptable salt thereof or the above composition for the manufacture of a medicament related to a thyroxine receptor- β agonist.

In some embodiments of the present invention, the above-mentioned use, wherein the thyroxine receptor- β agonist related medicament is a medicament for the treatment of nonalcoholic steatohepatitis.

Definitions and explanations

As used herein, the following terms and phrases are intended to have the following meanings, unless otherwise indicated. A particular term or phrase, unless specifically defined, should not be considered as indefinite or unclear, but rather construed according to ordinary meaning. When a trade name appears herein, it is intended to refer to its corresponding commodity or its active ingredient. The term "pharmaceutically acceptable" as used herein is intended to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

The term "pharmaceutically acceptable salts" refers to salts of the compounds of the present invention, prepared from the compounds of the present invention found to have particular substituents, with relatively nontoxic acids or bases. When compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of a base in neat solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic ammonia or magnesium salts or similar salts. When compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in neat solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, phosphorous acid, and the like; and salts of organic acids including acids such as acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-toluenesulfonic, citric, tartaric, methanesulfonic, and the like; also included are salts of amino acids such as arginine and the like, and salts of organic acids such as glucuronic acid and the like. Certain specific compounds of the invention contain both basic and acidic functionalities and can thus be converted to any base or acid addition salt.

The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound, which contains an acid or base, by conventional chemical methods. In general, such salts are prepared by the following method: prepared by reacting these compounds in free acid or base form with a stoichiometric amount of the appropriate base or acid, in water or an organic solvent or a mixture of the two.

In addition to salt forms, the compounds provided herein also exist in prodrug forms. Prodrugs of the compounds described herein readily undergo chemical changes under physiological conditions to convert to the compounds of the present invention. In addition, prodrugs can be converted to the compounds of the present invention in an in vivo environment by chemical or biochemical means.

Certain compounds of the present invention may exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms are equivalent to unsolvated forms and are intended to be encompassed within the scope of the present invention.

The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-) -and (+) -enantiomers, (R) -and (S) -enantiomers, diastereomers, (D) -isomers, (L) -isomers, as well as racemic and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers, as well as mixtures thereof, are included within the scope of the present invention.

Unless otherwise indicated, the terms "enantiomer" or "optical isomer" refer to stereoisomers that are mirror images of each other.

Unless otherwise indicated, the term "cis-trans isomer" or "geometric isomer" results from the inability of a double bond or a single bond to rotate freely within a ring-forming carbon atom.

Unless otherwise indicated, the term "diastereomer" refers to a stereoisomer in which the molecules have two or more chiral centers and a non-mirror image relationship between the molecules.

Unless otherwise indicated, "(D)" or "(+)" means dextrorotation, "(L)" or "(-) -means levorotation," (DL) "or" (±) "means racemization.

Using solid wedge keys, unless otherwise indicated

And wedge dotted bond

Represents oneAbsolute configuration of solid center, using straight solid keys

And straight dotted line bond

Showing the relative configuration of the centres of solids, by wavy lines

Representing solid-line keys of wedge shape

Or wedge dotted bond

Or by wavy lines

Indicating straight solid-line keys

And straight dotted line bond

The compounds of the invention may be present specifically. Unless otherwise indicated, the term "tautomer" or "tautomeric form" means that at room temperature, the isomers of different functional groups are in dynamic equilibrium and can be rapidly interconverted. If tautomers are possible (e.g., in solution), then the chemical equilibrium of the tautomers can be reached. For example, proton tautomers (prototropic tautomers), also known as proton transfer tautomers (prototropic tautomers), include interconversions by proton transfer, such as keto-enol isomerization and imine-enamine isomerization. Valence isomers (valencetatomer) include interconversion by recombination of some of the bonding electrons. A specific example of where keto-enol tautomerism is the interconversion between two tautomers of pentane-2, 4-dione and 4-hydroxypent-3-en-2-one.

Unless otherwise indicated, the terms "enriched in one isomer", "isomer enriched", "enantiomer enriched" or "enantiomeric enrichment" refer to a content of one isomer or enantiomer of less than 100%, and the content of the isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.

Unless otherwise indicated, the term "isomeric excess" or "enantiomeric excess" refers to the difference between the relative percentages of two isomers or enantiomers. For example, if the content of one isomer or enantiomer is 90%, and the content of the other isomer or enantiomer is 10%, the isomer or enantiomer excess (ee value) is 80%.

Optically active (R) -and (S) -isomers as well as D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one of the enantiomers of a compound of the invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (e.g., amino) or an acidic functional group (e.g., carboxyl), diastereomeric salts are formed with an appropriate optically active acid or base, followed by diastereomeric resolution by conventional methods known in the art, and the pure enantiomers are recovered. Furthermore, separation of enantiomers and diastereomers is typically accomplished by using chromatography employing a chiral stationary phase, optionally in combination with chemical derivatization (e.g., carbamate formation from amines).

The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds may be labelled with radioactive isotopes, such as tritium (A), (B), (C) and C)³H) Iodine-125 (¹²⁵I) Or C-14(¹⁴C) In that respect For example, deuterium can be used to replace hydrogen to form a deuterated drug, the bond formed by deuterium and carbon is stronger than the bond formed by common hydrogen and carbon, and compared with an undeuterated drug, the deuterated drug has the advantages of reducing toxic and side effects, increasing the stability of the drug, enhancing the curative effect, prolonging the biological half-life period of the drug and the like. All isotopic variations of the compounds of the present invention, whether radioactive or not, are intended to be encompassed within the scope of the present invention.

The terms "optional" or "optionally" mean that the subsequently described event or circumstance may, but need not, occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

The term "substituted" means that any one or more hydrogen atoms on a particular atom is replaced with a substituent, and may include variations of deuterium and hydrogen, so long as the valency of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., ═ O), it means that two hydrogen atoms are substituted. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it may or may not be substituted, and unless otherwise specified, the kind and number of substituents may be arbitrary on the basis of chemical realizability.

When any variable (e.g., R) occurs more than one time in the composition or structure of a compound, its definition in each case is independent. Thus, for example, if a group is substituted with 0-2R, the group may optionally be substituted with up to two R, and there are separate options for R in each case. Furthermore, combinations of substituents and/or variants thereof are permissible only if such combinations result in stable compounds.

When the number of one linking group is 0, e.g. - (CRR)₀-, represents that the linking group is a single bond.

When one of the variables is selected from a single bond, it means that the two groups to which it is attached are directly connected, for example, where L represents a single bond in A-L-Z means that the structure is actually A-Z.

When a substituent is absent, it indicates that the substituent is absent, e.g., when X is absent in A-X, it indicates that the structure is actually A. When no atom through which a substituent is attached to a substituted group is indicated in the listed substituents, such substituents may be bonded through any atom thereof, for example, a pyridyl group as a substituent may be attached to a substituted group through any one of carbon atoms on the pyridine ring. When the listed linking groups do not indicate their direction of attachment, the direction of attachment is arbitrary, for example,

wherein the linking group L is-M-W-, in which case-M-W-can be formed by connecting the ring A and the ring B in the same direction as the reading sequence from left to right

The ring A and the ring B may be connected in the reverse direction of the reading sequence from left to right

Combinations of the linking groups, substituents, and/or variants thereof are permissible only if such combinations result in stable compounds.

Unless otherwise specified, the term "C_1-6Alkyl "is intended to mean a straight or branched saturated hydrocarbon group consisting of 1 to 6 carbon atoms. Said C is_1-6The alkyl group comprising C_1-5、C _1-4、C _1-3、C _1-2、C _2-6、C _2-4、C ₆And C₅Alkyl, etc.; it may be monovalent (e.g., methyl), divalent (e.g., methylene), or multivalent (e.g., methine). C_1-6Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, s-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl, and the like.

Unless otherwise specified, the term "C_1-3Alkyl "is intended to mean a straight or branched saturated hydrocarbon group consisting of 1 to 3 carbon atoms. Said C is_1-3The alkyl group comprising C_1-2And C_2-3Alkyl, etc.; it may be monovalent (e.g., methyl), divalent (e.g., methylene), or multivalent (e.g., methine). C_{1- 3}Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), and the like.

Unless otherwise specified, the term "C_1-3Alkoxy "denotes those alkyl groups containing 1 to 3 carbon atoms which are attached to the rest of the molecule through an oxygen atom. Said C is_1-3Alkoxy radicals comprising C_1-2、C _2-3、C ₃And C₂Alkoxy, and the like. C_1-3Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), and the like.

Unless otherwise specified, "C" is_3-8Cycloalkenyl "denotes partially unsaturated cyclic hydrocarbon groups consisting of 3 to 8 carbon atoms containing at least one carbon-carbon double bond, including monocyclic and bicyclic ring systems, wherein bicyclic ring systems include spirocyclic, fused and bridged rings, any ring of which is non-aromatic. Said C is_3-8Cycloalkenyl radicals including C_3-6、C _3-5、C _4-10、C _4-8、C _4-6、C _4-5、C _5-8Or C _5-6Cycloalkenyl groups and the like; it may be monovalent, divalent or polyvalent. C_3-8Examples of cycloalkenyl groups include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, and the like.

Unless otherwise specified, "C" is_6-8Cycloalkenyl "denotes a partially unsaturated cyclic hydrocarbon group consisting of 6 to 8 carbon atoms containing at least one carbon-carbon double bond, including monocyclic and bicyclic ring systems, wherein bicyclic ring systems include spirocyclic, fused and bridged rings, any ring of which is non-aromatic. Said C is_6-8Cycloalkenyl radicals including C_6-7Or C_6-8Cycloalkenyl groups and the like; it may be monovalent, divalent or polyvalent. C_6-8Examples of cycloalkenyl groups include, but are not limited to, cyclohexenyl, cycloheptenyl, cyclooctenyl, and the like.

Unless otherwise specified, the term "3-8 membered heterocycloalkenyl" by itself or in combination with other terms, means a partially unsaturated cyclic group of 3 to 8 ring atoms containing at least one carbon-carbon double bond, 1,2,3, or 4 of the ring atoms being heteroatoms independently selected from O, S and N, the remainder being carbon atoms, wherein the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms can be optionally oxidized (i.e., NO and S (O))_pAnd p is 1 or 2). They include monocyclic, bicyclic and tricyclic ring systems, wherein bicyclic and tricyclic ring systems include spirocyclic, bicyclic and bridged rings, any ring of which is non-aromatic. Further, with respect to the "3-8 membered heterocycloalkenyl", the heteroatom may occupy the position of attachment of the heterocycloalkenyl to the rest of the molecule. The 3-8 membered heterocycloalkenyl group includes 3-6 membered, 3-5 membered, 4-6 membered, 4-5 membered, 5-6 membered, 4 membered, 5 membered, and 6 membered heterocycloalkenyl and the like. Examples of 3-8 membered heterocycloalkenyl include, but are not limited to

Unless otherwise specified, the term "3-8 membered heterocycloalkyl" by itself or in combination with other terms denotes a saturated cyclic group consisting of 3 to 8 ring atoms,wherein 1,2,3, or 4 ring atoms are heteroatoms independently selected from O, S and N, the remainder being carbon atoms, wherein the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms may be optionally oxidized (i.e., NO and S (O))_pAnd p is 1 or 2). It includes monocyclic and bicyclic ring systems, wherein bicyclic ring systems include spiro, fused and bridged rings. Furthermore, with respect to the "3-8 membered heterocycloalkyl", the heteroatom may occupy the position of the heterocycloalkyl linkage to the rest of the molecule. The 3-8 membered heterocycloalkyl group includes 3-6 membered, 3-5 membered, 4-6 membered, 5-6 membered, 4 membered, 5-membered, and 6-membered heterocycloalkyl groups and the like. Examples of 3-8 membered heterocycloalkyl include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothienyl (including tetrahydrothien-2-yl and tetrahydrothien-3-yl, and the like), tetrahydrofuranyl (including tetrahydrofuran-2-yl, and the like), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl, and 3-piperidinyl, and the like), piperazinyl (including 1-piperazinyl, and 2-piperazinyl, and the like), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1, 2-oxazinyl, 1, 2-thiazinyl, hexahydropyridazinyl, homopiperazinyl, homopiperidinyl, dioxepanyl, and the like.

Unless otherwise specified, C_n-n+mOr C_n-C _n+mIncluding any one particular case of n to n + m carbons, e.g. C_1-12Comprising C₁、C ₂、C ₃、C ₄、C ₅、C ₆、C ₇、C ₈、C ₉、C ₁₀、C ₁₁And C₁₂Also included are any ranges of n to n + m, e.g. C_1-12Comprising C_{1- 3}、C _1-6、C _1-9、C _3-6、C _3-9、C _3-12、C _6-9、C _6-12And C_9-12Etc.; similarly, n-to n + m-members represent n to n + m ring atoms, and the 3-to 12-membered ring includes 3-membered ring, 4-membered ring, 5-membered ring, 6-membered ring, 7-membered ring,8-membered, 9-membered, 10-membered, 11-membered, and 12-membered rings, and also any one of n to n + m, for example, 3-12-membered rings include 3-6-membered rings, 3-9-membered rings, 5-6-membered rings, 5-7-membered rings, 6-8-membered rings, 6-10-membered rings, and the like.

The term "leaving group" refers to a functional group or atom that can be substituted by another functional group or atom through a substitution reaction (e.g., an affinity substitution reaction). For example, representative leaving groups include triflate; chlorine, bromine, iodine; sulfonate groups such as methanesulfonate, toluenesulfonate, p-bromobenzenesulfonate, p-toluenesulfonate and the like; acyloxy groups such as acetoxy, trifluoroacetyloxy, and the like.

The term "protecting group" includes, but is not limited to, "amino protecting group," hydroxyl protecting group, "or" thiol protecting group. The term "amino protecting group" refers to a protecting group suitable for use in preventing side reactions at the amino nitrogen position. Representative amino protecting groups include, but are not limited to: a formyl group; acyl, for example alkanoyl (such as acetyl, trichloroacetyl or trifluoroacetyl); alkoxycarbonyl such as tert-butoxycarbonyl (Boc); arylmethoxycarbonyl groups such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); arylmethyl groups such as benzyl (Bn), trityl (Tr), 1-bis- (4' -methoxyphenyl) methyl; silyl groups, such as Trimethylsilyl (TMS) and t-butyldimethylsilyl (TBS), and the like. The term "hydroxy protecting group" refers to a protecting group suitable for use in preventing side reactions of a hydroxy group. Representative hydroxy protecting groups include, but are not limited to: alkyl groups such as methyl, ethyl and tert-butyl; acyl groups, such as alkanoyl (e.g., acetyl); arylmethyl groups such as benzyl (Bn), p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl (benzhydryl, DPM); silyl groups, such as Trimethylsilyl (TMS) and t-butyldimethylsilyl (TBS), and the like.

The compounds of the present invention may be structurally confirmed by conventional methods well known to those skilled in the art, and if the present invention relates to the absolute configuration of the compound, the absolute configuration may be confirmed by means of conventional techniques in the art. For example, single crystal X-ray diffraction method (SXRD), the single crystal is cultured by Bruker D8 ventuThe re diffractometer collects diffraction intensity data, a light source is CuK alpha radiation, and the scanning mode is as follows:

after scanning and collecting relevant data, the crystal structure is further analyzed by a direct method (Shelxs97), so that the absolute configuration can be confirmed.

The compounds of the present invention may be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combinations thereof with other chemical synthetic methods, and equivalents thereof known to those skilled in the art, with preferred embodiments including, but not limited to, examples of the present invention.

The solvent used in the present invention can be commercially available. The invention employs the following abbreviations: aq represents water; HATU represents O- (7-azabenzotriazol-1-yl) -N, N' -tetramethyluronium hexafluorophosphate; EDC stands for N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride; m-CPBA represents 3-chloroperoxybenzoic acid; eq represents equivalent, equivalent; CDI represents carbonyldiimidazole; DCM represents dichloromethane; PE represents petroleum ether; DIAD represents diisopropyl azodicarboxylate; DMF represents N, N-dimethylformamide; DMSO represents dimethyl sulfoxide; EtOAc for ethyl acetate; EtOH stands for ethanol; MeOH represents methanol; CBz represents benzyloxycarbonyl, an amine protecting group; BOC represents tert-butoxycarbonyl as an amine protecting group; HOAc represents acetic acid; NaCNBH₃Represents sodium cyanoborohydride; r.t. represents room temperature; O/N stands for overnight; THF represents tetrahydrofuran; boc₂O represents di-tert-butyl dicarbonate; t formic acid represents trifluoroacetic acid; DIPEA stands for diisopropylethylamine; SOCl₂Represents thionyl chloride; CS₂Represents carbon disulfide; TsOH represents p-toluenesulfonic acid; NFSI represents N-fluoro-N- (phenylsulfonyl) benzenesulfonamide; NCS represents 1-chloropyrrolidine-2, 5-dione; n-Bu₄NF represents tetrabutyl ammonium fluoride; iPrOH represents 2-propanol; mp represents melting point; LDA represents lithium diisopropylamide; LiHMDS represents lithium hexamethyldisilazide; xantphos stands for 4,5-bis diphenylphosphino-9, 9-dimethylxanthene; LiAlH₄Represents lithium aluminum hydride; pd (dba)₂Represents tris (dibenzylideneacetone) dipalladium; pd (dppf) Cl₂Represents [1,1' -bis (diphenylphosphino) ferrocene]Palladium dichloride; DIEA represents N, N-diisopropylethylamine; pd (PPh)₃) ₄Represents palladium tetratriphenylphosphine; IPA stands for isopropanol; DEA stands for diethylamine.

The compounds are used according to the conventional naming principle in the field

The software names, and the commercial compounds are under the supplier catalog name.

Technical effects

The compound has obvious THR alpha/beta activity and THR alpha selectivity, has no inhibiting effect or weak inhibiting effect on CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4-M, shows higher protein binding rate in human plasma, and has higher exposure and better oral bioavailability.

Detailed Description

The present invention is described in detail below by way of examples, but is not meant to be limited to any of the disadvantages of the present invention. Having described the invention in detail and having disclosed specific embodiments thereof, it will be apparent to one skilled in the art that various changes and modifications can be made therein without departing from the spirit and scope thereof.

Example 1

The synthetic route is as follows:

step 1: synthesis of Compound WX001-2

To a pre-dried reaction flask was added hydrazine hydrate (3.29g,65.73mmol,3.19mL) followed by AcOH (10mL), H₂O (30mL) was dissolved, and then WX001-1(10g,65.73mmol) was added, and the reaction was refluxed at 100 ℃ for 3 hours. After completion of the reaction, cooled to room temperature (25 ℃), filtered, and the filter cake was washed with water (20mL × 3) and then spun dry with a water pump at 45 ℃ and replaced once with toluene (50mL) to give compound WX 001-2.¹H NMR(400MHz,DMSO-d ₆)δ:11.27(br s,2H),2.34(br s,4H),1.64(br s,4H)。

Step 2: synthesis of Compound WX001-3

WX001-2(9.16g,55.12mmol) was added to a pre-dried reaction flask, then dissolved with phosphorus oxychloride (45mL), and the reaction was stirred up to 110 ℃ for 1 hour. After completion of the reaction, it was cooled to room temperature (25 ℃ C.), and then the reaction solution was slowly added to room temperature water (600mL), stirred for 30 minutes, filtered, and the filter cake was concentrated under reduced pressure to give compound WX 001-3.¹H NMR(400MHz,DMSO-d ₆)δ:2.68(s,4H),1.77(s,4H)。

And step 3: synthesis of Compound WX001-5

WX001-3(6g,29.55mmol), WX001-4(6.31g,35.46mmol) were charged into a reaction flask dried in advance, and then dissolved with N, N-dimethylacetamide (30mL), followed by addition of cesium carbonate (11.07g,33.98mmol), and the reaction was stirred at 65 ℃ for 16 hours. The reaction was allowed to cool to room temperature (20 deg.C), supplemented with WX001-4(1.05g,5.91mmol), and slowly raised to 65 deg.C with stirring for 4 hours. After completion of the reaction, the reaction mixture was cooled to room temperature (25 ℃ C.), ethyl acetate (30mL) was added to dilute the reaction mixture, the reaction mixture was passed through a funnel covered with celite, the cake was washed with ethyl acetate (3X 30mL), the filtrate was diluted with water (100mL), the liquid was separated, the aqueous phase was extracted with ethyl acetate (100mL X2), the organic phases were combined, the mixture was washed with saturated saline (200mL 4), dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the solvent was removed under reduced pressure to obtain a crude product, which was separated by an automatic column chromatography (eluent: petroleum ether: ethyl acetate 1:0 to 1:1) to obtain a combined eluentSubstance WX 001-5.¹H NMR(400MHz, DMSO-d ₆)δ6.70(s,2H),5.65(s,2H),2.71-2.65(m,4H),1.77-1.81(m,4H)。

And 4, step 4: synthesis of Compound WX001-6

WX001-5(5g,14.51mmol), benzoic anhydride (3.94g,17.41mmol,3.28mL) were added to a pre-dried reaction flask, then dissolved with acetic acid (50mL), and the reaction was stirred at 70 ℃ for 4.5 hours. After the reaction is completed, the reaction solution of WX001-6 is directly used for the next reaction. MS-ESI M/z 447.9[ M + H ]] ⁺,449.9[M+H+2] ⁺。

And 5: synthesis of Compound WX001-7

To a solution of WX001-6(6.51g,14.51mmol) and acetic acid (50mL) was added sodium acetate (2.38g,29.02mmol), and the reaction was stirred at 110 ℃ for 15 hours. After completion of the reaction, it was cooled to room temperature (25 ℃), water (80mL) was added, solids were found to precipitate, which were not suitable for filtration, ethyl acetate (100mL) was added for dilution, the aqueous phase was extracted with ethyl acetate (100mL _ 3), the organic phases were combined, most of the solvent was removed under reduced pressure, then acetic acid was quenched with sodium bicarbonate solids until pH reached around 7, water (200mL) was added for dilution, followed by extraction with ethyl acetate (100mL _ 3), the organic phases were combined and washed with saturated brine (100mL _ 2), dried over anhydrous sodium sulfate. After the drying agent was removed by filtration, the solvent was removed under reduced pressure. The crude product was separated by high performance liquid chromatography (water (0.1% TFA) -acetonitrile), neutralized with sodium bicarbonate solid to pH about 7, diluted with ethyl acetate (100mL), the organic phase was separated, the aqueous phase was extracted with ethyl acetate (100mL x 2), the organic phases were combined and washed with saturated brine (100mL x 2), dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the solvent was removed under reduced pressure to give compound WX 001-7. MS-ESI M/z 430.1[ M + H ]] ⁺,432.0[M+H+2] ⁺。

Step 6: synthesis of Compound WX001-8

A pre-dried reaction flask was charged with WX001-7(0.5g,1.16mmol) followed by THF (7.5mL), H₂O (7.5mL) was dissolved and potassium hydroxide (260.78mg,4.65mmol,4eq) was added and the reaction stirred for 18 h slowly increasing to 85 ℃. Cooling the reaction system to room temperature(20 ℃ C.), potassium hydroxide (130.39mg,2.32mmol) was added and the reaction stirred slowly to 85 ℃ for 6 hours. After completion of the reaction, the reaction mixture was diluted with water (10mL) and ethyl acetate (20mL), separated, the aqueous phase was extracted with ethyl acetate (10mL × 2), the organic phases were combined, washed with saturated brine (10mL × 2), dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the solvent was removed under reduced pressure to obtain a crude product, which was purified by thin layer chromatography on a silica gel plate (developing solvent: petroleum ether: ethyl acetate ═ 1:1) to obtain a crude compound WX 001-8. MS-ESI M/z 326.1[ M + H ]] ⁺,328.1[M+H+2] ⁺。

And 7: synthesis of Compound WX001-10

WX001-8(0.09g, 275.92. mu. mol) was added to a pre-dried reaction flask, followed by dissolution with AcOH (2mL), followed by addition of hydrochloric acid (83.68mg, 849.22. mu. mol, 82.04. mu.L, 37% purity), cooling the reaction to 5 ℃ and addition of sodium nitrite (20.94mg, 303.52. mu. mol) in H₂O (1mL) solution was stirred at 5 ℃ for 0.5 hour, and after completion of the reaction of the starting materials was detected, WX001-9(47.39mg, 303.52. mu. mol) was added and stirred at 5 ℃ for 15 minutes. After completion of the reaction, a solution of sodium acetate (0.1g) in water (0.5mL) was added to the reaction system, and a solid was found to precipitate, followed by filtration, neutralization of the filtrate with a saturated sodium bicarbonate solution to a pH of about 7, dilution with dichloromethane (20mL), liquid separation, extraction of the aqueous phase with dichloromethane (20mL × 2), combination of the organic phases, drying with anhydrous sodium sulfate, filtration, removal of the drying agent by filtration, and removal of the solvent under reduced pressure to obtain compound WX 001-10.

And 8: synthesis of Compound WX001

WX001-10(0.066g, 133.79. mu. mol) was added to a pre-dried reaction flask, dissolved in N, N-dimethylacetamide (1mL), followed by potassium acetate (19.70mg, 200.69. mu. mol), slowly raised to 115 ℃ and stirred for 2 hours. After completion of the reaction, the reaction solution was filtered and separated by HPLC (column: Waters Xbridge 150 mM. times.25 mM,5 μm; mobile phase: [ water (10mM NH. sub.H.) ]₄HCO ₃) -acetonitrile](ii) a Acetonitrile percent of 10 percent to 40 percent, and 7min) to obtain the target compound WX 001.¹H NMR (400MHz, deuterated methanol) delta 7.74(s,2H),2.77-2.79(m,2H),2.57-2.59(m,2H),1.87-1.88(m,4H)。。MS-ESI m/z:447.0[M+H] ⁺,449.0[M+H+2] ⁺。

Example 2

The synthetic route is as follows:

step 1: synthesis of Compound WX002-3

WX002-1(33g,221.51mmol,1eq) and acetonitrile (32mL) were added in three portions to three reaction bottles dried in advance, silver nitrate (18.81g,110.75mmol,0.5eq) and WX002-2(21.47g,243.66mmol,22.60mL,1.1eq) were added, a mixed solution of sulfolane (197.51g,1.64mol,156.75mL,7.42eq) and water (363mL) was added at 25 ℃, the temperature was raised to 55 ℃, concentrated sulfuric acid (64.73g,659.96mmol,35.18mL,2.98eq) and water (111mL) were added, ammonium persulfate (50.55g,221.51mmol,48.14mL,1eq) and water (111mL) were slowly added dropwise and the mixed solution was stirred at 70 ℃ for 0.5 hour and at 25 ℃ for 23.5 hours. After the reaction was completed, ammonia water (500mL) was slowly added dropwise to the reaction solution at 0 ℃, pH was adjusted to 8, water (1500mL) was added to the reaction solution, the mixture was filtered through celite, the filter cake was washed with ethyl acetate (1000mL), the filtrate was collected, the filtrate was extracted with ethyl acetate (100mL x3), the organic phases were combined, washed with saturated brine (1000mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was chromatographed on silica gel (petroleum ether: ethyl acetate: 20:1 to 10:1 to 5:1 to 3:1) to give compound WX 002-3.¹H NMR (400MHz, deuterated chloroform) δ ppm 1.31-1.32 (d, J ═ 6.8Hz,6H)3.22-3.32(m,1H)7.38(s, 1H). MS-ESI M/z 191.1[ M + H ]] ⁺。

Step 2: synthesis of Compound WX002-5

WX002-4(60g,248.04mmol,1eq) was dissolved in N, N-dimethylacetamide (90mL) followed by additionPotassium tert-butoxide (33.40g,297.65mmol,1.2eq) was charged and heated to 100 ℃ with stirring for 1 hour, then a solution of WX002-3(49.76g,260.44mmol,1.05eq) in N, N-dimethylacetamide (30mL) was added and heated to 130 ℃ for 70 hours. After completion of the reaction, the reaction was allowed to cool to room temperature, then poured into water (500mL), extracted with methyl tert-butyl ether (500mL x3), all organic phases were combined, washed with saturated sodium chloride solution (500mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate 50:1 to 10:1) and then separated by high performance liquid chromatography (column: Phenomenex luna c 18250 mm × 100mm 10 μm; mobile phase: [ water (0.1% TFA) -acetonitrile](ii) a Acetonitrile accounting for 70 percent to 95 percent for 20min) to obtain a compound WX 002-5. MS-ESI M/z:397.0[ M + H ]] ⁺。

And step 3: synthesis of Compound WX002-6

WX002-5(8g,20.18mmol,1eq) and glacial acetic acid (240mL) were added to a pre-dried reaction flask, sodium acetate (3.31g,40.35mmol,2eq) was added, and nitrogen was replaced three times and the mixture was stirred at 110 ℃ for 12 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure, poured into 80mL of water, the pH of the solution was adjusted to 7 to 8 with solid sodium bicarbonate, and then a mixed solution of ethyl acetate and tetrahydrofuran was added for extraction (ethyl acetate: tetrahydrofuran ═ 3:1, 200mL × 3), all organic phases were combined and washed with saturated brine (50mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound WX 002-6. MS-ESI M/z 378.9[ M + H ]] ⁺。

And 4, step 4: synthesis of Compound WX002-7

WX002-6(7.6g,20.10mmol,1eq), potassium carbonate (8.34g,60.31mmol,3eq) and N, N-dimethylformamide (60mL) were charged into a pre-dried reaction flask, and p-methoxybenzyl chloride (4.72g,30.15mmol,4.11mL,1.5eq) was added last, the nitrogen was replaced three times, and the mixture was stirred at 25 ℃ for 16 hours. After the reaction is completed. Water (50mL) was added to the reaction solution, the aqueous phase (30mL x3) was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine (40mL x 2), the organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate: 20:1 to 10:1 to 5:1 to 3)1) obtaining the compound WX 002-7.¹H NMR (400MHz, deuterated chloroform) δ 1.23-1.25(d, J ═ 6.8Hz,6H)3.17-3.27(m,1H)3.79(s,3H)4.95(s,2H)6.78-6.80(d, J ═ 8.8Hz,2H)7.01(s,1H)7.17-7.19(d, J ═ 8.8Hz,2H)7.55(s, 2H). MS-ESI M/z 499.1[ M + H ]] ⁺。

And 5: synthesis of Compound WX002-9

WX002-7(100mg, 197.88. mu. mol,1eq) and acetonitrile (71mL) were charged to a pre-dried reaction flask, tributyl (1-ethoxyethylene) tin (WX002-8, 10.29g,28.50mmol,9.62mL,2eq) was added, nitrogen was replaced three times, dichlorobis (triphenylphosphine) palladium (1.50g,2.14mmol,0.15eq) and cuprous iodide (271.42mg,1.43mmol,0.1eq) were added, nitrogen was replaced again three times, and stirring was continued at 90 ℃ for 16 hours. After the reaction is completed. Directly cooling the reaction liquid to obtain a compound WX 002-9. MS-ESI M/z 489.2[ M + H ]] ⁺。

Step 6: synthesis of Compound WX002-10

Hydrochloric acid (1M,42.73mL,3eq) was added to a reaction solution of WX002-9(6.97g,14.24mmol,1eq) and stirred at 25 ℃ for 4 hours. After completion of the reaction, the reaction solution was adjusted to pH 7-8 with solid sodium bicarbonate, then extracted with ethyl acetate (200mL × 3), all organic phases were combined, washed with saturated sodium chloride solution (200mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to give crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate: 1:0 to 20:1 to 10:1 to 5:1) to give compound WX 002-10.¹H NMR (400MHz deuterated chloroform) δ 1.24-1.26(d, J ═ 6.8Hz,6H)2.66(s,3H)3.20-3.27(m,1H)3.78(s,3H)4.93(s,2H)6.75-6.77(d, J ═ 8.8Hz,2H)7.04(s,1H)7.14-7.16(d, J ═ 8.8Hz,2H)7.97(s, 2H). MS-ESI M/z 461.1[ M + H ]] ⁺。

And 7: synthesis of Compound WX002-11

WX002-10(5.5g,11.92mmol,1eq) and pyridine (55mL) were charged into a pre-dried reaction flask, selenium dioxide (2.65g,23.84mmol,2.59mL,2eq) was added, nitrogen was replaced three times, and the mixture was stirred at 100 ℃ for 16 hours. After the reaction is finished, adjusting the pH of the reaction solution to 2-3 by 2N hydrochloric acid solution (about 500mL), extracting the aqueous phase (400mL x3) by ethyl acetate, combining the organic phases, and saturatingWashed with brine (300mL × 2), the organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compound WX 002-11. MS-ESI M/z 491.2[ M + H ]] ⁺。

And 8: synthesis of Compound WX002-12

WX002-11(6g,12.21mmol,1eq) and methanol (60mL) were added to a reaction flask which had been dried in advance, sulfoxide chloride (2.91g,24.42mmol,1.77mL,2eq) was slowly added dropwise at 0 ℃ while stirring at 25 ℃ for 0.5 hour, and the mixture was heated to 70 ℃ and stirred for 15.5 hours. After the reaction is finished, directly concentrating the reaction solution under reduced pressure, adjusting the pH of the concentrate to 7-8 by using a saturated sodium bicarbonate solution (50mL), extracting an aqueous phase (50mL x3) by using ethyl acetate, combining organic phases, washing the organic phases by using saturated saline solution (30mL x 2), collecting the organic phases, drying the organic phases by using anhydrous sodium sulfate, filtering, and concentrating under reduced pressure to obtain a crude product. The crude product was purified by thin layer chromatography (petroleum ether: ethyl acetate: 20:1 to 10:1 to 5:1 to 2:1) to give compound WX 002-12.¹H NMR (400MHz, deuterated chloroform) δ 1.24-1.26(d, J ═ 6.8Hz,6H)3.18-3.28(m,1H)3.78(s,3H)4.04(s,3H)4.93(s,2H)6.75-6.78(m,2H)7.04(m,1H)7.13-7.16(m,2H)8.13(s, 2H). MS-ESI M/z 505.1[ M + H ]] ⁺。

And step 9: synthesis of Compound WX002-13

WX002-12(100mg, 197.88. mu. mol,1eq) and methanol (15mL) were added to a pre-dried reaction flask, placed at-78 ℃ and ammonia (33.70mg,1.98mmol,10eq) was bubbled through for 10 minutes, and the temperature was raised to 25 ℃ and stirring was continued for 15 hours and 50 minutes. After the reaction is finished, the reaction solution is decompressed and concentrated to obtain a compound WX 002-13.¹H NMR (400MHz, deuterated chloroform) δ 1.24-1.25(d, J ═ 6.8Hz,6H)3.24-3.27(d, J ═ 6.72Hz,1H)3.78(s,3H)4.93(s,2H)6.76-6.79(m,2H)7.01(s,1H)7.04-7.16(m,2H)8.46(s, 2H). MS-ESI M/z 490.3[ M + H ]] ⁺。

Step 10: synthesis of Compound WX002-14

WX002-13(2.3g,4.69mmol,1eq) and dichloroethane (50mL) were added to a pre-dried reaction flask, nitrogen was replaced three times, oxalyl chloride (773.98mg,6.10mmol, 533.78. mu.L, 1.3eq) was slowly added dropwise at 0 ℃, stirring was carried out at 25 ℃ for 1 hour, and stirring was continued at 70 ℃ for 15 hours. After the reaction is completed, the reaction solution is addedDirectly concentrating under reduced pressure to obtain the compound WX 002-14. MS-ESI M/z 548.1[ M + MeOH + H ]] ⁺。

Step 11: synthesis of Compound WX002-15

WX002-14(2.4g,4.65mmol,1eq) and dichloromethane (50mL) were added to a pre-dried reaction flask, ethanol (78.80g,1.71mol,100mL,367.99eq) was added, and the mixture was stirred at 25 ℃ for 16 hours. After the reaction is finished, directly carrying out reduced pressure concentration on the reaction liquid to obtain a crude product. The crude product was purified by thin layer chromatography silica gel plate (petroleum ether: ethyl acetate 1:1) to give compound WX 002-15.¹H NMR (400MHz, deuterated chloroform) δ 1.24-1.26(d, J ═ 6.8Hz,6H)1.36(t, J ═ 7.13Hz,3H)3.21-2.27(m,1H)3.78(s,3H)4.30-4.35(m,2H)4.93(s,2H)6.77(d, J ═ 8.63Hz,2H)7.04(s,1H)7.13-7.15(d, J ═ 8.63Hz,2H)8.29(s, 2H). MS-ESI M/z 562.1[ M + H ]] ⁺。

Step 12: synthesis of Compound WX002-16

WX002-15(750mg,1.33mmol,1eq) and glacial acetic acid (75mL) were added to a pre-dried reaction flask, hydroxylamine hydrochloride (111.20mg,1.60mmol,1.2eq) was added, nitrogen was replaced three times, and the mixture was stirred at 120 ℃ for 16 hours. Hydroxylamine hydrochloride (185.34mg,2.67mmol,2eq) was added and the mixture was heated to 125 ℃ and stirred for 16 hours. After the reaction is finished, directly decompressing and concentrating the reaction liquid to obtain a crude product. The crude product was purified by thin layer chromatography silica gel plate (petroleum ether: ethyl acetate ═ 0:1) to afford compound WX 002-16. MS-ESI M/z 531.1[ M + H ]] ⁺。

Step 13: synthesis of Compound WX002

WX002-16(80mg, 150.56. mu. mol,1eq) and acetonitrile (8mL) were added to a pre-dried reaction flask, nitrogen was replaced three times, the temperature was reduced to 0 ℃, ceric ammonium nitrate (247.62mg, 451.68. mu. mol, 225.11. mu.L, 3eq) and water (3.2mL) were added slowly, the temperature was raised slowly to room temperature of 25 ℃ and stirring was continued for 5 hours. After the reaction was completed, the reaction solution was extracted with ethyl acetate (15mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by thin layer chromatography on silica gel (dichloromethane: methanol 5:1) to give crude product, which was separated by high performance liquid chromatography (column: Waters Xbridge BEH C18100 × 25mm 5 μm; mobile phase)Water (10mM NH)₄HCO ₃) -acetonitrile](ii) a Acetonitrile accounting for 20 percent to 50 percent for 8min) to obtain a compound WX 002.¹H NMR (400MHz, deuterated methanol) δ 1.29-1.28(d, J ═ 6.88Hz,6H)3.20-3.13(m,1H)7.37(s,1H)8.01(s, 2H). MS-ESI M/z 411.0[ M + H ]] ⁺。

Example 3

The synthetic route is as follows:

step 1: synthesis of Compound WX003-3

To a reaction flask were added compound WX003-1(2g, 4.63mmol), compound WX003-2(1.43g, 9.25mmol), potassium carbonate (1.92g, 13.88mmol), water (15mL) and 1, 4-dioxane (45mL), and after purging with nitrogen, stirring was carried out for 20 minutes, followed by addition of Pd (dppf) Cl₂(338.54mg, 462.68. mu. mol), and the mixture was reacted at 80 ℃ for 6 hours. After completion of the reaction, the reaction mixture was diluted with water (10mL), extracted with dichloromethane (10mL × 3), and the organic phase was washed with saturated brine (10mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified using silica gel plates (petroleum ether: ethyl acetate 4:1) by thin layer chromatography. The compound WX003-3 is obtained.

Step 2: synthesis of Compound WX003-4

The compound WX003-3(1.0g, 2.64mmol), t-butanol (20mL), water (20mL) and acetonitrile (20mL) were charged into a reaction flask, and after purging nitrogen and cooling to 0 ℃ C, a mixture of sodium periodate (1.13g, 5.27mmol) and osmium tetroxide (67.01mg, 263.57. mu. mol) was added thereto and reacted at 25 ℃ for 3 hours. After completion of the reaction, the reaction mixture was filtered through celite, washed with ethyl acetate (20mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. Without further purification, crude compound WX003-4 is obtained,used directly in the next step.¹H NMR(400MHz,DMSO-d ₆)δppm 9.73(s,1H),7.27-7.30(m,7H),6.91-6.95(m,2H),5.32(s,2H),5.14(s,2H),4.60(s,2H),3.74(s,3H)。

And step 3: synthesis of Compound WX003-5

The compound WX003-4(500mg, 1.31mmol) and dichloromethane (20mL) were charged into a reaction flask, the temperature was lowered to 0 ℃ after purging nitrogen, and then diethylaminosulfur trifluoride (422.65mg, 2.62mmol) was added, and the mixture was reacted at 25 ℃ for 5 hours under nitrogen protection. After completion of the reaction, the reaction mixture was poured into ice water (10mL), dichloromethane (10mL × 3) was extracted, the organic phase was washed with saturated brine (10mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified using silica gel plates (petroleum ether: ethyl acetate ═ 3:1) by thin layer chromatography to afford compound WX 003-5.¹H NMR (400MHz, deuterated chloroform) delta 7.27-7.32(m,7H),6.69-6.97(m,3H),5.31(s, 2H),5.04(s,2H),4.61(s,2H),3.74(s, 3H).

And 4, step 4: synthesis of Compound WX003-6

A reaction flask was charged with WX003-5(80mg, 198.32. mu. mol) and acetonitrile (1.4mL), the temperature was lowered to 0 ℃ after purging nitrogen, a solution of ceric ammonium nitrate (326.18mg, 594.97. mu. mol) in water (0.6mL) was added, and the mixture was reacted at 27 ℃ for 16 hours. After the reaction is finished, directly spin-drying the reaction solution. The crude product was purified using silica gel plates (petroleum ether: ethyl acetate ═ 2:1) by thin layer chromatography to afford compound WX 003-6.¹H NMR (400MHz, deuterated chloroform) δ 13.00(s,1H),7.24-7.40(m,5H),6.61-7.00(t, J ═ 52.8Hz,1H),5.28(s,2H),4.61(s, 2H).

And 5: synthesis of Compound WX003-7

Compound WX003-6(53.14mg, 114.75. mu. mol), Compound WX004-9(25mg, 88.27. mu. mol), pyridine (17.45mg, 220.67. mu. mol) and 1, 2-dichloroethane (1mL) were added to a reaction flask, oxygen was purged and then copper acetate (24.05mg, 132.40. mu. mol) was added, and the mixture was reacted at 40 ℃ for 16 hours. And after the reaction is finished, spin-drying the reaction solution. The crude product was purified using silica gel plates (petroleum ether: ethyl acetate ═ 2:1) by thin layer chromatography to afford compound WX 003-7. MS-ESI M/z 700.1[ M + H ]] ⁺，702.1[M+H+2] ⁺。

Step 6: synthesis of Compound WX003-8

A reaction flask was charged with WX003-7(30mg, 42.83. mu. mol) and acetonitrile (0.5mL), the temperature was lowered to 0 ℃ after purging nitrogen, a solution of ammonium ceric nitrate (70.43mg, 128.49. mu. mol) in water (0.25mL) was slowly added dropwise, and the mixture was reacted at 25 ℃ for 16 hours. After completion of the reaction, the reaction mixture was diluted with water (5mL), extracted with dichloromethane (10mL × 3), and the organic phase was washed with saturated brine (2mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified using silica gel plates (petroleum ether: ethyl acetate ═ 1:1) by thin layer chromatography to give compound WX 003-8. MS-ESI M/z 580.1[ M + H ]] ⁺，582.1[M+H+2] ⁺。

And 7: synthesis of Compound WX003

Compound WX003-8(20mg, 34.46. mu. mol) and methylene chloride (0.5mL) were charged into a reaction flask, and after purging nitrogen and cooling to 0 ℃ C, boron tribromide (17.27mg, 68.92. mu. mol) was added, and the mixture was reacted at 0 ℃ for 20 minutes. After completion of the reaction, the reaction mixture was diluted with water (2mL), filtered, extracted with dichloromethane (10mL × 3), and the organic phase was washed with saturated brine (2mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was subjected to preparative high performance liquid chromatography (column: Luna C18100 mm × 30mm,5 μm; mobile phase: [ water (0.225% FA) -acetonitrile](ii) a Acetonitrile percent of 40-90 percent and 12 min). The target compound WX003 is obtained.¹H NMR (400MHz, deuterated dimethyl sulfoxide) δ 12.84(s,1H),12.21(s,1H),7.81(s,2H),7.44(s,1H),6.74-7.05(t, J ═ 52Hz,1H),3.02-3.08(m,1H),1.15-1.25(d, J ═ 6.8Hz, 6H). MS-ESI M/z 460.1[ M + H ]] ⁺，462.0[M+H+2] ⁺。

Example 4

Synthesis of intermediate WX004-9

The synthetic route is as follows:

step 1: synthesis of Compound WX004-12

In a pre-dried reaction flask were added bis-pinacolato borate (6.35g,24.99mmol,1.5eq), WX002-7(8.3g,16.66mmol,1eq) and dioxane (83mL), potassium acetate (3.27g,33.32mmol,2eq) was added, nitrogen was replaced three times, pd (dppf) Cl was added₂(1.22g,1.67mmol,0.1eq), the nitrogen was again replaced by three times, and the mixture was stirred at 70 ℃ for 16 hours. After the reaction is finished, filtering the reaction solution by using diatomite, and concentrating the filtrate under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate: 20:1 to 10:1 to 5:1 to 3:1) to afford compound WX 004-12.¹H NMR (400MHz, deuterated chloroform) δ 7.81(s,2H),7.16-7.23(m,2H),7.02(d, J ═ 0.86Hz,1H),6.75-6.81(m,2H),4.92(s,2H),3.78(s,3H),3.16-3.26(m,1H),1.38(s,12H),1.22-1.24(d, J ═ 6.97Hz, 6H). MS-ESI M/z 545.3[ M + H ]] ⁺。

Step 2: synthesis of Compound WX004-9

In a pre-dried reaction flask, WX004-12(7.6g,13.94mmol,1eq) and acetone (152mL), H₂O (76mL), sodium periodate (2.98g,13.94mmol, 772.35. mu.L, 1eq) and ammonium acetate (7.74g,100.36mmol,7.2eq) were added thereto, the nitrogen gas was replaced three times, and the mixture was stirred at 35 ℃ for 16 hours. After completion of the reaction, saturated sodium thiosulfate solution (80mL) was added, the pH was adjusted to 5 with 1M hydrochloric acid, and the crude product was filtered. And (3) freeze-drying the reaction crude product to obtain a compound WX 004-9.¹H NMR (400MHz, deuterated chloroform) delta 7.81(s,2H),7.16-7.22(m,2H),7.04(s,1H),6.76-6.80(m,2H),4.93(s,2H),3.78(s,3H),3.18-3.27(m,1H),1.22-1.25(m, 6H). MS-ESI M/z 463.1[ M + H ]] ⁺。

The synthetic route is as follows:

step 1: synthesis of Compound WX004-3

WX004-1(10g,64.08mmol,6.54mL,1eq), WX004-2(5.84g,64.08mmol,1eq) and ethanol (140mL) were charged into a pre-dried reaction flask, reacted at 100 ℃ for 16 hours, water (140mL) was added to the reaction mixture, sodium hydroxide (5.13g,128.16mmol,2eq) was added, and reacted at 100 ℃ for 1 hour. After the reaction is finished, adding ethyl acetate (100mL x 2) into the reaction solution for extraction, cooling the water phase to 0 ℃, adding glacial acetic acid to adjust the pH value to 4, extracting with ethyl acetate (100mL x3), combining the organic phases, washing with saturated saline (100mL x 2), separating the organic phase, drying with anhydrous sodium sulfate, filtering, and spin-drying to obtain a crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate: 20:1 to 10:1 to 5:1 to 2:1) to afford compound WX 004-3.¹H NMR(400MHz,DMSO-d ₆)δ14.01(s,1H)，13.66(s,1H)。MS-ESI m/z:197.9[M+H] ⁺。

Step 2: synthesis of Compound WX004-4

WX004-3(5.6g,28.41mmol,1eq) and sodium hydroxide (1M,85.22mL,3eq) were charged into a reaction flask which had been dried in advance, nitrogen gas was replaced three times, and a hydrogen peroxide solution (16.10g,142.03mmol,13.65mL, 30% purity, 5eq) was slowly added dropwise thereto at 0 ℃ to react at 25 ℃ for 2 hours. After completion of the reaction, the reaction mixture was adjusted to pH 3 with 1M hydrochloric acid solution (50mL), and the aqueous phase (150mL × 3) was extracted (ethyl acetate: tetrahydrofuran ═ 3:1), the organic phases were combined, washed with saturated brine (150mL × 2), the organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound WX 004-4.¹H NMR(400MHz,DMSO-d ₆)δ12.99(s,1H)，12.45(s,1H)。MS-ESI m/z:182.0[M+H] ⁺。

And step 3: synthesis of Compound WX004-6

WX004-4(5.6g,30.93mmol,1eq), WX004-5(6.29g,30.93mmol,7.64mL,1eq) and acetonitrile (120mL) were charged into a pre-dried reaction flask, the nitrogen was replaced three times, and the mixture was stirred at 80 ℃ for 2 hours. P-methoxybenzyl chloride (5.81g,37.11mmol,5.05mL,1.2eq) and sodium iodide (463.57mg,3.09mmol,0.1eq) were added, nitrogen was replaced three times, and stirring was continued at 80 ℃ for 14 hours. After the reaction is finished, directly carrying out reduced pressure concentration on the reaction liquid to obtain a crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate: 20:1 to 10:1 to 5:1 to 2:1) to afford compound WX 004-6.¹H NMR(400MHz,DMSO-d ₆)δ12.70(br,1H)，7.27-7.29(d,J＝8.68Hz,2H),6.91-6.93(d,J＝8.80Hz,2H),5.01(s,2H),3.74(s,3H)。MS-ESI m/z:300.2[M+H] ⁺。

And 4, step 4: synthesis of Compound WX004-7

WX004-6(4.1g,13.61mmol,1eq) and N, N-dimethylformamide (40mL) were charged to a pre-dried reaction flask, nitrogen was purged, then sodium hydrogen (816.60mg,20.42mmol, 60% purity, 1.5eq) was added slowly at 0 deg.C, stirring was carried out for 0.5 hr at 0 deg.C, benzyl chloromethyl ether (2.56g,16.33mmol,2.26mL,1.2eq) was added, and stirring was carried out for 3 hr at 25 deg.C. The reaction was quenched with water (40mL), the aqueous phase (40mL x3) was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine (40mL x 2), the organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate: 20:1 to 10:1 to 5:1) to afford compound WX 004-7.¹H NMR(400MHz,DMSO-d ₆)δ7.28-7.32(m,7H),6.90-6.95(m,2H),5.31(s,2H),5.07(s,2H),4.62(s,2H),3.74(s,3H)。MS-ESI m/z:439.2[M+18] ⁺。

And 5: synthesis of Compound WX004-8

WX004-7(3.54g,8.40mmol,1eq) and acetonitrile (54mL) were charged into a reaction flask which had been dried in advance, and a mixed solution of ammonium ceric nitrate (13.82g,25.20mmol,12.56mL,3eq) and water (27mL) was slowly added dropwise thereto at 0 ℃ while replacing nitrogen three times, and the mixture was stirred at 25 ℃ for 16 hours. After completion of the reaction, the reaction mixture was quenched with water (50mL) and acetic acidThe aqueous phase was extracted with ethyl ester (50mL x3), the organic phases were combined, washed with saturated brine (50mL x 2), the organic phase was collected, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate: 20:1 to 10:1 to 5:1 to 2:1) to afford compound WX 004-8.¹H NMR (400MHz, deuterated chloroform) delta 9.89(s,1H), 7.29-7.39(M,5H), 5.49(s,2H), 4.74(s,2H) MS-ESI M/z 319.1[ M +18 ]] ⁺。

Step 6: synthesis of Compound WX004-10

In a pre-dried reaction flask, WX004-8(325.21mg,1.08mmol,1eq), WX004-9(500mg,1.08mmol,1eq) and 1, 2-dichloroethane (50mL), pyridine (213.50mg,2.70mmol, 217.86. mu.L, 2.5eq) and copper acetate (294.15mg,1.62mmol,1.5eq) were added and stirred at 40 ℃ for 12 hours under oxygen protection. After the reaction is finished, filtering the reaction solution by using diatomite, and concentrating the filtrate under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate: 20: 1: 10: 1: 2:1) to afford compound WX 004-10.¹H NMR (400MHz, deuterated chloroform) δ ppm 1.24-1.26(d, J ═ 6.85Hz,6H)3.19-3.29(m,1H)3.77(s,3H)4.79(s,2H)4.97(s,2H)5.61(s,2H) 6.78-6.80(d, J ═ 8.56Hz,2H)7.04(s,1H) 7.18-7.20 (d, J ═ 8.44Hz,2H)7.31-7.40(m,5H)7.58(s, 2H). MS-ESI M/z 718.2[ M + H ]] ⁺。

And 7: synthesis of Compound WX004-11

WX004-10(205mg, 285.31. mu. mol,1eq) and acetonitrile (20mL) were added to a pre-dried reaction flask, and a mixed solution of ceric amine nitrate (469.25mg, 855.94. mu. mol, 426.59. mu.L, 3eq) and water (8mL) was slowly added thereto at 0 ℃ and stirred at 25 ℃ for 16 hours. After completion of the reaction, the reaction mixture was quenched with water (10mL), the aqueous phase (20mL × 3) was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine (20mL × 2), the organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a crude product. The crude product was purified by thin layer chromatography silica gel plate (petroleum ether: ethyl acetate 1:1) to afford compound WX 004-11.¹H NMR (400MHz, deuterated chloroform) δ ppm 1.29-1.31(d, J ═ 6.85Hz,6H), 3.20-3.27(m,1H),4.78(s,2H),5.59(s,2H),7.15(s,1H),7.31-7.38(m,5H),7.55(s,2H),9.64(br s,1H). MS-ESI m-z:598.1[M+H] ⁺。

And 8: synthesis of Compound WX004

WX004-11(140mg, 233.97. mu. mol,1eq) and dichloromethane (14mL) were added to a reaction flask which had been dried in advance, and boron tribromide (117.23mg, 467.95. mu. mol, 45.09. mu.L, 2eq) was slowly added dropwise thereto at 0 ℃ and stirred at 0 ℃ for 15 minutes. After the reaction was completed, the reaction solution was quenched with saturated sodium bicarbonate solution (2mL), the aqueous phase (5mL × 3) was extracted with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product. The crude product was separated by high performance liquid chromatography (column: Waters Xbridge BEH C18100 30mM 10 μm; mobile phase: [ water (10mM NH)₄HCO ₃) -acetonitrile](ii) a 30-70 percent of acetonitrile for 8min) to obtain the target compound WX 004.¹H NMR (400MHz, deuterated methanol) δ 1.28-1.30(d, J ═ 6.85Hz,6H)3.14-3019(m,1H)7.36(s,1H)7.75(s, 2H). MS-ESI M/z 478.0[ M + H ]] ⁺。

Example 5

The synthetic route is as follows:

step 1: synthesis of Compound WX005-2

WX002-4(6.92g,28.61mmol) was added to a pre-dried reaction flask, followed by dissolution with N, N-dimethylacetamide (31mL), followed by addition of potassium tert-butoxide (3.85g,34.33mmol), heating to 100 deg.C and stirring for 1 hour, followed by addition of a solution of WX001-3(6.1g,30.04mmol) and N, N-dimethylacetamide (15.5mL), heating to 130 deg.C and stirring for 64 hours. After completion of the reaction, the reaction system was cooled to room temperature (20 ℃ C.), diluted with water (150mL) and ethyl acetate (100mL), the organic phase was collected by liquid separation, the aqueous phase was extracted with ethyl acetate (100 mL. times.2), the organic phases were combined, and the mixture was usedThe extract was washed with saturated brine (200mL × 4), dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the filtrate was concentrated under reduced pressure to obtain a residue. Methyl tert-butyl ether (15mL) was added, stirred for 10min, filtered, the filter cake rinsed with methyl tert-butyl ether (10mL x3) and the filter cake was dried to give compound WX 005-2.¹H NMR(400MHz,DMSO-d ₆)δ7.99(s,2H),2.77-2.79(m,2H),2.71-2.68(m,2H),1.81-1.84(m,4H)。

Step 2: synthesis of Compound WX005-3

WX005-2(6.19g,15.15mmol) was added to a pre-dried reaction flask, dissolved in acetic acid (62mL), followed by sodium acetate (2.49g,30.31mmol), and the reaction was stirred slowly increasing to 110 ℃ for 16 h. After the reaction was completed, the reaction was allowed to cool to room temperature (25 ℃), most of the acetic acid was spun off with a water pump, then diluted with water (60mL), filtered, and the filter cake was rinsed with ethyl acetate (20mL × 3) and spin-dried to give compound WX 005-3.¹H NMR (400MHz, deuterated chloroform) delta 11.08(s,1H),7.53(s,2H),2.70-2.77(m,2H),2.60-2.64(m,2H),1.88-1.82(m, 4H).

And step 3: synthesis of Compound WX005-4

WX005-3(4.4g,11.28mmol) was charged into a reaction flask dried in advance, and then dissolved in N, N-dimethylformamide (44mL), potassium carbonate (4.68g,33.84mmol) and p-methoxybenzyl chloride (2.65g,16.92mmol,2.30mL) were added, and the reaction was stirred at room temperature (20 ℃ C.) for 16 hours. After completion of the reaction, water (100mL) was added for dilution, ethyl acetate (200mL) was added, the organic phase was collected by liquid separation, the aqueous phase was extracted with ethyl acetate (100mL × 3), the organic phases were combined, and then washed with saturated brine (100mL × 4), dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the solvent was removed under reduced pressure to obtain compound WX 005-4. 1H NMR (400MHz, DMSO-d)₆)δ7.96(s,2H),7.01-7.03(d,J＝8.4Hz,2H),6.77-6.80(d,J＝8.4Hz,2H),4.81(s,2H),3.71(s,3H),2.61(s,2H),2.44(s,2H),1.72(s,4H)。

And 4, step 4: synthesis of Compound WX005-5

A pre-dried reaction flask was charged with WX005-4(4.27g,8.37mmol), bis-pinacolato borate (3.19g,12.55mmol) and dioxane (64mL), potassium acetate (1.64g,16.74mmol,2eq) and replacedNitrogen three times, Pd (dppf) Cl₂(612.38mg, 836.92. mu. mol), the nitrogen was again replaced three times, and the mixture was stirred at 70 ℃ for 16 hours. After completion of the reaction, the reaction was passed through a funnel covered with celite, the filter cake was washed with dichloromethane (50mL × 3), the combined filtrates were concentrated under reduced pressure, the residue was dissolved again in dichloromethane (100mL), washed with water (50mL × 3) and saturated brine (50mL × 3) in this order, dried over anhydrous sodium sulfate, filtered to remove the drying agent, the solvent was removed under reduced pressure, and the obtained residue was separated by a chromatography column (eluent: petroleum ether: ethyl acetate ═ 1:0 to 20:80) to obtain compound WX 005-5.¹H NMR(400MHz,DMSO-d ₆)δ7.72(s,2H),6.97-7.00(d,J＝8.4Hz,2H),6.77-6.81(d,J＝8.4Hz,2H),4.79(s,2H),3.71(s,3H),2.63(s,2H),2.44(s,2H),1.73(s,4H),1.32(s,12H)。

And 5: synthesis of Compound WX005-6

WX005-5(1.2g,2.15mmol) was added to a pre-dried reaction flask, followed by dissolution in acetone (24mL), water (12mL), followed by addition of sodium periodate (460.58mg,2.15mmol, 119.32. mu.L), ammonium acetate (1.20g,15.50mmol), nitrogen substitution three times, and stirring at 35 ℃ for 16 hours. After the reaction, the reaction system was cooled to room temperature (20 ℃), 50mL of a saturated sodium sulfite solution was added, the mixture was stirred for 20 minutes, the mixture was checked to have not changed orange blue by a starch potassium iodide paper, the pH was neutralized to about 5 with 1N hydrochloric acid, a solid was found to precipitate, acetone was removed by vacuum concentration at 45 ℃ with a water pump, the solid was found to increase, the filtration was carried out, and the filter cake was dried by spin-drying to obtain compound WX 005-6.¹H NMR(400MHz,DMSO-d ₆)δ8.51(s,2H),7.90(s,2H),6.98-7.01(d,J＝8.8Hz,2H),6.75-6.81(d,J＝8.8Hz,2H),4.79(s,2H),3.70(s,3H),2.63(s,2H),2.44(s,2H),1.73-1.76(m,4H)。

Step 6: synthesis of Compound WX005-7

WX003-6(0.1g, 353.07. mu. mol), WX005-6(218.08mg, 458.99. mu. mol), followed by dissolution with dichloroethane (5mL) and addition of copper acetate (96.19mg, 529.60. mu. mol), pyridine (69.82mg, 882.67. mu. mol, 71.24. mu.L) were put into a reaction flask dried in advance, and after completion of the addition, the reaction was stirred at 40 ℃ under oxygen (15psi) for 15 hours. After the reaction is completed, water is added(15mL) and ethyl acetate (15mL), the organic phases were collected by separation, the aqueous phase was extracted with ethyl acetate (15mL × 2), the organic phases were combined, and then washed with saturated saline (10mL × 2), dried over anhydrous sodium sulfate, the drying agent was removed by filtration, the solvent was removed under reduced pressure, and the residue was purified by thin layer chromatography silica gel plate (petroleum ether: ethyl acetate ═ 1:1) to give compound WX 005-7. MS-ESI M/z 712.2[ M + H ]] ⁺,714.1[M+H+2] ⁺。

And 7: synthesis of Compound WX005-8

WX005-7(0.22g, 308.76. mu. mol) was added to a reaction flask dried in advance, followed by dissolution in acetonitrile (7mL), bringing the reaction system to 0 ℃ and then adding a solution of ceric ammonium nitrate (507.81mg, 926.28. mu. mol, 461.64. mu.L) in water (3mL), and the reaction was stirred while slowly rising to room temperature (25 ℃) for 2 hours. After completion of the reaction, water (10mL) and ethyl acetate (10mL) were added for dilution, the organic phase was separated, the aqueous phase was extracted with ethyl acetate (10mL × 2), the organic phases were combined and washed with saturated brine (10mL × 2), dried over anhydrous sodium sulfate, filtered to remove the drying agent, the solvent was removed under reduced pressure, and the resulting residue was purified by thin layer chromatography silica gel plate (petroleum ether: ethyl acetate ═ 1:1) to give compound WX 005-8. MS-ESI M/z 592.2[ M + H ]] ⁺,594.1[M+H+2] ⁺。

And 8: synthesis of Compound WX005

WX005-8(0.081g, 136.74. mu. mol) was charged into a reaction flask dried in advance, followed by dissolution in methylene chloride (2.4mL), the reaction system was cooled to 0 ℃, boron tribromide (68.51mg, 273.47. mu. mol, 26.35. mu.L) was added, and the reaction was stirred at 0 ℃ for 0.5 hour. After completion of the reaction, the reaction mixture was neutralized with a saturated sodium bicarbonate solution to about pH 7, diluted with ethyl acetate (5mL), the organic phase was separated, the aqueous phase was extracted with ethyl acetate (5 mL. times.2), the organic phases were combined and washed with a saturated saline solution (5 mL. times.2), dried over anhydrous sodium sulfate, filtered, the drying agent was removed by filtration, the solvent was removed under reduced pressure, and the residue was separated by high performance liquid chromatography (column: Phenomenex Luna C18200. times.40 mm. times.10 μm; mobile phase: [ water (0.2% FA) -acetonitrile](ii) a 30-70 percent of acetonitrile for 10min) to obtain the target compound WX 005.¹H NMR(400MHz,DMSO-d ₆)δ12.85(br s,1H),12.07(s,1H),7.80(s,2H),6.76-7.05(t,J＝52.4Hz,1H),2.66-2.67(m,2H),2.43-2.45(m,2H),1.75-1.78(m,4H)。MS-ESI m/z:472.1[M+H] ⁺,474.0[M+H+2] ⁺。

Example 6

The synthetic route is as follows:

step 1: synthesis of Compound WX006-3

Sodium hydride (22.18g,554.52mmol, 60% purity, 3.11eq) and tetrahydrofuran (200mL) were added to a reaction flask which had been dried in advance, nitrogen was replaced three times, WX006-2(40.15g,445.75mmol,37.53mL,2.5eq) was slowly added dropwise, the mixture was stirred at 80 ℃ for 1.5 hours, the temperature was reduced to 25 ℃, a mixed solution of WX006-1(20g,178.30mmol,21.65mL,1eq) and tetrahydrofuran (200mL) was slowly added dropwise, and stirring was continued at 70 ℃ for 4 hours. After the reaction is finished, slowly pouring the reaction bottle into an ice water bath (100mL), slowly adding a 2M hydrochloric acid solution (200mL) to adjust the pH value to 5-6, dropwise adding concentrated hydrochloric acid (10mL) to adjust the pH value to 1, extracting an aqueous phase (300mL x3) by methyl tert-butyl ether, combining organic phases, washing with saturated saline solution (300mL x 2), filtering, decompressing and concentrating to obtain a crude product. The crude product was purified by silica gel chromatography (petroleum ether: ethyl acetate: 1: 0: 80: 1: 50:1) to give compound WX 006-3.¹H NMR (400MHz, deuterated chloroform) δ ppm 1.18-1.20 (d, J ═ 7.13Hz,3H)1.38-1.45(m,1H)1.47-1.55(m,1H)1.65-1.74(m,1H)1.78-1.87(m,1H)2.19-2.26(m,2H)2.40-2.51(m,1H)3.76(s, 3H). MS-ESI M/z 171.1[ M + H ]] ⁺。

Step 2: synthesis of Compound WX006-4

Diethyl ether (260mL) was added to each of the two pre-dried reaction vials, sodium hydride (15.28g,381.89mmol, 60% purity, 5eq) was added slowly,a mixed solution of WX006-3(13g,76.38mmol,1eq) and diethyl ether (195mL) was slowly added dropwise thereto at 0 ℃ and nitrogen was replaced three times. After the reaction was completed, the reaction mixture was slowly added to stirred ice water (500mL), the aqueous phase (600mL × 3) was extracted with dichloromethane, the organic phases were combined, washed with saturated brine (500mL × 2), the organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel chromatography (petroleum ether: ethyl acetate: 1: 0: 50: 1: 20: 1: 10:1) to give compound WX 006-4.¹H NMR (400MHz, deuterated chloroform) δ ppm 1.20-1.22 (d, J ═ 7.00Hz,3H)1.46-1.54(m,1H)1.58-1.78(m,2H)1.91-2.01(m,1H)2.39-2.49(m,1H)2.50-2.67(m,2H)3.81(s, 3H). MS-ESI M/z 303.0[ M + H ]] ⁺。

And step 3: synthesis of Compound WX006-6

WX006-4(2g,6.62mmol,1eq) and dried N, N-dimethylformamide (40mL) were charged to a predried reaction flask, molecular sieve (50mg) was added, sodium formate (1.35g,19.85mmol,1.07mL,3eq) was added, anhydrous lithium chloride (841.52mg,19.85mmol, 406.53. mu.L, 3eq) was added and nitrogen was replaced three times, WX006-5(1.35g,13.23mmol,1.24mL,2eq) and diisopropylethylamine (1.71g,13.23mmol,2.31mL,2eq) were added dropwise in sequence at 0 deg.C, palladium acetate (148.55mg, 661.67. mu. mol,0.1eq) was added and nitrogen was replaced three times again and stirring was continued at 25 deg.C for 16 hours. After the reaction was completed, the reaction solution was quenched with 2M hydrochloric acid solution (20mL), the aqueous phase (30mL × 3) was extracted with methyl tert-butyl ether, the organic phases were combined, washed with saturated brine (30mL × 3), the organic phase was collected, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain compound WX 006-6. MS-ESI M/z 167.1[ M + H ]] ⁺。

And 4, step 4: synthesis of Compound WX006-7

A reaction flask dried in advance was charged with WX006-6(1.1g,6.62mmol,1eq) and glacial acetic acid (20mL), hydrazine hydrate (338.14mg,6.62mmol, 328.29. mu.L, 98% pure, 1eq) and water (2mL) were added, nitrogen was replaced three times, and the mixture was stirred at 100 ℃ for 16 hours. After the reaction is finished, directly decompressing and concentrating the reaction liquid to obtain a crude product. The crude product was slurried with methyl tert-butyl ether (10mL) to give compound WX 006-7.¹H NMR(400MHz,DMSO-d ₆)δppm 1.11–1.13(d,J＝7.00Hz,3H)1.58-1.72(m,4H)1.85-1.88(m,1H)2.14-2.25(m,1H)2.77–2.78(br d,J＝4.13Hz,1H)9.67(br s,2H)。MS-ESI m/z:181.1[M+H] ⁺。

And 5: synthesis of Compound WX006-8

WX006-7(1.13g,6.27mmol,1eq) and phosphorus oxychloride (26.64g,173.71mmol,16.14mL,27.70eq) were charged in a pre-dried reaction flask, replaced with nitrogen three times, and stirred at 100 ℃ for 3 hours. After the reaction is finished, directly concentrating the reaction solution under reduced pressure, adding ice water (50mL) into the concentrated solution, adjusting the pH value to 7-8 by using sodium bicarbonate solid, extracting an aqueous phase (50mL x3) by using ethyl acetate, combining organic phases, washing the organic phases by using saturated saline (50mL x 2), collecting the organic phases, drying the organic phases by using anhydrous sodium sulfate, filtering, and concentrating under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate: 1: 0: 20: 1: 10: 1: 5:1) to give compound WX 006-8.¹H NMR (400MHz, deuterated chloroform) δ ppm 1.31-1.32 (d, J ═ 7.00Hz,3H)1.72-1.81(m,1H)1.85-2.03(m,3H)2.53-2.66(m,1H)2.91(dd, J ═ 19.26,5.50Hz,1H)3.19(t, J ═ 6.38Hz, 1H). MS-ESI M/z 217.1[ M + H ]] ⁺。

Step 6: synthesis of Compound WX006-9

WX001-4(983.99mg,5.53mmol,1.5eq) and dimethyl sulfoxide (16mL) were charged into a reaction flask which had been dried in advance, WX006-8(800mg,3.69mmol,1eq) and potassium carbonate (2.04g,14.74mmol,4eq) were added, nitrogen was replaced three times, cuprous iodide (421.09mg,2.21mmol,0.6eq) was added, nitrogen was replaced three times again, and the mixture was stirred at 90 ℃ for 16 hours. After the reaction is finished, cooling the reaction solution to room temperature, adjusting the pH value to 7-8 by using 2N hydrochloric acid solution (20mL), extracting an aqueous phase (30mL x3) by using ethyl acetate, washing by using saturated saline, drying by using anhydrous sodium sulfate, filtering, and concentrating under reduced pressure to obtain a crude product. Purifying the crude product by chromatography (petroleum ether: ethyl acetate: 1: 0: 20: 1: 10: 1: 5:1) to obtain a solid product, pulping the solid product by methyl tert-butyl ether, collecting the filtrate, and concentrating the filtrate under reduced pressure to obtain the compound WX 006-9. MS-ESI M/z 358.0[ M + H ]] ⁺。

WX006-9 structural identification:

WX006-9(0.1g, 278.82. mu. mol,1eq) and methanol (1mL) were charged to a pre-dried reaction flask, wet palladium on carbon (0.1g, 139.41. mu. mol, 10% pure, 0.5eq) was added, the hydrogen was replaced three times, and the mixture was stirred under a pressure of hydrogen balloon (15psi) at 25 ℃ for 2 hours. After the reaction is finished. Filtering the reaction solution with diatomaceous earth, concentrating the filtrate under reduced pressure to obtain crude product, and separating the crude product by high performance liquid chromatography (chromatographic column: Waters Xbridge BEH C18100 mM 30mM 10 μm; mobile phase: [ water (10mM ammonium bicarbonate) -acetonitrile](ii) a 25 to 45 percent of acetonitrile for 10 minutes) to obtain WX 006-9A. NOE (400MHz, deuterated chloroform) detects that the hydrogen (1.34) of the methyl on the methyl six-membered ring is related to the hydrogen (8.69) on the pyridazine ring, and the structure of WX006-9 is correct.¹HNMR (400MHz, deuterated chloroform) δ 8.69(s,1H),6.99(d, J ═ 8.6Hz,2H),6.66-6.77(d, J ═ 8.6Hz,2H),3.61(br s,2H),2.91(d, J ═ 6.50Hz,1H),2.74(d, J ═ 4.00Hz,2H),1.91-2.01(m,2H),1.79-1.89(m,1H),1.61-1.65(m,1H),1.34(d, J ═ 7.13Hz, 3H).

And 7: synthesis of Compound WX006-10

WX006-9(500mg,1.39mmol,1eq) and glacial acetic acid (20mL) were added to a pre-dried reaction flask, and sodium acetate (228.73mg,2.79mmol,2eq) was added and stirred at 110 ℃ for 16 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure, poured into water (30mL), the pH of the solution was adjusted to 7 to 8 with solid sodium bicarbonate, and then extracted with (ethyl acetate: tetrahydrofuran ═ 3:1, 40mL × 3), all the organic phases were combined, and the organic phases were washed with a saturated sodium chloride solution (30mL × 2), dried over anhydrous sodium sulfate, poured over, and concentrated to give compound WX 006-10. MS-ESI M/z 382.1[ M + H ]] ⁺。

And 8: synthesis of Compound WX006-11

WX006-10(532mg,1.39mmol,1eq) and ethanol (11mL) were added to a pre-dried reaction flask, diluted hydrochloric acid (2M,10.44mL,15eq) was added, nitrogen was replaced three times, and the mixture was stirred at 90 ℃ for 16 hours. After the reaction is finished, directly decompressing and concentrating the reaction liquid,adjusting the pH of the concentrate to 7-8 by using solid sodium bicarbonate, extracting an aqueous phase (ethyl acetate: tetrahydrofuran: 3:1, 40 mL. times.3), combining organic phases, washing with saturated saline, filtering, and concentrating under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate: 1: 0: 20: 1: 10: 1: 5: 1: 2:1) to give compound WX 006-11.¹H NMR(400MHz,DMSO-d ₆)δppm 1.15–1.17(d,J＝7.00Hz,3H)1.63-1.82(m,4H)2.63-2.72(m,1H)2.83-2.92(m,1H)3.32(s,1H)5.60(s,2H)6.65(s,2H)11.95(s,1H)。MS-ESI m/z:340.1[M+H] ⁺。

And step 9: synthesis of Compound WX006-12

WX006-11(330mg, 970.01. mu. mol,1eq) and glacial acetic acid (2mL), concentrated hydrochloric acid (12M, 249.78. mu.L, 3.09eq) were charged in a pre-dried reaction flask, and a mixed solution of sodium nitrite (70.94mg,1.03mmol,1.06eq) and water (2mL) was added slowly at 0 ℃ and stirred at 0 ℃ for 0.5 hour. WX001-9(166.60mg,1.07mmol,1.1eq) was added at 0 ℃ and stirred at 0 ℃ for 1.5 hours. After the reaction was completed, the reaction solution was quenched with an aqueous solution (3mL) of sodium acetate (3eq) to produce a large amount of solid, and the compound WX006-12 was obtained by filtration. MS-ESI M/z 507.1[ M + H ]] ⁺。

Step 10: synthesis of Compound WX006

WX006-12(400mg, 788.45. mu. mol,1eq) and N, N-dimethylacetamide (5mL) were charged into a pre-dried reaction flask, potassium acetate (85.12mg, 867.29. mu. mol,1.1eq) was added thereto, nitrogen was replaced three times, and the mixture was stirred at 115 ℃ for 2 hours. After the reaction, water (30mL) was added to the reaction mixture, and a large amount of solid was formed, which was filtered to obtain the crude product. Separating the crude product by high performance liquid chromatography (chromatographic column: Phenomenex luna C)₁₈250 x 50mm x 10 μm; mobile phase [ water (0.05% HCl) -acetonitrile](ii) a Acetonitrile%: 45% -75% for 10min) to obtain the target compound WX 006.¹H NMR(400MHz,DMSO-d ₆)δppm 1.17–1.19(d,J＝6.8Hz,3H)1.66-1.83(m,4H)2.56–2.58(m,1H)2.68-2.79(m,1H)2.90(s,1H)7.78(s,2H)12.06(s,1H)13.27(s,1H)。MS-ESI m/z:461.0[M+H] ⁺。

Example 8

The synthetic route is as follows:

step 1: synthesis of Compound WX008-2

WX008-1(5g,26.05mmol,1eq), acetonitrile (50mL), N, O-bis trimethylsilyl acetamide (10.60g,52.09mmol,12.88mL,2eq) were added to a reaction flask, replaced with nitrogen three times, and then reacted at 80 ℃ for 2 hours, followed by addition of sodium iodide (3.90g,26.05mmol,1eq), p-methoxybenzyl chloride (4.89g,31.25mmol,4.26mL,1.2eq), and reacted at 80 ℃ for 14 hours, after completion of the reaction, filtration, spin-drying of the filtrate to obtain a crude product, which was purified by a silica gel column (eluent: petroleum ether: ethyl acetate ═ 0:1 to 3:1) to obtain compound WX 008-2. MS M/z 309.8[ M-H ]] ⁺,311.8[M-H+2] ⁺。

Step 2: synthesis of Compound WX008-3

WX008-2(3g,9.61mmol,1eq), N-dimethylformamide (20mL) was added to the reactor followed by addition of sodium hydride (576.65mg,14.42mmol, 60% purity, 1.5eq) at 0 ℃, stirring for 0.5 h, followed by addition of benzyl chloromethyl ether (1.81g,11.53mmol,1.60mL,1.2eq) and gradual warming to 20 ℃ for 11.5 h. After the reaction, the reaction solution was added dropwise to 30mL of ice water, followed by addition of methyl tert-butyl ether (50 mL. times.3) for extraction, and the organic phases were combined and spin-dried to give the crude product. And purifying the crude product by a silica gel column (eluent: ethyl acetate: petroleum ether: 3:1) to obtain a compound WX 008-3.

And step 3: synthesis of Compound WX008-4

WX008-3(3.3g,7.63mmol,1eq), acetonitrile (30mL) were added to the reaction flask, then cooled to 0 deg.C, a solution of ceric ammonium nitrate (12.56g,22.90mmol,11.41mL,3eq) in water (10mL) was added dropwise, and the temperature was gradually raised to 20 deg.C for 12 hours of reaction. After the reaction is finished, the reaction liquid is dried in a spinning mode to obtain a crude product, and the crude product is obtainedThe product was dissolved in 50mL of ethyl acetate, washed with saturated brine (50mL), and the organic phase was spin-dried. The crude product was purified by slurrying with methyl tert-butyl ether (10mL) to give the compound WX 008-4. MS M/z 310.0[ M-H ]] ⁺,312.0[M-H+2] ⁺。

And 4, step 4: synthesis of Compound WX008-5

WX008-4(1g,2.56mmol,1eq), WX004-9(2.37g,5.13mmol,2eq), 1, 2-dichloroethane (30mL), copper acetate (931.10mg,5.13mmol,2eq), pyridine (405.49mg,5.13mmol, 413.76. mu.L, 2eq) were added to a reaction flask and reacted at 60 ℃ for 12 hours. After the reaction, the reaction solution was filtered, the filter cake was washed with 15mL of 1, 2-dichloroethane, and the filtrate was spin-dried to obtain a crude product. The crude product was purified by thin layer chromatography silica gel plate (ethyl acetate: petroleum ether ═ 3:1) to afford compound WX 008-5. MS M/z 728.1[ M + H ]] ⁺,730.0[M+H+2] ⁺。

And 5: synthesis of Compound WX008-6

WX008-5(0.48g, 658.07. mu. mol,1eq) and acetonitrile (5mL) were added to a reaction flask, the temperature was reduced to 0 ℃, then a solution of ceric ammonium nitrate (1.08g,1.97mmol, 983.92. mu.L, 3eq) in water (2.5mL) was added, and the temperature was gradually raised to 20 ℃ for 12 hours. And (3) carrying out spin drying on the reaction liquid to obtain a crude product, and purifying the crude product by a thin-layer chromatography silica gel plate (ethyl acetate: petroleum ether ═ 1:1) to obtain a compound WX 008-6. MS M/z 608.0[ M + H ]] ⁺,610.0[M+H+2] ⁺。

Step 6: synthesis of Compound WX008-7

After WX008-6(0.2g, 328.27. mu. mol,1eq) and dichloromethane (5mL) were added to a reaction flask, the temperature was then reduced to 0 ℃, boron tribromide (246.72mg, 984.81. mu. mol, 94.89. mu.L, 3eq) was added dropwise, and the temperature was gradually increased to 20 ℃ for reaction for 0.5 hour. The reaction solution is slowly dripped into saturated sodium bicarbonate solution for quenching, then ethyl acetate (50mL multiplied by 2) is added for extraction, and the organic phase is dried in a spinning mode to obtain the compound WX 008-7.¹H NMR (400MHz, deuterated methanol) delta 7.78(s,2H), 7.39(s,1H),3.14-3.17(m,1H),1.29(s,3H),1.31(s, 3H). MS M/z 487.9[ M + H ]] ⁺, 489.9[M+H+2] ⁺。

And 7: synthesis of compound WX008

WX008-7(0.08g, 163.56. mu. mol,1eq), methanol (5mL), and sodium methoxide (26.51mg, 490.69. mu. mol,3eq) were added to a reaction flask and reacted at 60 ℃ for 12 hours. Spin-drying the reaction solution to obtain crude product, and separating by high performance liquid chromatography (chromatographic column: Phenomenex Luna C18150 mm 30mm 5 μm; mobile phase: [ water (0.04% HCl) -acetonitrile: [ acetonitrile ]](ii) a 35 percent to 60 percent of acetonitrile for 10min) to obtain the target compound WX 008.¹H NMR(400MHz,DMSO-d ₆)δ:ppm 1.21(d,J＝6.88Hz,6H),3.01-3.11(m,1H),3.86(s,3H),7.45(s,1H),7.87(s,2H),12.23(s,1H),12.52(s,1H)。MS m/z:440.1[M+H] ⁺,442.1[M+H+2] ⁺。

Example 9

The synthetic route is as follows:

step 1: synthesis of Compound WX009

Adding WX006(790mg,1.71mmol,1eq) into a pre-dried reaction flask, and performing supercritical chromatography (chromatography column (s, s) WHELK-O1(250mm 30mm,5 μm); mobile phase: [ neutral-methanol ]](ii) a 55% -55% of methanol for 10 minutes) to obtain a crude product, and performing supercritical chromatography (chromatographic column (s, s) WHELK-O1(250mm 30mm,5 μm); mobile phase of 0.1% ammonia methanol](ii) a 50 percent to 50 percent of methanol and 15min) to obtain WX 009.¹H NMR(400MHz,DMSO-d ₆)δ:ppm 1.17–1.19(d,J＝7.00Hz,3H),1.67-1.82(m,4H),2.58(s,1H),2.69-2.78(m,1H),2.90(s,1H),7.77(s,2H),12.04(s,1H)。MS m/z:461.0[M+H] ⁺. Retention time 2.36 min (instrument: Thar analytical SFC; column: S, S _ wolf _ 013.5 μm,0.46cm id x 5cm L; mobile phase: a: food grade supercritical carbon dioxide; B: methanol (0.05% isopropylamine, vol.); gradient: B content from 4 min5% to 50%; the flow rate is 4.0 mL/min; column temperature: 35 ℃; detection wavelength: 220 nm; the back pressure of the system is 100 bar).

Example 11

The synthetic route is as follows:

step 1: synthesis of Compound WX011-2

A predried three-necked flask was charged with WX011-1(15g,151.32mmol,1eq) and tetrahydrofuran (200mL) while displacing nitrogen three times, and while cooling to-65 ℃ with dry ice ethanol, n-butyllithium (2M,83.22mL,1.1eq) was added slowly, held at-65 ℃ and stirred for 30 minutes, followed by-65 ℃. Methyl iodide (22.55g,158.88mmol,9.89mL,1.05eq) was added and after the addition was complete the ice bath was removed, allowed to warm slowly to room temperature and stirred at room temperature (18-20 ℃) for 2 hours. After completion of the reaction, 100mL of water and ethyl acetate (100mL × 3) were added to the reaction, followed by extraction, and the organic phases were combined and washed with saturated brine (100mL × 2), separated, dried over anhydrous sodium sulfate, filtered to remove the drying agent, and then dried under reduced pressure to obtain WX 011-2.¹HNMR (400MHz, deuterated chloroform) δ 3.24-3.27(t, J ═ 4.8Hz,2H),2.92(s,3H),2.34-2.37(t, J ═ 6.4Hz,2H),1.76-1.80(m, 4H).

Step 2: synthesis of Compound WX011-3

A single neck flask which had been previously dried was charged with WX011-2(14.5g,128.14mmol,1eq) and toluene (200mL), added with Lawson's reagent (51.83g,128.14mmol,1eq), and heated to 80 ℃ with stirring for 2 hours. After completion of the reaction, 200mL of water was added to the reaction mixture, ethyl acetate (200mL × 3) was added for extraction, the organic phases were combined, washed with 200mL of saturated brine, separated, dried over anhydrous sodium sulfate, filtered, and the filtrate was spin-dried under reduced pressure to obtain a crude product. Purifying the crude product by using an automatic column-passing machine, and carrying out gradient leaching:petroleum ether: ethyl acetate 10:1 to 5:1 to 1:1 to give WX 011-3.¹H NMR (400MHz, deuterated chloroform) δ 3.83-3.88(t, J ═ 7.2Hz,2H),3.46(s,3H),2.98-3.01(t, J ═ 6Hz,2H),1.88-1.91(m,2H),1.71-1.74(m, 2H).

And step 3: synthesis of Compound WX011-4

In a single neck flask which had been previously dried, WX011-3(15g,116.08mmol,1eq) and ethanol (200mL) were charged, iodomethane (24.71g,174.12mmol,10.84mL,1.5eq) was added, and after completion of the addition, the reaction mixture was warmed to 80 ℃ and stirred for 1 hour. And after the reaction is finished, directly carrying out reduced pressure spin-drying on the reaction liquid, adding 100mL of tetrahydrofuran into the spin-dried residue, stirring at room temperature for 10 minutes, separating out solids, filtering, washing a filter cake with 20mL of tetrahydrofuran, and carrying out reduced pressure spin-drying on the filter cake to obtain WX 011-4.¹H NMR (400MHz, deuterated dimethyl sulfoxide) δ 3.75-3.78(t, J ═ 5.6Hz,2H),3.41(s,3H),3.03-3.06(m, 2H),2.77(s,3H),1.76-1.85(m, 4H).

And 4, step 4: synthesis of Compound WX011-6

Adding WX011-4(10g,68.84mmol,1eq, HI) and tetrahydrofuran (100mL) into a pre-dried single-neck bottle, adding potassium tert-butoxide (11.59g,103.26mmol,1.5eq), stirring at room temperature (25 ℃) for 10 minutes, filtering, spin-drying the filtrate under reduced pressure, adding toluene (10mL), filtering, preparing a pre-dried three-neck bottle, adding WX007-1(6.23g,41.30mmol,0.6eq) and toluene (80mL), replacing with nitrogen three times, cooling to 0 ℃, dropping the toluene solution just filtered, controlling the temperature to be kept at 0 ℃, heating to 25 ℃ after dropping, and stirring for 40 minutes. After the reaction is finished, adding 100mL of water into the reaction solution, adding ethyl acetate (100mL 4) for extraction, combining organic phases, washing the organic phases by using saturated saline solution (200mL), separating the liquid, drying the organic phases by using anhydrous sodium sulfate, filtering, performing reduced pressure spin drying on the filtrate, purifying a crude product by using an automatic column chromatography machine, and performing gradient elution: petroleum ether: ethyl acetate 10% to 20% to 30% gave WX 011-6.¹H NMR (400MHz, deuterated chloroform) δ 3.25-3.30(m,5H),2.73-2.76(t, J ═ 6.4Hz,2H),1.95-2.01(m, 2H).

And 5: synthesis of Compound WX011-8

A pre-dried reaction flask was charged with WX011-6(0.6g,2.75 mmol)1eq) and WX001-4(734.65mg,4.13mmol,1.5eq), dimethyl sulfoxide (10mL) and potassium carbonate (1.52g,11.00mmol,4eq) were added, nitrogen was bubbled, cuprous iodide (314.39mg,1.65mmol,0.6eq) was added, the reaction was warmed to 90 ℃ and stirred for 16 hours. After the reaction, 10mL of water was added to the reaction solution, ethyl acetate was added for extraction (10mL × 3), the organic phases were combined, washed with saturated brine (20mL), separated, dried over anhydrous sodium sulfate, filtered, the filtrate was spin-dried under reduced pressure, and the crude product was purified by an automatic column chromatography, gradient elution: petroleum ether: ethyl acetate 20% to give WX 011-8.¹H NMR (400MHz, deuterated dimethyl sulfoxide) delta 6.69(s,2H),5.64(s,2H),3.33-3.02(m,5H),2.65-2.61(m,2H),1.96-1.87(m, 2H).

And (3) identifying the structure of WX 011-8:

WX011-8(0.05g, 139.03. mu. mol,1eq) and methanol (10mL) were charged in a 50mL single-neck flask, wet palladium on carbon (0.01g, 139.03. mu. mol, 10% purity, 1eq) was added, hydrogen was replaced three times, and then reaction was carried out at 20 ℃ for 2 hours under the pressure of a hydrogen balloon (15 psi). After the reaction, the reaction solution was directly filtered with celite, and the filtrate was spin-dried under reduced pressure to give a crude product, which was then purified by high performance liquid chromatography (conditions: column: Phenomenex Luna C18150 × 30mm × 5 μm; mobile phase: [ water (0.04% hydrochloric acid) -acetonitrile: [ acetonitrile ] -acetonitrile ]](ii) a 20 to 45 percent of acetonitrile) and 10 minutes), and freeze-drying to obtain the WX 011-8A. NOE (400MHz, deuterated dimethyl sulfoxide) shows that hydrogen at the (2.74-2.77) position on the N-methylpiperidine ring has correlation with hydrogen at the (8.62) position on the pyrazine ring, and the structure of WX011-8 is correct.¹HNMR (400MHz, deuterated dimethyl sulfoxide) delta 1.93-1.96(m,2H), 2.74-2.77(m,2H), 3.60-3.65(m,5H), 6.74(s,2H), 8.62(s, 1H).

Step 6: synthesis of Compound WX011-9

WX011-8(300.00mg, 834.17. mu. mol,1eq) was charged into a reaction flask dried in advance, glacial acetic acid (5mL) and sodium acetate (684.30mg,8.34mmol,10eq) were added, and the reaction solution was warmed to 120 ℃ and stirred for 12 hours.After the reaction was completed, glacial acetic acid in the reaction solution was removed by spinning under reduced pressure, 10mL of water was added to the residue, sodium bicarbonate was added to adjust pH to 7, ethyl acetate (10mL × 4) was added to extract, the organic phases were combined, washed with saturated brine (10mL), separated, dried over anhydrous sodium sulfate, filtered, and the filtrate was spin-dried under reduced pressure to give WX 011-9.¹H NMR (400MHz, deuterated dimethyl sulfoxide) delta 7.78(s,2H),3.17(s,3H),2.34-2.36(m,4H),2.07(s,3H),1.80-1.83(m, 2H).

And 7: synthesis of Compound WX011-10

WX011-9(250.00mg, 652.35. mu. mol,1eq) was charged into a reaction flask which had been dried in advance, ethanol (2mL) and 2M hydrochloric acid (2mL) were added, and the reaction mixture was stirred at 80 ℃ for 12 hours. After completion of the reaction, a saturated aqueous solution of sodium bicarbonate was added to the reaction mixture to adjust pH to 7, ethyl acetate (10 mL. times.3) was added for extraction, the organic phases were combined, the mixture was dried under reduced pressure, the crude product was dissolved in 1mL of methanol, the mixture was filtered, and the filtrate was subjected to high performance liquid chromatography (column: Phenomenex luna C18250 mm. times.50 mm. times.10 μm; mobile phase: [ water (0.05% HCl) -acetonitrile](ii) a 20 to 50 percent of acetonitrile) for 10 minutes) to obtain the target compound WX 011-10.¹HNMR (400MHz, deuterated dimethyl sulfoxide) delta 11.56(s,1H),6.78(s,2H),3.16-3.20(m,5H),2.35-2.38(m,2H),1.78-1.81(m, 2H).

And 8: synthesis of Compound WX011-12

WX011-10(0.2g, 586.18. mu. mol,1eq) was charged into a pre-dried bottle, glacial acetic acid (2mL) and hydrochloric acid (177.91mg,1.81mmol, 174.42. mu.L, 37% purity, 3.08eq) were added, the temperature was reduced to 5 ℃, sodium nitrite (44.49mg, 644.80. mu. mol,1.1eq) was dissolved in water (1mL), the reaction system was dropped, and then stirred at 5 ℃ for 0.5 hour, WX001-9(100.68mg, 644.80. mu. mol,1.1eq) was added, and stirred at 5 ℃ for 0.5 hour. After the reaction, saturated aqueous sodium acetate (5mL) was added to the reaction mixture, a solid precipitated, filtered, the filter cake washed with 2mL of water, and the filter cake was spin-dried under reduced pressure to give WX 011-12.¹H NMR (400MHz, deuterated dimethyl sulfoxide) δ 12.10(s,1H),11.54(s,1H),10.81(s,1H),7.96(s,2H),4.22-4.17(m,2H),3.19(m,5H),2.49-2.38(m,2H),1.82-1.81(m,2H),1.28-1.24(t, J ═ 7.2Hz, 3H).

And 8: synthesis of compound WX011

WX011-12(200.00mg, 402.15. mu. mol,1eq) was added to a pre-dried reaction flask, N-dimethylacetamide (2mL) was added to dissolve the substrate, potassium acetate (39.47mg, 402.15. mu. mol,1eq) was added, and the mixture was stirred at 115 ℃ for 1 hour. After completion of the reaction, the reaction solution was directly filtered, and the filtrate was subjected to high performance liquid chromatography (column: Welch Xtimate C18150 mm 25mm 5 μm; mobile phase: [ water (0.04% HCl) -acetonitrile](ii) a Acetonitrile percent of 40 percent to 70 percent for 10 minutes) to obtain the target compound WX 011.¹H NMR (400MHz, deuterated dimethyl sulfoxide) δ 13.27(s,1H),11.57(s,1H),7.78(s,2H),3.27-3.22(m,5H),2.41-2.36(t, J ═ 6Hz,2H),1.84-1.81(m, 2H). MS-ESI M/z 462.0[ M + H ]] ⁺,464.0[M+H+2] ⁺。

Example 12

The synthetic route is as follows:

step 1: synthesis of Compound WX012-4

WX008-3(4.66g,10.78mmol,1eq) was charged to a pre-dried reaction flask, then dissolved in acetonitrile (93mL), followed by tri-tert-butyl-1-ethoxyethylene (7.79g,21.56mmol,7.28mL,2eq), nitrogen purged three times, bis triphenylphosphine palladium dichloride (1.14g,1.62mmol,0.15eq), cuprous iodide (205.31mg,1.08mmol,0.1eq), nitrogen purged three times again, slowly raised to 90 ℃ and stirred for 15 hours. After the reaction is completed, the reaction solution of WX012-4 is directly used for the next reaction.

Step 2: synthesis of Compound WX012-5

Hydrochloric acid (6M,30mL,16.68eq) was added to a solution of WX012-4(4.57g,10.79mmol,1eq) in acetonitrile (93mL) at room temperature (25 deg.C)) The reaction was stirred for 30 minutes. After completion of the reaction, 200mL of a saturated potassium fluoride solution was added, followed by dilution with ethyl acetate (100mL), the organic phase was collected by liquid separation, the aqueous phase was extracted with ethyl acetate (100mL × 2), the organic phases were combined, washed with a saturated saline solution (100mL × 2), dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the solvent was removed under reduced pressure to obtain a crude product. The crude product was purified by automatic column chromatography (gradient elution: petroleum ether: ethyl acetate: 1:0 to 3:7) to afford WX 012-5.¹H NMR (400MHz, deuterated chloroform) δ 7.33-7.29(d, J ═ 8.8Hz,2H),7.26-7.22(m,2H),7.19-7.15(m,3H),6.84-6.81(d, J ═ 8.8Hz,2H),5.39(s,2H),5.03(s,2H),4.61(s,2H),3.74(s,3H),2.43(s, 3H). MS-ESI M/z 396.1[ M + H ]] ⁺。

And step 3: synthesis of Compound WX012-6

WX012-5(4g,10.12mmol,1eq) was charged into a pre-dried reaction flask, then dissolved in dichloromethane (80mL), the reaction was cooled to 0 deg.C, then diethylaminosulfur trifluoride (8.15g,50.58mmol,6.68mL,5eq) was added, the temperature was slowly raised to room temperature (25 deg.C) and the reaction was stirred for 2 hours, the reaction was cooled to 0 deg.C, then diethylaminosulfur trifluoride (8.15g,50.58mmol,6.68mL,5eq) was added, and the reaction was slowly raised to room temperature (25 deg.C) and stirred for 17 hours. After completion of the reaction, the reaction mixture was slowly added to room temperature water (200mL), followed by neutralization of the reaction mixture to about pH 7 with saturated sodium bicarbonate solution, dilution with dichloromethane (200mL), liquid separation to collect the organic phase, extraction of the aqueous phase with dichloromethane (200mL × 2), combination of the organic phases, washing with saturated brine (200mL × 2), drying over anhydrous sodium sulfate, filtration to remove the drying agent, and removal of the solvent under reduced pressure to give crude product WX 012-6.

And 4, step 4: synthesis of Compound WX012-7

WX012-6(3.46g,8.29mmol,1eq) was added to a pre-dried reaction flask, then dissolved in acetonitrile (73mL), the reaction was brought to 0 deg.C, and a solution of ammonium ceric nitrate (13.63g,24.87mmol,12.39mL,3eq) in water (31mL) was added, and the reaction was stirred for 2 hours while slowly increasing to room temperature (25 deg.C). Diluting with water (100mL) and ethyl acetate (100mL), separating, collecting organic phase, extracting aqueous phase with ethyl acetate (100mL × 2), combining organic phases, and diluting with saturated waterAnd brine (100mL × 2), dried over anhydrous sodium sulfate, filtered to remove the drying agent, and then the solvent was removed under reduced pressure to obtain a crude product. The crude product was purified by automatic column chromatography (gradient elution: petroleum ether: ethyl acetate: 1:0 to 16:84) to afford WX 012-7.¹H NMR (400MHz, deuterated dimethyl sulfoxide) δ 12.92(s,1H),7.41-7.18(m,5H),5.29(s,2H),4.62(s,2H), 1.98-1.84 (t, J ═ 19.2Hz, 3H).

And 5: synthesis of Compound WX012-8

WX012-7(328.46mg,1.10mmol,1.05eq), WX005-6(0.5g,1.05mmol,1eq) were charged into a pre-dried reaction flask, then dissolved in dichloroethane (25mL), followed by addition of copper acetate (286.71mg,1.58mmol,1.5eq), pyridine (208.10mg,2.63mmol, 212.35. mu.L, 2.5eq), and after completion of the charge, the reaction was stirred at 40 ℃ under oxygen 15psi for 12 hours. After completion of the reaction, water (50mL) and dichloromethane (50mL) were added for dilution, the organic phase was collected by separation, the aqueous phase was extracted with dichloromethane (50mL × 2), the organic phases were combined and washed with saturated brine (50mL × 2), dried over anhydrous sodium sulfate, the drying agent was removed by filtration, and the solvent was removed under reduced pressure to obtain a crude product. The crude product was purified by automatic column chromatography (gradient elution: petroleum ether: ethyl acetate: 1:0 to 20:80) to afford WX 012-8.¹H NMR (400MHz, deuterated dimethyl sulfoxide) δ 7.82(s,2H),7.36-7.34(m,2H),7.32-7.27(m,1H),7.06-7.00(m,3H),6.83-6.77(m,3H),5.41(s,2H),4.84(s,2H),4.69(s,2H),3.69(s,3H),2.67-2.64(m,2H),2.47-2.45(m,2H),2.06-1.95(t, J ═ 19.2Hz,3H),1.75-1.71(d, J ═ 5.1Hz, 4H).

Step 6: synthesis of Compound WX012-9

WX012-8(0.43g, 591.84. mu. mol,1eq) was added to a pre-dried reaction flask, then dissolved in acetonitrile (14mL), the reaction was brought to 0 ℃ and a solution of ammonium ceric nitrate (973.37mg,1.78mmol, 884.88. mu.L, 3eq) in water (6mL) was added, and the reaction was stirred for 1 hour while slowly increasing to room temperature (25 ℃). After completion of the reaction, water (20mL) and ethyl acetate (20mL) were added for dilution, the organic phase was collected by separation, the aqueous phase was extracted with ethyl acetate (20 × 3mL), the organic phases were combined and washed with saturated brine (20mL × 2), dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the solvent was removed under reduced pressure to obtain a crude product. The crude product was purified by thin layer chromatography on silica gel plate (petroleum ether: ethyl acetate 1:1) to give WX 012-9. MS-ESI M/z 606.1[ M + H ]] ⁺,608.0[M+H+2] ⁺。

And 7: synthesis of Compound WX012

WX012-9(0.12g, 197.89. mu. mol,1eq) is added into a reaction flask dried in advance, then dichloromethane (1.2mL) is dissolved, the reaction system is cooled to 0 ℃, boron tribromide (99.15mg, 395.78. mu. mol, 38.13. mu.L, 2eq) is added, stirring is carried out at 0 ℃ for reaction for 0.5 hour, boron tribromide (99.15mg, 395.78. mu. mol, 38.13. mu.L, 2eq) is added, after the reaction is finished, the pH of the reaction solution is neutralized by saturated sodium bicarbonate solution at 0 ℃, dichloromethane (3mL) is added for dilution, liquid separation is carried out to obtain an organic phase, an aqueous phase is extracted by dichloromethane (3mL 2), the organic phases are combined, anhydrous sodium sulfate is dried, drying agents are filtered, and then the solvents are removed under reduced pressure to obtain a crude product. The crude product was separated by high performance liquid chromatography (column: Welch XTimate C18150 mm 25mm 5 μm; mobile phase: [ water (0.04% HCl) -acetonitrile](ii) a 35 percent to 55 percent of acetonitrile for 10min) and purifying to obtain the WX 012.¹H NMR (400MHz, deuterated dimethyl sulfoxide) δ 12.78(s,1H),12.06(s,1H),7.81(s,2H),2.68-2.64(m,2H),2.45-2.42(m,2H),2.02-1.89(t, J ═ 19.2Hz,3H),1.81-1.69(m, 4H). MS-ESI M/z 486.0[ M + H ]] ⁺,488.0[M+H+2] ⁺。

Example 13

The synthetic route is as follows:

step 1: synthesis of Compound WX013-2

WX013-1(25g,175.92mmol,1eq) was added to the flask, heated to 100 deg.C, followed by addition of cyclopentadiene (69.77g,527.76 mmol,3eq) and the temperature was raised to 180 deg.CThe reaction was carried out for 2 hours. After the reaction is finished, the reaction solution is concentrated in vacuum by an oil pump, a crude product is beaten and purified by (methyl tert-butyl ether: n-hexane: 1:3, total 120mL), and the obtained product is filtered, and filtrate is dried in a spinning mode to obtain WX 013-2.¹H NMR (400MHz, deuterated dimethyl sulfoxide) δ 2.00-2.06(m,1H)2.16-2.22(m,1H)3.71(s,6H)3.89-3.93(m,2H)6.97(t, J ═ 1.88Hz, 2H).

Step 2: synthesis of Compound WX013-3

WX013-2(4.5g,21.61mmol,1eq), tetrahydrofuran (20mL), palladium on carbon (0.4g, 10% purity) was added to the reaction flask, replaced with hydrogen three times, and reacted at 25 ℃ under hydrogen pressure 15psi for 5 min. And after the reaction is finished, adding diatomite into the reaction solution for filtering, and spin-drying the filtrate to obtain WX 013-3.¹H NMR (400MHz, deuterated dimethyl sulfoxide) delta 1.14-1.20(M,3H)1.32-1.40(M,1H)1.80-1.87(M,2H)3.23(s,2H)3.70(s,6H), MS-ESI M/z 211.1[ M + H ]] ⁺。

And step 3: synthesis of Compound WX013-4

WX013-3(2g,9.51mmol,1eq), tetrahydrofuran (10mL), water (3mL), methanol (3mL), sodium hydroxide (951.29mg,23.78mmol,2.5eq) were added to the reaction flask and reacted at 25 ℃ for 12 hours. After the reaction is finished, adding hydrochloric acid into the reaction liquid to adjust the pH value to 5, then spin-drying, soaking with methanol, filtering and spin-drying to obtain WX 013-4. MS-ESI M/z 181.0[ M-H ]] ⁺。

And 4, step 4: synthesis of Compound WX013-5

WX013-4(1.5g,8.23mmol,1eq), trifluoroacetic anhydride (15mL) was added to the reaction flask and reacted at 65 ℃ for 12 h. And after the reaction is finished, spin-drying the reaction liquid to obtain WX 013-5. MS-ESI M/z 165.1[ M + H ]] ⁺。

And 5: synthesis of Compound WX013-6

WX013-5(1.25g,7.61mmol,1eq), acetic acid (10mL), water (5mL) were added to the reaction flask, followed by hydrazine hydrate (972.42mg,19.04mmol, 944.10. mu.L, 98% purity 2.5eq) with stirring and reacted at 100 ℃ for 12 hours. After the reaction is finished, directly spin-drying the reaction solution to obtain a crude product. The crude product is slurried with methyl tert-butyl ether (20mL) to obtain WX 013-6. MS-ESI M/z 179.1[ M + H ]] ⁺。

Step 6: synthesis of Compound WX013-7

WX013-6(1g,5.61mmol,1eq), phosphorus oxychloride (10mL) was added to the reaction flask and reacted at 100 ℃ for 4 hours. After the reaction is finished, dropwise adding the reaction liquid into 30mL warm water to quench the reaction, then adding sodium bicarbonate solid to adjust the pH value to about 7, then adding ethyl acetate (30mL x3) for extraction, merging organic phases, and carrying out spin drying on the organic phases to obtain a crude product. The crude product was purified by silica gel column (petroleum ether: ethyl acetate 3:1, gradient elution to 1:1) to afford WX 013-7.¹HNMR (400MHz, deuterated dimethyl sulfoxide) delta 1.15-1.17(m,2H)1.63-1.66(m,1H)1.86-1.95(m,1H)2.02-2.14(m,2H)3.52-3.66(m, 2H). MS-ESI M/z 215.0[ M + H ]] ⁺,217.0[M+H+2] ⁺。

And 7: synthesis of Compound WX013-8

WX013-7(0.165g, 767.16. mu. mol,1eq), WX001-4(341.42mg,1.92mmol,2.5eq), N, N-dimethylacetamide (5mL), cesium carbonate (624.89mg,1.92mmol,2.5eq) were added to a reaction flask and reacted at 130 ℃ for 4 hours. After the reaction was completed, the reaction solution was filtered, ethyl acetate (50mL) was added to the filtrate and washed with saturated brine (100 mL. times.3), the organic phase was dried by spinning to obtain a crude product, which was separated by high performance liquid chromatography (column: Phenomenex Luna C18150. times.30 mm. times.5 μm; mobile phase: [ water (0.04% hydrochloric acid) -acetonitrile:.)/solvent)](ii) a 55 to 85 percent of acetonitrile for 10 minutes) to obtain WX 013-8. MS-ESI M/z 355.9[ M + H ]] ⁺,357.9[M+H+2] ⁺。

And 8: synthesis of Compound WX013-9

WX013-8(0.135g, 378.54. mu. mol,1eq), acetic acid (3mL), sodium acetate (77.63mg, 946.35. mu. mol,2.5eq) were added to the reaction flask, followed by reaction at 110 ℃ for 12 hours. And after the reaction is finished, spin-drying the reaction liquid to obtain a crude product, dissolving the crude product with ethyl acetate (30mL), adjusting the pH of the solution to be 7-8 with a sodium carbonate solution, separating an organic phase, and spin-drying the organic phase to obtain WX 013-9. MS-ESI M/z 380.0[ M + H ]] ⁺,382.0[M+H+2] ⁺。

And step 9: synthesis of Compound WX013-10

Mixing WX013-9(0.13g, 341.90. mu. mol,1eq) with ethanol (5mL)) Hydrochloric acid (12M, 142.46. mu.L, 5eq) was added to the reaction, followed by reaction at 100 ℃ for 12 hours. And after the reaction is finished, spin-drying the reaction liquid to obtain WX 0012-11.¹HNMR (400MHz, deuterated dimethyl sulfoxide) δ 1.05-1.14(m,1H)1.20-1.22(m,1H)1.53-1.56(m,1H)1.77-1.79(d, J ═ 8.88Hz,1H)1.98-2.12(m,2H)3.51(s,1H)3.61(s,1H)5.68(s,2H)6.72(s,2H)12.07(s, 1H). MS-ESI M/z 338.0[ M + H ]] ⁺,340.0[M+H+2] ⁺。

Step 10: synthesis of Compound WX013-11

WX013-10(0.05g, 147.85. mu. mol,1eq), acetic acid (1.5mL), hydrochloric acid (1mL) were added to the reaction flask, followed by dropwise addition of a solution of sodium nitrite (13.26mg, 192.20. mu. mol,1.3eq) in water (1mL) at 0 ℃ and reaction at 0 ℃ for 0.5 hour, followed by addition of WX001-9(26.55mg, 170.02. mu. mol,1.15eq) and reaction at 0 ℃ for 1.5 hour. After the reaction is finished, sodium acetate solid of about 200mg is added, solid is separated out, then filtration is carried out, and filter cake is dried in a spinning mode to obtain WX 013-11.¹HNMR (400MHz, deuterated dimethyl sulfoxide) delta 1.03-1.18(m,2H)1.25-1.27(m,3H)1.52-1.54(m,1H)1.76-1.78(m,1H)1.97-2.07(m,2H)3.31-3.33(m,2H)3.49(s,1H)3.61(s,1H)4.16-4.20(m,2H)7.81(s,2H)11.94-12.13(m, 2H). MS-ESI M/z 505.1[ M + H ]] ⁺,507.1[M+H+2] ⁺。

Step 11: synthesis of Compound WX013-12

WX013-11(0.085g, 168.54. mu. mol,1eq), N, N-dimethylacetamide (1mL), potassium acetate (82.71mg, 842.72. mu. mol,5eq) were added to the reaction flask, followed by reaction at 115 ℃ for 8 hours. After completion of the reaction, the reaction solution was filtered. Separating the filtrate by high performance liquid chromatography (chromatographic column: Phenomenex Luna C18150: 30 mm: 5 μm; mobile phase: [ water (0.04% hydrochloric acid) -acetonitrile](ii) a Acetonitrile percent of 40 percent to 70 percent and 10min) to obtain WX 013-12.¹H NMR (400MHz, deuterated dimethyl sulfoxide) delta 1.04-1.14(m,1H)1.20-1.30(m,1H)1.53-1.55(m,1H)1.78-1.81(m,1H)2.01-2.08(m,2H)3.50(s,1H)3.64(s,1H)7.80(s,2H)12.14(s,1H)13.31(s, 1H). MS-ESI M/z 459.0[ M + H ]] ⁺,461.0[M+H+2] ⁺。

Step 12: synthesis of Compounds WX013 and WX014

WX013-12 was subjected to supercritical chromatography (column: DAICEL CHIRALCEL OJ (250 mm. times.30 mm,10 μm), mobile phase: [ 0.1% ammonia methanol ], methanol%: 25% -25%, 15min) to give WX013 and WX 014.

WX013： ¹H NMR (400MHz, deuterated methanol) δ 1.18-1.26(m,1H),1.33-1.38(m,1H),1.63-1.66(d, J ═ 9.26Hz,1H),1.86-1.89(d, J ═ 9.26Hz,1H),2.02-2.20(m,2H),3.62-3.76(d, J ═ 29.2Hz,2H),7.75(s, 2H). MS-ESI M/z 459.0[ M + H ]] ⁺,461.0[M+H+2] ⁺. Retention time 2.15 min (instrument: CAS-TJ-ANA-SFC-A (Waters SFC-MS); chromatographic column: Chiralcel OJ-33 μm,0.46cm id x 10cm L; mobile phase: A: food grade supercritical carbon dioxide; B: methanol (0.05% diethylamine, volume ratio); gradient: B content increased from 10% to 40% within 5min of A content, flow rate: 4.0 mL/min; column temperature: 35 ℃; detection wavelength: 220 nm; system back pressure: 100 bar).

WX014： ¹H NMR (400MHz, deuterated methanol) δ 1.18-1.26(m,1H),1.33-1.38(m,1H),1.63-1.66(d, J ═ 9.26Hz,1H),1.86-1.89(d, J ═ 9.26Hz,1H),2.02-2.20(m,2H),3.62-3.76(d, J ═ 29.2Hz,2H),7.75(s, 2H). MS-ESI M/z 459.0[ M + H ]] ⁺,461.0[M+H+2] ⁺. Retention time: 2.32 minutes (instrument: CAS-TJ-ANA-SFC-A (Waters SFC-MS); chromatographic column: Chiralcel OJ-33 μm,0.46cm id x 10cm L; mobile phase: A: food grade supercritical carbon dioxide; B: methanol (0.05% diethylamine, volume ratio); gradient: B content rises from 10% to 40% within 5 minutes of A content; flow rate: 4.0 mL/min; column temperature: 35 ℃; detection wavelength: 220 nm; system back pressure: 100 bar).

Example 15

The synthetic route is as follows:

step 1: synthesis of Compound WX015-2

Potassium tert-butoxide (14.67g,130.77mmol,1.1eq) and n-hexane (40mL) were added to a reaction flask, the temperature was then lowered to-30 ℃ and tetramethylethylenediamine (15.20g,130.77mmol,19.74mL,1.1eq) and n-butyllithium (2.5M,52.31mL,1.1eq) were added to the reaction, respectively, followed by WX015-1(10g,118.88mmol,10.87mL,1eq) being added to the reaction, the temperature being maintained at-40 ℃ and the reaction being stirred at-20 ℃ for a further 15 minutes after the addition was complete, and then at-10 ℃ for 5 minutes. After the addition was complete, tetrahydrofuran (50mL) was added to the reaction and the temperature was reduced to-50 deg.C, then dimethyl disulfide (11.76g,124.83mmol,11.20mL,1.05eq) was added to the reaction in one portion, then the temperature was raised to 25 deg.C and stirred for 2 hours. After completion of the reaction, the reaction mixture was added to a saturated aqueous solution of ammonium chloride (100mL), followed by extraction with methyl tert-butyl ether (200mL × 2), and the organic phase was washed with a saturated brine (50mL × 2), dried over anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure to give WX 015-2.¹HNMR (400MHz, deuterated chloroform) delta 4.85-4.86(m,1H),4.00-4.026(m,2H),2.15(s,3H),1.99-2.03(m,2H),1.75-1.78(m, 2H).

Step 2: synthesis of Compound WX015-3

WX007-1(3g,19.87mmol,1eq) and toluene (30mL) were added to a pre-dried reaction flask, nitrogen was purged and stirred at 0 deg.C, then WX015-2(3.62g,27.82mmol,1.4eq) was dissolved in toluene (30mL) and added dropwise to the reaction system, stirring was continued at 0 deg.C for 10 minutes, then warmed to 30 deg.C and stirred for 20 minutes. The reaction was then heated at 50 ℃ for 5.5 hours. After the reaction, the reaction mixture was concentrated under reduced pressure at 40 ℃ by a water pump, 60mL of water was added, ethyl acetate (50mL × 2) was added for extraction, the organic phases were combined, the organic phase was washed with saturated brine (70mL), the organic phase was dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure at 40 ℃ by a water pump to give a crude product, and the crude product was purified by automatic column chromatography (gradient elution: petroleum ether: ethyl acetate 1:0 to 4:1) to give compound WX 015-3.¹HNMR (400MHz, deuterated chloroform) delta 4.41-4.43(m,2H),2.75-2.78(m,2H),2.13-2.18(m, 2H).

And step 3: synthesis of Compound WX015-4

WX015-3(2.35g,11.46mmol,1eq) was charged to a pre-dried reaction flask, after which it was dissolved in dimethyl sulfoxide (90mL), then WX001-4(3.06g,17.19mmol,1.5eq), potassium carbonate (6.34g,45.84mmol,4eq), nitrogen was purged three times, cuprous iodide (1.31g,6.88mmol,0.6eq) was added, nitrogen was again purged three times, and the reaction was stirred at 90 ℃ under nitrogen for 17 hours. After the reaction is finished, the reaction liquid is filtered by using kieselguhr, filtrate is washed by using saturated ammonium chloride (200mL), a 1:1 mixed solution of ethyl acetate and petroleum ether (150mL × 5) is added for extraction, organic phases are combined, the organic phase is washed by using saturated saline solution (200mL), dried and decompressed and concentrated to obtain a crude product, and the crude product is separated and purified by using a rapid column chromatography (gradient elution: petroleum ether: ethyl acetate: 1:0 to 2:1) to obtain compounds WX015-4 and WX 015-4A. And (3) respectively hydrogenating WX015-4 and WX015-4A under Pd/C and hydrogen conditions, wherein chlorine at the ortho position of a nitrogen atom of a pyridazine ring is replaced by a hydrogen atom, and the structure is judged to be shown by two-dimensional nuclear magnetism.

And 4, step 4: synthesis of Compound WX015-5

WX015-4(470mg,1.36mmol,1eq) and hydrogen bromide (8mL) were charged into a reaction flask which had been previously dried, and placed at 0 ℃ followed by addition of a mixed solution of sodium nitrite (112.27mg,1.63mmol,1.2eq) and water (4mL), stirring at 0 ℃ for 0.5 hour, addition of a mixed solution of cuprous bromide (213.98mg,1.49mmol, 45.43. mu.L, 1.1eq), water (4mL) and hydrogen bromide (8mL) dropwise to the above system, and stirring was continued for 15.5 hours while naturally warming to 25 ℃. After the reaction was completed, the reaction solution was quenched with water (30mL), extracted with ethyl acetate (50mL × 2), the combined organic phases were washed with saturated sodium bicarbonate (60mL), the organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure to give a crude product, and the crude product was separated and purified by an automatic column chromatography (gradient elution: petroleum ether: ethyl acetate 1:0 to 4:1) to give compound WX 015-5.

And 5: synthesis of Compound WX015-6

WX015-5(400mg, 974.47. mu. mol,1eq) was added to a reaction flask which had been dried beforehand, followed by dissolution with glacial acetic acid (12mL), followed by addition of sodium acetate (159.88mg,1.95mmol,2eq) and stirring at 110 ℃ for 15 hours. After the reaction is finished, saturated sodium bicarbonate is added into the reaction liquid to adjust the pH value to 7, and ethyl is usedExtracting with ethyl acetate (80 mL. times.2), washing the organic phase with saturated sodium bicarbonate solution (100mL) and saturated brine solution (100mL), drying the organic phase with anhydrous sodium sulfate, concentrating under reduced pressure to obtain crude product, and pulping the crude product with a mixed solution (10mL) of ethyl acetate and petroleum ether 1:1 to obtain the compound WX 015-6.¹HNMR (400MHz, deuterated chloroform) delta 10.15(s,1H),7.55(s,2H),4.41-4.44(m,2H),2.76-2.79(m,2H),2.14-2.16(m, 2H).

Step 6: synthesis of Compound WX015-7

To a previously dried reaction flask were added p-methoxybenzyl chloride (221.71mg,1.42mmol, 192.79. mu.L, 1.5eq), potassium carbonate (391.32mg,2.83mmol,3eq) and N, N-dimethylformamide (9mL), followed by addition of WX015-6(370.00mg, 943.80. mu. mol,1eq), nitrogen gas was purged, and the reaction was stirred at 25 ℃ for 16 hours. After the reaction was completed, a saturated ammonium chloride solution (100mL) was added to the reaction mixture, extraction was performed with ethyl acetate (80mL × 2), the organic phases were combined, the organic phase was washed with saturated brine (100mL × 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure at 40 ℃ using a water pump to obtain a crude product, which was separated and purified by an automatic column chromatography (gradient elution: petroleum ether: ethyl acetate 1:0 to 5:2) to obtain compound WX 015-7.

And 7: synthesis of Compound WX015-8

WX015-7(170.00mg, 331.91. mu. mol,1eq) was charged into a pre-dried reaction flask, dioxane (5mL) was added to dissolve it, then the bis pinacolato borate (126.43mg, 497.87. mu. mol,1.5eq) and potassium acetate (65.15mg, 663.83. mu. mol,2eq) were added and replaced with nitrogen 3 times, then 1,1' -bis (diphenylphosphino) ferrocene palladium chloride (24.29mg, 33.19. mu. mol,0.1eq) was added and replaced with nitrogen 3 times, and the temperature was raised to 100 ℃ for reaction for 2 hours. After the reaction is finished, the reaction solution is filtered by using kieselguhr, the filtrate is concentrated under reduced pressure and dried, and the crude product is separated and purified by using a preparation silica gel large plate (petroleum ether: ethyl acetate: 4:5) to obtain a compound WX 015-8.

And 8: synthesis of Compound WX015-9

WX015-8(160mg, 286.10. mu. mol,1eq) was charged into a reaction flask dried in advance, and then dissolved in acetone (3mL), water (1.5mL), followed by addition of sodium periodate (61.19mg, 286.10. mu. mol, 15.85. mu.L, 1eq), ammonium acetate (158.78mg,2.06mmol,7.2eq) and nitrogen substitution three times, and the reaction was stirred at room temperature (25 ℃ C.) for 15 hours. After the reaction, the reaction solution was concentrated with a water pump to remove acetone, then concentrated under reduced pressure with an oil pump to remove water to obtain a crude product, and the crude product was separated by high performance liquid chromatography (column: Luna Omega 5u Polar C18100A; mobile phase: [ water (0.04% hydrochloric acid) -acetonitrile ];% acetonitrile: 45% -65%, 7 min.) to obtain compound WX 015-9.

And step 9: synthesis of Compound WX015-10

WX003-6(31.17mg, 110.04. mu. mol,1.05eq) and WX015-9(50mg, 104.80. mu. mol,1eq) were charged into a reaction flask which had been dried in advance, and then dissolved in 1, 2-dichloroethane (3mL), followed by addition of copper acetate (28.55mg, 157.20. mu. mol,1.5eq) and pyridine (20.72mg, 262.00. mu. mol, 21.15. mu.L, 2.5eq), and after completion of the charge, the reaction was stirred at 40 ℃ under oxygen protection for 12 hours. After the reaction, the reaction solution was washed with 1N hydrochloric acid solution (30mL) and extracted with ethyl acetate (30mL × 2), the organic phases were combined and washed with saturated brine (40mL), the organic phase was dried and concentrated to obtain a crude product, and the crude product was separated and purified with a preparative silica gel large plate (petroleum ether: ethyl acetate 1:1) to obtain compound WX 015-10.¹HNMR (400MHz, deuterated chloroform) δ 7.51(s,2H),7.23-7.31(m,5H),7.08-7.10(m,2H),6.68-6.70(m,2H),6.44-6.57(t, J ═ 52.4Hz,1H),5.51(s,2H),4.88(s,2H),4.70(s,2H),4.29-4.31(m,2H),3.68(s,3H),2.62-2.66(m,2H),1.99-2.02(m, 2H).

Step 10: synthesis of Compound WX015-11

WX015-10(40.00mg, 55.98. mu. mol,1eq) was added to a predried storage bottle, followed by dissolution in acetonitrile (1.6mL), stirring at 0 ℃ and then a mixed solution of ceric ammonium nitrate (92.07mg, 167.95. mu. mol, 83.70. mu.L, 3eq) and water (0.8mL) was slowly added dropwise, followed by raising to 25 ℃ and stirring for 2 hours. After the reaction was completed, the reaction mixture was washed with water (20mL), extracted with ethyl acetate (20mL × 2), the organic phases were combined, washed with saturated brine (30mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give a crude product, which was separated and purified with a preparative thin layer chromatography silica gel plate (petroleum ether: ethyl acetate: 3:5) to give compound WX 015-11.¹HNMR (400MHz, deuterated chloroform) delta 10.28(s,1H),7.48(s,2H),7.28-7.23(m,5H),6.65-6.39(t, J ═ 52.4Hz,1H),5.49(s,2H),4.69(s,2H),4.69-4.35(m,2H),2.65-2.61(m,2H),2.07-1.99(m,2H)。

Step 11: synthesis of Compound WX015

WX015-11(30mg, 50.48. mu. mol,1eq) was added to a predried storage bottle, followed by dissolution in methylene chloride (0.5mL), nitrogen substitution 3 times, stirring at 0 ℃ and slow dropwise addition of a mixed solution of boron tribromide (25.29mg, 100.95. mu. mol, 9.73. mu.L, 2eq) and methylene chloride (0.5mL), followed by reaction at 0 ℃ for 0.5 hour with stirring. After the reaction, the reaction solution was neutralized to pH 7 with saturated sodium bicarbonate solution, extracted with ethyl acetate (15 mL. multidot.3), the organic phases were combined, the organic phase was concentrated under reduced pressure to give crude product, which was separated by HPLC (column: Luna Omega 5u Polar C18100A; mobile phase: [ water (0.04% hydrochloric acid) -acetonitrile](ii) a 27% -53% of acetonitrile, 7min) to obtain a compound WX 015.¹HNMR (400MHz, deuterated dimethyl sulfoxide) δ 12.83(s,1H),12.10(s,1H),7.79(s,2H),6.90-6.77(t, J ═ 52.4Hz,1H),4.34-4.32(m,2H),2.67-2.66(m,2H),2.03-2.01(m, 2H). MS-ESI M/z 473.9[ M + H ]]+,475.9[M+H+2] ⁺。

Example 16

The synthetic route is as follows:

step 1: synthesis of Compound WX016-1

WX015-4A (440.00mg,1.27mmol,1eq) and hydrobromic acid (4mL) were charged into a pre-dried reaction flask, the mixture was left at 0 ℃ and a mixed solution of sodium nitrite (175.18mg,2.54mmol,2eq) and water (2mL) was added, the mixture was stirred at 0 ℃ for 0.5 hour, the reaction solution was added dropwise to a stirred mixed solution of cuprous bromide (273.16mg,1.90mmol, 58.00. mu.L, 1.5eq), water (2mL) and hydrobromic acid (4mL), and the mixture was naturally warmed to 25 ℃ and stirred for 15.5 hours. After the reaction is finished, the reaction is carried outThe reaction mixture was added with water (30mL), extracted with ethyl acetate (30mL × 2), the organic phases were combined, washed with saturated sodium bicarbonate (50mL × 2), washed with saturated brine (50mL × 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give WX 016-1.¹HNMR (400MHz, deuterated chloroform) delta 7.43(s,2H),4.42-4.39(m,2H),2.74-2.71(m,2H),2.16-2.13(m, 2H).

Step 2: synthesis of Compound WX016-2

WX016-1(520.00mg,1.27mmol,1eq) was added to a pre-dried reaction flask, followed by dissolution with glacial acetic acid (12mL), followed by addition of sodium acetate (207.84mg,2.53mmol,2eq) and reaction stirred at 110 ℃ for 15 h. After the reaction, adding saturated sodium bicarbonate (50mL), extracting with ethyl acetate (100mL × 2), combining organic phases, washing the organic phases with saturated sodium bicarbonate (100mL), washing with saturated saline (100mL), drying with anhydrous sodium sulfate, concentrating under reduced pressure to obtain a crude product, and pulping the crude product with a mixed solution (10mL) of ethyl acetate and petroleum ether 1:1 to obtain WX 016-2.¹HNMR (400MHz, deuterated chloroform) delta 9.95(s,1H),7.46(s,2H),4.34-4.32(m,2H),2.56-2.53(m,2H),2.07-2.04(m, 2H).

And step 3: synthesis of Compound WX016-3

A pre-dried reaction flask was charged with WX016-2(520.00mg,1.33mmol,1eq), N, N-dimethylformamide (15mL) and potassium carbonate (549.96mg,3.98mmol,3eq), followed by p-methoxybenzyl chloride (311.59mg,1.99mmol, 270.95. mu.L, 1.5eq), purged with nitrogen and stirred at 25 ℃ for 16 hours. After the reaction is finished, adding saturated ammonium chloride solution (70mL), adding ethyl acetate (70mL × 2) for extraction, combining organic phases, washing the organic phase with saturated salt solution (100mL × 4), drying with anhydrous sodium sulfate, filtering, concentrating the filtrate under reduced pressure at 40 ℃ by using a water pump to obtain a crude product, and separating and purifying the crude product by using an automatic column chromatography machine (gradient washing: petroleum ether: ethyl acetate 1:0 to 1:1) to obtain WX 016-3.¹H NMR (400MHz, deuterated chloroform) delta 7.47(s,2H),7.08-7.06(m,2H),6.71-6.69(m,2H),4.83(s,2H),4.28-4.25(m,2H),3.71(s,3H),2.53(s,2H),2.01-1.98(m, 2H).

And 4, step 4: synthesis of Compound WX016-4

Adding WX016-3 into a pre-dried reaction flask(550mg,1.07mmol,1eq) and dioxane (11mL), bis-pinacolato borate (409.03mg,1.61mmol,1.5eq) and potassium acetate (210.77mg,2.15mmol,2eq) were added, nitrogen was replaced three times, 1-bis (diphenylphosphino) ferrocene palladium dichloride (78.57mg, 107.38. mu. mol,0.1eq) was added, nitrogen was replaced three times again, and the mixture was stirred at 100 ℃ for 16 hours. After the reaction is finished, the reaction solution is filtered by diatomite, the filtrate is decompressed and concentrated to obtain a crude product, and the crude product is purified by column chromatography (gradient elution: petroleum ether: ethyl acetate: 10:1 to 5:1 to 1:1) to obtain WX 016-4.¹HNMR (400MHz, deuterated chloroform) δ 1.38(s,12H)2.06(d, J ═ 7.46Hz,2H)2.57-2.63(m,2H)3.78(s,3H)4.32-4.36(d, J ═ 4.9Hz,2H)4.88(s,2H)6.76-6.67(d, J ═ 8.6Hz,2H)7.12-7.16(d, J ═ 8.4Hz,2H)7.81(s, 2H). MS-ESI M/z 559.2[ M + H ]] ⁺。

And 5: synthesis of Compound WX016-5

A predried reactor was charged with WX016-4(600mg,1.07mmol,1eq) and acetone (6mL), water (2.5mL), sodium periodate (229.48mg,1.07mmol, 59.45. mu.L, 1eq), ammonium acetate (595.42mg,7.72mmol,7.2eq) was added, nitrogen was replaced three times, and stirring was carried out at 35 ℃ for 16 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and a saturated sodium thiosulfate solution (20mL) was added to the remaining solution, and the pH was adjusted to 5 with 1M hydrochloric acid, and the filter cake was collected by filtration. Concentrating the filter cake under reduced pressure with oil pump to obtain crude product, and separating by high performance liquid chromatography (chromatographic column: Luna Omega 5u Polar C18100A; mobile phase: [ water (0.04% hydrochloric acid) -acetonitrile](ii) a 28 to 58 percent of acetonitrile for 7min) to obtain the WX 016-5.¹H NMR (400MHz, deuterated dimethyl sulfoxide) δ 1.93-2.00(m,2H)2.42-2.45(t, J ═ 6.07Hz,2H)3.70(s,2H)4.30-4.35(m,2H)4.77(s,2H)6.77-6.79(d, J ═ 8.4Hz,2H)6.97-6.99(d, J ═ 8.4Hz,2H)7.89(s,2H)8.51(s, 2H). MS-ESI M/z 477.1[ M + H ]] ⁺。

Step 6: synthesis of Compound WX016-6

WX016-5(260mg, 544.96. mu. mol,1.1eq) and dichloroethane (10mL) were charged into a pre-dried reaction flask, WX003-6(140.32mg, 495.42. mu. mol,1eq) was added, then copper acetate (134.97mg, 743.12. mu. mol,1.5eq) and pyridine (97.97mg,1.24mmol, 99.97. mu.L, 2.5eq) were added in that order, oxygen was replaced three times, the flask was left at 40 ℃ for 15psi was stirred for 16 hours. After the reaction was completed, the reaction mixture was quenched with water (50mL), extracted with ethyl acetate (50mL × 3), the organic phases were combined, washed with saturated brine (50mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a crude product, which was purified by thin layer chromatography silica gel plate (petroleum ether: ethyl acetate 1:1) to give WX 016-6.¹HNMR (400MHz, deuterated chloroform) δ 2.07-2.13(m,2H)2.60-2.65(m,2H)3.77(s,3H)4.34-4.37(m,2H)4.79(s,2H)4.93(s,2H)5.59(s,2H)6.51-6.74(t, J ═ 52.4Hz,1H)6.77-6.80(m,2H)7.15-7.17(m,2H)7.29-7.39(m,5H)7.61(s, 2H). MS-ESI M/z 714.1[ M + H ]] ⁺。

And 7: synthesis of Compound WX016-7

WX016-6(120mg, 167.95. mu. mol,1eq) and acetonitrile (4mL) were added to a pre-dried reaction flask, a mixed solution of ammonium ceric nitrate (276.22mg, 503.85. mu. mol, 251.11. mu.L, 3eq) and water (2mL) was slowly added dropwise at 0 ℃, and the temperature was slowly raised to 25 ℃ and stirring was continued for 2 hours. After the reaction was completed, the reaction mixture was quenched with water (50mL), extracted with ethyl acetate (55mL × 3), the organic phases were combined, the organic phase was washed with saturated brine (55mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product, which was purified by thin layer chromatography silica gel plate (petroleum ether: ethyl acetate 1:1) to obtain WX 016-7.¹HNMR (400MHz, deuterated chloroform) δ 2.10-2.18(m,2H)2.63-2.66(t, J ═ 6.38Hz,2H)4.38-4.44(m,2H)4.77(s,2H)5.58(s,2H)6.44-6.77(t, J ═ 52.4Hz,1H)7.29-7.41(m,5H)7.58(s,2H)9.39(s, 1H). MS-ESI M/z 594.2[ M + H ]] ⁺。

And 8: synthesis of Compound WX016

WX016-7(90mg, 151.43. mu. mol,1eq) and dichloromethane (3mL) were charged into a reaction flask dried in advance, nitrogen was replaced three times, and boron tribromide (75.87mg, 302.85. mu. mol, 29.18. mu.L, 2eq) was slowly added dropwise thereto at 0 ℃ and stirred at 0 ℃ for 0.5 hour. After the reaction, the pH of the reaction solution was adjusted to about 7 with a saturated sodium bicarbonate solution, ethyl acetate (20mL) was added to dilute the reaction solution, the organic phase was separated, the aqueous phase was extracted with ethyl acetate (20mL 2), the organic phases were combined and washed with a saturated saline solution (20mL 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure with a water pump at 40 ℃ to obtain a crude productThe crude product was separated by HPLC (column: Phenomenex Luna C18200 x 40mm x 10 μm; mobile phase: [ water (0.2% formic acid) -acetonitrile](ii) a 20-50% of acetonitrile for 8min) to obtain the WX 016.¹HNMR (400MHz, deuterated dimethyl sulfoxide) δ 1.95-2.06(m,2H)2.42-2.45(t, J ═ 6.32Hz,2H)4.35-4.37(t, J ═ 4.94Hz,2H)6.70-7.10(t, J ═ 52.4Hz,1H)7.81(s,2H)11.88(s,1H)12.85(s, 1H). MS-ESI M/z 474.0[ M + H ]] ⁺。

Example 17

The synthetic route is as follows:

step 1: synthesis of Compound WX017-1

WX013-1(50g,351.84mmol,1eq), xylene (400mL), sulfolane (207.86g,1.76mol,5eq) was added to the reaction flask and reacted at 135 ℃ for 12 h. And after the reaction is finished, spin-drying the reaction liquid to obtain a crude product. And (3) carrying out gradient elution and purification on the crude product by a silica gel column (petroleum ether: ethyl acetate ═ 3:1 to 1:1) to obtain WX 017-1.

Step 2: synthesis of Compound WX017-2

WX017-1(4.4g,22.43mmol,1eq) was added to a pre-dried reaction flask, then dissolved in tetrahydrofuran (44mL), followed by the addition of sodium iodide (1.68g,11.21mmol,0.5eq), trifluoromethyl trimethylsilane (7.97g,56.07mmol,2.5eq), the reaction was stirred slowly up to 65 ℃ for 4 hours, and trifluoromethyl trimethylsilane (7.97g,56.07mmol,2.5eq) was added, and the reaction was stirred further at 65 ℃ for 15 hours. After the reaction, the reaction solution was spun at 45 ℃ with a water pump, then dissolved in dichloromethane (100mL), washed with water (100mL), 0.1N sodium thiosulfate (100mL) and saturated brine (100mL), and then concentrated under reduced pressure at 45 ℃ with a water pump to give a crude product, which was separated by an automatic column chromatography (gradient elution)Washing: petroleum ether ethyl acetate 1:0 to 9:1) to give WX 017-2.¹HNMR (400MHz, deuterated chloroform) delta 3.77(s,6H),2.77-2.53(m,4H),1.80-1.90(m, 2H).

And step 3: synthesis of Compound WX017-3

WX017-2(3.59g,14.58mmol,1eq) was added to a pre-dried reaction flask, followed by dissolution in methanol (17mL), followed by addition of sodium hydroxide solution (5.83g,14.58mmol,17mL, 10% purity, 1eq), and the reaction stirred at room temperature (25 ℃ C.) for 14 hours. After the reaction is finished, neutralizing the pH value of the reaction solution to about 6 by using 1N hydrochloric acid, and directly concentrating under reduced pressure to obtain WX 017-3.

And 4, step 4: synthesis of Compound WX017-4

WX017-3(3.18g,14.58mmol,1eq) was added to a reaction flask dried in advance, and then dissolved in trifluoroacetic anhydride (32mL), and the reaction was stirred at room temperature (25 ℃ C.) for 14 hours. After the reaction, the reaction solution was concentrated under reduced pressure at 45 ℃ using a water pump to obtain WX 017-4.

And 5: synthesis of Compound WX017-5

WX017-4(2.92g,14.59mmol,1eq) was added to a reaction flask dried in advance, followed by dissolution with acetic acid (58mL), followed by addition of hydrazine hydrate (1.03g,17.51mmol,1.00mL,1.2eq), raising to 100 ℃ and stirring for reaction for 3 hours. After the reaction is finished, cooling the reaction liquid to room temperature, spinning out most of the solvent at 60 ℃ by using a water pump, adding water (20mL) to precipitate a solid, filtering, collecting a filter cake, and spinning the filter cake at 45 ℃ by using an oil pump to obtain WX 017-5.¹H NMR (400MHz, deuterated dimethyl sulfoxide) delta 11.82(s,1H),11.01(s,1H),2.71-2.57(m,4H),2.08-2.17(m, 2H).

Step 6: synthesis of Compound WX017-6

WX017-5(1.28g,5.98mmol,1eq) was added to a previously dried reaction flask, after which it was dissolved in phosphorus oxychloride (12mL), and the reaction was stirred at 110 ℃ for 2 hours. After the reaction was completed, the reaction system was cooled to room temperature, and then it was slowly added to room temperature water (100mL), petroleum ether (50mL) was added to dilute, the organic phase was separated, the aqueous phase was extracted with petroleum ether (50mL × 2), the organic phases were combined and washed with saturated brine (50mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was pumped with a water pump at 45 ℃ under reduced pressureAnd (3) concentrating to obtain a crude product, and purifying the crude product by an automatic column chromatography (gradient elution: petroleum ether: ethyl acetate: 1:0 to 2:8) to obtain WX 017-6.¹HNMR (400MHz, deuterated methanol) delta 3.24-3.01(m,4H),2.19-2.28(m, 2H).

And 7: synthesis of Compound WX017-8

WX017-6(300mg,1.19mmol,1eq), WX017-7(365.58mg,1.31mmol,1.1eq) and toluene (10mL) were charged into a reaction flask dried in advance, potassium phosphate (507.30mg,2.39mmol,2eq) was added, nitrogen was replaced three times, 2-di-t-butylphosphine-2 ', 4', 6 ' -triisopropylbiphenyl (101.48mg, 238.99. mu. mol,0.2eq) and palladium acetate (53.65mg, 238.99. mu. mol,0.2eq) were added, nitrogen was replaced again three times, the mixture was stirred at 50 ℃ for 16 hours, and the temperature was raised to 100 ℃ and stirring was continued for 16 hours. After the reaction, the reaction system was cooled to room temperature, petroleum ether (30mL) was added to the reaction solution, filtered through celite, the filtrate was concentrated under reduced pressure to obtain a crude product, and the crude product was purified by a thin layer chromatography silica gel plate (petroleum ether: petroleum ether ═ 5:1) to obtain WX 017-8.¹HNMR (400MHz, deuterated chloroform) δ 1.53(s,9H)2.12(d, J ═ 11.6Hz,2H)3.05(s,2H)3.19(s,2H)6.53(s,1H)7.47(s, 2H). MS-ESI M/z 492.0[ M + H ]] ⁺。

And 8: synthesis of Compound WX017-9

WX017-8(120mg, 243.54. mu. mol,1eq) and acetic acid (2mL) were added to a reaction flask dried in advance, sodium acetate (39.96mg, 487.08. mu. mol,2eq) was added thereto, nitrogen gas was replaced three times, and the mixture was stirred at 110 ℃ for 16 hours. After the reaction is finished, cooling the reaction liquid to room temperature, and concentrating under reduced pressure to obtain WX 017-9. MS-ESI M/z 416.0[ M + H ]] ⁺。

And step 9: synthesis of Compound WX017-10

WX017-9(100mg, 240.27. mu. mol,1eq) and ethanol (2mL) were added to a pre-dried reaction flask, HCl (2M,2mL,16.65eq) was added, and the mixture was stirred at 90 ℃ for 16 hours. After the reaction is finished, concentrating the reaction solution under reduced pressure, adjusting the pH value to 7-8 by using solid sodium bicarbonate, extracting an aqueous phase (15mL x3) by using ethyl acetate, combining organic phases, washing the organic phase by using saturated salt water (20mL x 2), drying by using anhydrous sodium sulfate, filtering, concentrating under reduced pressure to obtain a crude product, and passing the crude product through a thin-layer chromatography silica gel platePurification (petroleum ether: ethyl acetate ═ 1:5) afforded WX 017-10.¹H NMR (400MHz, deuterated chloroform) delta 1.99-2.10(m,2H)2.92(s,2H)3.05(s,2H)6.68(s,2H)9.24(br s,1H). MS-ESI M/z 374.9[ M + H ]] ⁺。

Step 10: synthesis of Compound WX017-11

WX017-10(48mg, 128.28. mu. mol,1eq) and glacial acetic acid (1mL) were added to a pre-dried reaction flask, concentrated hydrochloric acid (12M, 33.03. mu.L, 3.09eq) was slowly added at 0 ℃, sodium nitrite (9.38mg, 135.98. mu. mol,1.06eq) and water (1mL) and the mixed solution were slowly added, the mixture was stirred at 0 ℃ for 1.5 hours, then WX001-9(22.03mg, 141.11. mu. mol,1.1eq) was added, and stirring was continued at 0 ℃ for 2.5 hours. After the reaction is finished, slowly adding a mixed solution of sodium acetate (3eq) and water (3mL) into the reaction solution at 0 ℃ to generate a large amount of solid, filtering, collecting a filter cake, and concentrating the filter cake under reduced pressure to obtain WX 017-11. MS-ESI M/z 541.1[ M + H ]] ^+,543.1[M+H+2] ⁺。

Step 11: synthesis of Compound WX017

WX017-11(68mg, 125.63. mu. mol,1eq) and N, N-dimethylacetamide (1mL) were charged into a reaction flask which had been dried in advance, potassium acetate (14.18mg, 144.47. mu. mol,1.15eq) was added thereto, nitrogen gas was replaced three times, and the mixture was stirred at 115 ℃ for 6 hours. After the reaction, the reaction solution was directly filtered, and the filtrate was separated by high performance liquid chromatography (column: Phenomenex Luna C18200: 40 mm: 10 μm; mobile phase: [ water (0.2% formic acid) -acetonitrile](ii) a 25 percent to 55 percent of acetonitrile for 8min) to obtain WX 017.¹HNMR (400MHz, deuterated methanol) δ 2.18-2.22(d, J ═ 13.4Hz,2H)2.89(s,2H)3.06-3.21(m,2H)7.77(s, 2H). MS-ESI M/z of 495.0[ M + H ]] ⁺。

Example 18

The synthetic route is as follows:

step 1: synthesis of Compound WX018-1

WX008-3(3g,6.94mmol,1eq) and vinyltri-n-butyltin (3.30g,10.41mmol,3.03mL,1.5eq) were charged in a 50mL single-neck flask, 1, 4-dioxane (30mL) was added, nitrogen was replaced three times, triphenylphosphine (546.09mg,2.08mmol,0.3eq) and palladium acetate (233.72mg,1.04mmol,0.15eq) were added, nitrogen was replaced three times, and the reaction was heated to 80 ℃ and stirred for 12 hours. After completion of the reaction, 20mL of a saturated aqueous solution of potassium fluoride was added to the reaction solution, stirred at room temperature for 1 hour, filtered, the filtrate was extracted with ethyl acetate (50mL × 2), and the combined organic phases were directly concentrated under reduced pressure to give WX018-1, which was not purified and easily broken, and used in the next step.

Step 2: synthesis of Compound WX018-2

WX018-1(5.2g,13.71mmol,1eq), tert-butanol (20mL), water (70mL) and acetonitrile (20mL) were charged into a reaction flask, the nitrogen gas was purged and then cooled to 0 ℃ followed by addition of osmium tetroxide (348.43mg,1.37mmol, 71.11. mu.L, 0.1eq) and sodium periodate (5.86g,27.41mmol,1.52mL,2eq) and reaction of a mixture at 25 ℃ for 3 hours. After completion of the reaction, a saturated aqueous sodium sulfite solution (30mL) was added to the reaction solution, stirred at room temperature for 2 hours, and then extracted with ethyl acetate (50mL × 2), the organic phases were combined, concentrated under reduced pressure to give a crude product, which was purified by a column chromatography (eluent polarity, petroleum ether: ethyl acetate: 20% -30%) to give WX 018-2.¹H NMR (400MHz, deuterated chloroform) δ 9.82(s,1H),7.40-7.42(d, J ═ 8.8Hz,2H),7.24-7.32(m,5H),6.90-6.92(d, J ═ 8.8Hz,2H),5.48(s,2H),5.16(s,2H),4.70(s,2H),3.82(s, 3H).

And step 3: synthesis of Compound WX018-3

WX018-2(1.6g,4.20mmol,1eq) and 1, 2-dichloroethane (20mL) were charged in a 100mL three-necked flask, the temperature was reduced to 0 ℃ and sodium borohydride acetate (1.38g,6.50mmol,1.55eq) was added and after the addition was complete, the mixture was stirred at 25 ℃ for 3 hours. After the reaction is completed. The reaction mixture was slowly added to a saturated aqueous ammonium chloride solution (20mL), extracted with ethyl acetate (20mL × 3), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressureObtaining a crude product, and purifying the crude product by using a flash silica gel column (eluent polarity, petroleum ether: ethyl acetate is 10% -20% -30%) to obtain WX 018-3.¹H NMR (400MHz, deuterated chloroform) δ 7.27-7.38(m,7H),6.88-6.89(d, J ═ 6.4Hz,2H),5.47(s,2H),5.04(s,2H),4.69(s,2H),4.55(s,2H),3.81(s, 3H).

And 4, step 4: synthesis of Compound WX018-4

WX018-3(1.2g,3.13mmol,1eq) was charged into a 40mL storage flask, 1, 2-dichloromethane (20mL) was added, the temperature was reduced to 0 ℃ and diethylaminosulfur trifluoride (504.51mg,3.13mmol, 413.53. mu.L, 1eq) was added, and the mixture was warmed to room temperature (27 ℃ C.) and stirred for 3 hours. After the reaction is completed. Water (10mL) was added to the reaction mixture, extracted with ethyl acetate (10mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a crude product, which was purified on a flash silica gel column (eluent polarity, petroleum ether: ethyl acetate 10% -20% -30%) to give WX 018-4.¹H NMR (400MHz, deuterated chloroform) δ 7.18-7.30(m,7H),6.80-6.82(d, J ═ 8.8Hz,2H),5.39(s,2H),5.20(s,1H),5.08(s,1H),4.97(s,2H),4.61(s,2H),3.73(s, 3H).

And 5: synthesis of Compound WX018-5

WX018-4(0.5g,1.30mmol,1eq) and acetonitrile (10mL) were charged into a 40mL predried storage flask, the temperature was lowered to 0 ℃, ceric ammonium nitrate (1.42g,2.59mmol,1.29mL,2eq) was dissolved in water (5mL), the reaction mixture was added dropwise, and after completion of the addition, the mixture was warmed to room temperature (25 ℃) and stirred for 3 hours. After the reaction is finished, directly concentrating the reaction solution under reduced pressure to obtain a crude product, and purifying the crude product by using a quick silica gel column (eluent polarity, petroleum ether: ethyl acetate is 10% -20% -30%) to obtain WX 018-5.¹HNMR (400MHz, deuterated chloroform) delta 9.54(s,1H),7.31-7.38(m,5H),5.48(s,2H),5.32(s,1H),5.20(s,1H),4.73(s, 2H).

Step 6: synthesis of Compound WX018-6

Adding WX018-5(0.29g,1.09mmol,1eq) and WX005-6(675.33mg,1.42mmol,1.3eq), 1, 2-dichloroethane (2mL) to a 50mL single-neck flask, purging oxygen three times, adding copper acetate (297.88mg,1.64mmol,1.5eq) and pyridine (216.21mg,2.73mmol, 220.62. mu.L, 2.5eq), continuing purging oxygen three times, heating to 40 deg.C, stirring for 12 hr. After the reaction is finished, the reaction liquid is filtered by diatomite, the filtrate is dried by spinning under reduced pressure, and the crude product is purified by a quick silica gel column (the polarity of an eluent, petroleum ether and ethyl acetate account for 10% -20% -30%) to obtain WX 018-6.¹HNMR (400MHz, deuterated chloroform) δ 7.34-7.43(m,7H),7.24-7.26(d, J ═ 8.8Hz,2H),6.79-6.81(d, J ═ 8.8Hz,2H),5.63(s,2H),5.34-4.46(d, J ═ 46.4Hz,2H),4.98(s,2H),4.82(s,2H),3.79(s,3H),2.64-2.73(m,4H),1.65(s, 4H).

And 7: synthesis of Compound WX018-7

WX018-6(0.4g, 575.92. mu. mol,1eq) was charged into a 50mL single-neck flask, dissolved in acetonitrile (12mL), cooled to 0 ℃ and ceric ammonium nitrate (947.20mg,1.73mmol, 861.09. mu.L, 3eq) dissolved in water (4mL) was added dropwise to the reaction mixture, the temperature was maintained at 0 ℃ and after completion of the addition, the mixture was slowly warmed to room temperature (25 ℃) and stirred for 3 hours. After completion of the reaction, water (10mL) was added to the reaction mixture, ethyl acetate (10mL × 3) was added thereto for extraction, and the organic phases were combined, washed with saturated brine (10mL), dried over anhydrous sodium sulfate, filtered, and dried under reduced pressure. The crude product was purified using thin layer chromatography silica gel plates (petroleum ether: ethyl acetate ═ 1:1) to afford WX 018-7.¹HNMR (400MHz, deuterated chloroform) delta 7.30-7.38(m,7H),5.58(s,2H),5.38(s,1H),5.27(s,1H),4.77(s,2H),2.64-2.74(m,4H),1.87(s, 4H).

And 8: synthesis of Compound WX018-8

WX018-7(0.18g, 313.38. mu. mol,1eq) and dichloromethane (2mL) were added to a reaction flask, nitrogen was purged and the temperature was reduced to 0 ℃ followed by addition of boron tribromide (157.02mg, 626.76. mu. mol, 60.39. mu.L, 2eq) and reaction of the mixture at 0 ℃ for 20 minutes. After completion of the reaction, the reaction mixture was quenched by pouring into water (10mL), extracted with dichloromethane (10mL × 5), and the organic phases were combined, washed with saturated brine (20mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give WX 018-8.

And step 9: synthesis of Compound WX018

WX018-8(0.04g, 77.65. mu. mol,1eq), potassium carbonate (21.46mg, 155.30. mu. mol,2eq), dichloromethane (2mL) and water (2mL) were charged into an 8mL thumb bottle, nitrogen was purged three times, the temperature was reduced to 0 ℃ and tetrabutylammonium bromide (375.47. mu.g, 1.16. mu. mol,0.015eq) and trimethylsilyl cyanide were added(15.41mg, 155.30. mu. mol, 19.43. mu.L, 2eq), nitrogen was purged three times, the temperature was raised to 25 ℃ and stirring was carried out for 12 hours. After completion of the reaction, the reaction mixture was directly spin-dried without any post-treatment, dissolved in 1mL of dimethyl sulfoxide, filtered, and the filtrate was separated by high performance liquid chromatography (conditions: column: Luna Omega 5u Polar C18100A; mobile phase: [ water (0.04% hydrochloric acid) -acetonitrile](ii) a Acetonitrile% 33% -57%, 7min) to obtain WX 018.¹H NMR (400MHz, deuterated dimethyl sulfoxide) delta 12.71(s,1H),12.05(s,1H),7.80(s,2H),4.05(s,2H),2.67(s,2H),2.44(s,2H),1.75-1.76(m, 4H). MS-ESI M/z 461.1[ M + H ]] ⁺,463.0[M+H+2] ⁺。

Example 19

The synthetic route is as follows:

step 1: synthesis of Compound WX019-1

WX018-8(100mg, 194.12. mu. mol,1eq), water (0.5mL) and dimethyl sulfoxide (1mL) were added to a pre-dried reaction flask, the nitrogen was purged three times, the temperature was raised to 50 ℃ and stirred for 24 hours. After completion of the reaction, the reaction solution was directly filtered, and the filtrate was separated and purified by high performance liquid chromatography (conditions: column: Welch Xitinate C18150 × 25mm × 5 μm; mobile phase: [ water (0.04% hydrochloric acid) -acetonitrile ]; acetonitrile%: 30% -45%, 10 minutes) to obtain WX 019-1.

Step 2: synthesis of Compound WX019

WX019-1(0.02g, 44.22. mu. mol,1eq) was charged in an 8mL storage flask, methylene chloride (2mL) was added, the temperature was lowered to 0 ℃, diethylaminosulfur trifluoride (10.69mg, 66.34. mu. mol, 8.76. mu.L, 1.5eq) was added, and the temperature was raised to 27 ℃ and stirred for 1 hour. After completion of the reaction, water (10mL) was added to the reaction mixture, extracted with ethyl acetate (10mL × 3), the organic phases were combined,drying the organic phase with anhydrous sodium sulfate, filtering, concentrating under reduced pressure to obtain crude product, separating and purifying by high performance liquid chromatography (conditions: chromatographic column: Phenomenex luna C1880: 40mm × 3 μm; mobile phase: [ water (0.04% hydrochloric acid) -acetonitrile](ii) a 32 percent to 56 percent of acetonitrile for 7 minutes) to obtain WX 019.¹HNMR (400MHz, deuterated dimethyl sulfoxide) δ 12.69(s,1H),12.08(s,1H),7.82(s,2H),5.25-5.36(d, J ═ 46.8Hz,2H),2.67(s,2H),2.44(s,2H),1.77(s, 4H). MS-ESI M/z 454.0[ M + H ]] ⁺,456.0[M+H+2] ⁺。

Example 20

The synthetic route is as follows:

step 1: synthesis of Compound WX020-1

Tetrahydrofuran (1500mL) and WX011-1(25g,252.19mmol,1eq) were charged in a predried three-necked flask, followed by sodium hydride (13.11g,327.85mmol, 60% purity, 1.3eq) at 0 ℃ and stirred for 1 hour, p-methoxybenzyl chloride (51.34g,327.85mmol,44.65mL,1.3eq) was added, stirring was continued for 1 hour, and then the mixture was heated to 80 ℃ and stirred for 12 hours. After completion of the reaction, the reaction solution was quenched by carefully adding ice water (300mL), followed by extraction with ethyl acetate (100mL × 3), the organic phase was washed with a saturated ammonium chloride solution (100mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a crude product, which was then purified by column chromatography (petroleum ether: ethyl acetate ═ 3:1 to 1:1) to give WX 020-1.¹H NMR (400MHz, deuterated dimethyl sulfoxide) delta 7.18-7.15(m,2H)6.90-6.88(m,2H)4.42(s,2H)3.74(s,3H)3.14-3.13(m,2H)2.28-2.27(m,2H)1.66-1.70(m, 4H).

Step 2: synthesis of Compound WX020-2

To a dry 1L round bottom flask was added WX020-1(48 g)218.90mmol,1eq), then toluene (550mL) and Lawson's reagent (89.19g,218.90mmol,1eq) were added and the reaction stirred at 80 ℃ for 2 hours. After the reaction is finished, the reaction solution is directly concentrated under reduced pressure to be dry and then purified by column chromatography (petroleum ether: ethyl acetate: 3:1 to 1:1) to obtain WX 020-2.¹H NMR (400MHz, deuterated dimethyl sulfoxide) delta 7.24-7.19(m,2H)6.79-6.81(m,2H)5.19(s,2H)3.73(s,3H)3.26-3.25(m,2H)3.01-3.00(m,2H)1.66-1.75(m, 4H).

And step 3: synthesis of Compound WX020-3

To a dry 1L round bottom flask was added WX020-2(55g,233.70mmol,1eq), followed by absolute ethanol (550mL) and iodomethane (49.76g,350.55mmol,21.82mL,1.5eq), and the reaction was stirred at 80 ℃ for 1.5 h. After the reaction is finished, the reaction solution is concentrated to be dry, and then tetrahydrofuran (100mL) is added for pulping to obtain WX 020-3.¹H NMR (400MHz, deuterated dimethyl sulfoxide) delta 7.37-7.35(m,2H)7.01-6.98(m,2H)5.03(s,2H)3.78(s,3H)3.75-3.72(m,2H)3.18(s,2H)2.81(s,3H)1.84-1.82(m, 4H).

And 4, step 4: synthesis of Compound WX020-4

WX020-3(10g,26.36mmol,1eq) and tetrahydrofuran (100mL) were added to a 250mL single-neck flask, potassium tert-butoxide (4.44g,39.55mmol,1.5eq) was added, stirring was carried out at room temperature (25 ℃ C.) for 30 minutes, filtration was carried out, the filtrate was spin-dried under reduced pressure, toluene (50mL) was added, filtration was carried out, a dry 250mL three-neck flask was prepared, WX007-1(1.99g,13.18mmol,0.5eq) and toluene (30mL) were added, nitrogen was replaced three times, the temperature was reduced to 0 ℃ C., the toluene solution just filtered was added dropwise, the temperature was maintained at 0 ℃ C., and after completion of the dropwise addition, the temperature was raised to 25 ℃ C. and stirring was carried out for 12 hours. After the reaction is finished, adding water (100mL), extracting with ethyl acetate (100 mL. times.5), combining organic phases, washing the organic phases with saturated saline (200mL), drying with anhydrous sodium sulfate, filtering, concentrating under reduced pressure to obtain a crude product, and separating and purifying the crude product by using a flash silica gel column (eluent polarity, petroleum ether: ethyl acetate is 10% -20% -30%) to obtain WX 020-4.¹HNMR (400MHz, deuterated chloroform) δ 1.87-1.92(m,2H)2.75-2.78(m,2H)3.11-3.14(m,2H)3.83(s,3H)4.61(s,2H)6.91-6.93(d, J ═ 8.4Hz,2H)7.30-7.33(d, J ═ 8.8Hz, 2H).

And 5: synthesis of Compound WX020-5

WX020-4(3g,9.25mmol,1eq) and dichloromethane (50mL) were added to a previously dried reaction flask, nitrogen was replaced three times, trifluoroacetic acid (23.10g,202.59mmol,15.00mL,21.89eq) was slowly added dropwise, and stirring was continued at 25 ℃ for 3 hours. After the reaction is finished, concentrating the reaction solution under reduced pressure, adjusting the pH value to 7-8 with a saturated sodium bicarbonate solution, extracting with ethyl acetate (100mL × 3), combining organic phases, drying with anhydrous sodium sulfate (100mL × 2), collecting the organic phases, drying with anhydrous sodium sulfate, filtering, concentrating under reduced pressure to obtain a crude product, and purifying the crude product by column chromatography (petroleum ether: ethyl acetate: 10: 1: 5: 1: 2:1) to obtain WX 020-5.¹H NMR (400MHz, deuterated chloroform) δ 1.98-2.05(m,2H)2.75-2.78(t, J ═ 6.4Hz,2H)3.48-3.51(m,2H)5.17(s, 1H).

Step 6: synthesis of Compound WX020-6

In a previously dried reaction flask, WX020-5(500mg,2.45mmol,1eq) and dichloromethane (10mL) were added di-tert-butyl carbonate (802.16mg,3.68mmol, 844.38. mu.L, 1.5eq) and 4-dimethylaminopyridine (598.70mg,4.90mmol,2eq) and the nitrogen was replaced three times, and the mixture was stirred at 25 ℃ for 16 hours. After the reaction, the reaction solution was cooled to room temperature, dichloromethane (30mL) and water (30mL) were added, liquid separation was performed, the organic phase was collected, the aqueous phase was extracted with dichloromethane (30mL × 3), the organic phases were combined, the organic phase was washed with saturated brine (50mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (petroleum ether: ethyl acetate: 10: 1: 5:1) to obtain WX 020-6. MS-ESI M/z 304.1[ M + H ]] ⁺。

And 7: synthesis of Compound WX020-7

WX020-6(400mg,1.32mmol,1eq), WX001-4(280.92mg,1.58mmol,1.2eq) and N, N-dimethylacetamide (4mL) were added to a pre-dried reaction flask, cesium carbonate (535.59mg,1.64mmol,1.25eq) was added and nitrogen was replaced three times, and stirring was continued at 65 ℃ for 16 hours. After completion of the reaction, the reaction mixture was cooled to room temperature, ethyl acetate (250mL) and water (250mL) were added thereto, the mixture was separated, the organic phase was collected, the aqueous phase was extracted with ethyl acetate (250 mL. times.3), the organic phases were combined, the organic phase was washed with saturated brine (400 mL. times.3), dried over anhydrous sodium sulfate, filtered, and reduced pressure was applied theretoConcentrating to obtain crude product, and purifying by column chromatography (petroleum ether: ethyl acetate: 10: 1: 5: 1: 2:1) to obtain WX 020-7. MS-ESI M/z 445.0[ M + H ]] ⁺。

And (3) identifying the structure of WX 020-7:

WX20-7(160mg, 358.96. mu. mol,1 eq.) and hydrobromic acid (3mL) were added to a pre-dried reaction flask, the flask was left at 0 ℃ and NaNO was added₂A mixed solution of (29.72mg, 430.76. mu. mol,1.2eq) and water (1.5mL) was stirred at 0 ℃ for 0.5 hour, a mixed solution of cuprous bromide (56.64mg, 394.86. mu. mol, 12.03. mu.L, 1.1eq) and hydrobromic acid (3mL) was added dropwise to the above reaction mixture, and the mixture was naturally warmed to 25 ℃ and stirred for 15.5 hours. After the reaction was completed, the reaction was quenched with water (10mL), the aqueous phase (15mL × 3) was extracted with dichloromethane, the organic phases were combined, washed with saturated brine (15mL × 2), the organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a crude product, which was purified by thin layer chromatography silica gel plate (petroleum ether: ethyl acetate ═ 2:1) to give WX 020-7A.

WX020-7A (40.00mg, 97.68. mu. mol,1eq) and methanol (3mL) were charged to a pre-dried reaction flask, wet palladium on carbon (20mg, 976.82. mu. mol, 10% pure, 10eq) was added to replace hydrogen three times, and the mixture was placed under a pressure of a hydrogen balloon (15psi) at 25 ℃ for 1 hour. After the reaction is finished. And directly filtering the reaction solution through diatomite, and concentrating the filtrate under reduced pressure to obtain a crude product. The crude product was separated by HPLC (column: Welch XTimate C18150 mm 25mm 5 μm; mobile phase: [ water (0.04% hydrochloric acid) -acetonitrile](ii) a 30 to 50 percent of acetonitrile) for 10 minutes) to obtain WX020-7B, and detecting hydrogen H on carbon (160) connected with oxygen and a nitrogen six-membered ring on the pyridazine ring and detecting hydrogen H on HMBC (400MHz, deuterated chloroform)₁(2.96) and the correct structure of WX020-7 is proved.¹H NMR (400MHz, deuterated methanol) δ 8.46(s,1H),7.56(d, J ═ 8.13Hz,2H),7.32-7.41(m,1H),3.50-3.57(m,2H),2.96(m,2H),2.05(m, 2H).

And 8: synthesis of Compound WX020-8

WX020-7(150.00mg, 336.53. mu. mol,1eq) and glacial acetic acid (2mL) were added to a pre-dried reaction flask, sodium acetate (138.03mg,1.68mmol,5eq) was added, the mixture was replaced with nitrogen three times, and the mixture was stirred at 120 ℃ for 48 hours. After the reaction is finished, cooling the reaction liquid to room temperature, and concentrating under reduced pressure to obtain WX 020-8. MS-ESI M/z 369.0[ M + H ]] ⁺。

And step 9: synthesis of Compound WX020-9

WX020-8(120.00mg, 325.03. mu. mol,1eq) and ethanol (2mL) were added to a pre-dried reaction flask. Dilute hydrochloric acid (2M,2mL,12.31eq) was added, nitrogen was replaced three times, and the mixture was stirred at 90 ℃ for 16 hours. After the reaction was completed, the reaction mixture was cooled to room temperature, water (20mL) was added, ethyl acetate was used for extraction (20mL × 3), the organic phases were combined, the organic phase was washed with saturated brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product, which was purified by thin layer chromatography silica gel plate (petroleum ether: ethyl acetate: 0:1) to obtain WX 020-9. MS-ESI M/z 327.0[ M + H ]] ⁺。

Step 10: synthesis of Compound WX020-10

WX020-9(70.00mg, 213.96. mu. mol,1eq) and glacial acetic acid (1mL) were added to a pre-dried reaction flask, concentrated hydrochloric acid (12M, 55.09. mu.L, 3.09eq) was added at 0 ℃, a solution of sodium nitrite (15.65mg, 226.80. mu. mol,1.06eq) in water (1mL) was slowly added, the mixture was stirred at 0 ℃ for 1.5 hours, WX001-9(36.75mg, 235.35. mu. mol,1.1eq) was added, and then stirring was continued at 0 ℃ for 2.5 hours. After the reaction is finished, slowly adding a mixed solution of sodium acetate (3eq) and water (3mL) into the reaction solution at 0 ℃, generating a large amount of solid, filtering, collecting a filter cake, and concentrating the filter cake under reduced pressure to obtain WX 020-10. MS-ESI M/z 493.9[ M + H ]] ⁺。

Step 11: synthesis of Compound WX020

WX020-10(100.00mg, 202.31. mu. mol,1eq) and N, N-dimethylacetamide (2mL) were added to a pre-dried reaction flask, potassium acetate (22.83mg, 232.66. mu. mol,1.15eq) was added thereto, nitrogen was replaced three times, and stirring was continued at 115 ℃ for 6 hours. After the reaction was completed, the reaction solution was directly filtered, and the filtrate was separated by high performance liquid chromatography (column: Welch Xitinate C18150 mm 25mm 5 μm; mobile phase)[ water (0.04% hydrochloric acid) -acetonitrile](ii) a 30-50 percent of acetonitrile for 10min) to obtain WX 020.¹H NMR (400MHz, deuterated dimethyl sulfoxide) delta 1.83-1.86(m,2H)2.62-2.65(m,2H)3.37-3.40(m,2H)6.97(s,1H)7.73(s,2H)11.77(s,1H)13.25(s, 1H). MS-ESI M/z 448.0[ M + H ]] ⁺。

Example 21

The synthetic route is as follows:

step 1: synthesis of Compound WX021-1

WX001-3(10g,49.24mmol,1eq) and 1.2-dichloroethane (150mL) were charged to a pre-dried reaction flask, bromosuccinimide (7.89g,44.32mmol,0.9eq) and azobisisobutyronitrile (808.63mg,4.92mmol,0.1eq) were added, nitrogen was replaced three times, and the mixture was stirred in an oil bath at 70 ℃ for 3 hours. After the reaction was completed, the reaction mixture was diluted with water (50mL), and the organic phase was collected, the aqueous phase was extracted with dichloromethane (50mL × 2), the organic phases were combined and washed with saturated brine (50mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (petroleum ether: ethyl acetate: 1: 0: 40: 1: 20:1), and slurried with methyl tert-butyl ether (10mL) to obtain WX 021-1.¹HNMR (400MHz, deuterated chloroform) delta 1.88-1.91(m,1H)2.06-2.10(m,2H)2.47-2.51(m,1H)2.76-2.78(m,1H)3.06-3.12(m,1H)5.25-5.44(m, 1H).

Step 2: synthesis of Compound WX021-2

WX021-1(1.2g,4.26mmol,1eq) and methanol (10mL) were added to a reaction flask, followed by the addition of sodium methoxide (574.79mg,10.64mmol,2.5eq), after which nitrogen was replaced three times and stirring was continued at 25 ℃ for 8 hours. After the reaction is completed, the reaction solution is decompressed and concentrated to obtain a crude product, and the crude product is purified by column chromatography (gradient elution: petroleum ether: ethyl acetate: 1: 0: 40: 1: 6:1) to obtain WX 021-2.

And step 3: synthesis of Compound WX021-3

WX021-2(2.1g,9.01mmol,1eq) and WX001-4(1.92g,10.81mmol,1.2eq) and N, N-dimethylacetamide (18mL) were charged to a pre-dried reaction flask, then replaced three times with nitrogen, followed by addition of cesium carbonate (4.40g,13.51mmol,1.5eq), and after the addition was complete, stirring was continued at 70 ℃ for 8 hours. After completion of the reaction, the reaction solution was poured into ice water (80mL), and then the pH of the solution was adjusted to 5 with diluted hydrochloric acid 2N, extracted with methyl tert-butyl ether (100mL × 3), washed with saturated brine (30mL × 5), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give a crude product, which was purified by flash column chromatography (petroleum ether: ethyl acetate ═ 50:1 to 20:1 to 1:1) to give a crude product. Then, separation and purification were carried out by high performance liquid chromatography (purification conditions: column: Phenomenex luna C18250 x 50mm x 10 μm; mobile phase: water (0.1% trifluoroacetic acid) -acetonitrile; acetonitrile%: 55% -55%, 20 minutes) to obtain WX 021-3. 1HNMR (400MHz, deuterated chloroform) delta 1.49-1.51(m,1H)1.85-1.89(m,2H)2.28-2.32(m,1H)2.53-2.56(m,1H)2.95-3.00(m,1H)3.52(s,3H)3.66(s,2H)4.31(s,1H)6.57(s, 2H).

And (3) identifying the structure of WX 021-3:

WX021-3(0.05g, 133.46. mu. mol,1eq) and methanol (2mL) were charged in a 50mL single-neck flask, wet palladium on carbon (0.01g, 133.46. mu. mol, 10% purity, 1eq) was added, hydrogen was replaced three times, and then a reaction was carried out at 20 ℃ for 2 hours under a pressure of a hydrogen balloon (15 psi). After completion of the reaction, the reaction mixture was directly filtered with celite, and the filtrate was spin-dried under reduced pressure to give a crude product, which was purified by high performance liquid chromatography (column: Welch Xitimate C18150 mm 25mm 5 μm; mobile phase: [ water (0.04% hydrochloric acid) -acetonitrile](ii) a 30 to 50 percent of acetonitrile for 10 minutes) to obtain WX 021-3A. NOE (400MHz, deuterated dimethyl sulfoxide) shows hydrogen and H on methoxy (3.43)₁(8.94) there is a correlation, which proves that the structure of WX021-3 is correct.¹HNMR (400MHz, deuterated dimethyl sulfoxide) delta 1.84-1.94(m,4H),2.69-2.82(m,2H),3.43(s,3H),4.42-4.44(m,1H),6.67(br s,2H),6.91(s,2H), 8.94(s, 1H).

And 4, step 4: synthesis of Compound WX021-4

WX021-3(600mg,1.60mmol,1eq) and sodium acetate (262.74mg,3.20mmol,2eq) and acetic acid (12mL) were added to a pre-dried reaction flask, and after the addition was complete, stirring was continued at 110 ℃ for 8 hours. After the raw materials are completely reacted, directly carrying out reduced pressure concentration on the reaction solution to obtain WX 021-4.

And 5: synthesis of Compound WX021-5

WX021-4(1.60g,4.02mmol,1eq) and ethanol (10mL) and hydrochloric acid (10mL) were added to a pre-dried reaction flask, and after the addition was complete, stirring was continued at 90 ℃ for 8 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, ethyl acetate (20mL) and water (30mL) were added, the pH of the solution was adjusted to 6 to 7 with solid sodium bicarbonate, the aqueous phase was separated, the aqueous phase was extracted with ethyl acetate (20mL × 2), the organic phases were combined, washed with saturated brine (20mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give WX 021-5. 1HNMR (400MHz, deuterated chloroform) delta 1.45-1.48(m,1H)1.82-1.86(m,2H)2.20-2.21(m,1H)2.24-2.48(m,1H)2.85-2.86(m,1H)3.53(s,3H)4.42(s,1H)6.61(s,2H)9.96(s, 1H).

Step 6: synthesis of Compound WX021-6

WX021-5(0.18g, 505.33. mu. mol,1eq), water (4mL), acetic acid (2mL) and concentrated hydrochloric acid (12M, 130.54. mu.L, 3.1eq) were charged to a reaction flask, a solution of sodium nitrite (38.35mg, 555.86. mu. mol,1.1eq) in water (2mL) was added dropwise at 0 ℃ after the completion of the charge, and then stirring was continued at 0 ℃ for 1 hour, then WX001-9(86.79mg, 555.86. mu. mol,1.1eq) was added, and after the completion of the charge, stirring was continued at 0 ℃ for 1 hour. After the reaction of the raw materials was completed, a saturated sodium acetate solution (10mL) was added to the reaction solution, at which time a large amount of solid precipitated, and then filtered with a five-hole funnel, and the filter cake was collected and concentrated under reduced pressure with an oil pump to obtain WX 021-6.

And 7: synthesis of Compound WX021-7

WX021-6(0.19g, 363.06. mu. mol), potassium acetate (71.26mg, 726.13. mu. mol,2eq) and N, N-dimethylacetamide (2mL) were added to a pre-dried reaction flask, and stirring was continued at 115 ℃ for 4 hours after the addition. After the reaction was completed, the reaction solution was cooled to room temperature, followed by filtration, and the filtrate was purified by preparative high performance liquid chromatography (column: Phenomenex Luna C18100: 30 mm: 5 μm; mobile phase: [ water (0.2% trifluoroacetic acid) -acetonitrile ];% acetonitrile: 35% -55%, 9 minutes) to obtain WX 021-7.

And 8: synthesis of Compounds WX021 and WX022

WX021-7 was separated by supercritical chromatography (column: DAICEL CHIRALPAK AD (250 mm. about.30 mm,10 μm), mobile phase: [ carbon dioxide-isopropanol, 0.1% ammonia ], isopropanol:% 53% -53%, 7min ] to obtain the desired compounds WX021 and WX 022.

WX021： ¹HNMR (400MHz, deuterated methanol) δ 1.56-1.61(m,1H),1.91-1.93(m,2H),2.29-2.33(d, J ═ 14.4Hz,1H),2.59-2.66(m,1H),2.92-2.97(d, J ═ 19.2Hz,1H),3.54(s,3H),4.43(s,1H),7.77(s, 2H). Retention time 1.46 min (instrument: Waters UPCC with PDA stripper; chromatography column: Chiralpak AD-3,50 x 4.6mm, i.d.,3 μm; mobile phase: a: food grade supercritical carbon dioxide; B: isopropanol (0.1% diethylamine, volume ratio); gradient: B content rises from 5% to 50% in 1.2 min and stays for 1 min, then rises from 50% to 5% in 0.8 min; flow rate: 3.4 mL/min; column temperature: 35 ℃; detection wavelength: 220 nm; system back pressure: 100 bar).

WX022： ¹HNMR (400MHz, deuterated methanol) δ 1.56-1.61(m,1H),1.91-1.93(m,2H),2.29-2.33(d, J ═ 14.4Hz,1H),2.59-2.66(m,1H),2.92-2.97(d, J ═ 19.2Hz,1H),3.54(s,3H),4.43(s,1H),7.77(s, 2H). Retention time 1.71 min (instrument: Waters UPCC with PDA separator; chromatography column: Chiralpak AD-3,50 x 4.6mm, i.d.,3 μm; mobile phase: a: food grade supercritical carbon dioxide; B: isopropanol (0.1% diethylamine, volume ratio); gradient: B content rises from 5% to 50% in 1.2 min and stays for 1 min, then rises from 50% to 5% in 0.8 min; flow rate: 3.4 mL/min; column temperature: 35 ℃; detection wavelength: 220 nm; system back pressure: 100 bar).

Example 23

The synthetic route is as follows:

step 1: synthesis of Compound WX023-3

To a pre-dried reaction flask was added dimethyl sulfoxide (50mL), followed by sodium hydride (2.29g,57.22mmol, 60% purity, 1.1eq), followed by the slow addition of WX023-2(12.59g,57.22mmol,1.1eq), the reaction was stirred at room temperature (20 ℃) for 1 hour, followed by the addition of a solution of WX023-1(5g,52.01mmol,5.04mL,1eq) in dimethyl sulfoxide (10mL), followed by the slow increase to 50 ℃ and the reaction stirred for 2 hours. After completion of the reaction, water (200mL) and petroleum ether (100mL) were added for dilution, the organic phase was separated, the aqueous phase was extracted with petroleum ether (100mL × 2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give WX 023-3.¹H NMR (400MHz, deuterated chloroform) delta 2.15-2.30 (m,1H),1.85-1.90(m,1H),1.71-1.62(m,2H),1.60-1.52(m,1H),1.20-1.13(m,2H),1.06-1.00(m,1H),0.85-0.74(m, 2H).

Step 2: synthesis of Compound WX023-4

WX023-3(2.5g,22.70mmol,1eq) was added to a pre-dried reaction flask, then dissolved with tetrahydrofuran (37.5mL), the reaction was cooled to-78 ℃, lithium diisopropylamide (2M,14.18mL,1.25eq) was added, the reaction was stirred at 78 ℃ for 0.5 hour, then chlorotrimethylsilane (2.96g,27.23mmol,3.46mL,1.2eq) was added, and the reaction was slowly raised to room temperature (25 ℃) and stirred for 1 hour. After completion of the reaction, it was slowly added to ice water (50mL), followed by dilution with ethyl acetate (50mL), the organic phase was separated, the aqueous phase was extracted with ethyl acetate (50mL × 2), the organic phases were combined and washed with saturated brine (50mL × 2), dried over anhydrous sodium sulfate, filtered to remove the drying agent, and concentrated under reduced pressure to give WX 023-4.

And step 3: synthesis of Compound WX023-5

In advance of dryingAfter adding WX007-1(1.66g,10.97mmol,0.5eq) and dissolving in toluene (40mL), WX023-4(4g,21.94mmol,1eq) was added, and the reaction was stirred for 1 hour after heating to 60 ℃. After completion of the reaction, the reaction system was cooled to room temperature (25 ℃ C.), diluted with water (50mL) and ethyl acetate (50mL), the organic phase was separated, the aqueous phase was extracted with ethyl acetate (10 mL. times.2), the organic phases were combined and washed with saturated brine (50 mL. times.2), dried over anhydrous sodium sulfate, filtered to remove the drying agent, and concentrated under reduced pressure to give a crude product, which was purified by automatic column chromatography (gradient elution: PE: EA: 1:0 to 80:20) to give WX 023-5.¹HNMR (400MHz, deuterated chloroform) delta 3.08-2.95(m,1H),2.38-2.19(m,3H),1.97-1.85(m,1H),1.76-1.69(m,1H),1.36-1.26(m,1H),1.04-0.97(m, 1H).

And 4, step 4: synthesis of Compounds WX023-6 and WX023-7

WX023-5(1.00g,4.65mmol,1eq), WX017-7(1.42g,5.12mmol,1.1eq) and toluene (25mL) were added to a pre-dried reaction flask, potassium phosphate (1.97g,9.30mmol,2eq) was added to displace nitrogen three times, then 2-di-tert-butylphosphine-2 ', 4', 6 ' -triisopropylbiphenyl (394.87mg, 930.00. mu. mol,0.2eq) and palladium acetate (208.77mg, 930.00. mu. mol,0.2eq) were added to displace nitrogen three times again, and stirred at 50 ℃ for 16 hours. The reaction was then stirred for 16 hours at 100 ℃. After completion of the reaction, water (100mL) and ethyl acetate (100mL) were added to dilute the mixture, insoluble matter was present, the mixture was filtered through a funnel packed with celite, the filtrate was separated into an organic phase, the aqueous phase was extracted with ethyl acetate (100 mL. times.2), the organic phases were combined and washed with saturated brine (100 mL. times.2), dried over anhydrous sodium sulfate, filtered to remove the drying agent, and concentrated under reduced pressure to give a crude product, which was purified by automatic column chromatography (gradient elution: PE: EA: 1:0 to 10:90) to give a mixture of WX023-6 and WX 023-7. Then separating by high performance liquid chromatography column (chromatographic column: Phenomenex luna C18250 mm 10 μm; mobile phase: [ water (0.05% hydrochloric acid) -acetonitrile](ii) a 65-95 percent of acetonitrile for 10min) to obtain WX023-6 and WX 023-7. And after the WX023-6 and the WX023-7 are respectively subjected to Pd/C hydrogenation, chlorine adjacent to a nitrogen atom of a pyridazine ring is replaced by a hydrogen atom, and the structure is judged to be shown by two-dimensional nuclear magnetism. WX 023-6:¹HNMR (400MHz, deuterated chlorine)Imitation) delta 7.46(s,2H),6.66(br s,1H),3.07-2.94(m,1H),2.41-2.49(m,1H),2.37-2.24(m,2H),1.92-1.75(m,2H),1.52(s,9H),1.34-1.28(m,1H),1.07-1.00(m, 1H). WX 023-7:¹HNMR (400MHz, deuterated chloroform) delta 7.43(s,2H),6.68(br s,1H),3.22-3.10(m,1H),2.37-2.24(m,3H), 1.84-1.94(m, 1H),1.83-1.72(m,1H),1.51(s,9H),1.31-1.26(m,1H),1.09-1.00(m, 1H).

Structural identification of WX 023-6:

WX023-6(25.00mg, 54.73. mu. mol,1eq) was added to a pre-dried reaction flask, after which it was dissolved with methanol (1mL), hydrogen was purged three times, Pd/C (20mg, 10% purity) was added, hydrogen was purged three times again, and placed under pressure of hydrogen balloon (15psi) at 20 ℃ for 2 hours. After the reaction is finished, the reaction liquid is filtered and decompressed and concentrated to obtain a crude product. The crude product was separated by high performance liquid chromatography (column: Phenomenex luna C1880 x 40mm x3 μm; mobile phase: [ water (0.04% hydrochloric acid) -acetonitrile](ii) a Acetonitrile percent 46-72 percent and 7min) to obtain WX 023-6A. NOE (400MHz, deuterated methanol) for H detection₁Hydrogen and H₂Hydrogen was correlated, confirming that WX023-6 was structurally correct. WX 023-6A:¹h NMR (400MHz, deuterated methanol) delta 8.91(s,1H),7.71-7.79(m,1H),7.44-7.37(m,1H),7.32-7.28(m,1H),2.99-2.89(m,1H),2.70-2.59(m,2H),2.44-2.35(m,1H),2.20-2.12(m,1H),1.83-1.71(m,1H),1.59-1.55(m,1H),1.53(s,9H),1.49-1.41(m, 1H).

Structural identification of WX 023-7:

WX023-7(50.00mg, 109.47. mu. mol,1eq) was added to a pre-dried reaction flask, after which it was dissolved with methanol (2mL), hydrogen was exchanged three times, wet Pd/C (20mg, 10% pure) was added, hydrogen was exchanged three times again, and placed under pressure of hydrogen balloon (15psi) at 20 ℃ for 2 hours.After the reaction is finished, the reaction solution is filtered, and the filtrate is decompressed and concentrated to obtain a crude product. The crude product was separated by high performance liquid chromatography (column: Welch Xtimate C18150 mm 25mm 5 μm; mobile phase: [ water (0.04% hydrochloric acid) -acetonitrile](ii) a 50-80% of acetonitrile for 8min) to obtain WX 023-7A. NOE (400MHz, deuterated methanol) for H detection₁Hydrogen and H₂Hydrogen was correlated, confirming that WX023-7 was structurally correct. WX023-7A¹H NMR (400MHz, deuterated methanol) delta 9.07(s,1H),7.61(s,2H),3.26-3.14(m,1H),2.54-2.36(m,2H),2.25-2.17(m,1H),2.04-1.96(m,1H),1.82-1.71(m,1H),1.53(s,9H),1.38-1.27(m,1H),1.27-1.19(m, 1H).

And 5: synthesis of Compound WX023-8

WX023-6(0.097g, 212.37. mu. mol,1eq) was added to a pre-dried reaction flask, then dissolved with acetic acid (1mL), then sodium acetate (34.84mg, 424.74. mu. mol,2eq) was added, and the reaction was stirred slowly rising to 110 ℃ for 12 hours. After the reaction was completed, the reaction solution was cooled to room temperature, the pH of the reaction solution was adjusted to about 7 with a saturated sodium bicarbonate solution, ethyl acetate (20mL) was added to dilute the reaction solution, the organic phase was separated, the aqueous phase was extracted with ethyl acetate (20mL × 2), the organic phases were combined and washed with a saturated saline solution (20mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the objective compound WX 023-8.

Step 6: synthesis of Compound WX023-9

WX023-8(80.00mg, 210.40. mu. mol,1eq) was added to a pre-dried reaction flask, then dissolved with ethanol (1.2mL), then hydrochloric acid (2M,1.26mL,11.95eq) was added, and the reaction was stirred for 15 hours slowly raising to 95 ℃. After the reaction was completed, the reaction solution was centrifuged out of ethanol by a water pump at 50 ℃, the pH of the reaction solution was adjusted to about 7 with a saturated sodium bicarbonate solution, ethyl acetate (20mL) was added for dilution, the organic phase was separated, the aqueous phase was extracted with ethyl acetate (20mL × 2), the organic phases were combined and dried with anhydrous sodium sulfate, and after removing the drying agent by filtration, the reaction solution was concentrated under reduced pressure to obtain WX 023-9.

And 7: synthesis of Compound WX023-10

WX023-9(70.00mg,206.99 mu mol,1eq) is added into a reaction bottle which is dried in advance, then dissolved by acetic acid (1mL), then hydrochloric acid (62.82mg,637.52 mu mol,61.59 mu L, 37% purity, 3.08eq) is added, the reaction system is cooled to 5 ℃, a solution of sodium nitrite (15.71mg,227.68 mu mol,1.1eq) in water (0.5mL) is added, the reaction is stirred at 5 ℃ for 0.5 hour, WX001-9(35.55mg,227.68 mu mol,1.1eq) is added after the detection of the reaction of the raw materials is finished, and then the stirring is continued at 5 ℃ for 0.5 hour. After completion of the reaction, a solution of sodium acetate (51mg) in water (1.5mL) was added to the reaction mixture to find a solid precipitated, water (10mL) was added thereto, filtration was carried out to collect a cake, the cake was washed with water (10 mL. times.3), and the cake was spin-dried to give WX 023-10.

And 8: synthesis of Compound WX023-11

WX023-10(73.00mg, 144.47. mu. mol,1eq) was added to a pre-dried reaction flask, then dissolved with N, N-dimethylacetamide (0.5mL), followed by addition of potassium acetate (21.27mg, 216.70. mu. mol,1.5eq) and the reaction stirred at 115 ℃ for 2 hours. After the reaction was completed, the reaction mixture was filtered and purified by high performance liquid chromatography (column: Phenomenex luna C1880: 40 mm: 3 μm; mobile phase: water (0.04% hydrochloric acid) -acetonitrile; acetonitrile%: 35% -55%, 7min) to obtain WX 023-11.

And 8: synthesis of Compounds WX023 and WX024

WX023-11 WX023 and WX024 were obtained by supercritical chromatography (chromatography column: Chiralpak AS,250 x 30mm i.d.10 μm; mobile phase: a carbon dioxide B methanol (0.1% ammonia), Gradient: methanol% ═ 50% isocratic elution mode; flow rate: 70 g/min; column temperature: 35 ℃; system back pressure: 150 bar).

WX023： ¹H NMR (400MHz, deuterated methanol) delta 7.75(s,2H),2.90-2.97(m,1H),2.26-2.34(m,2H),1.99-2.11(m,1H),1.85-1.92(m,1H),1.64-1.73(m,1H), 1.27-1.32 (m,1H),1.10-1.15(m, 1H). MS-ESI M/z 459.1[ M + H ]] ⁺,461.1[M+H+2] ⁺. Retention time 1.478 min (instrument: Waters UPCC with PDA separator; chromatography column: Chiralpak AS-3,50 x 4.6mm, i.d.,3 μm; mobile phase: a: food grade supercritical carbon dioxide; B: methanol (0.05% diethylamine, volume ratio); gradient: B content rises from 5% to 50% in 1.2 min, 50% is maintained for 1 min, then falls from 50% to 5% in 0.8 min; flow rate: 3.4 mL/min; column temperature: 35 ℃; detection wavelength: 220 nm; system back pressure: 100 bar).

WX024： ¹H NMR (400MHz, deuterated methanol) delta 7.75(s,2H),2.90-2.96(m,1H),2.27-2.32(m,2H), 1.99-2.05 (m,1H),1.85-1.92(m,1H),1.68-1.70(m,1H), 1.32-1.27 (m,1H),1.10-1.14(m, 1H). MS-ESI M/z 459.1[ M + H ]] ⁺,461.1[M+H+2] ⁺. Retention time 1.696 minutes (instrument: Waters UPCC with PDA separator; column: Chiralpak AS-3,50 x 4.6mm, i.d.,3 μm; mobile phase: a: food grade supercritical carbon dioxide; B: methanol (0.05% diethylamine, volume ratio); gradient: B content rises from 5% to 50% in 1.2 minutes, 50% is maintained for 1 minute, then falls from 50% to 5% in 0.8 minutes; flow rate: 3.4 mL/min; column temperature: 35 ℃; detection wavelength: 220 nm; system back pressure: 100 bar).

Example 25

The synthetic route is as follows:

step 1: synthesis of Compound WX025-1

WX023-7(0.27g, 591.13. mu. mol,1eq) was added to a pre-dried reaction flask, after which it was dissolved with acetic acid (4mL), then sodium acetate (96.98mg,1.18mmol,2eq) was added and the reaction was stirred slowly raising to 110 ℃ for 12 hours. After the reaction was completed, the reaction solution was cooled to room temperature, the pH of the reaction solution was neutralized with a saturated sodium bicarbonate solution to about 7, ethyl acetate (20mL) was added to dilute the reaction solution, the organic phase was separated, the aqueous phase was extracted with ethyl acetate (20mL × 2), the organic phases were combined and washed with a saturated saline solution (20mL × 2), dried over anhydrous sodium sulfate, filtered to remove the drying agent, and concentrated under reduced pressure to obtain the objective compound WX 025-1.

Step 2: synthesis of Compound WX025-2

WX025-1(0.22g, 578.61. mu. mol,1eq) was added to a pre-dried reaction flask, then dissolved in ethanol (3.3mL), followed by hydrochloric acid (2M,3.30mL,11.41eq) and the reaction was stirred slowly to 95 ℃ for 15 hours. After the reaction is finished, ethanol is spun out of the reaction solution at 50 ℃ by a water pump, the pH of the reaction solution is neutralized to about 7 by saturated sodium bicarbonate solution, 20mL of ethyl acetate is added for dilution, the organic phase is collected by liquid separation, the aqueous phase is extracted by ethyl acetate (20mL by 2), the organic phase is combined and dried by anhydrous sodium sulfate, and after a drying agent is removed by filtration, the mixture is concentrated under reduced pressure to obtain WX 025-2.

And step 3: synthesis of Compound WX025-3

WX025-2(130.00mg, 384.40. mu. mol,1eq) was charged into a pre-dried reaction flask, followed by dissolution with acetic acid (6.5mL), followed by addition of hydrochloric acid (116.67mg,1.18mmol, 114.38. mu.L, 37% purity, 3.08eq), cooling the reaction system to 5 ℃, addition of a solution of sodium nitrite (29.18mg, 422.84. mu. mol,1.1eq) in water (3.25mL), reaction with stirring at 5 ℃ for 0.5 hour, detection of completion of the reaction of the starting materials, addition of WX001-9(66.02mg, 422.84. mu. mol,1.1eq), and stirring at 5 ℃ for 0.5 hour. After completion of the reaction, a solution of sodium acetate (100mg) in water (5mL) was added to the reaction mixture to find a solid precipitated, water (10mL) was further added thereto, the mixture was filtered, and the filter cake was collected, washed with water (10mL × 3), and the filter cake was spin-dried to obtain WX 025-3.

And 4, step 4: synthesis of Compound WX025-4

WX025-3(114.00mg, 225.60. mu. mol,1eq) was added to a pre-dried reaction flask, then dissolved in N, N-dimethylacetamide (1mL), followed by addition of potassium acetate (33.21mg, 338.41. mu. mol,1.5eq) and the reaction stirred at 115 ℃ for 2 hours. After the reaction, the reaction solution was filtered and purified by HPLC (column: Phenomenex luna C1880: 40 mm: 3 μm; mobile phase: water (0.04% hydrochloric acid) -acetonitrile; acetonitrile%: 35% -55%, 7min) to obtain WX 025-4.

And 4, step 4: synthesis of Compounds WX025 and WX026

WX025-4 was separated by supercritical chromatography (column: regi (s, s) WHELK-O1(250mm x 30mm, id.10 μ), mobile phase: a carbon dioxide B methanol (0.1% ammonia), gradient: methanol%: 55% isocratic elution mode; flow rate: 80 g/min; column temperature: 40 ℃; system back pressure: 100bar) to give WX025 and WX 026.

WX025： ¹HNMR (400MHz, deuterated dimethyl sulfoxide) delta 12.07(s,1H),7.76(s,2H), 2.77-2.87 (m,1H),2.32-2.25(m,1H),2.24-2.17(m,2H), 1.73-1.80 (m,1H),1.66-1.55(m,1H),1.17-1.09(m,1H),1.08-1.01(m, 1H). MS-ESI M/z 459.1[ M + H ]] ⁺,461.1[M+H+2] ⁺. Retention time 2.182 min (Instrument: Waters Acquity UPC)²(ii) a Chromatographic column (S, S) -WHELK-O1,3.5 μm,0.46cm id x 5cm L; mobile phase: a, food-grade supercritical carbon dioxide; b methanol (0.05% diethylamine, volume ratio); gradient: the B content increased from 5% to 50% in 1.2 minutes, 50% was maintained for 1 minute, and then decreased from 50% to 5% in 0.8 minute; the flow rate is 3.4 mL/min; column temperature: 35 ℃; detection wavelength: 220 nm; system backpressure 1800 psi).

WX026： ¹HNMR (400MHz, deuterated dimethyl sulfoxide) delta 12.07(s,1H),7.76(s,2H),2.87-2.77(m,1H),2.32-2.25(m,1H),2.24-2.17(m,2H), 1.73-1.80 (m,1H),1.66-1.55(m,1H), 1.15-1.09(m,1H), 1.08-1.01(m, 1H). MS-ESI M/z 459.1[ M + H ]] ⁺,461.0[M+H+2] ⁺. Retention time 2.507 min (Instrument: Waters Acquity UPC)²(ii) a Chromatographic column (S, S) -WHELK-O1,3.5 μm,0.46cm id x 5cm L; mobile phase: a, food-grade supercritical carbon dioxide; b methanol (0.05% diethylamine, volume ratio); gradient: the B content increased from 5% to 50% in 1.2 minutes, 50% was maintained for 1 minute, and then decreased from 50% to 5% in 0.8 minute; the flow rate is 3.4 mL/min; column temperature: 35 ℃; detection wavelength: 220 nm; system backpressure 1800 psi).

Example 27

The synthetic route is as follows:

step 1: synthesis of Compound WX027-2

WX027-1(10g,57.62mmol,1eq) and acetic acid (100mL) were added to a reaction flask, and after nitrogen was purged, liquid bromine (20.26g,126.76mmol,6.53mL,2.2eq) was added and the mixture was reacted at 70 ℃ for 2 hours. After completion of the reaction, the reaction solution was diluted with water (1000mL) to precipitate a solid, which was filtered, and the cake was collected, dissolved in methylene chloride (300mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give WX 027-2.¹HNMR (400MHz, deuterated chloroform) δ 8.38-8.39(d, J ═ 2.4Hz,1H),8.28-8.29(d, J ═ 2.4Hz,1H),6.47(s, 1H).

Step 2: synthesis of Compound WX027-3

WX027-2(5g,19.81mmol,1eq), hydrochloric acid (12M,50.00mL,30.29eq) and ethanol (100mL) were charged into a reaction flask, and after nitrogen gas was purged, stannous chloride dihydrate (17.88g,79.22mmol,4eq) was added, and the mixture was reacted at 80 ℃ for 3 hours. After the reaction is finished, the reaction solution is firstly dried in a spinning mode, then the pH value is adjusted to 10 by using saturated sodium bicarbonate aqueous solution, a solid is separated out, the filtration is carried out, the filtrate is extracted by using ethyl acetate (50mL by 3), an organic phase is dried by using anhydrous sodium sulfate, the filtration is carried out, and the filtrate is concentrated under reduced pressure to obtain WX 027-3.¹HNMR (400MHz, deuterated dimethyl sulfoxide) δ 8.70(s,1H),6.71-6.72(d, J ═ 2.6Hz,1H),6.59-6.60(d, J ═ 2.6Hz,1H),5.00(br s, 2H).

And step 3: synthesis of Compound WX027-4

WX027-3(657.32mg,2.95mmol,1.2eq), WX001-3(0.5g,2.46mmol,1eq) were added to a pre-dried reaction flask, then dissolved in dimethyl sulfoxide (10mL), followed by the addition of potassium carbonate (510.44mg,3.69mmol,1.5eq), nitrogen purged 3 times, and the reaction was stirred slowly to 100 ℃ for 15 hours. After completion of the reaction, the reaction solution was cooled to room temperature (25 ℃ C.), filtered through a funnel packed with celite, the filtrate was washed with ethyl acetate (20 mL. times.3), water (100mL) was added to the filtrate, the organic phase was separated, the aqueous phase was extracted with ethyl acetate (50 mL. times.2), the organic phases were combined and washed with saturated brine (100 mL. times.4), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure at 45 ℃ with a water pump to give a crude product, and the crude product was separated by an automatic column chromatography (gradient elution: petroleum ether: ethyl acetate: 1:0 to 1:1) to purify to obtain WX 027-4.¹HNMR (400MHz, deuterated)Dimethylsulfoxide) δ 6.80-6.85(d, J ═ 2.4Hz,1H),6.67-6.72(d, J ═ 2.4Hz,1H),3.79(br s,2H),2.81-2.91(m,2H), 2.66-2.77 (m,2H),1.80-1.97(m, 4H).

And 4, step 4: synthesis of Compound WX027-5

WX027-4(0.3g, 771.06. mu. mol,1eq) was added to a pre-dried reaction flask, then dissolved with acetic acid (3mL), followed by addition of sodium acetate (126.50mg,1.54mmol,2eq) and the reaction stirred at 110 ℃ for 14 h. After completion of the reaction, the reaction liquid was cooled to room temperature (25 ℃), neutralized with a saturated sodium bicarbonate solution to about 7 degrees, diluted with ethyl acetate (10mL), separated, the organic phase was extracted, the aqueous phase was extracted with ethyl acetate (10mL × 2), the organic phases were combined and washed with a saturated saline solution (20mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure at 45 ℃ with a water pump to give WX 027-5.

And 5: synthesis of Compound WX027-6

WX027-5(0.16g, 387.72. mu. mol,1eq) was charged in a 40mL storage bottle, ethanol (2mL) and 2M dilute hydrochloric acid (2mL) were added, nitrogen was replaced three times, and the mixture was stirred at 90 ℃ for 12 hours. After the reaction, saturated aqueous sodium bicarbonate solution was added to the reaction solution to adjust pH to 7, ethyl acetate (10mL × 3) was added for extraction, the organic phases were combined and concentrated under reduced pressure to obtain a crude product, which was purified by automatic column chromatography (gradient elution: petroleum ether: ethyl acetate: 0:1) to obtain WX 027-6.

Step 6: synthesis of Compound WX027-7

WX027-6(0.045g, 121.42. mu. mol,1eq) was charged in a 40mL storage bottle, acetic acid (2mL) and concentrated hydrochloric acid (12M, 31.16. mu.L, 3.08eq) were added, the temperature was reduced to 5 ℃, a solution of sodium nitrite (9.22mg, 133.56. mu. mol,1.1eq) in water (1mL) was added, then stirring was carried out at 5 ℃ for 0.5 hour, WX001-9(20.85mg, 133.56. mu. mol,1.1eq) was added, and stirring was carried out at 5 ℃ for 0.5 hour. After the reaction was completed, 10mL of a saturated aqueous solution of sodium acetate was added to the reaction solution, a large amount of solid was precipitated, filtered, the filter cake was washed with 2mL of water, and the filter cake was concentrated under reduced pressure to obtain WX 027-7.

And 7: synthesis of Compound WX027

WX027-7(0.03g, 55.79. mu. mol,1eq) was added to a 40mL storage bottle, and N, N-dimethyl was addedAcetamide (2mL) dissolves the substrate, potassium acetate (6.02mg, 61.37. mu. mol,1.1eq) is added, and the mixture is stirred at 115 ℃ for 3 hours. After completion of the reaction, the reaction mixture was filtered and separated by HPLC (column: Phenomenex Luna C18100: 30 mm: 5 μm; mobile phase: [ water (0.04% hydrochloric acid) -acetonitrile](ii) a 35 percent to 55 percent of acetonitrile, 9min) to obtain WX 027.¹HNMR (400MHz, deuterated dimethyl sulfoxide) δ 1.76-1.78(m,4H),2.45-2.46(m,2H),2.53-2.67(m,2H),7.81-7.82(d, J ═ 3.6Hz,1H),7.88(s,1H),12.08(s,1H),13.25(s,1H). MS-ESI M/z 490.9[ M + H ]] ⁺,492.9[M+H+2] ⁺。

Example 28

The synthetic route is as follows:

step 1: synthesis of Compound WX028-2

WX001-2(2g,12.04mmol,1eq) and WX028-1(3.13g,12.04mmol,1eq) were charged to a pre-dried reaction flask, N-dimethylacetamide (20mL) and cesium carbonate (5.88g,18.05mmol,1.5eq) were added, nitrogen was bubbled, and the reaction was charged to a 65 ℃ oil pan and stirred for 1 hour. After the reaction, 10mL of water is added into the reaction solution, ethyl acetate (10mL × 3) is added for extraction, organic phases are combined, the organic phases are washed by saturated saline solution (10mL), the organic phases are dried by anhydrous sodium sulfate after separation, filtration is carried out, filtrate is dried by decompression and spin-drying to obtain a crude product, and the crude product is purified by an automatic column chromatography machine (gradient elution: petroleum ether: ethyl acetate: 10:1 to 5:1 to 3:1) to obtain WX 028-2.

Step 2: synthesis of Compound WX028-3

Adding WX028-2(0.7g,1.80mmol,1eq) to a 40mL storage bottle, adding ethanol (6mL) and hydrochloric acid (3mL), adding stannous chloride dihydrate (2.03g,8.98mmol,5eq), and heating to 80 deg.CStirring at deg.C for 2 hours. After completion of the reaction, the reaction mixture was spin-dried under reduced pressure, water (10mL) and ethyl acetate (10mL) were added, the spin-evaporated residue was dissolved, a saturated aqueous sodium bicarbonate solution was added to adjust pH to 7-8, liquid separation was performed, the aqueous phase was extracted with ethyl acetate (10mL × 3), the organic phases were combined, washed with a saturated saline solution (20mL), liquid separation was performed, and the organic phase was dried over anhydrous sodium sulfate, filtered, and spin-dried under reduced pressure to obtain WX 028-3.¹HNMR (400MHz, deuterated dimethyl sulfoxide) δ 1.72-1.74(m,4H),2.39-2.42(m,2H),2.52-2.56(m,2H),5.79(s,2H),6.87-6.88(d, J ═ 2.8Hz,1H),6.94(s,1H),11.97(s, 1H).

And step 3: synthesis of Compound WX028-4

WX028-3(0.4g,1.11mmol,1eq) was charged to a 40mL storage bottle, glacial acetic acid (4mL) and hydrochloric acid (12M,285.40 μ L,3.08eq) were added, the temperature was reduced to 5 deg.C, a solution of sodium nitrite (84.40mg,1.22mmol,1.1eq) in water (2mL) was added, then stirring was carried out at 5 deg.C for 0.5 hour, WX001-9(190.98mg,1.22mmol,1.1eq) was added, and stirring was carried out at 5 deg.C for 0.5 hour. After the reaction is finished, 10mL of saturated sodium acetate aqueous solution is added into the reaction solution, a large amount of solid is separated out, the mixture is filtered, the filter cake is washed by 2mL of water, and the filter cake is decompressed and dried to obtain WX 028-4.¹HNMR (400MHz, deuterated dimethyl sulfoxide) δ 1.25-1.28(m,3H),1.74-1.77(s,4H),2.42-2.47(m,2H),2.51(m,2H),4.12-4.23(m,2H),8.01-8.02(d, J ═ 2Hz,1H),8.39(s,1H),10.84(s,2H),12.08(s, 1H).

And 4, step 4: synthesis of Compound WX028

WX028-4(0.25g, 474.52. mu. mol,1eq) was added to a 40mL storage bottle, N-dimethylacetamide (2mL) was added to dissolve the substrate, potassium acetate (51.23mg, 521.97. mu. mol,1.1eq) was added, the temperature was raised to 115 ℃ and stirred for 1 hour. After the reaction, the reaction solution was directly filtered, and the filtrate was subjected to high performance liquid chromatography (column: Phenomenex luna C1880: 40 mm: 3 μm; mobile phase: [ water (0.04% hydrochloric acid) -acetonitrile: [ Ach ])](ii) a Acetonitrile percent of 37 percent to 57 percent and 7min) to obtain WX 028.¹HNMR (400MHz, deuterated dimethyl sulfoxide) δ 1.75-1.77(m,4H),2.32-2.44(m,2H),2.63-2.67(m,2H),7.96-7.97(d, J ═ 2.4Hz,1H),8.11-8.12(d, J ═ 2.0Hz,1H),12.12(s,1H),13.31(s, 1H). MS-ESI M/z 481.1[ M + H ]] ⁺，483.1[M+H+2] ⁺。

Example 29

The synthetic route is as follows:

step 1: synthesis of Compound WX029-1

Deuterated methanol (2.56g,70.93mmol,2.88mL,20eq) is added into a reaction bottle which is dried in advance, nitrogen is replaced three times, the temperature is reduced to 0 ℃, sodium hydride (354.62mg,8.87mmol, 60% purity, 2.5eq) is added, the temperature is kept at 0 ℃ and stirring is carried out for 0.5 hour, another dried reaction bottle is taken, WX021-1(1g,3.55mmol,1eq) and tetrahydrofuran (1mL) are added, the reaction solution is slowly dropped at the temperature of 0 ℃, and the reaction solution is kept at the temperature of 0 ℃ and stirred for 1 hour. After the reaction, the reaction solution was poured into 20mL of water, ethyl acetate (20mL × 3) was added for extraction, the organic phases were combined, washed with saturated brine (20mL), separated, dried over anhydrous sodium sulfate, filtered, and the filtrate was spin-dried under reduced pressure to obtain a crude product. The crude product was purified by automatic column chromatography (eluent polarity, petroleum ether: ethyl acetate 10% -20%) to yield WX 029-1.¹HNMR (400MHz, deuterated dimethyl sulfoxide) delta 4.39-4.40(m,1H),2.34-2.38(m,2H),1.92-1.94(m,2H),1.50-1.53(m, 2H).

Step 2: synthesis of Compound WX029-2

WX029-1(1.8g,7.62mmol,1eq) and WX001-4(2.71g,15.25mmol,2eq) were charged into a pre-dried 40mL storage bottle, N-dimethylacetamide (20mL) and cesium carbonate (4.97g,15.25mmol,2eq) were added, nitrogen was bubbled through, the reaction was added to a 70 ℃ oil pan and stirred for 8 hours. After completion of the reaction, 100mL of water was added to the reaction mixture, ethyl acetate (100mL × 3) was added for extraction, the organic phases were combined, and the organic phase was washed with saturated brine (100mL), dried over anhydrous sodium sulfate, filtered, and spin-dried under reduced pressure to obtain a crude product. Quick silica gel column for crude productPurification (eluent polarity: petroleum ether: ethyl acetate 10% to 20%), followed by further separation by high performance liquid chromatography (column: Phenomenex luna C18(250 x 70mm,15 μm); mobile phase: [ water (0.1% trifluoroacetic acid) -acetonitrile](ii) a 50 percent to 60 percent of acetonitrile, 20 minutes) to obtain WX 029-2.¹HNMR (400MHz, deuterated chloroform) delta 6.91(s,2H),4.41(s,1H),3.07-3.12(m,1H),2.60-2.70(m,1H),2.39-2.43(m,1H),1.99(s,2H),1.61-1.64(m, 1H).

And (3) identifying the structure of WX 029-2:

WX029-2(0.05g, 132.39. mu. mol,1eq) and ethyl acetate (10mL) were charged into a 100mL single-neck flask, wet palladium on carbon (0.05g, 132.39. mu. mol, 10% purity, 1eq) was added, hydrogen was replaced three times, and then the reaction was carried out at 20 ℃ under the pressure of a hydrogen balloon (15psi) for 2 hours, after completion of the reaction, the reaction solution was filtered with celite, the filtrate was dried under reduced pressure, and the crude product was subjected to a high performance liquid chromatography column (Waters Xbridge BEC 18100. about.25 mM. mu.5 μm; mobile phase: [ water (10mM ammonium bicarbonate) -acetonitrile](ii) a Acetonitrile percent of 1 percent to 25 percent and 10 minutes) to obtain WX 029-2A. NOE (400Hz, deuterated chloroform) showed a correlation between H1(4.178) and H2(7.849), demonstrating that WX029-2 has the correct structure.¹HNMR (400MHz, deuterated chloroform) delta 11.17(s,1H),7.85(s,1H),4.18-4.20(m,1H),2.55-2.67(m,2H),1.89-1.95(m, 2H),1.71-1.76(m, 2H).

And step 3: synthesis of Compound WX029-3

WX029-2(0.4g,1.06mmol,1eq) and glacial acetic acid (2mL) were added to a pre-dried reaction flask followed by sodium acetate (260.64mg,3.18mmol,3eq) and stirring continued at 110 ℃ for 12h after the addition. After completion of the reaction, the reaction solution was concentrated under reduced pressure to dryness, then a saturated sodium bicarbonate solution was added to adjust the pH of the solution to 6-7, followed by extraction with ethyl acetate (100mL × 3), the organic phases were combined, the organic phase was washed with a saturated saline solution (100mL × 2), dried over anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure to give WX 029-3.

And 4, step 4: synthesis of Compound WX029-4

WX029-3(0.35g, 872.26. mu. mol,1eq) was charged into a 40mL storage bottle, 2N hydrochloric acid (10mL) and ethanol (10mL) were added, and the mixture was heated to 80 ℃ and stirred for 4 hours. After completion of the reaction, a saturated aqueous sodium bicarbonate solution was added to the reaction mixture to adjust pH to 7, ethyl acetate (10mL × 3) was added for extraction, and the organic phases were combined and spin-dried under reduced pressure to obtain WX 029-4.

And 5: synthesis of Compound WX029-5

Adding WX029-4(0.2g,556.76 mu mol and 1eq) into a 40mL storage bottle, adding water (1mL) and hydrochloric acid (12M,142.90 mu L and 3.08eq), cooling to 5 ℃, adding a glacial acetic acid (2mL) solution of sodium nitrite (42.26mg,612.43 mu mol and 1.1eq), stirring at 5 ℃ for 0.5 hour, adding WX001-9(95.62mg,612.43 mu mol and 1.1eq), after the reaction is finished, adding 10mL of saturated sodium acetate aqueous solution into the reaction solution, precipitating a large amount of solid, filtering, washing the filter cake with 10mL of water, and carrying out reduced pressure spin-drying to obtain WX 029-5.

Step 6: synthesis of Compound WX029-6

WX029-5(0.2g, 379.98. mu. mol,1eq) was added to a 40mL storage bottle, N-dimethylacetamide (2mL) was added to dissolve the substrate, potassium acetate (74.58mg, 759.96. mu. mol,2eq) was added, and the mixture was stirred at 115 ℃ for 1 hour. After completion of the reaction, the reaction solution was directly filtered, and the filtrate was purified by high performance liquid chromatography (column: Waters Xbridge Prep OBD C18150 mM 40mM 10 μm; mobile phase: [ water (10mM ammonium bicarbonate) -acetonitrile ]; acetonitrile%: 25% -55%, 8 min.) to obtain WX 029-6.

And 7: synthesis of Compounds WX029 and WX030

WX029-6 was separated by supercritical chromatography (instrument: Waters SFC150AP preparative SFC; column: DAICEL CHIRALCEL OJ (250 mm. about.30 mm,10 μm); mobile phase: A carbon dioxide B methanol [ neutral ]; gradient: methanol%: 35% isocratic elution mode for 10 minutes; flow rate: 70 g/min; column temperature: 35 ℃; system back pressure: 150bar) to give WX029 and WX 030.

WX029： ¹HNMR (400MHz, deuterated methanol) δ 7.74-7.75(d, J ═ 4Hz,2H),4.40-4.42(m,1H),2.90-2.95(m,1H),2.64-2.89(m,1H),2.28-2.31(m,1H),1.90-1.94(m,2H), 1.55-1.57(m, 1H). MS-ESI m/z:480.2[M+H] ⁺,482.2[M+H+2] ⁺. Retention time 1.67 min (instrument: per analytical SFC; chromatographic column: Chiralpak AS-33 μm,0.46cm id x 5cm L; mobile phase: a: food grade supercritical carbon dioxide; B: methanol (0.05% isopropanol, volume ratio), gradient: B content increases from 10% to 40% in 3 min; flow rate: 4.0 mL/min; column temperature: 35 ℃; detection wavelength: 220 nm; system back pressure: 100 bar).

WX030： ¹H NMR (400MHz, deuterated methanol) δ 7.72-7.73(d, J ═ 4Hz,2H),4.38-4.40(m,1H),2.88-2.93(m,1H),2.62-2.88(m,1H),2.26-2.87(m,1H),1.88-1.91(m,2H), 1.53-1.55(m, 1H). MS-ESI M/z 480.2[ M + H ]]+,482.2[M+H+2] ⁺. Retention time 1.63 minutes (instrument: per analytical SFC; column: Chiralpak AS-33 μm,0.46cm id x 5cm L; mobile phase: a: food grade supercritical carbon dioxide; B: methanol (0.05 isopropanol, volume ratio); gradient: B content rises from 10% to 40% in 3 minutes; flow rate: 4.0 mL/min; column temperature: 35 ℃; detection wavelength: 220 nm; system back pressure: 100 bar).

Example 31

The synthetic route is as follows:

step 1: synthesis of Compound WX031-1

WX021-1(5g,17.73mmol,1eq) and ethanol (100mL) were charged into a pre-dried reaction flask, placed at 0 ℃, sodium ethoxide (1.81g,26.60mmol,1.5eq) was added slowly in portions, nitrogen was replaced three times, and stirring was carried out at 0 ℃ for 1.5 hours. After the reaction, adding water (20mL), diluting, separating, collecting organic phase, adjusting pH to about 6 with 2N hydrochloric acid, concentrating methanol in the system under reduced pressure, extracting the residual solution with dichloromethane (20mL 2), combining organic phasesAnd washed with saturated brine (30mL × 2), dried over anhydrous sodium sulfate, and spin-dried to give a crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate: 1: 0: 20: 1: 10: 1: 5: 1: 2:1) to afford WX 031-1.¹H NMR (400MHz, deuterated chloroform) δ 4.50(t, J ═ 2.57Hz,1H),3.64-3.87(m,2H),2.93(dd, J ═ 4.27,18.20Hz,1H),2.47-2.60(m,1H),2.34(dd, J ═ 2.95,14.49Hz,1H),1.93-2.06(m,2H),1.47-1.54(m,1H),1.23-1.28(m, 3H). MS-ESI M/z 247.0[ M + H ]] ⁺,249.0[M+H+2] ⁺。

Step 2: synthesis of Compound WX031-2

WX031-1(2.7g,10.93mmol,1eq), WX001-4(2.14g,12.02mmol,1.1eq) and DMF (30mL) were charged in a pre-dried reaction flask, cesium carbonate (4.27g,13.11mmol,1.2eq) was added, nitrogen was replaced three times, and the mixture was stirred at 65 ℃ for 16 hours. After the reaction is finished. Diluting the reaction solution with ethyl acetate (200mL), filtering with diatomite, diluting the filtrate with water (200mL), separating, extracting the aqueous phase (200mL x3) with ethyl acetate, combining the organic phases, washing with saturated saline (300mL x3), collecting the organic phase, drying with anhydrous sodium sulfate, filtering, and concentrating under reduced pressure to obtain a crude product. Purifying the crude product by column chromatography (petroleum ether: ethyl acetate: 1: 0: 20: 1: 10: 1: 5: 1: 2:1) to obtain crude product, separating the crude product by high performance liquid chromatography (chromatographic column: Phenomenex luna C18250: 50 mm: 15 μm; mobile phase: [ water (0.1% trifluoroacetic acid) -acetonitrile)](ii) a Acetonitrile accounting for 50 percent to 70 percent for 20min) to obtain WX 031-2.¹H NMR (400MHz, deuterated chloroform) δ 6.70(s,2H),4.49(s,1H),4.09(s,2H),3.67-3.89(m,2H),3.06(dd, J ═ 4.82,19.32Hz,1H),2.62(m,1H),2.36(d, J ═ 14.01Hz,1H),1.89-2.05(m,2H),1.59(tt, J ═ 3.36,13.84Hz,1H),1.27-1.33(m, 3H). MS-ESI M/z 390.0[ M + H ]] ⁺,392.0[M+H+2] ⁺。

And step 3: synthesis of Compound WX031-3

WX031-2(400mg,1.03mmol,1eq) and glacial acetic acid (8mL) were added to a pre-dried reaction flask, and sodium acetate (168.84mg,2.06mmol,2eq) was added, replaced with nitrogen three times, and stirred at 120 ℃ for 16 hours. After the reaction is finished. The reaction mixture was concentrated under reduced pressure, poured into water (10mL), and the pH of the solution was adjusted to 7-8 with solid sodium bicarbonate, thenThen extracted with (ethyl acetate: tetrahydrofuran 3:1, 30mL 3), all organic phases were combined, the organic phase was washed with saturated sodium chloride solution (30mL 2), dried over anhydrous sodium sulfate, filtered, and concentrated to give WX 031-3. MS-ESI M/z 412.0[ M + H ]] ⁺,414.0[M+H+2] ⁺。

And 4, step 4: synthesis of Compound WX031-4

WX031-3(420mg,1.02mmol,1eq) and ethanol (6mL) were added to a pre-dried reaction flask. Hydrochloric acid (2M,6mL,11.78eq) was added, nitrogen was replaced three times, and the mixture was stirred at 90 ℃ for 3 hours. After the reaction, the reaction solution was cooled to room temperature, water (20mL) was added, the aqueous phase (20mL × 3) was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine (30mL × 1), the organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain WX 031-4. MS-ESI M/z 370.0[ M + H ]] ⁺,372.0[M+H+2] ⁺。

And 5: synthesis of Compound WX031-5

WX031-4(377mg,1.02mmol,1eq) and glacial acetic acid (5mL), hydrochloric acid (12M, 262.21. mu.L, 3.09eq) were added to a reaction flask which had been dried in advance, a mixed solution of sodium nitrite (74.48mg,1.08mmol,1.06eq) and water (5mL) was added dropwise slowly at 0 ℃, and stirring was continued at 0 ℃ for 0.5 hour, WX001-9(158.99mg,1.02mmol,1eq) was added at 0 ℃ and stirring was continued at 0 ℃ for 1.5 hours. After the reaction is finished. The reaction was quenched with aqueous sodium acetate (3eq) (9mL) to yield a large amount of solid, which was filtered to yield WX 031-5. MS-ESI M/z 537.1[ M + H ]] ⁺,539.0[M+H+2] ⁺。

Step 6: synthesis of Compound WX031-6

WX031-5(547mg,1.02mmol,1eq) and N, N-dimethylacetamide (8mL) were charged into a reaction flask which had been dried in advance, potassium acetate (109.89mg,1.12mmol,1.1eq) was added thereto, nitrogen was replaced three times, and the mixture was stirred at 115 ℃ for 5 hours. After the reaction is finished, directly filtering the reaction solution, and separating the filtrate by high performance liquid chromatography. Separating by high performance liquid chromatography (column: Phenomenex luna C18250 mm 10 μm; mobile phase: [ water (0.04% hydrochloric acid) -acetonitrile](ii) a Acetonitrile accounting for 40 percent to 60 percent for 10min) to obtain WX 031-6.¹H NMR (400MHz, deuterated methanol) delta 7.75(s,2H),4.52(s,1H),3.71-3.85(m,2H),2.93(dd,J＝4.31,19.70Hz,1H),2.61(m,1H),2.22-2.32(m,1H),1.86-2.00(m,2H),1.52-1.63(m,1H),1.22(t,J＝7.00Hz,3H)。MS-ESI m/z＝491.0[M+H] ⁺,493.0[M+H+2] ⁺. Carbonyl carbon C detected by HMBC nuclear magnetism₁(156.29) and H₂(4.53) correlation, confirmed to be the structure shown.

And 7: synthesis of Compounds WX031 and WX032

WX031-6(0.25g,508.87umol,1eq) was added to a pre-dried reaction flask. WX031 and WX032 were obtained by supercritical chromatography (instrument: Waters SFC150AP preparative SFC; column: DAICEL CHIRALCEL OJ (250mm 30mm,10um), mobile phase: A carbon dioxide B ethanol [ neutral ]; gradient: ethanol%: 55% isocratic elution mode, 10 min; flow rate: 65 g/min; column temperature: 35 ℃ C.; system back pressure: 150 bar).

WX031: ¹H NMR (400MHz, deuterated methanol) δ 7.75(s,2H),4.52(s,1H),3.71-3.85(m,2H),2.88-3.00(m,1H),2.54-2.68(m,1H),2.24-2.28(m,1H),1.91-1.97(m,2H),1.51-1.62(m,1H),1.22(t, J ═ 7.02Hz, 3H). Retention time 1.035 minutes (instrument: Waters UPCC with PDA separator; chromatographic column: Chiralcel OJ-3,50 × 4.6mm, i.d.,3 um; mobile phase: a: food grade supercritical carbon dioxide; B: ethanol (0.05% diethylamine, volume ratio); gradient: B content rises from 5% to 50% in 1.2 minutes and stays for 1 minute, then decreases from 50% to 5% in 0.8 minutes; flow rate: 3.4 mL/min; column temperature: 35 ℃; detection wavelength: 220 nm; system back pressure: 100bar), MS-esim/z 491.0[ M + H ] 491.0[ M + H] ⁺,492.9[M+H+2] ⁺。

WX032： ¹H NMR (400MHz, deuterated methanol) δ 7.75(s,2H),4.52(s,1H),3.71-3.85(m,2H),2.89-2.95(m,1H),2.56-2.66(m,1H),2.24-2.28(m,1H),1.89-2.00(m,2H),1.51-1.60(m,1H),1.21(t, J ═ 7.02Hz, 3H). Retention time 1.151 minutes (instrument: Waters UPCC with PDA detactor; chromatographic column: Chiralcel OJ-3, 50)4.6mm, i.d.,3 um; mobile phase: a, food-grade supercritical carbon dioxide; b, ethanol (0.05 percent of diethylamine by volume); gradient: b content increased from 5% to 50% in 1.2 minutes and held for 1 minute, then decreased from 50% to 5% in 0.8 minute; the flow rate is 3.4 mL/min; column temperature: 35 ℃; detection wavelength: 220 nm; system back pressure 100bar), MS-ESI M/z 491.0[ M + H ]] ⁺,492.9[M+H+2] ⁺。

Example 33

The synthetic route is as follows:

step 1: synthesis of Compound WX033-1

WX021-1(5g,17.73mmol,1eq) and isopropanol (50mL) were added to a previously dried reaction flask, and the mixture was stirred at 90 ℃ for 2 hours while displacing nitrogen. After the reaction is finished. Cooling the reaction solution to 25 ℃, adding solid sodium bicarbonate to adjust the pH value to 6-7, then concentrating under reduced pressure to obtain a solid, adding water (40mL) and ethyl acetate (40mL) to the solid for separating, collecting an organic phase, extracting an aqueous phase with ethyl acetate (3X 40mL), combining the organic phases, sequentially washing with saturated saline (100mL), drying with anhydrous sodium sulfate, filtering, and concentrating under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (gradient elution: petroleum ether: ethyl acetate ═ 10:1 to 3:1 to 1:1) to afford WX 033-1.¹HNMR (400MHz, deuterated chloroform) δ 1.17-1.36(m,6H), 1.43-1.60(m,2H), 1.98-2.16(m,1H), 2.33(dd, J ═ 14.32,2.81Hz,1H), 2.43-2.63(m,1H), 2.83-3.00(m,1H), 3.91-4.06(m,1H), 4.60-4.73(m, 1H). MS-ESI M/z 261.2[ M + H ]] ⁺,263.0[M+H+2] ⁺。

Step 2: synthesis of Compound WX033-2

A pre-dried reaction flask was charged with WX033-1 (1)5g,5.74mmol,1eq), WX001-4(1.02g,5.74mmol,1eq) and dimethyl sulfoxide (150mL), potassium carbonate (3.18g,22.98mmol,4eq) was added, nitrogen was replaced three times, copper iodide (656.35mg,3.45mmol,0.6eq) was added, nitrogen was replaced again, and the mixture was stirred at 90 ℃ for 12 hours. After the reaction is finished. The reaction was cooled to 25 ℃, diluted with water (300mL), filtered through celite, and ethyl acetate (300mL) was added for liquid separation, the organic phases were collected, the aqueous phase was extracted with ethyl acetate (3 × 300mL), the organic phases were combined, washed successively with saturated brine (3 × 1000mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude product. The crude product was purified by column chromatography (gradient elution: petroleum ether: ethyl acetate: 5:1 to 1:1) followed by high performance liquid chromatography (separation conditions: column: Phenomenex Luna C18200: 40mm 10 μm; mobile phase: [ water (0.2% formic acid) -acetonitrile)](ii) a Acetonitrile percent of 50 percent to 90 percent and 8 minutes) to obtain WX 033-2.¹H NMR (400MHz, deuterated chloroform) δ 1.30(dd, J ═ 18.39,6.25Hz,6H), 1.54-1.62(m,1H), 1.94(s,1H), 2.01-2.14(m,1H), 2.29-2.41(m,1H), 2.59-2.68(m,1H), 3.06(dd, J ═ 19.45,5.44Hz,1H), 3.97-4.04(m,1H), 4.66(s,1H), 6.66(s, 2H). MS-ESI M/z 402.2[ M + H ]] ⁺,404.0[M+H+2] ⁺。

And (3) identifying the structure of WX 033-2:

detection of WX033-2 by (HMBC) showed carbon atom C₂(153) With hydrogen H₁(4.66) related, carbon atom C₄(163) With hydrogen H₂(2.59-2.68, 3.06) and the correct structure of WX033-2 was confirmed.

And step 3: synthesis of Compound WX033-3

WX033-2(350mg, 869.13. mu. mol,1eq), sodium acetate (142.60mg,1.74mmol,2eq) and glacial acetic acid (4mL) were added to a pre-dried reaction flask, and the mixture was stirred at 110 ℃ for 12 hours while displacing nitrogen. After the reaction is finished. The reaction solution was cooled to 25 ℃ and directly concentrated under reduced pressure to give WX 033-3. MS-ESI M/z 462.2[ M + H ]] ⁺,464.0[M+H+2] ⁺。

And 4, step 4: synthesis of Compound WX033-4

WX033-3(0.3g, 703.74. mu. mol,1eq), hydrochloric acid (2M,7.04mL,20eq) and ethanol (3mL) were charged into a pre-dried reaction flask, and the mixture was stirred at 90 ℃ for 12 hours while replacing nitrogen. After the reaction is finished. The reaction was cooled to 25 ℃, diluted with water (15mL), adjusted to pH 6-7 by the addition of solid sodium bicarbonate, separated by the addition of ethyl acetate (15mL), the organic phase collected, the aqueous phase extracted with ethyl acetate (3 × 15mL), the organic phases combined, washed successively with saturated brine (50mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude product. The reaction was used directly in the next step without purification. WX033-4 was obtained. MS-ESI M/z 384.2[ M + H ]] ⁺,386.0[M+H+2] ⁺。

And 5: synthesis of Compound WX033-5

WX033-4(350mg, 910.85. mu. mol,1eq) was charged into a pre-dried reaction flask, followed by dissolution with glacial acetic acid (2mL), then hydrochloric acid (276.45mg,2.81mmol, 271.03. mu.L, 37% purity, 3.08eq) was added, the reaction system was cooled to 5 ℃, a solution of sodium nitrite (69.13mg,1.00mmol,1.1eq) in water (1mL) was added, the reaction was stirred at 5 ℃ for 0.5 hour, WX011-9(156.44mg,1.00mmol,1.1eq) was added, and the reaction was stirred at 5 ℃ for 0.5 hour. After the reaction was complete, sodium acetate (82mg) and water (2mL) were added, solids were found to precipitate, the filter cake was collected by filtration, the filter cake was washed with water (10mL x3), and the filter cake was spin dried to give WX 033-5. MS-ESI M/z 551.3[ M + H ]] ⁺,553.0[M+H+2] ⁺。

Step 6: synthesis of Compound WX033-6

After adding WX033-5(320mg, 580.36. mu. mol,1eq) to a pre-dried reaction flask, it was dissolved in N, N-dimethylacetamide (3mL), and then potassium acetate (85.44mg, 870.55. mu. mol,1.5eq) was added thereto, followed by stirring at 115 ℃ for 4 hours. After the reaction, the reaction solution was cooled to 25 ℃ and directly filtered, and the filtrate was sent to high performance liquid chromatography. The crude product was purified by high performance liquid chromatography (column: 3. about. Phenomenex Luna C1875. about. 30 mm. about. 3 μm; mobile phase: [ water (0.2% formic acid) -acetonitrile](ii) a Acetonitrile percent of 40 percent to 70 percent and 9 minutes) to obtain WX 033-6. MS-ESI m/z 505.2[M+H] ⁺,507.0[M+H+2] ⁺。

And 7: synthesis of Compounds WX033 and WX034

7(0.1g, 197.90. mu. mol,1eq) was subjected to supercritical chromatography. Supercritical chromatography was carried out by (apparatus: Waters SFC150AP preparative SFC; column: DAICEL CHIRALPAK AD (250mm x 30mm,10 μm), mobile phase: a carbon dioxide B isopropanol [ neutral ], gradient: isopropanol%: 50% isocratic elution mode, 30min, flow rate: 80g/min, column temperature: 35 ℃ and system back pressure: 150bar) to give WX033 and WX 034.

WX033： ¹H NMR (400MHz, deuterated methanol) δ 1.23(dd, J ═ 6.00,3.88Hz,6H), 1.54-1.62(m,1H), 1.84-2.07(m,2H), 2.20(d, J ═ 12.38Hz,1H), 2.54-2.68(m,1H), 2.92(dd, J ═ 19.32,4.69Hz,1H), 4.05-4.11(m,1H), 4.64(s,1H), 7.75(s, 2H). Retention time 1.295 min (instrument: Waters UPCC with PDA separator; chromatography column: Chiralcel OJ-3,50 × 4.6mm, i.d.,3 μ M; mobile phase: a: food grade supercritical carbon dioxide; B: isopropanol (0.05% diethylamine, volume ratio); gradient: B content rises from 5% to 50% in 1.2 min and stays for 1 min, then decreases from 50% to 5% in 0.8 min; flow rate: 3.4 mL/min; column temperature: 35 ℃; detection wavelength: 220 nm; system back pressure: 100bar), MS-esim/z 491.0[ M + H ] bar] ⁺,492.9[M+H+2] ⁺。

WX034： ¹H NMR (400MHz, deuterated methanol) δ 1.23(dd, J ═ 6.00,4.00Hz,6H), 1.52-1.64(m,1H), 1.87-2.07(m,2H), 2.20(d, J ═ 13.63Hz,1H), 2.53-2.68(m,1H), 2.92(dd, J ═ 19.45,4.94Hz,1H), 4.05-4.11(m,1H), 4.64(s,1H), 7.75(s, 2H). Retention time 1.517 min (instrument: Waters UPCC with PDA stripper; chromatography column: Chiralcel OJ-3,50 × 4.6mm, i.d.,3 μm; mobile phase: a: food grade supercritical carbon dioxide; B: isopropanol (0.05% diethylamine, volume ratio); gradient: B content rises from 5% to 50% in 1.2 min and stays for 1 min, then decreases from 50% to 5% in 0.8 min; flow rate: 3.4 mL/min; column temperature: 35 ℃; detection wavelength: 220 nm; system back pressure: 100 bar). MS-ESI M/z 491.0[ M + H ]] ⁺,492.9[M+H+2] ⁺。

Example 35

The synthetic route is as follows:

step 1: synthesis of Compound WX035-1

WX021-5(287mg, 805.72. mu. mol,1eq) was charged into a reaction flask dried in advance, followed by dissolution with dichloromethane (6mL), cooling the reaction system to 0 ℃ and then adding boron tribromide (403.70mg,1.61mmol, 155.27. mu.L, 2eq), stirring the reaction at 0 ℃ for 1 hour, raising to 40 ℃ and stirring the reaction for 12 hours. After completion of the reaction, the reaction system was cooled to room temperature (25 ℃), the reaction solution pH was neutralized with saturated sodium bicarbonate to neutral, then dichloromethane (20mL), tetrahydrofuran (10mL) was added, the organic phase was dissolved, the organic phase was separated, the aqueous phase was extracted with ethyl acetate (20mL _ 3) (a little tetrahydrofuran was added to aid dissolution), the organic phases were combined and washed with saturated brine (20mL _ 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give WX 035-1.

Step 2: synthesis of Compound WX035-3

WX035-2(140.22mg,2.41mmol,10mL,3eq) was added to a pre-dried reaction flask, the reaction was cooled to 0 ℃ and then sodium hydride (48.29mg,1.21mmol, 60% purity, 1.5eq) was added and the reaction was stirred at 0 ℃ for 10 minutes, after which the reaction was slowly added to a solution of WX035-1(326mg, 804.79. mu. mol,1eq) in WX035-2(10mL) (which was warmed during the addition) and stirred at 0 ℃ for 30 minutes. After completion of the reaction, the reaction solution was neutralized to about pH 6 with 1N hydrochloric acid at 0 ℃, water (30mL) and ethyl acetate (30mL) were added, the organic phase was separated, the aqueous phase was extracted with ethyl acetate (30mL × 2), the organic phases were combined and washed with saturated brine (30mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give WX 035-3.

And step 3: synthesis of Compound WX035-4

WX035-3(300mg, 784.85. mu. mol,1eq) was charged into a reaction flask dried in advance, followed by dissolution with glacial acetic acid (6mL), addition of hydrochloric acid (238.20mg,2.42mmol, 233.53. mu.L, 37% purity, 3.08eq), cooling of the reaction system to 5 ℃ and addition of a solution of sodium nitrite (59.57mg, 863.33. mu. mol,1.1eq) in water (3mL), and reaction was stirred at 5 ℃ for 0.5 hour. WX001-9(134.80mg, 863.33. mu. mol,1.1eq) was added and stirred at 5 ℃ for 0.5 hour. After completion of the reaction, sodium acetate (200mg) in water (10mL) was added at 0 ℃ to find a solid precipitated, water (10mL) was added to increase the solid, the mixture was filtered, the filter cake was washed with water (5 mL. times.2), and the filter cake was spin-dried to give WX 035-4.

And 4, step 4: synthesis of Compound WX035-5

WX035-4(430mg, 782.73. mu. mol,1eq) was added to a pre-dried reaction flask, then dissolved with N, N-dimethylacetamide (4mL), followed by potassium acetate (115.23mg,1.17mmol,1.5eq) and the reaction was stirred slowly raising to 115 ℃ for 1 hour. Potassium acetate (115.23mg,1.17mmol,1.5eq) was added and the reaction was continued with stirring at 115 ℃ for 0.5 h. After completion of the reaction, the reaction mixture was filtered and subjected to high performance liquid chromatography (separation conditions: column: Welch Xitimate C18150 × 25mm × 5 μm; mobile phase: water (0.04% hydrochloric acid) -acetonitrile; acetonitrile%: 50% -60%, 8 minutes) to obtain WX 035-5.

And 5: synthesis of Compounds WX035 and WX036

WX035-5 was separated by supercritical chromatography (instrument: Waters SFC150AP preparative SFC, column: DAICEL CHIRALCEL OJ (250 mm. about.30 mm,10 μm), mobile phase: A carbon dioxide B methanol [ neutral ], gradient: methanol%: 35% isocratic elution mode, 15min, flow rate: 70g/min, column temperature: 35 ℃ C., system back pressure: 150bar) to give WX035 and WX 036.

WX035： ¹HNMR (400MHz, deuterated methanol) delta 7.75(s,2H),4.71s (m,1H),3.66-3.70(m,1H),2.90-2.95(m,1H),2.62-2.67(m,1H),2.36-2.40(m,1H),1.91-1.93(m,2H),1.60-1.61(m,1H),0.82-0.85(m,1H),0.65-0.66(m,1H),0.47-0.54(m, 2H). MS-ESI M/z 503.0[ M + H ]] ⁺,504.9[M+H+2] ⁺. Retention time 1.143 min (Instrument: Waters UPCC w)ith PDA detactor; chromatographic column chromatography, Chiralpak AS-3,50 × 4.6mm, i.d.,3 μm; mobile phase: a, food-grade supercritical carbon dioxide; b methanol (0.05% diethylamine, volume ratio); gradient: the B content increased from 5% to 50% in 1.2 minutes, 50% was maintained for 1 minute, and then decreased from 50% to 5% in 0.8 minute; the flow rate is 3.4 mL/min; column temperature: 35 ℃; detection wavelength: 220 nm; system backpressure 1800 psi).

WX036： ¹HNMR (400MHz, deuterated methanol) delta 7.75(s,2H),4.75-4.67(m,1H),3.71-3.66(m,1H),2.98-2.87(m,1H),2.68-2.55(m,1H),2.44-2.32(m,1H),1.99-1.88(m,2H),1.66-1.54(m,1H),0.89-0.79(m,1H),0.71-0.61(m,1H),0.59-0.50(m,1H),0.50-0.40(m, 1H). MS-ESI M/z 503.0[ M + H ]]+,504.9[M+H+2] ⁺. Retention time 1.328 min (instrument: Waters UPCC with PDA separator; chromatography column: Chiralpak AS-3,50 x 4.6mm, i.d.,3 μm; mobile phase: a: food grade supercritical carbon dioxide; B: methanol (0.05% diethylamine, volume ratio); gradient: B content rises from 5% to 50% in 1.2 min, 50% is maintained for 1 min, then falls from 50% to 5% in 0.8 min; flow rate: 3.4 mL/min; column temperature: 35 ℃; detection wavelength: 220 nm; system back pressure: 1800 psi).

Biological assay

Experiment one: nuclear hormone receptor activity assay

The NHR protein interaction (Pro) and Nuclear Translocation (NT) experimental techniques, which use a technique developed by discover x called Enzyme Fragment Complementation (EFC), allow the extent of nuclear hormone receptor activation to be monitored in a homogeneous, non-imaging format. The principle of the NHR Pro experiment is to detect protein interactions between activated full-length NHR proteins and nuclear fusion proteins containing a Steroid Receptor Coactivator Peptide (SRCP) domain of one or more classical LXXLL interaction motifs. The NHR is labeled with the ProLinkTM component of the EFC detection system, and the SRCP domain is fused to an enzyme receptor component (EA) expressed in the nucleus. When bound by a ligand, NHR will migrate to the nucleus and recruit the SRCP domain, whereby complementation occursOne unit of active beta-galactosidase (beta-Gal) is produced and a chemiluminescent signal is produced. Benefits associated with this approach include reduced compound incubation time, direct measurement of NHR targets, use of full-length human NHR sequences, and the ability to pick new classes of compounds that break the limitations of protein-protein interactions. NHR NT experiments monitored NHR movement between the cytoplasmic and nuclear compartments. ProLabel of this receptor with our EFC assay System^TMThe components are labeled and EA is fused to a nuclear localization sequence, limiting EA expression to the nucleus. Migration of the NHR to the nucleus results in complementation with EA, production of active β -Gal units and production of a chemiluminescent signal.

Experiment design:

cell processing

1. PathHunter NHR cells were revived and expanded according to standard procedures.

2. mu.L of cells were seeded into a white wall 384 microwell plate, incubated at 37 ℃ for an appropriate period of time, and then tested. The test medium used charcoal dextran filtered serum to reduce hormone levels.

Agonist testing

1. For agonist assays, cells are incubated with compounds to induce a response.

2. Compound stock was diluted 5-fold with system buffer.

3. mu.L of the 5-fold diluted compound was added to the cells and incubated at 37 ℃ or room temperature for 3-16 hours. The final concentration of the solvent is 1%.

Signal detection:

1. assay signals were generated by adding 12.5 or 15 μ L (50% v/v) of PathHunter detection reagent and incubating for 1 hour at room temperature.

2. The chemiluminescent signal generated in the microplate was read using envision (tm) by PerkinElmer.

And (3) data analysis:

1. compound activity was analyzed using CBIS data analysis software (chemlinnovation, CA).

2. For agonist testing, the percent activity was calculated using the following formula:

% activity is 100% × (average RLU for test sample-average RLU for vehicle control)/(average maximum RLU for control ligand-average RLU for vehicle control).

TABLE 1 THR α and THR β activities of the compounds of the examples

Examples	Compound (I)	THRαEC 50(μM)	THRβEC 50(μM)	Selectivity (THR alpha/THR beta)
Reference compound	MGL-3196	4.18	2.9	1.4
1	WX001	>100	2.46	>40.7
2	WX002	0.76	1.75	0.4

3	WX003	1.6	0.24	6.7
4	WX004	7.45	2.13	3.5
5	WX005	10.11	0.49	20.6
6	WX006	>100	12.11	>8.3
8	WX008	27.23	9.94	2.7
9	WX009	6.51	26.4	0.2
11	WX011	>100	4.7	>21.3
12	WX012	22.2	13.6	1.6
13	WX013	>100	5.02	>19.9
13	WX014	>100	3.79	>26.4
15	WX015	19.46	2.68	7.2
16	WX016	20.91	3.78	5.5
17	WX017	>100	3.97	>25.2
18	WX018	>100	21.75	>4.6
19	WX019	3.27	0.076	43
20	WX020	>100	3.36	>29.8
21	WX021	>100	6.76	>14.8
21	WX022	>100	10.27	>9.7
23	WX023	>100	1.58	>63.4
23	WX024	>100	5.13	>19.5
25	WX025	>100	4.25	>23.5
25	WX026	3.62	1.09	3.3
27	WX027	>100	1.3	>76.9
28	WX028	>100	1.3	>76.9
29	WX029	>100	37.39	>2.7
29	WX030	>100	2.27	>44.1
31	WX031	>100	6.14	>16.3
31	WX032	7.61	1.35	5.6
33	WX033	>100	4.61	>21.7
33	WX034	35.81	5.53	6.5

35	WX035	36.26	11.25	3.2
35	WX036	2.58	0.57	4.5

And (4) conclusion: the compound of the invention has obvious THR alpha/beta activity, and the THR alpha selectivity of partial compound is superior to MGL-3196.

Experiment two: cytochrome P450 isoenzyme inhibitory study

Purpose of the experiment: the inhibitory effect of the test compounds on the activity of human liver microsomal cytochrome P450 isozymes (CYP1a2, CYP2C9, CYP2C19, CYP2D6 and CYP3a4) was measured.

Experimental procedure 1: firstly, diluting a tested compound (10.0mM) to prepare a working solution (100 multiplied by the final concentration), wherein the concentration of the working solution is 1.00mM, and simultaneously preparing working solutions of P450 isozymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4) positive inhibitors and specific substrate mixtures thereof (5 in 1); human liver microsomes stored in a refrigerator at a temperature lower than-60 ℃ are unfrozen on ice, and diluted by PB after the human liver microsomes are completely dissolved to prepare working solution (0.253mg/mL) with a certain concentration. Adding 20.0 μ L of substrate mixture into the reaction plate (adding 20.0 μ L PB into blank hole), adding 158 μ L of human liver microsome working solution into the reaction plate, and placing the reaction plate on ice for later use; at this time, 2.00 μ L of test compound (N ═ 2) and specific inhibitor (N ═ 2) were added to the corresponding wells, the group without inhibitor (test compound or positive inhibitor) was added to the corresponding organic solvent, and the organic phases of the test compound control sample and positive control sample were both 1:9DMSO: MeOH; after pre-incubation in water bath at 37 ℃ for 10min, adding 20.0 mu L of coenzyme factor (NADPH) solution into the reaction plate, and placing the reaction plate in water bath at 37 ℃ for incubation reaction for 10 min; the reaction was stopped by adding 400. mu.L of a pre-chilled acetonitrile solution (containing 200ng/mL of Tolbutamide and Labetalol as internal standards); placing the reaction plate in a shaking table, and shaking and mixing for 10 min; then centrifuging for 20min at the conditions of 4 ℃ and 4000 rpm; adding 200 mu L of supernatant into 100 mu L of water for sample dilution; finally, closing the plate, oscillating, shaking up and carrying out LC/MS/MS detection. The results of the experiment are shown in table 2:

TABLE 2 results of inhibition of human liver microsomal cytochrome P450 isozyme activity by test compounds

Experimental operation 2: the test compound (10.0mM) was first diluted in a gradient to prepare working solutions (100 × final concentration) at respective concentrations: 5.00,1.50,0.500,0.150,0.0500,0.0150 and 0.00500mM, and simultaneously preparing working solution of each positive inhibitor of P450 isozyme (CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4) and a specific substrate mixture (5 in 1); human liver microsomes stored in a refrigerator at a temperature lower than-60 ℃ are unfrozen on ice, and diluted by PB after the human liver microsomes are completely dissolved to prepare working solution (0.253mg/mL) with a certain concentration. Adding 20.0 μ L of substrate mixture into the reaction plate (adding 20.0 μ L PB into blank hole), adding 158 μ L of human liver microsome working solution into the reaction plate, and placing the reaction plate on ice for later use; at this time, 2.00 μ L of each concentration of the test compound (N ═ 1) and the specific inhibitor (N ═ 2) was added to the corresponding well, and the group without the inhibitor (test compound or positive inhibitor) was added with the corresponding organic solvent as a control sample (test compound control sample was 1:1DMSO: MeOH, and positive control samples were all 1:9DMSO: MeOH); after pre-incubation in water bath at 37 ℃ for 10min, adding 20.0 mu L of coenzyme factor (NADPH) solution into the reaction plate, and placing the reaction plate in water bath at 37 ℃ for incubation reaction for 10 min; the reaction was stopped by adding 400. mu.L of a pre-chilled acetonitrile solution (containing 200ng/mL of Tolbutamide and Labetalol as internal standards); placing the reaction plate in a shaking table, and shaking and mixing for 10 min; then centrifuging for 20min at the conditions of 4 ℃ and 4000 rpm; adding 200 mu L of supernatant into 100 mu L of water for sample dilution; finally, closing the plate, oscillating, shaking up and carrying out LC/MS/MS detection. The results of the experiment are shown in table 3:

TABLE 3 results of the inhibition of the activity of human liver microsomal cytochrome P450 isozyme by test compounds

And (4) experimental conclusion: the compound of the invention has no inhibition effect on CYP2C19, CYP2D6 and CYP3A4, and has moderate or weak inhibition effect on CYP1A2 and CYP2C 9.

Experiment three: plasma protein binding Rate Studies

Purpose of the experiment: determination of the protein binding Rate of test Compounds in CD-1 mice, SPRAGE-DAWLEY rats, beagle dogs, cynomolgus monkeys and human plasma

And (3) experimental operation: 995. mu.L of blank plasma from CD-1 mice, SPRAGUE-DAWLEY rats, beagle dogs, cynomolgus monkeys and humans were taken, and 5. mu.L of a test compound working solution (400. mu.M) or a warfarin working solution (400. mu.M) was added to make the final concentrations of the test compound and warfarin 2. mu.M in the plasma samples. The samples were mixed thoroughly. The final concentration of organic phase DMSO is 0.5%; 50 μ L of test compound and warfarin plasma samples were pipetted into sample receiving plates (three in parallel) and corresponding volumes of blank plasma or buffer were immediately added such that the final volume of each sample well was 100 μ L, plasma: the volume ratio of dialysis buffer was 1:1, and 500. mu.L of stop solution was added to these samples, which were to be designated as T₀Samples were used for recovery determinations. Will T₀Storing the sample at 2-8 deg.C, and waiting for subsequent treatment with other dialyzed samples; mu.L of test compound and warfarin plasma samples were added to the administration end of each dialysis well, and 150. mu.L of blank dialysis buffer was added to the receiving end of the corresponding dialysis well. The dialysis plates were then placed in a humidified, 5% CO₂The incubator of (4) was incubated at 37 ℃ for 4 hours with shaking at about 100 rpm. After the dialysis was completed, 50. mu.L of the buffer sample after dialysis and the plasma sample after dialysis were removed to prepare new samplesA product receiving plate. Corresponding blank plasma or buffer was added to the samples in corresponding volumes such that the final volume of each sample well was 100 μ L, plasma: the volume ratio of the dialysis buffer was 1: 1. All samples were subjected to LC/MS/MS analysis after protein precipitation and analyzed by the formula: % unbound ═ 100 × F/T,% bound ═ 100 [% unbound,% recovered ═ 100 × (F + T)/T₀The protein binding rate and recovery rate were calculated (where F is the peak area ratio of the compound in the dialysate after 4h dialysis; T is the peak area ratio of the compound in the plasma after 4h dialysis)₀Is the peak area ratio of the compound in the plasma sample at time zero). The results of the experiment are shown in table 4:

TABLE 4 plasma protein binding rates of test compounds in CD-1 mice, SPRAGEE-DAWLEY rats, beagle dogs, cynomolgus monkeys, and humans

Note: "/" indicates no detection.

And (4) experimental conclusion: the compounds of the invention show higher protein binding rates in CD-1 mice, SPRAGUE-DAWLEY rats, beagle dogs, cynomolgus monkeys and human plasma.

Experiment four: in vivo pharmacokinetic Studies

Pharmacokinetics studies of oral and intravenous injections of the Compounds of the invention in C57BL/6 mice

The test compound was mixed with 2% N, N-dimethylformamide/10% polyethylene glycol 400/44% 0.1M phosphate buffer/44% 40% aqueous cyclodextrin solution, vortexed and sonicated to prepare a 1.5mg/mL clear solution. C57BL/6 male mice 7 to 10 weeks old were selected and administered the candidate compound solution Intravenously (IV) at a dose of 3 mg/kg.

The test compound was mixed with 2% hydroxypropylmethylcellulose/0.1% tween 80 aqueous solution, vortexed and sonicated to prepare a 1mg/mL homogeneous suspension for use. C57BL/6 male mice 7 to 10 weeks old were selected and the candidate compound solution was administered orally (PO) at a dose of 5 mg/kg.

Whole blood was collected for a certain period of time, plasma was prepared, drug concentration was analyzed by LC-MS/MS method, and drug parameters were calculated using Phoenix WinNonlin software (Pharsight, USA).

Whole blood was collected for a certain period of time, plasma was prepared, drug concentration was analyzed by LC-MS/MS method, and drug parameters were calculated using Phoenix WinNonlin software (Pharsight, USA).

The results of the experiment are shown in table 5:

TABLE 5 pharmacokinetic results for the Compounds of the invention

Note: IV: intravenous injection; PO: orally taking; DNAUC ═ AUC/dose administered; N/A: not applicable; and (4) conclusion: the compound of the invention has higher exposure and better oral bioavailability.

Claims (20)

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof,

   

   wherein,

   

   is selected fromSingle and double bonds;

   T ₁is NR_a；

   T ₂Selected from C, CH and N;

   T ₃selected from the group consisting of CR_bAnd N;

   T ₄selected from the group consisting of CR_cAnd O;

   R _aselected from H and C_1-3An alkyl group;

   R _band R_cEach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl and C_1-3Alkoxy radical, said C_1-3Alkyl and C_{1- 3}Alkoxy is optionally substituted with 1,2 or 3R;

   and when R is_cSelected from F, Cl, Br, I, OH, NH₂And C optionally substituted by 1,2 or 3R_1-3Alkyl and C_1-3At alkoxy radical, R₁And R₂Each independently selected from H and optionally substituted by 1,2 or 3R_dSubstituted C_1-6Alkyl, or, R₁、R ₂And the atoms to which they are attached together form thienyl, C_3-8Cycloalkenyl or 3-8 membered heterocycloalkenyl, said thienyl, C_3-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

   when R is_cSelected from H and CN, R₁、R ₂And the atoms to which they are attached together form thienyl, C_6-8Cycloalkenyl or 3-8 membered heterocycloalkenyl, said thienyl, C_6-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

   R ₃and R₄Each independently selected from H, F, Cl, Br, I, OH, NH₂CN and optionally substituted by 1,2 or 3R_fSubstitutionC of (A)_1-3An alkyl group;

   R _dand R_eEach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl radical, C_1-3Alkoxy and-O-C_3-5Cycloalkyl radical, said C_1-3Alkyl radical, C_1-3Alkoxy and-O-C_3-5Cycloalkyl is optionally substituted with 1,2 or 3R';

   R _findependently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl and C_1-3An alkoxy group;

   each R is independently selected from H, F, Cl, Br, I, OH and NH₂CN and C_1-3An alkyl group;

   each R' is independently selected from D, F, Cl, Br and I;

   the 3-8 membered heterocycloalkenyl contains 1,2,3, or 4 heteroatoms or groups of heteroatoms independently selected from-NH-, -O-, -S-, and N.
2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, selected from

   

   Wherein,

   

   selected from single and double bonds;

   T ₂selected from C and N;

   T ₄selected from the group consisting of CR_cAnd O;

   R _cselected from F, Cl, Br, I, OH, NH₂、C _1-3Alkyl and C_1-3Alkoxy radical, said C_1-3Alkyl and C_1-3Alkoxy is optionally substituted with 1,2 or 3R;

   R ₁and R₂Each independently selected from H and optionally substituted by 1,2 or 3R_dSubstituted C_1-6Alkyl, or, R₁、R ₂And the atoms to which they are attached together form thienyl, C_3-8Cycloalkenyl or 3-8 membered heterocycloalkenyl, said thienyl, C_3-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

   R ₃and R₄Each independently selected from H, F, Cl, Br, I, OH, NH₂CN and optionally substituted by 1,2 or 3R_fSubstituted C_1-3An alkyl group; r_dAnd R_eEach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl radical, C_1-3Alkoxy and-O-C_3-5Cycloalkyl radical, said C_1-3Alkyl radical, C_1-3Alkoxy and-O-C_3-5Cycloalkyl is optionally substituted with 1,2 or 3R';

   R _findependently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl and C_1-3An alkoxy group;

   each R is independently selected from H, F, Cl, Br, I, OH and NH₂CN and C_1-3An alkyl group;

   each R' is independently selected from D, F, Cl, Br and I;

   the 3-8 membered heterocycloalkenyl contains 1,2,3, or 4 heteroatoms or groups of heteroatoms independently selected from-NH-, -O-, -S-, and N.
3. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, selected from

   

   Wherein,

   R _cselected from H and CN;

   R ₁、R ₂and the atoms to which they are attached together form thienyl, C_6-8Cycloalkenyl and 3-8 membered heterocycloalkenyl, said thienyl, C_6-8Cycloalkenyl and 3-8 membered heterocycloalkenyl optionally substituted with 1,2 or 3R_eSubstitution;

   R ₃and R₄Each independently selected from H, F, Cl, Br, I, OH, NH₂CN and optionally substituted by 1,2 or 3R_fSubstituted C_1-3An alkyl group; r_eEach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl radical, C_1-3Alkoxy and-O-C_3-5Cycloalkyl radical, said C_1-3Alkyl radical, C_1-3Alkoxy and-O-C_3-5Cycloalkyl is optionally substituted with 1,2 or 3R';

   R _findependently selected from H, F, Cl, Br, I, OH, NH₂、CN、C _1-3Alkyl and C_1-3An alkoxy group;

   each R' is independently selected from D, F, Cl, Br and I;

   the 3-8 membered heterocycloalkenyl contains 1,2,3, or 4 heteroatoms or groups of heteroatoms independently selected from-NH-, -O-, -S-, and N.
4. A compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein R_bAnd R_cEach independently selected from H, F, Cl, Br, I, OH, NH₂、CN、CH ₃、CH ₂F、CH ₂CN、CHF ₂、CF ₃、CH ₂CH ₃、CF ₂CH ₃、

   
5. A compound according to claim 2, or a pharmaceutically acceptable salt thereof, wherein R_cSelected from F, Cl, Br, I, OH, NH₂、CH ₃、CH ₂F、CHF ₂、CH ₂CN、CF ₃、CH ₂CH ₃、CF ₂CH ₃、

   
6. The compound according to any one of claims 1,2, 4 or 5, or a pharmaceutically acceptable salt thereof, wherein R₁And R₂Each independently selected from H, CH₃、CH ₂CH ₃、CH(CH ₃) ₂And C (CH)₃) ₃。
7. A compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, wherein R₁、R ₂And the atoms to which they are attached together form a cyclohexenyl, bicyclo [2.2.1 ]]Hept-2-enyl, 1,2,3, 4-tetrahydropyridinyl, bicyclo [4.1.0]Hept-3-enyl, 3, 6-dihydro-2H-pyranyl, 7-oxabicyclo [2.2.1]Hept-2-enyl, thienyl, 3, 4-dihydro-2H-pyranyl and bicyclo [4.1.0]Hept-2-enyl, said cyclohexenyl, bicyclo [2.2.1]Hept-2-enyl, 1,2,3, 4-tetrahydropyridinyl, bicyclo [4.1.0]Hept-3-enyl, 3, 6-dihydro-2H-pyranyl, 7-oxabicyclo [2.2.1]Hept-2-enyl, thienyl, 3, 4-dihydro-2H-pyranyl and bicyclo [4.1.0]Hept-2-enyl optionally substituted by 1,2 or 3R_eAnd (4) substitution.
8. A compound according to claim 7, or a pharmaceutically acceptable salt thereof, wherein the building block

   

   Is selected from

   

   
9. A compound according to claim 8, or a pharmaceutically acceptable salt thereof, wherein the building block

   

   Is selected from

   

   
10. A compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, wherein R₃And R₄Each independently selected from H, F, Cl, Br, I, OH, NH₂、CN、CH ₃And CH₂CH ₃Said CH₃And CH₂CH ₃Optionally substituted by 1,2 or 3R_fAnd (4) substitution.
11. A compound according to claim 10, or a pharmaceutically acceptable salt thereof, wherein R₃And R₄Each independently selected from F, Cl, Br, I, OH, NH₂And CF₃。
12. A compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein the building block

    

    Is selected from

    

    
13. A compound according to claim 4 or 12, or a pharmaceutically acceptable salt thereof, wherein the building block

    

    Is selected from

    

    
14. A compound according to any one of claims 1,3, 4, 10 or 11, or a pharmaceutically acceptable salt thereof, selected from

    

    Wherein,

    Z ₁and Z₂Are each independently selected from CH (R)_e) O and N (R)_e)；

    Z ₃Is selected from CH₂And O;

    Z ₄is selected from O;

    Z ₅is selected from C (R)_e) And N;

    Z ₆is selected from N (R)_e) And O;

    R _cselected from H and CN;

    R ₃、R ₄and R_eAs defined in any one of claims 1,3, 4, 10 or 11.
15. A compound according to any one of claims 1,2, 5, 10 or 11, or a pharmaceutically acceptable salt thereof, selected from

    

    Wherein,

    R ₁、R ₂、R ₃、R ₄and R_cAs defined in any one of claims 1,2, 5, 10 or 11.
16. A compound of the formula or a pharmaceutically acceptable salt thereof,

    

    

    
17. the compound of claim 16, an isomer thereof, or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

    

    
18. a pharmaceutical composition comprising as an active ingredient a therapeutically effective amount of a compound according to any one of claims 1 to 17 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.
19. Use of a compound according to any one of claims 1 to 17 or a pharmaceutically acceptable salt thereof or a composition according to claim 18 for the manufacture of a medicament associated with a thyroxine receptor- β agonist.
20. The use of claim 19, wherein the thyroxine receptor- β agonist related medicament is a medicament for the treatment of non-alcoholic steatohepatitis.