RU2832735C2 - Oestrogen receptor antagonist - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to a compound of formula (II), an isomer thereof or a pharmaceutically acceptable salt thereof, (II), where X is NH; Y is selected from a group consisting of N and CR₇; Y₁ is CH and Y₂ is N; or Y₁ is N and Y₂ is CH or CF; or Y₁ is CH and Y₂ is CH; L is or ; ring A is selected from the group consisting of phenyl, thienyl and pyridinyl; R₁ is selected from the group consisting of H, halogen and C_1-6 alkyl; R₂ is C_1-3 alkyl, optionally substituted with 1, 2 or 3 R_b; R₃ is selected from the group consisting of H and C_1-3 alkyl; R₁₃ is H or ; R₄ is selected from a group consisting of COOH, , , -NHCH₃ and -N(CH₃)₂; R₅ is H; each of R₆, R₇, R₈ and R₉ independently is H; each of R₁₀, R₁₁ and R₁₂ is independently selected from the group consisting of H, halogen, CN, C_1-3alkyl and -OC_1-3 alkyl; and R_b is selected from the group consisting of H, F, Cl, Br and I, as well as to a pharmaceutical composition based thereon and use in preparing a medicament for treating oestrogen receptor-positive breast cancer.

EFFECT: novel compounds which can be used as oestrogen receptor antagonists for treating oestrogen receptor-positive breast cancer are obtained.

29 cl, 4 tbl, 31 ex

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit and priority of Chinese Patent Application No. 201811545117.2 filed with the National Intellectual Property Administration of the People's Republic of China on December 17, 2018, Chinese Patent Application No. 201910611070.3 filed with the National Intellectual Property Administration of the People's Republic of China on July 8, 2019, and Chinese Patent Application No. 201910877339.2 filed with the National Intellectual Property Administration of the People's Republic of China on September 17, 2019, which are incorporated herein by reference in their entireties.

AREA OF TECHNOLOGY

The present application relates to new indole compounds and, in particular, discloses a compound of formula (II), its isomer or a pharmaceutically acceptable salt thereof, or their use as an estrogen receptor antagonist in the preparation of a medicament for the treatment of estrogen receptor-positive breast cancer.

BACKGROUND OF THE INVENTION

According to WHO statistics, breast cancer is the second most common cancer in the world population and the most common cancer in women. Over the years, research has identified the role of the estrogen-estrogen receptor signaling pathway in the development of breast cancer; the estrogen receptor (ER) has become the most important biomarker for breast cancer. Based on estrogen receptor expression, breast cancers can be classified into estrogen receptor-positive breast cancer and estrogen receptor-negative breast cancer. Of these, estrogen receptor-positive breast cancer accounts for more than 70% of breast cancer patients.

Endocrine therapy (ET) of the estrogen-estrogen receptor (ER) signaling pathway in breast cancer cells has become the first choice for the treatment of ER-positive breast cancer due to its minimal adverse effects and exceptional efficacy. Typically, the first-line endocrine therapy is an aromatase inhibitor (AI). Although letrozole, an aromatase inhibitor, has shown good efficacy in the treatment of ER-positive breast cancer in two marketed drugs, the problem of ER-positive breast cancer resistance to AI is becoming more pronounced. A large number of studies have shown that in the presence of AI, the ER gene can mutate, predominantly at Y537X, resulting in the production of a mutant ER that can remain in the excited conformation in the absence of estrogen and continue to function as a receptor, promoting proliferation of breast cancer cells. The only marketed selective estrogen receptor suppressor is fulvestrant, which has demonstrated good results in the treatment of hormone-resistant breast cancer. However, fulvestrant has many drawbacks in the treatment of AI-resistant, ER-mutant breast cancer. Due to the poor pharmacokinetics (PK) of fulvestrant, it is characterized by a lack of oral bioavailability with a high rate of renal clearance. For the two reasons mentioned above, this drug can only be administered by intramuscular injection. However, due to its pronounced lipophilicity, fulvestrant also has serious tissue distribution problems when administered by intramuscular injection, resulting in a clinical response rate of approximately 50% in patients with breast cancer. Also, due to unsatisfactory PK properties, currently approved doses of fulvestrant cannot completely destroy ER, especially mutant ER, when concentrated in tissues. Therefore, the treatment of AI-resistant ER-mutant breast cancer is far from optimal. Therefore, the development of drugs with better PK aimed at treating ER-mutant breast cancer remains an unmet need in the field of medicine.

Patent No. US20160347717A1 reports an orally administered covalent estrogen receptor antagonist, H3B-6545. Phase I/II clinical trials of this molecule for the treatment of ER-positive breast cancer are ongoing.

BRIEF DESCRIPTION OF THE INVENTION

The present application provides a compound of formula (II), its isomer or a pharmaceutically acceptable salt thereof,

,

Where

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

Y is selected from the group consisting of N and CR ₇ ;

Y ₁ is CH, and Y ₂ is N;

or Y ₁ is N and Y ₂ is CH or CF;

or Y ₁ is CH and Y ₂ is CH;

L represents or ;

ring A is selected from the group consisting of C _6-10 aryl and 5-10 membered heteroaryl;

R ₁ is selected from the group consisting of H, halogen, CN, COOH, NH ₂ , C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl, wherein C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _a ;

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

R ₃ is selected from the group consisting of H, halogen, CN, NO ₂ , OH, COOH, NH ₂ , C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl, wherein C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _c ;

R ₁₃ is H or ;

R ₄ is selected from the group consisting of COOH, NH ₂ , C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl, 3-6-membered heterocycloalkyl, phenyl and 5-6-membered heteroaryl, wherein C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl, 3-6-membered heterocycloalkyl, phenyl and 5-6-membered heteroaryl are optionally substituted with 1, 2 or 3 R _d ;

R ₅ is selected from the group consisting of H, C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl, wherein C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _e ;

each of R ₆ , R ₇ , R ₈ and R ₉ is independently selected from the group consisting of H, halogen, CN, NO ₂ , OH, COOH, NH ₂ , C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl, wherein the C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _f ;

each of R ₁₀ , R ₁₁ and R ₁₂ is independently selected from the group consisting of H, halogen, CN, NO ₂ , OH, COOH, NH ₂ , C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl, wherein the C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _g ;

each of R _a , R _b , R _c , R _d , _Re , R _f and R _g is independently selected from the group consisting of H, F, Cl, Br, I, OH, CN, NH ₂ , COOH, C(=O)NH ₂ , C _1-8 alkyl, C _1-8 heteroalkyl, C _3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl and C _3-6 cycloalkyl-C _1-3 alkyl-, wherein the C _1-8 alkyl, C _1-8 heteroalkyl, C _3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl and C _3-6 cycloalkyl-C _1-3 alkyl- are optionally substituted with 1, 2 or 3 R;

R is independently selected from the group consisting of F, Cl, Br, I, OH, CN, _NH2 , COOH, Me, Et, _CF3 , _CHF2 , _CH2F , _NHCH3, and N( _CH3 ) ₂ ;

each of the C _1-6 heteroalkyl, 3-6-membered heterocycloalkyl and 5-6-membered heteroaryl contains 1, 2, 3 or 4 heteroatoms or heteroatomic groups, each of which is independently selected from the group consisting of -NH-, -O-, -S-, -ON=, -C(=O)-O-, -C(=O)-S-, -C(=O)-, -C(=S)-, -S(=O)-, -S(=O) _2- and N.

In some embodiments of the present application, X is selected from the group consisting of NH, O, and S; Y is CR ₇ ; Y ₁ is CH, and Y ₂ is N, or Y ₁ is N, and Y ₂ is CH; L is ; R ₁₃ is ; other variables are as defined in this application.

In some embodiments of the present application, X is NH; Y is CR ₇ ; Y ₁ is N, and Y ₂ is CH; L is ; R ₁₃ is ; other variables are as defined in this application.

In some embodiments of the present application, X is NH; Y is CR ₇ ; Y ₁ is CH, and Y ₂ is N; L is ; R ₁₃ is ; other variables are as defined in this application.

In some embodiments of the present application, X is NH; Y is N; Y ₁ is N, and Y ₂ is CH; L is ; R ₁₃ is ; other variables are as defined in this application.

In some embodiments of the present application, R is independently selected from the group consisting of F, Cl, Br, and I.

In some embodiments of the present application, each of R _a , R _b , R _c , R _d , _Re , R _f and R _g is independently selected from the group consisting of H, F, Cl, Br, I, OH, CN, NH ₂ , COOH, C(=O)NH ₂ , C _1-6 alkyl, C _1-6 heteroalkyl and cyclopropyl, wherein C _1-6 alkyl, C _1-6 heteroalkyl and cyclopropyl are optionally substituted with 1, 2 or 3 R; other variables are as defined herein.

In some embodiments of the present application, each of R _a , R _b , R _c , R _d , _Re , R _f and R _g is independently selected from the group consisting of H, F, Cl, Br, I, OH, CN, NH ₂ , COOH, C(=O)NH ₂ , Me, Et, CF ₃ , CHF ₂ , CH ₂ F, NHCH ₃ and N(CH ₃ ) ₂ ; other variables are as defined herein.

In some embodiments of the present application, each of R _a , R _b , R _c , R _d , _Re , R _f and R _g is independently selected from the group consisting of F, Cl, Br, I, Me, Et, CF ₃ , CHF ₂ and CH ₂ F; other variables are as defined herein.

In some embodiments of the present application, R _b is selected from the group consisting of F, Cl, Br, and I; other variables are as defined herein.

In some embodiments of the present application, R _d is C _1-6 alkyl, wherein the C _1-6 alkyl is optionally substituted with 1, 2, or 3 R selected from the group consisting of F, Cl, Br, and I; other variables are as defined herein.

In some embodiments of the present application, R _d is methyl, wherein methyl is optionally substituted with 1, 2, or 3 R, wherein R is F; other variables are as defined herein.

In some embodiments of the present application, R _d is selected from the group consisting of CF ₃ , CHF _{2 ,} and CH ₂ F; other variables are as defined herein.

In some embodiments, R _d is CH ₂ F; other variables are as defined herein.

In some embodiments, ring A is selected from the group consisting of phenyl and 5-6 membered heteroaryl; other variables are as defined herein.

In some embodiments, ring A is selected from the group consisting of phenyl, 5-membered sulfur-containing heteroaryl, and 6-membered nitrogen-containing heteroaryl; other variables are as defined herein.

In some embodiments, ring A is selected from the group consisting of phenyl, thienyl, and pyridinyl; other variables are as defined herein.

In some embodiments, ring A is selected from the group consisting of phenyl, And ; other variables are as defined in this application.

In some embodiments of the present application, ring A is selected from the group consisting of phenyl and pyridinyl; other variables are as defined herein. In some embodiments of the present application, R ₁ is selected from the group consisting of H, F, Cl, Br, I, CN, COOH, NH ₂ , C _1-3 alkyl, and C _1-3 heteroalkyl, wherein the C _1-3 alkyl and C _1-3 heteroalkyl are optionally substituted with 1, 2, or 3 _Ra ; other variables are as defined herein. In some embodiments of the present application, R ₁ is selected from the group consisting of H, F, Cl, Br, I, CN, COOH, NH ₂ , Me, Et, CF ₃ , CHF ₂ , CH ₂ F, -NHCH ₃ , and -N(CH ₃ ) ₂ ; other variables are as defined in this application.

In some embodiments of the present application, R ₁ is selected from the group consisting of H, F, Cl, Br, I, and C _1-6 alkyl; other variables are as defined herein.

In some embodiments of the present application, R ₁ is selected from the group consisting of H, F, Cl, Br, I, and C _1-3 alkyl; other variables are as defined herein.

In some embodiments of the present application, R ₁ is selected from the group consisting of H, Cl, Br, I and Me; other variables are as defined herein.

In some embodiments of the present application, R ₂ is selected from the group consisting of C _1-3 alkyl and C _1-3 heteroalkyl, wherein the C _1-3 alkyl and C _1-3 heteroalkyl are optionally substituted with 1, 2, or 3 R _b ; other variables are as defined herein. In some embodiments of the present application, R ₂ is selected from the group consisting of CF ₃ , CH ₂ CF ₃ , CHF ₂ , CH ₂ F, -NHCH ₃ and -N(CH ₃ ) ₂ ; other variables are as defined herein.

In some embodiments of the present application, R ₂ is C _1-6 alkyl, wherein the C _1-6 alkyl is optionally substituted with 1, 2, or 3 R _b ; other variables are as defined herein.

In some embodiments of the present application, R ₂ is C _1-6 alkyl, wherein the C _1-6 alkyl is optionally substituted with 1, 2, or 3 R _b , wherein R _b is selected from the group consisting of F, Cl, Br, and I; other variables are as defined herein.

In some embodiments of the present application, R ₂ is C _1-3 alkyl, wherein the C _1-3 alkyl is optionally substituted with 1, 2, or 3 R _b , wherein R _b is F; other variables are as defined herein.

In some embodiments of the present application, R ₂ is ethyl, wherein ethyl is optionally substituted with 3 R _b , wherein R _b is F; other variables are as defined herein.

In some embodiments of the present application, R ₂ is selected from the group consisting of ethyl and CH ₂ CF ₃ ; other variables are as defined herein.

In some embodiments of the present application, R ₃ is selected from the group consisting of H and C _1-6 alkyl; other variables are as defined herein.

In some embodiments of the present application, R ₃ is selected from the group consisting of H and C _1-3 alkyl; other variables are as defined herein.

In some embodiments of the present application, R ₃ is selected from the group consisting of H and methyl; other variables are as defined herein.

In some embodiments of the present application, R ₃ is H; other variables are as defined herein.

In some embodiments of the present application, R ₄ is selected from the group consisting of COOH, NH ₂ , C _1-6 alkyl, -NH-C _1-6 alkyl, -N(C _1-6 alkyl) ₂ , -C(=O)-OC _1-6 alkyl, -C(=O)-SC _1-6 alkyl, C _3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, and phenyl, wherein C _1-6 alkyl, -NH-C _1-6 alkyl, -N(C _1-6 alkyl) ₂ , -C(=O)-OC _1-6 alkyl, -C(=O)-SC _1-6 alkyl, C _3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, and phenyl are optionally substituted with 1, 2, or 3 R _d ; other variables are as defined herein. In some embodiments of the present application, said R ₄ is selected from the group consisting of COOH, NH ₂ , C _1-3 alkyl, -NH-C _1-3 alkyl, -N(C _1-3 alkyl) ₂ , -C(=O)-OC _1-3 alkyl, -C(=O)-SC _1-3 alkyl, cyclopropyl, cyclopentyl, cyclohexyl, azetidinyl and phenyl, wherein C _1-3 alkyl, -NH-C _1-3 alkyl, -N(C _1-3 alkyl) ₂ , -C(=O)-OC _1-3 alkyl, -C(=O)-SC _1-3 alkyl, cyclopropyl, cyclopentyl, cyclohexyl, azetidinyl and phenyl are optionally substituted with 1, 2 or 3 R _d ; other variables are as defined herein.

In some embodiments of the present application, R ₄ is selected from the group consisting of -NH-C _1-6 alkyl, -N(C _1-6 alkyl) ₂ , and 3-6 membered heterocycloalkyl, wherein -NH-C _1-6 alkyl, -N(C _1-6 alkyl) ₂ , and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2, or 3 R _d ; other variables are as defined herein.

In some embodiments of the present application, R ₄ is selected from the group consisting of -NH-C _1-3 alkyl, -N(C _1-3 alkyl) ₂ and 4-membered heterocycloalkyl, wherein the -NH-C _1-3 alkyl, -N(C _1-3 alkyl) ₂ and 4-membered heterocycloalkyl groups are optionally substituted with 1, 2 or 3 R _d ; other variables are as defined herein.

In some embodiments of the present application, R ₄ is selected from the group consisting of -NH-C _1-3 alkyl, -N(C _1-3 alkyl) ₂ and , while optionally substituted with 1, 2 or 3 R _d ; other variables are as defined in this application.

In some embodiments of the present application, R ₄ is selected from the group consisting of -NH-C _1-3 alkyl, -N(C _1-3 alkyl) ₂ and , while optionally substituted with 1, 2 or 3 R _d , wherein R _d is selected from the group consisting of CF ₃ , CHF ₂ and CH ₂ F; other variables are as defined herein.

In some embodiments of the present application, R ₄ is selected from the group consisting of , , -NHCH ₃ and -N(CH ₃ ) ₂ ; other variables are as defined in this application.

In some embodiments of the present application, R ₅ is selected from the group consisting of H, C _1-3 alkyl, and C _1-3 heteroalkyl, wherein the C _1-3 alkyl and C _1-3 heteroalkyl are optionally substituted with 1, 2, or 3 R _e ; other variables are as defined herein.

In some embodiments of the present application, R ₅ is H; other variables are as defined herein.

In some embodiments of the present application, R ₆ , R ₇ , R ₈ and R ₉ are H; other variables are as defined herein.

In some embodiments of the present application, each of R ₁₀ , R ₁₁ , and R ₁₂ is independently selected from the group consisting of H, F, Cl, Br, I, CN, NO ₂ , OH, COOH, NH ₂ , C _1-3 alkyl, and C _1-3 heteroalkyl, wherein the C _1-3 alkyl and C _1-3 heteroalkyl are optionally substituted with 1, 2, or 3 R _g ; other variables are as defined herein. In some embodiments of the present application, each of R ₁₀ , R ₁₁ , and R ₁₂ is independently selected from the group consisting of H, F, Cl, Br, I, CN, NO ₂ , OH, COOH, NH ₂ , Me, Et, CF ₃ , OMe, CHF ₂ , CH ₂ F, NHCH ₃ , and N(CH ₃ ) ₂ ; other variables are as defined in this application.

In some embodiments of the present application, each of R ₁₀ , R ₁₁ and R ₁₂ is independently selected from the group consisting of H, F, Cl, Br, I, CN, C _1-6 alkyl and C _1-6 heteroalkyl, wherein the C _1-6 alkyl and C _1-6 heteroalkyl are optionally substituted with 1, 2 or 3 R _g ; other variables are as defined herein.

In some embodiments of the present application, each of R ₁₀ , R ₁₁ and R ₁₂ is independently selected from the group consisting of H, F, Cl, Br, I, CN, C _1-3 alkyl and C _1-3 heteroalkyl.

In some embodiments of the present application, each of R ₁₀ , R ₁₁ and R ₁₂ is independently selected from the group consisting of H, F, Cl, Br, I, CN, Me and OMe; other variables are as defined herein.

In some embodiments of the present application, X is NH; other variables are as defined herein.

In some embodiments of the present application, Y is selected from the group consisting of N and CH; other variables are as defined herein.

In some embodiments of the present application, Y ₁ is N and Y ₂ is CH or CF; other variables are as defined herein.

In some embodiments of the present application, Y ₁ is CH and Y ₂ is CH; other variables are as defined herein.

In some embodiments of the present application, the structural unit selected from the group consisting of And ; other variables are as defined in this application.

In some embodiments of the present application, the structural unit selected from the group consisting of And ; other variables are as defined in this application.

In some embodiments of the present application, the structural unit selected from the group consisting of , , And ; other variables are as defined in this application.

In some embodiments of the present application, the structural unit selected from the group consisting of , , , , And ; other variables are as defined in this document.

In some embodiments of the present application, the structural unit selected from the group consisting of , , , , , , , , And ; other variables are as defined in this application.

In some embodiments of the present application, R ₁₃ is H, , , or ; other variables are as defined in this application.

In some embodiments of the present application, R ₁₃ is H, , or ; other variables are as defined in this application.

In some embodiments of the present application, the structural unit selected from the group consisting of , , , And ; other variables are as defined in this application.

In some embodiments of the present application, the structural unit selected from the group consisting of , , And ; other variables are as defined in this application.

In some embodiments of the present application, there is provided a compound, an isomer thereof, or a pharmaceutically acceptable salt thereof selected from the group consisting of:

, , , And ,

Where

R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ , R ₉ , R ₁₀ , R ₁₁ and R ₁₂ are as defined herein;

each of R ₄₁ and R ₄₂ is independently selected from H and C _1-6 alkyl, wherein C _1-6 alkyl is optionally substituted with 1, 2 or 3 R _d ; R _d is as defined herein.

In some embodiments of the present application, each of R ₄₁ and R ₄₂ is independently selected from the group consisting of H and C _1-3 alkyl; other variables are as defined herein.

In some embodiments of the present application, each of R ₄₁ and R ₄₂ is independently selected from the group consisting of H and methyl; other variables are as defined herein.

In some embodiments of the present application, R ₄₁ is H and R ₄₂ is methyl; other variables are as defined herein.

In some embodiments of the present application, there is provided a compound, an isomer thereof, or a pharmaceutically acceptable salt thereof selected from the group consisting of:

, , , And ,

Where

R ₁ is selected from the group consisting of H, Cl, Br, I and Me,

R ₂ is selected from the group consisting of ethyl and CH ₂ CF ₃ ,

R ₃ is selected from the group consisting of H and methyl, and

R ₁₀ , R ₁₁ , R ₁₂ , R _d , R ₄₁ and R ₄₂ are as defined in this application.

The present application further provides a compound of the following formulas, its isomer or a pharmaceutically acceptable salt thereof:

The present application further provides a compound of the following formulas, its isomer or a pharmaceutically acceptable salt thereof:

The present application provides a compound of formula (I'), its isomer or a pharmaceutically acceptable salt thereof,

,

Where

represents a single bond or a double bond;

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

Y is selected from the group consisting of N and CR ₇ ;

Y ₁ is CH, and Y ₂ is N;

or Y ₁ is N and Y ₂ is CH;

ring A is selected from the group consisting of C _6-10 aryl and 5-10 membered heteroaryl;

R ₁ is selected from the group consisting of H, halogen, CN, COOH, NH ₂ , C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl, wherein C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _a ;

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

R ₃ is selected from the group consisting of H, halogen, CN, NO ₂ , OH, COOH, NH ₂ , C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl, wherein C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _c ;

R ₄ is selected from the group consisting of COOH, NH ₂ , C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl, 3-6-membered heterocycloalkyl, phenyl and 5-6-membered heteroaryl, wherein C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl, 3-6-membered heterocycloalkyl, phenyl and 5-6-membered heteroaryl are optionally substituted with 1, 2 or 3 R _d ;

R ₅ is selected from the group consisting of H, C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl, wherein C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _e ;

each of R ₆ , R ₇ , R ₈ and R ₉ is independently selected from the group consisting of H, halogen, CN, NO ₂ , OH, COOH, NH ₂ , C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl, wherein the C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _f ;

each of R ₁₀ , R ₁₁ and R ₁₂ is independently selected from the group consisting of H, halogen, CN, NO ₂ , OH, COOH, NH ₂ , C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl, wherein the C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _g ;

each of R _a , R _b , R _c , R _d , _Re , R _f and R _g is independently selected from the group consisting of H, F, Cl, Br, I, OH, CN, NH ₂ , COOH, C(=O)NH ₂ , C _1-8 alkyl, C _1-8 heteroalkyl, C _3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl and C _3-6 cycloalkyl-C _1-3 alkyl-, wherein the C _1-8 alkyl, C _1-8 heteroalkyl, C _3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl and C _3-6 cycloalkyl-C _1-3 alkyl- are optionally substituted with 1, 2 or 3 R;

R is independently selected from the group consisting of F, Cl, Br, I, OH, CN, _NH2 , COOH, Me, Et, _CF3 , _CHF2 , _CH2F , _NHCH3, and N( _CH3 ) ₂ ;

each of the C _1-6 heteroalkyl, 3-6-membered heterocycloalkyl and 5-6-membered heteroaryl contains 1, 2, 3 or 4 heteroatoms or heteroatomic groups, each of which is independently selected from the group consisting of -NH-, -O-, -S-, -ON=, -C(=O)-O-, -C(=O)-S-, -C(=O)-, -C(=S)-, -S(=O)-, -S(=O) _2- and N.

In some embodiments of the present application, each of R _a , R _b , R _c , R _d , _Re , R _f and R _g in formula (I') is independently selected from the group consisting of H, F, Cl, Br, I, OH, CN, NH ₂ , COOH, C(=O)NH ₂ , C _1-6 alkyl, C _1-6 heteroalkyl and cyclopropyl, wherein the C _1-6 alkyl, C _1-6 heteroalkyl and cyclopropyl are optionally substituted with 1, 2 or 3 R.

In some embodiments of the present application, each of R _a , R _b , R _c , R _d , _Re , R _f and R _g in formula (I') is independently selected from the group consisting of H, F, Cl, Br, I, OH, CN, NH ₂ , COOH, C(=O)NH ₂ , Me, Et, CF ₃ , CHF ₂ , CH ₂ F, NHCH ₃ and N(CH ₃ ) ₂ .

In some embodiments of the present application, R ₁ in formula (I') is selected from the group consisting of H, F, Cl, Br, I, CN, COOH, NH ₂ , C _1-3 alkyl, and C _1-3 heteroalkyl, wherein the C _1-3 alkyl and C _1-3 heteroalkyl are optionally substituted with 1, 2, or 3 R _a .

In some embodiments of the present application, R ₁ in formula (I') is selected from the group consisting of H, F, Cl, Br, I, CN, COOH, NH ₂ , Me, Et, CF ₃ , CHF ₂ , CH ₂ F, -NHCH ₃ and -N(CH ₃ ) ₂ .

In some embodiments of the present application, R ₂ in formula (I') is selected from the group consisting of C _1-3 alkyl and C _1-3 heteroalkyl, wherein the C _1-3 alkyl and C _1-3 heteroalkyl are optionally substituted with 1, 2, or 3 R _b .

In some embodiments of the present application, R ₂ in formula (I') is selected from the group consisting of Me, Et, CF ₃ , CH ₂ CF ₃ , CHF ₂ , CH ₂ F, -NHCH ₃ and -N(CH ₃ ) ₂ .

In some embodiments of the present application, R ₃ in formula (I') is H.

In some embodiments of the present application, R ₄ in formula (I') is selected from the group consisting of COOH, NH ₂ , C _1-6 alkyl, -NH-C _1-6 alkyl, -N(C _1-6 alkyl) ₂ , -C(=O)-OC _1-6 alkyl, -C(=O)-SC _1-6 alkyl, C _3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, and phenyl, wherein the C _1-6 alkyl, -NH-C _1-6 alkyl, -N(C _1-6 alkyl) ₂ , -C(=O)-OC _1-6 alkyl, -C(=O)-SC _1-6 alkyl, C _3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, and phenyl are optionally substituted with 1, 2, or 3 R _d .

In some embodiments of the present application, R ₄ in formula (I') is selected from the group consisting of COOH, NH ₂ , C _1-6 alkyl, -NH-C _1-6 alkyl, -N(C _1-6 alkyl) ₂ , -C(=O)-OC _1-6 alkyl, -C(=O)-SC _1-6 alkyl, cyclopropyl, cyclopentyl, cyclohexyl, azetidinyl, and phenyl.

In some embodiments of the present application, R ₄ in formula (I') is selected from the group consisting of -NHCH ₃ and -N(CH ₃ ) ₂ .

In some embodiments of the present application, R ₅ in formula (I') is selected from the group consisting of H, C _1-3 alkyl, and C _1-3 heteroalkyl, wherein the C _1-3 alkyl and C _1-3 heteroalkyl are optionally substituted with 1, 2, or 3 R _e .

In some embodiments of the present application, R ₅ in formula (I') is H.

In some embodiments of the present application, R ₆ , R ₇ , R ₈ and R ₉ in formula (I') are H.

In some embodiments of the present application, each of R ₁₀ , R ₁₁ , and R ₁₂ in formula (I') is independently selected from the group consisting of H, F, Cl, Br, I, CN, NO ₂ , OH, COOH, NH ₂ , C _1-3 alkyl, and C _1-3 heteroalkyl, wherein the C _1-3 alkyl and C _1-3 heteroalkyl are optionally substituted with 1, 2, or 3 R _g .

In some embodiments of the present application, each of R ₁₀ , R ₁₁ , and R ₁₂ in formula (I') is independently selected from the group consisting of H, F, Cl, Br, I, CN, NO ₂ , OH, COOH, NH ₂ , Me, Et, CF ₃ , OMe, CHF ₂ , CH ₂ F, NHCH ₃ and N(CH ₃ ) ₂ .

In some embodiments of the present application, ring A in formula (I') is phenyl.

In some embodiments of the present application, the structural unit in formula (I') is selected from the group consisting of And .

In some embodiments of the present application, the structural unit in formula (I') is selected from the group consisting of And .

In some embodiments of the present application, the structural unit in formula (I') is selected from the group consisting of And .

In some embodiments of the present application, the structural unit in formula (I') is selected from the group consisting of

In some embodiments of the present application, each of R _a , R _b , R _c , R _d , _Re , R _f and R _g in formula (I') is independently selected from the group consisting of H, F, Cl, Br, I, OH, CN, NH ₂ , COOH, C(=O)NH ₂ , C _1-6 alkyl, C _1-6 heteroalkyl and cyclopropyl, wherein the C _1-6 alkyl, C _1-6 heteroalkyl and cyclopropyl are optionally substituted with 1, 2 or 3 R; other variables are as defined herein.

In some embodiments of the present application, each of R _a , R _b , R _c , R _d , _Re , R _f and R _g in formula (I') is independently selected from the group consisting of H, F, Cl, Br, I, OH, CN, NH ₂ , COOH, C(=O)NH ₂ , Me, Et, CF ₃ , CHF ₂ , CH ₂ F, NHCH ₃ and N(CH ₃ ) ₂ ; other variables are as defined herein.

In some embodiments of the present application, R ₁ in formula (I') is selected from the group consisting of H, F, Cl, Br, I, CN, COOH, NH ₂ , C _1-3 alkyl, and C _1-3 heteroalkyl, wherein the C _1-3 alkyl and C _1-3 heteroalkyl are optionally substituted with 1, 2, or 3 R _a ; other variables are as defined herein.

In some embodiments of the present application, R ₁ in formula (I') is selected from the group consisting of H, F, Cl, Br, I, CN, COOH, NH ₂ , Me, Et, CF ₃ , CHF ₂ , CH ₂ F, -NHCH ₃ and -N(CH ₃ ) ₂ ; other variables are as defined herein.

In some embodiments of the present application, R ₂ in formula (I') is selected from the group consisting of C _1-3 alkyl and C _1-3 heteroalkyl, wherein the C _1-3 alkyl and C _1-3 heteroalkyl are optionally substituted with 1, 2, or 3 R _b ; other variables are as defined herein.

In some embodiments of the present application, R ₂ in formula (I') is selected from the group consisting of Me, Et, CF ₃ , CH ₂ CF ₃ , CHF ₂ , CH ₂ F, -NHCH ₃ and -N(CH ₃ ) ₂ .

In some embodiments of the present application, R ₃ in formula (I') is H; other variables are as defined herein.

In some embodiments of the present application, R ₄ in formula (I') is selected from the group consisting of COOH, NH ₂ , C _1-6 alkyl, -NH-C _1-6 alkyl, -N(C _1-6 alkyl) ₂ , -C(=O)-OC _1-6 alkyl, -C(=O)-SC _1-6 alkyl, C _3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, and phenyl, wherein the C _1-6 alkyl, -NH-C _1-6 alkyl, -N(C _1-6 alkyl) ₂ , -C(=O)-OC _1-6 alkyl, -C(=O)-SC _1-6 alkyl, C _3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, and phenyl are optionally substituted with 1, 2, or 3 R _d ; other variables are as defined in this application.

In some embodiments of the present application, R ₄ in formula (I') is selected from the group consisting of COOH, NH ₂ , C _1-6 alkyl, -NH-C _1-6 alkyl, -N(C _1-6 alkyl) ₂ , -C(=O)-OC _1-6 alkyl, -C(=O)-SC _1-6 alkyl, cyclopropyl, cyclopentyl, cyclohexyl, azetidinyl, and phenyl; other variables are as defined herein.

In some embodiments of the present application, R ₄ in formula (I') is selected from the group consisting of -NHCH ₃ and -N(CH ₃ ) ₂ ; other variables are as defined herein.

In some embodiments of the present application, R ₅ in formula (I') is selected from the group consisting of H, C _1-3 alkyl, and C _1-3 heteroalkyl, wherein the C _1-3 alkyl and C _1-3 heteroalkyl are optionally substituted with 1, 2, or 3 R _e ; other variables are as defined herein.

In some embodiments of the present application, R ₅ in formula (I') is H; other variables are as defined herein.

In some embodiments of the present application, R ₆ , R ₇ , R ₈ and R ₉ in formula (I') are H; other variables are as defined herein.

In some embodiments of the present application, each of R ₁₀ , R ₁₁ , and R ₁₂ in formula (I') is independently selected from the group consisting of H, F, Cl, Br, I, CN, NO ₂ , OH, COOH, NH ₂ , C _1-3 alkyl, and C _1-3 heteroalkyl, wherein the C _1-3 alkyl and C _1-3 heteroalkyl are optionally substituted with 1, 2, or 3 R _g ; other variables are as defined herein.

In some embodiments of the present application, each of R ₁₀ , R ₁₁ , and R ₁₂ in formula (I') is independently selected from the group consisting of H, F, Cl, Br, I, CN, NO ₂ , OH, COOH, NH ₂ , Me, Et, CF ₃ , OMe, CHF ₂ , CH ₂ F, NHCH ₃ and N(CH ₃ ) ₂ ; other variables are as defined herein.

In some embodiments of the present application, ring A in formula (I') is phenyl; other variables are as defined herein.

In some embodiments of the present application, the structural unit in formula (I') is selected from the group consisting of And ; other variables are as defined in this application.

In some embodiments of the present application, the structural unit in formula (I') is selected from the group consisting of And ; other variables are as defined in this application.

In some embodiments of the present application, the structural unit in formula (I') is selected from the group consisting of other variables are as defined in this application.

In some embodiments of the present application, the structural unit in formula (I') is selected from the group consisting of other variables are as defined in this application.

In some embodiments of the present application, there is provided a compound, an isomer thereof, or a pharmaceutically acceptable salt thereof selected from the group consisting of:

, And ,

Where

R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ , R ₉ , R ₁₀ , R ₁₁ and R ₁₂ are as defined herein.

In some embodiments of the present application, there is provided a compound, an isomer thereof, or a pharmaceutically acceptable salt thereof selected from the group consisting of:

, And ,

Where

R ₁ , R ₂ , R ₃ , R ₅ , R ₆ , R ₇ , R ₈ , R ₉ , R ₁₀ , R ₁₁ and R ₁₂ are as defined in this application,

each of R ₄₁ and R ₄₂ is independently selected from H and C _1-6 alkyl, wherein C _1-6 alkyl is optionally substituted with 1, 2 or 3 R _d ,

R _d is as defined in this application.

The present application provides a compound of formula (I), its isomer or a pharmaceutically acceptable salt thereof,

,

Where

represents a single bond or a double bond;

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

Y ₁ is CH, and Y ₂ is N;

or Y ₁ is N and Y ₂ is CH;

ring A is selected from the group consisting of C _6-10 aryl and 5-10 membered heteroaryl;

R ₁ is selected from the group consisting of H, halogen, CN, COOH, NH ₂ , C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl, wherein C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _a ;

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

R ₃ is selected from the group consisting of H, halogen, CN, NO ₂ , OH, COOH, NH ₂ , C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl, wherein C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _c ;

R ₄ is selected from the group consisting of COOH, NH ₂ , C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl, 3-6-membered heterocycloalkyl, phenyl and 5-6-membered heteroaryl, wherein C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl, 3-6-membered heterocycloalkyl, phenyl and 5-6-membered heteroaryl are optionally substituted with 1, 2 or 3 R _d ;

R ₅ is selected from the group consisting of H, C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl, wherein C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _e ;

each of R ₆ , R ₇ , R ₈ and R ₉ is independently selected from the group consisting of H, halogen, CN, NO ₂ , OH, COOH, NH ₂ , C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl, wherein the C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _f ;

each of R ₁₀ , R ₁₁ and R ₁₂ is independently selected from the group consisting of H, halogen, CN, NO ₂ , OH, COOH, NH ₂ , C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl, wherein the C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _g ;

each of R _a , R _b , R _c , R _d , _Re , R _f and R _g is independently selected from the group consisting of H, F, Cl, Br, I, OH, CN, NH ₂ , COOH, C(=O)NH ₂ , C _1-8 alkyl, C _1-8 heteroalkyl, C _3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl and C _3-6 cycloalkyl-C _1-3 alkyl-, wherein the C _1-8 alkyl, C _1-8 heteroalkyl, C _3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl and C _3-6 cycloalkyl-C _1-3 alkyl- are optionally substituted with 1, 2 or 3 R;

R is independently selected from the group consisting of F, Cl, Br, I, OH, CN, _NH2 , COOH, Me, Et, _CF3 , _CHF2 , _CH2F , _NHCH3, and N( _CH3 ) ₂ ;

each of the C _1-6 heteroalkyl, 3-6-membered heterocycloalkyl and 5-6-membered heteroaryl contains 1, 2, 3 or 4 heteroatoms or heteroatomic groups, each of which is independently selected from the group consisting of -NH-, -O-, -S-, -ON=, -C(=O)-O-, -C(=O)-S-, -C(=O)-, -C(=S)-, -S(=O)-, -S(=O) _2- and N.

In some embodiments of the present application, each of R _a , R _b , R _c , R _d , _Re , R _f and R _g in formula (I) is independently selected from the group consisting of H, F, Cl, Br, I, OH, CN, NH ₂ , COOH, C(=O)NH ₂ , C _1-6 alkyl, C _1-6 heteroalkyl and cyclopropyl, wherein the C _1-6 alkyl, C _1-6 heteroalkyl and cyclopropyl are optionally substituted with 1, 2 or 3 R.

In some embodiments of the present application, each of R _a , R _b , R _c , R _d , _Re , R _f and R _g in formula (I) is independently selected from the group consisting of H, F, Cl, Br, I, OH, CN, NH ₂ , COOH, C(=O)NH ₂ , Me, Et, CF ₃ , CHF ₂ , CH ₂ F, NHCH ₃ and N(CH ₃ ) ₂ .

In some embodiments of the present application, R ₁ in formula (I) is selected from the group consisting of H, F, Cl, Br, I, CN, COOH, NH ₂ , C _1-3 alkyl, and C _1-3 heteroalkyl, wherein the C _1-3 alkyl and C _1-3 heteroalkyl are optionally substituted with 1, 2, or 3 R _a .

In some embodiments of the present application, R ₁ in formula (I) is selected from the group consisting of H, F, Cl, Br, I, CN, COOH, NH ₂ , Me, Et, CF ₃ , CHF ₂ , CH ₂ F, -NHCH ₃ and -N(CH ₃ ) ₂ .

In some embodiments of the present application, R ₂ in formula (I) is selected from the group consisting of C _1-3 alkyl and C _1-3 heteroalkyl, wherein the C _1-3 alkyl and C _1-3 heteroalkyl are optionally substituted with 1, 2, or 3 R _b .

In some embodiments of the present application, R ₂ in formula (I) is selected from the group consisting of Me, Et, CF ₃ , CH ₂ CF ₃ , CHF ₂ , CH ₂ F, -NHCH ₃ and -N(CH ₃ ) ₂ .

In some embodiments of the present application, R ₃ in formula (I) is H.

In some embodiments of the present application, R ₄ in formula (I) is selected from the group consisting of COOH, NH ₂ , C _1-6 alkyl, -NH-C _1-6 alkyl, -N(C _1-6 alkyl) ₂ , -C(=O)-OC _1-6 alkyl, -C(=O)-SC _1-6 alkyl, C _3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, and phenyl, wherein the C _1-6 alkyl, -NH-C _1-6 alkyl, -N(C _1-6 alkyl) ₂ , -C(=O)-OC _1-6 alkyl, -C(=O)-SC _1-6 alkyl, C _3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, and phenyl are optionally substituted with 1, 2, or 3 R _d .

In some embodiments of the present application, R ₄ in formula (I) is selected from the group consisting of COOH, NH ₂ , C _1-6 alkyl, -NH-C _1-6 alkyl, -N(C _1-6 alkyl) ₂ , -C(=O)-OC _1-6 alkyl, -C(=O)-SC _1-6 alkyl, cyclopropyl, cyclopentyl, cyclohexyl, azetidinyl, and phenyl.

In some embodiments of the present application, R ₄ in formula (I) is selected from the group consisting of -NHCH ₃ and -N(CH ₃ ) ₂ .

In some embodiments of the present application, R ₅ in formula (I) is selected from the group consisting of H, C _1-3 alkyl, and C _1-3 heteroalkyl, wherein the C _1-3 alkyl and C _1-3 heteroalkyl are optionally substituted with 1, 2, or 3 R _e .

In some embodiments of the present application, R ₅ in formula (I) is H.

In some embodiments of the present application, R ₆ , R ₇ , R ₈ and R ₉ in formula (I) are H.

In some embodiments of the present application, each of R ₁₀ , R ₁₁ , and R ₁₂ in formula (I) is independently selected from the group consisting of H, F, Cl, Br, I, CN, NO ₂ , OH, COOH, NH ₂ , C _1-3 alkyl, and C _1-3 heteroalkyl, wherein the C _1-3 alkyl and C _1-3 heteroalkyl are optionally substituted with 1, 2, or 3 R _g .

In some embodiments of the present application, each of R ₁₀ , R ₁₁ , and R ₁₂ in formula (I) is independently selected from the group consisting of H, F, Cl, Br, I, CN, NO ₂ , OH, COOH, NH ₂ , Me, Et, CF ₃ , OMe, CHF ₂ , CH ₂ F, NHCH ₃ and N(CH ₃ ) ₂ .

In some embodiments of the present application, ring A in formula (I) is phenyl.

In some embodiments of the present application, the structural unit in formula (I) represents .

In some embodiments of the present application, the structural unit in formula (I) represents .

In some embodiments of the present application, the structural unit in formula (I) is selected from the group consisting of And .

In some embodiments of the present application, the structural unit in formula (I) is selected from the group consisting of , , And .

In some embodiments of the present application, each of R _a , R _b , R _c , R _d , _Re , R _f and R _g in formula (I) is independently selected from the group consisting of H, F, Cl, Br, I, OH, CN, NH ₂ , COOH, C(=O)NH ₂ , C _1-6 alkyl, C _1-6 heteroalkyl and cyclopropyl, wherein the C _1-6 alkyl, C _1-6 heteroalkyl and cyclopropyl are optionally substituted with 1, 2 or 3 R; other variables are as defined herein.

In some embodiments of the present application, each of R _a , R _b , R _c , R _d , _Re , R _f and R _g in formula (I) is independently selected from the group consisting of H, F, Cl, Br, I, OH, CN, NH ₂ , COOH, C(=O)NH ₂ , Me, Et, CF ₃ , CHF ₂ , CH ₂ F, NHCH ₃ and N(CH ₃ ) ₂ ; other variables are as defined herein.

In some embodiments of the present application, R ₁ in formula (I) is selected from the group consisting of H, F, Cl, Br, I, CN, COOH, NH ₂ , C _1-3 alkyl and C _1-3 heteroalkyl, wherein the C _1-3 alkyl and C _1-3 heteroalkyl are optionally substituted with 1, 2 or 3 R _a , other variables are as defined herein.

In some embodiments of the present application, R ₁ in formula (I) is selected from the group consisting of H, F, Cl, Br, I, CN, COOH, NH ₂ , Me, Et, CF ₃ , CHF ₂ , CH ₂ F, -NHCH ₃ and -N(CH ₃ ) ₂ ; other variables are as defined herein.

In some embodiments of the present application, R ₂ in formula (I) is selected from the group consisting of C _1-3 alkyl and C _1-3 heteroalkyl, wherein the C _1-3 alkyl and C _1-3 heteroalkyl are optionally substituted with 1, 2, or 3 R _b ; other variables are as defined herein.

In some embodiments of the present application, R ₂ in formula (I) is selected from the group consisting of Me, Et, CF ₃ , CH ₂ CF ₃ , CHF ₂ , CH ₂ F, -NHCH ₃ and -N(CH ₃ ) ₂ .

In some embodiments of the present application, R ₃ in formula (I) is H; other variables are as defined herein.

In some embodiments of the present application, R ₄ in formula (I) is selected from the group consisting of COOH, NH ₂ , C _1-6 alkyl, -NH-C _1-6 alkyl, -N(C _1-6 alkyl) ₂ , -C(=O)-OC _1-6 alkyl, -C(=O)-SC _1-6 alkyl, C _3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, and phenyl, wherein the C _1-6 alkyl, -NH-C _1-6 alkyl, -N(C _1-6 alkyl) ₂ , -C(=O)-OC _1-6 alkyl, -C(=O)-SC _1-6 alkyl, C _3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, and phenyl are optionally substituted with 1, 2, or 3 R _d ; other variables are as defined in this application.

In some embodiments of the present application, R ₄ in formula (I) is selected from the group consisting of COOH, NH ₂ , C _1-6 alkyl, -NH-C _1-6 alkyl, -N(C _1-6 alkyl) ₂ , -C(=O)-OC _1-6 alkyl, -C(=O)-SC _1-6 alkyl, cyclopropyl, cyclopentyl, cyclohexyl, azetidinyl, and phenyl; other variables are as defined herein.

In some embodiments of the present application, R ₄ in formula (I) is selected from the group consisting of -NHCH ₃ and -N(CH ₃ ) ₂ ; other variables are as defined herein.

In some embodiments of the present application, R ₅ in formula (I) is selected from the group consisting of H, C _1-3 alkyl, and C _1-3 heteroalkyl, wherein the C _1-3 alkyl and C _1-3 heteroalkyl are optionally substituted with 1, 2, or 3 R _e ; other variables are as defined herein.

In some embodiments of the present application, R ₅ in formula (I) is H; other variables are as defined herein.

In some embodiments of the present application, R ₆ , R ₇ , R ₈ and R ₉ in formula (I) are H; other variables are as defined herein.

In some embodiments of the present application, each of R ₁₀ , R ₁₁ , and R ₁₂ in formula (I) is independently selected from the group consisting of H, F, Cl, Br, I, CN, NO ₂ , OH, COOH, NH ₂ , C _1-3 alkyl, and C _1-3 heteroalkyl, wherein the C _1-3 alkyl and C _1-3 heteroalkyl are optionally substituted with 1, 2, or 3 R _g ; other variables are as defined herein.

In some embodiments of the present application, each of R ₁₀ , R ₁₁ , and R ₁₂ in formula (I) is independently selected from the group consisting of H, F, Cl, Br, I, CN, NO ₂ , OH, COOH, NH ₂ , Me, Et, CF ₃ , OMe, CHF ₂ , CH ₂ F, NHCH ₃ and N(CH ₃ ) ₂ ; other variables are as defined herein.

In some embodiments of the present application, ring A in formula (I) is phenyl; other variables are as defined herein.

In some embodiments of the present application, the structural unit in formula (I) represents ; other variables are as defined in this application.

In some embodiments of the present application, the structural unit in formula (I) represents ; other variables are as defined in this application.

In some embodiments of the present application, the structural unit in formula (I) is selected from the group consisting of And ; other variables are as defined in this application.

In some embodiments of the present application, the structural unit in formula (I) is selected from the group consisting of , , And ; other variables are as defined in this application.

Certain other embodiments of the present application are obtained by any combination of the variables described above.

The present application also provides a pharmaceutical composition comprising a therapeutically effective amount of the compound, its isomer or its pharmaceutically acceptable salt as an active ingredient and a pharmaceutically acceptable carrier.

The present application also provides a method of treating an ER-associated disease in a mammal, comprising administering to a mammal, preferably a human, in need of such treatment, a therapeutically effective amount of the compound, its isomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

The present application also provides the use of the compound, its isomer or its pharmaceutically acceptable salt or a pharmaceutical composition based on them in the preparation of a medicinal product for the treatment of a disease associated with ER.

The present application also provides the use of the compound, its isomer or its pharmaceutically acceptable salt or a pharmaceutical composition based on them in the treatment of a disease associated with ER.

The present application also provides a compound, its isomer or its pharmaceutically acceptable salt or a pharmaceutical composition based on them for the treatment of a disease associated with ER.

The present application also provides the use of the compound, its isomer or a pharmaceutically acceptable salt thereof in the preparation of an ER inhibitor.

The present application also provides for the use of the composition in the preparation of an ER inhibitor.

In some embodiments of the present application, the ER-associated disease is breast cancer.

In some embodiments of the present application, the breast cancer is ER-positive breast cancer.

The present application also provides the use of the compound, its isomer or a pharmaceutically acceptable salt thereof in the preparation of a medicinal product for the treatment of ER-positive breast cancer.

Technical effects

The compound according to the present application has an excellent inhibitory effect on cytochrome P450 enzymes. This provides great potential for the development of combination therapies in clinical practice. The compounds according to the present application can be used in the treatment of diseases associated with the estrogen signaling pathway, such as breast cancer.

The compounds of the present application generally exhibit excellent antiproliferative activity against MCF7 breast cancer cells.

The compounds according to the present application are characterized by excellent ADME properties in vitro : they demonstrate excellent stability in the presence of liver microsomes (PPB) and minimal species differences; they are characterized by good inhibitory activity against CYP1A2, CYP2C9, CYP2C19, CYP2D6, etc. and significantly reduce the risk of drug interactions (DDI) in clinical practice; they are also characterized by good absorption.

With regard to PK properties in vivo , the compounds of the present application also exhibit excellent pharmacokinetic properties: the apparent volume of distribution (Vdss) in mice and rats indicates that the compounds of the present application may have a more extensive tissue distribution. The compounds of the present application exhibit good exposure and bioavailability when administered orally. In a study examining the efficacy in MCF7 breast cancer mice, the compounds of the present application demonstrated good efficacy in reducing tumor size.

Observation of suppression of uterine wet weight gain in immature rats demonstrated that the compounds of the present application significantly suppress uterine growth in rats, thereby reducing the risk of developing endometrial thickening or endometrial cancer, and are characterized by sufficient safety.

Definitions and description

Unless otherwise specified, the following terms and phrases, when used in this document, are intended to have the following meanings. A particular term or phrase, unless clearly defined otherwise, is not to be regarded as vague or unclear, but is to be understood in accordance with its generally accepted meaning. Reference to a trade name is intended to refer to its corresponding commercial product or its active ingredient.

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

The term "pharmaceutically acceptable salt" refers to a salt of a compound disclosed herein that is prepared from a compound containing the specific substituents disclosed herein and a relatively non-toxic acid or base. If a compound according to the present application contains a relatively acidic functional group, a base addition salt can be prepared by contacting such a compound with a sufficient amount of a base in neat solution or a suitable inert solvent.

Pharmaceutically acceptable salts according to the present application can be synthesized from the parent compound containing an acidic or basic group by conventional methods of chemical synthesis. In general, such salts are prepared by the following method: the compound in the form of a free acid or base is reacted with a stoichiometric amount of a suitable base or acid in water or in an organic solvent or in a mixture of both.

A compound of the present application may have a particular geometric or stereoisomeric form. All such compounds are contemplated herein, including cis and trans isomers, (-) and (+) enantiomers, ( R ) and ( S ) enantiomers, diastereoisomers, ( D ) isomers, ( L ) isomers and racemic mixtures thereof and other mixtures such as enantiomerically or diastereoisomerically enriched mixtures, all of which are encompassed within the scope of the present application. Substituents such as alkyl may have an additional asymmetric carbon atom. All such isomers and mixtures thereof are encompassed within the scope of the present application.

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

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

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

Unless otherwise noted, "(+)" denotes right-handed rotation, "(-)" denotes left-handed rotation, and "(±)" denotes racemization.

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

Unless otherwise stated, when a double bond structure such as a carbon-carbon double bond, a carbon-nitrogen double bond, and a nitrogen-nitrogen double bond is present in a compound, and each atom in the double bond is bonded to two different substituents (in a double bond involving a nitrogen atom, the lone pair of electrons on the nitrogen atom is considered as the substituent to which the nitrogen atom is bonded), if the atom in the double bond in the compound and its substituents are connected by a wavy line ( ), it means that the compound exists in the form of an ( Z )-type isomer, an ( E )-type isomer, or a mixture of the two isomers. For example, the following formula (A) means that the compound exists in the form of a single isomer of formula (A-1) or formula (A-2), or in the form of a mixture of both isomers of formula (A-1) or formula (A-2); the following formula (B) means that the compound exists in the form of a single isomer of formula (B-1) or formula (B-2), or in the form of a mixture of both isomers of formula (B-1) and formula (B-2); and the following formula (C) means that the compound exists in the form of a single isomer of formula (C-1) or formula (C-2), or in the form of a mixture of both isomers of formula (C-1) and formula (C-2).

(A) (A-1) (A-2)

(B) (B-1) (B-2)

(C) (C-1) (C-2)

The compound of the present application may be present in a particular form. Unless otherwise specified, the term "tautomer" or "tautomeric form" means that the different functional isomers are in dynamic equilibrium at room temperature and can be rapidly converted into each other. If tautomers are possible (e.g., in solution), a chemical equilibrium of the tautomers can be achieved. For example, the proton tautomer, also known as the prototropic tautomer, involves interconversion by proton transfer, such as keto-enol isomerization and imine-enamine isomerization. The valence isomer involves interconversion by recombination of some bonding electrons. A specific example of keto-enol tautomerization is the interconversion between the two tautomers pentane-2,4-dione and 4-hydroxypent-3-en-2-one.

Unless otherwise specified, the term "enriched in one isomer", "enriched in an isomer", "enriched in one enantiomer" or "enantiomer enriched" means that the content of one of the isomers or enantiomers is less than 100% and greater than or equal to 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8% or 99.9%.

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

Optically active (R)- And (S)-isomers, as well asD- And L-isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If a single enantiomer of a particular compound disclosed herein is to be obtained, the desired pure enantiomer can be obtained by asymmetric synthesis or derivatization using a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved. Alternatively, if the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), the compound is reacted with a suitable optically active acid or base to form a salt of the diastereoisomer, which is then subjected to diastereomeric resolution by conventional methods in the art to yield the pure enantiomer. In addition, the enantiomer and diastereoisomer are typically separated by chromatography using a chiral stationary phase, optionally in combination with chemical derivatization (e.g., carbamate derived from amines). The compound of the present application may contain a non-naturally occurring fraction of isotopes of atoms at one or more of the atoms that form the compound. For example, the compound may be labeled with a radioactive isotope such as tritium (³H), iodine-125 (¹²⁵I) or C-14 (¹⁴C) As another example, hydrogen can be replaced by deuterium to form a deuterated drug, and the bond formed between deuterium and carbon is stronger than that formed between normal hydrogen and carbon. Compared to a non-deuterated drug, a deuterated drug has the advantages of reduced toxic side effects, increased stability, increased efficacy, extended biological half-life, etc. All isotopic variations of a compound according to the present application, whether radioactive or not, are included within the scope of the present application. "Optional" or "optionally" means that the event or circumstance further described may, but does not necessarily, occur, wherein the description includes instances where the event or circumstance occurs and instances where it does not.

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

If any variable (e.g., R) occurs more than once in the composition or structure of a compound, then the variable is determined independently in each instance. Thus, for example, if a group is substituted with 0-2 R, then the group may optionally be substituted with up to two R, and the determination of R in each instance is independent. In addition, a combination of a substituent and/or a variant thereof is permissible only if the combination can result in a stable compound.

If the number of linking groups is 0, for example -(CRR) _0- , it means that the linking group is a single bond.

If one of the variants is a single bond, then the two groups linked by that variant are directly linked. For example, if the L in A-L-Z is a single bond, that means the structure is actually A-Z.

If a substituent is absent, it means that the substituent does not exist. For example, if X in A-X is absent, then the structure is actually A.

If the bond at a substituent is a cross-link between two or more atoms in a ring, then that substituent can be bonded to any atom in the ring. For example, the structural unit or reflects the fact that substitution of the R substituent can occur at any position on cyclohexyl or cyclohexadienyl.

If a substituent is listed without specifying the atom through which such substituent is bonded to the group to be substituted, then such substituent may be bonded through any atom in the substituent. For example, pyridinyl as a substituent may be bonded to the group to be substituted through any carbon atom on the pyridine ring.

If the listed linking group does not indicate a direction for linking, the direction for linking is arbitrary. For example, if the linking group L contained in , represents -MW-, -MW- can link ring A and ring B in the same direction, which is the same as the left-to-right reading order, to form , or link ring A and ring B in the opposite direction to form A combination of a linking group, a substituent and/or their variants is permissible only if the combination can result in a stable compound.

Unless otherwise stated, when a group has one or more connectable regions, any one or more such regions of the group may be attached to other groups by chemical bonds. A chemical bond that connects such a region to another group may be represented by a straight continuous bond ( ), a straight dotted line of connection ( ) or a wavy line ( ). For example, the solid straight line in -OCH ₃ indicates that the group is attached to another group through an oxygen atom; in a straight dotted line indicates that the group is attached to another group via a nitrogen atom at both ends; The wavy line indicates that the phenyl group is attached to another group via carbon atoms at positions 1 and 2.

Unless otherwise stated, the number of atoms in a ring is generally defined as the number of ring members. For example, a "5-7 membered ring" refers to a "ring" in which 5-7 atoms are arranged in a ring.

Unless otherwise specified, the term "C _1-8 alkyl" refers to a straight or branched chain saturated hydrocarbon group consisting of 1 to 8 carbon atoms. C _1-8 alkyl includes C _1-6 -, C _1-5 -, C _1-4 -, C _1-3 -, C _1-2 -, C _2-6 -, C _2-4 -, C ₈ -, C ₇ -, C ₆ -, C ₅ alkyl groups, etc., and may be monovalent (e.g. methyl), divalent (e.g. methylene), or multivalent (e.g. methenyl). Examples of C _1-8 alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n -propyl and isopropyl), butyl (including n -butyl, isobutyl, sec -butyl and tert -butyl), pentyl (including n -pentyl, isopentyl and neopentyl), hexyl, heptyl, octyl, etc.

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

Unless otherwise specified, the term "C _1-3 alkyl" refers to a straight or branched saturated hydrocarbon group consisting of 1 to 3 carbon atoms. C _1-3 alkyl includes, but is not limited to, C _1-2 -, C _2-3 alkyl, etc., and may be monovalent (e.g., methyl), divalent (e.g., methylene), or multivalent (e.g., methenyl). Examples of C _1-3 alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n -propyl and isopropyl), etc. Unless otherwise specified, "C _2-8 alkenyl" is used to mean a straight or branched hydrocarbon group containing 2 to 8 carbon atoms and at least one carbon-carbon double bond, which may be located anywhere in the group. _C2-8 alkenyl includes _C2-6- , _C2-4- , _C2-3- , _C4- , _C3- , _C2 alkenyl, etc., and may be monovalent, divalent, or multivalent. Examples of _C2-8 alkenyl include, but are not limited to, ethenyl, propenyl, butenyl, pentenyl, hexenyl, butadienyl, 1,3-pentadienyl, 1,3-hexadienyl, etc.

The term "heteroalkyl", alone or in combination with another term, refers to a stable linear or branched alkyl radical or combinations thereof, consisting of the stated number of carbon atoms and at least one heteroatom or heteroatomic group. In some embodiments, the heteroatom is selected from the group consisting of B, O, N and S, wherein the nitrogen and sulfur atoms are optionally oxidized and the nitrogen heteroatom is optionally quaternized. In other embodiments, the heteroatomic group is selected from the group consisting of -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O), -S(=O) ₂ -, -C(=O)N(H)-, -N(H)-, -C(=NH)-, -S(=O) ₂ N(H)- and -S(=O)N(H)-. In some embodiments, the heteroalkyl is C _1-6 heteroalkyl. In other embodiments, the heteroalkyl is C _1-3 heteroalkyl. The heteroatom or heteroatomic group may be located at any internal position of the heteroalkyl, including the position where the alkyl is bonded to the rest of the molecule. However, the terms "alkoxy", "alkylamino" and "alkylthio" (or thioalkoxy) are commonly used expressions and refer to those alkyl groups that are bonded to the rest of the molecule through an oxygen atom, an amino group, or a sulfur atom, respectively. Examples of heteroalkyl include, but are not limited to _{, -OCH3} , _-OCH2CH3 , _{-OCH2CH2CH3, -OCH2 ( CH3} ) ₂ , -CH2 _{- CH2} -O- _CH3 , _-NHCH3 , -N(CH3) _{2 , -NHCH2CH3} , _-N ( _CH3 )( _CH2CH3 ), _{-CH2 - CH2-} NH- _CH3 , -CH2 _-CH2 -N( _CH3 ) _-CH3 , _-SCH3 , _{-SCH2CH3 ,} -SCH2CH2CH3 _, -SCH2(CH3 _{)2 , -CH2} - _S - _{CH2 - CH3} , _-CH2 - _CH2 - _S (=O _{) -CH3 , and -CH2} - _CH2 -S(=O) ₂ -CH ₃ . No more than two heteroatoms can be located in a row, such as -CH ₂ -NH-OCH ₃ .

Unless otherwise specified, the term "halogen" or "halo", alone or as part of another substituent, refers to a fluorine, chlorine, bromine or iodine atom.

Unless otherwise specified, "C _3-6 cycloalkyl" refers to a saturated cyclic hydrocarbon group of 3 to 6 carbon atoms, including monocyclic and bicyclic ring systems. C _3-6 cycloalkyl includes C _3-5 cycloalkyl, C _4-5 cycloalkyl, C _5-6 cycloalkyl, and the like, and may be monovalent, divalent, or multivalent. Examples of C _3-6 cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

Unless otherwise specified, "C _3-5 cycloalkyl" means a saturated monocyclic hydrocarbon group consisting of 3 to 5 carbon atoms. C _3-5 cycloalkyl includes C _3-4 -, C _4-5 cycloalkyl, etc., and may be monovalent, divalent, or multivalent. Examples of C _3-5 cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, etc.

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

Unless otherwise specified, the terms "C _6-10 aromatic ring" and "C _6-10 aryl" are used interchangeably in this application. The term "C _6-10 aromatic ring" or "C _6-10 aryl" refers to a cyclic hydrocarbon group consisting of 6 to 10 carbon atoms and characterized by a conjugated pi-electron system. The group may be monocyclic, fused bicyclic or fused tricyclic system, wherein the rings are aromatic. It may be monovalent, divalent or multivalent, and C _6-10 aryl includes C _6-9 -, C ₉ -, C ₁₀ -, C ₆ aryl groups, etc. Examples of C _6-10 aryl include, but are not limited to, phenyl, naphthyl (including 1-naphthyl, 2-naphthyl, etc.).

Unless otherwise specified, the terms "5- to 10-membered heteroaromatic ring" and "5- to 10-membered heteroaryl" are used interchangeably herein. The term "5- to 10-membered heteroaryl" refers to a cyclic group of 5- to 10-ring atoms and characterized by a conjugated pi-electron system in which 1, 2, 3, or 4 of the ring atoms are heteroatoms independently selected from the group consisting of O, S, and N, while the remainder are carbon atoms. It may be a monocyclic, fused bicyclic, or fused tricyclic system wherein the rings are aromatic. The nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O) _p , where p is 1 or 2). The 5- to 10-membered heteroaryl may be attached to the remainder of the molecule through a heteroatom or a carbon atom. The 5 to 10-membered heteroaryl includes 5 to 8-membered, 5 to 7-membered, 5 to 6-membered, 5-membered, 6-membered heteroaryl groups, etc. Examples of the 5 to 10-membered heteroaryl include, but are not limited to, pyrrolyl (including N -pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl, 3-pyrazolyl, etc.), imidazolyl (including N -imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, etc.), triazolyl (including 1H- 1,2,3-triazolyl, 2H - 1,2,3-triazolyl, 1H -1,2,4-triazolyl, 4H- 1,2,4-triazolyl, etc.), tetrazolyl, isoxazolyl (including 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, etc.), thiazolyl (including 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, etc.), furanyl (including 2-furanyl, 3-furanyl, etc.), thienyl (including 2-thienyl, 3-thienyl, etc.), pyridinyl (including 2-pyridinyl, 3-pyridinyl, 4-pyridinyl, etc.), pyrazinyl, pyrimidinyl (including 2-pyrimidinyl, 4-pyrimidinyl, etc.), benzothiazolyl (including 5-benzothiazolyl, etc.), purinyl, benzimidazolyl (including 2-benzimidazolyl, etc.), benzoxazolyl, indolyl (including 5-indolyl, etc.), isoquinolinyl (including 1-isoquinolinyl, 5-isoquinolinyl, etc.), quinoxalinyl (including 2-quinoxalinyl, 5-quinoxalinyl, etc.) or quinolyl (including 3-quinolyl, 6-quinolyl, etc.).

Unless otherwise specified, the terms "5-6 membered heteroaromatic ring" and "5-6 membered heteroaryl" are used interchangeably herein. The term "5-6 membered heteroaryl" refers to a monocyclic group of 5-6 ring atoms and characterized by a conjugated pi electron system in which 1, 2, 3, or 4 ring atoms are heteroatoms independently selected from the group consisting of O, S, and N, while the others are carbon atoms. The nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O) _p , where p is 1 or 2). A 5-6 membered heteroaryl may be attached to the remainder of the molecule through a heteroatom or a carbon atom. A 5-6 membered heteroaryl includes a 5-membered and a 6-membered heteroaryl. Examples of the 5- to 6-membered heteroaryl include, but are not limited to, pyrrolyl (including N- pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl, 3-pyrazolyl, etc.), imidazolyl (including N -imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, etc.), triazolyl (including 1H- 1,2,3-triazolyl, 2H- 1,2,3-triazolyl, 1H - 1,2,4-triazolyl, 4H - 1,2,4-triazolyl, etc.), tetrazolyl, isoxazolyl (including 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, etc.), thiazolyl (including 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, etc.), furanyl (including 2-furanyl, 3-furanyl, etc.), thienyl (including 2-thienyl, 3-thienyl, etc.), pyridinyl (including 2-pyridinyl, 3-pyridinyl, 4-pyridinyl, etc.), pyrazinyl or pyrimidinyl (including 2-pyrimidinyl, 4-pyrimidinyl, etc.).

Unless otherwise specified, C _n-n+m or C _n -C _n+m includes any of the specified occurrences of carbon atoms from n to n+m. For example, C _1-12 includes C ₁ , C ₂ , C ₃ , C ₄ , C ₅ , C ₆ , C ₇ , C ₈ , C ₉ , C ₁₀ , C ₁₁ , and C ₁₂ . Any range between n and n+m may also be included. For example, C _1-12 includes C _1-3 , C _1-6 , C 1-9 , C _3-6 , C _3-9 , C _3-12 , C _6-9 , C _6-12 , C _9-12 , etc. Similarly, "n-n+m-membered" means that the number of atoms in the ring is from n to _n +m. For example, a 3-12 membered ring includes a 3-membered ring, a 4-membered ring, a 5-membered ring, a 6-membered ring, a 7-membered ring, an 8-membered ring, a 9-membered ring, a 10-membered ring, an 11-membered ring, and a 12-membered ring. “n-n+m-membered” also means any range within n to n+m. For example, a 3-12 membered ring includes a 3-6-membered ring, a 3-9-membered ring, a 5-6-membered ring, a 5-7-membered ring, a 6-7-membered ring, a 6-8-membered ring, a 6-10-membered ring, etc.

The term "leaving group" refers to a functional group or atom that can be replaced by another functional group or atom through a substitution reaction (e.g., nucleophilic substitution). For example, illustrative leaving groups include triflate; chlorine, bromine, and iodine; sulfonate groups such as mesylate, tosylate, p -bromobenzenesulfonate, p -toluenesulfonate, etc.; acyloxy groups such as acetoxy, trifluoroacetoxy, etc.

The term "protecting group" includes, but is not limited to, "amino protecting group", "hydroxy protecting group", or "sulfhydryl protecting group". The term "amino protecting group" refers to a protecting group suitable for preventing side reactions involving the nitrogen atom of the amino group. Exemplary amino protecting groups include, but are not limited to, formyl; acyl, such as alkanoyl (such as acetyl, trichloroacetyl, or trifluoroacetyl); alkoxycarbonyl, such as tert -butoxycarbonyl (Boc); arylmethyloxycarbonyl, such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethyloxycarbonyl (Fmoc); arylmethyl, such as benzyl (Bn), trityl (Tr), 1,1-di(4'-methoxyphenyl)methyl; and silyl, such as trimethylsilyl (TMS) and tert -butyldimethylsilyl (TBS). The term "hydroxyl protecting group" refers to a protecting group suitable for preventing side reactions involving a hydroxyl group. Exemplary hydroxyl protecting groups include, but are not limited to, alkyl, such as methyl, ethyl, and tert -butyl; acyl, such as alkanoyl (such as acetyl); arylmethyl, such as benzyl (Bn), p -methoxybenzyl (PMB), 9-fluorenylmethyl (Fm), and diphenylmethyl (DPM); and silyl, such as trimethylsilyl (TMS) and tert- butyldimethylsilyl (TBS).

The compounds of the present application can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments obtained by combining them with other chemical synthetic methods, and equivalents thereof known to those skilled in the art. Preferred embodiments include, but are not limited to, the examples disclosed herein.

Compounds are named according to generally accepted nomenclature rules in the field or using ChemDraw® software, and for commercially available compounds, names are given according to the manufacturer's catalogue.

SPECIFIC IMPLEMENTATION OPTIONS

The present invention is described in detail below with the aid of examples. However, this does not limit the scope of the present application in an unfavorable manner.

Those skilled in the art will recognize that the order of reactions in each reaction scheme may be different to obtain the compound of the present application, which is also within the scope of the present application.

Example 1

Step A: Under a nitrogen atmosphere at -75°C, n -butyl lithium (2.5 M, 428.40 mL, 1.05 equiv) was added dropwise and slowly (over 1 h) to a solution of 1-1 (100.00 g, 1.02 mol, 140.85 mL, 1.00 equiv) in tetrahydrofuran (500 mL). The reaction mixture was warmed to 0°C and stirred for 10 min, then cooled to -75°C. Hexamethylphosphoric triamide (201.06 g, 1.12 mol, 197.12 mL, 1.10 equiv) was added over 1 h. The reaction mixture was stirred at -75 °C for one hour and ethyl iodide (198.86 g, 1.27 mol, 101.98 mL, 1.25 equiv) was added (over one hour). The reaction mixture was warmed to 20 °C and allowed to react for 10 hours before 400 mL of water was added. The organic phase was separated and washed three times with 400 mL of water, dried over anhydrous sodium sulfate, filtered and separated by distillation to give compound 1-2 .

Step B: Dimethylaminopyridine (3.65 g, 29.88 mmol, 0.10 equiv) and Boc ₂ O (68.46 g, 313.70 mmol, 72.07 mL, 1.05 equiv) were added to a solution of compound 1-3 (35.00 g, 298.76 mmol, 1.00 equiv) in dichloromethane (400 mL). The reaction was then allowed to proceed at 20 °C for 12 h, and the reaction mixture was extracted and washed twice with 400 mL of ammonium chloride solution. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 1-4 .

Step C: Under a nitrogen atmosphere at -75 °C, lithium diisopropylamide (2 M, 75.95 mL, 1.10 equiv) was slowly added dropwise to a solution of compound 1-4 (30.00 g, 138.08 mmol, 1.00 equiv) in tetrahydrofuran (400 mL). The reaction mixture was stirred at -75 °C for 30 min and cyanogen bromide (55.40 g, 523.04 mmol, 38.47 mL, 3.79 equiv) was added. The reaction mixture was warmed to 15 °C and allowed to react for 12 h before 400 mL of water was added. The organic phase was separated and washed three times with 300 ml of water, dried over anhydrous sodium sulfate and filtered to obtain the crude product. The crude product was separated by silica gel column chromatography (petroleum ether:ethyl acetate = 1:0-50:1, v/v) to give compound 1-5 .

Step D: To a solution of compound 1-5 (39.00 g, 131.69 mmol, 1.00 equiv.) in N , N -dimethylacetamide (300 mL) were added cesium carbonate (85.81 g, 263.38 mmol, 2.00 equiv.), copper(I) iodide (1.25 g, 6.58 mmol, 0.05 equiv.), palladium acetate (1.48 g, 6.58 mmol, 0.05 equiv.), and 1,1'-bis(diphenylphosphino)ferrocene (3.65 g, 6.58 mmol, 0.05 equiv.). Then, compound 1-2 (33.26 g, 263.38 mmol, 2.00 equiv) was added under nitrogen atmosphere. The reaction mixture was allowed to react at 80°C for 12 hours, 1 L of ethyl acetate and 1 L of water were added, and the mixture was filtered and separated. The organic phase was extracted and washed three times with 1 L of water. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was separated by silica gel column chromatography (petroleum ether:ethyl acetate = 1:0-30:1, v/v) to give compound 1-6 .

Step E: Potassium carbonate (69.27 g, 501.25 mmol, 5.00 equiv) was added to a solution of compound 1-6 (27.00 g, 100.25 mmol, 1.00 equiv) in 300 mL of methanol and 15 mL of water. The reaction mixture was allowed to react at 70 °C for 12 h and then filtered and concentrated before 300 mL of ethyl acetate was added. The reaction mixture was extracted and washed twice with 300 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude product. The crude product was separated by silica gel column chromatography (petroleum ether:ethyl acetate = 100:1-30:1, v/v) to give compound 1-7 . MS [ESI, M+1]: 170.1.

Step F: Bis(pinacolato)diboron (600.25 mg, 2.36 mmol, 1.00 equiv) and tetrakis(triphenylphosphine)platinum(0) (58.82 mg, 47.28 μmol, 0.02 equiv) were added to a solution of 1-7 (400 mg, 2.36 mmol, 1.00 equiv) in 2-methyltetrahydrofuran (5 mL). The reaction mixture was allowed to react at 85 °C for 7 h under nitrogen atmosphere and then cooled to room temperature to afford 1-8 for direct use in the next reaction without purification. MS [ESI, M+1]: 424.3.

Step G. Compound 1-9 (791 mg, 1.66 mmol, 0.7 equiv, see US 20160347717A1 for the synthesis of the compound), cesium carbonate (1.08 g, 3.33 mmol, 2.00 equiv), and bis(triphenylphosphine)palladium(II) dichloride (58.41 mg, 83.21 μmol, 0.05 equiv) were added to a solution of compound 1-8 (1.0 g, 2.36 mmol, 1.00 equiv) in 2-methyltetrahydrofuran (5 mL) at room temperature. The reaction system was purged with nitrogen three times before adding water (0.2 mL). The reaction mixture was allowed to react at 30°C for 12 h under nitrogen atmosphere to afford compound 1-10 for direct use in the next reaction without purification. MS [ESI, M+1]: 645.5.

Step H. 2-Chloro-4-fluoroiodobenzene (510.81 mg, 1.99 mmol, 1.2 equiv) and aqueous potassium hydroxide solution (4 M, 2.9 mL, 7.00 equiv) were added to a solution of compound 1-10 (1.07 g, 1.66 mmol, 1.00 equiv) in 2-methyltetrahydrofuran (5 mL). The reaction system was purged with nitrogen three times. The reaction mixture was allowed to react at 85 °C for 6 h under nitrogen atmosphere, bis(triphenylphosphine)palladium(II) dichloride (58.25 mg, 83 μmol, 0.05 equiv) was added and the reaction was allowed to react at 85 °C for 16 h under nitrogen atmosphere. The reaction mixture was cooled to room temperature and 10 ml of water and 10 ml of ethyl acetate were added. The aqueous phase was extracted three times with 10 ml of ethyl acetate and the organic phases were combined and washed once with 50 ml of saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1-1:1, v/v) to give compound 1-11 . MS [ESI, M+1]: 647.2.

Step I: To a solution of 1-11 (694 mg, 1.07 mmol, 1.00 equiv) in dichloromethane (40 mL) was added N -chlorosuccinimide (171.83 mg, 1.29 mmol, 1.2 equiv). The reaction mixture was allowed to react at 25 °C for 1 h and then washed twice with 30 mL of saturated aqueous sodium sulfite solution. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give 1-12 . MS [ESI, M+1]: 681.1.

Step J. To a solution of compound 1-12 (621 mg, 911.06 μmol, 1.00 equiv.) in dichloromethane (8 mL) was added 8 mL of trifluoroacetic acid (TFA). The reaction mixture was allowed to react at 25 °C for 1 h and concentrated to give the crude product. The crude product was separated by preparative HPLC (water (0.1% trifluoroacetic acid, v/v)/acetonitrile) to give compound 1. MS [ESI, M+1]: 581.1.

¹ H NMR(400 MHz, CDCl₃): 9.19 (s, 1H), 7.77-7.69 (m, 1H), 7.69-7.61 (m, 1H), 7.35 (dd, J = 2.4, 8.8 Hz, 1H), 7.32-7.28 (m, 1H), 7.25-7.11 (m, 3H), 7.07 (dd, J = 2.4, 8.4 Hz, 1H), 6.95-6.90 (m, 1H), 6.60-6.55 (m, 2H), 6.51-6.44 (m, 1H ), 5.81 (br s, 2H), 4.39 (br s, 2H), 3.70 (br d, J = 4.4 Hz, 2H), 3.28 (br s, 2H), 2.97 (s, 3H), 2.92 (s, 3H), 2.68-2.48 (m, 2H), 0.96(t, J= 7.6 Hz, 3H).

Example 2

Step A. 2-Iodobenzonitrile (760.3 mg, 3.32 mmol, 2 equiv), aqueous potassium hydroxide (4 M, 2.90 mL, 7 equiv), and bis(triphenylphosphine)palladium(II) dichloride (58.25 mg, 83 μmol, 0.05 equiv) were added to a solution of compound 1-10 (1.07 g, 1.66 mmol, 1.00 equiv) in 2-methyltetrahydrofuran (5 mL). The reaction mixture was purged with nitrogen three times and the reaction was carried out at 85 °C for 8 h under nitrogen atmosphere. To the reaction mixture was then added bis(triphenylphosphine)palladium(II) dichloride (58.25 mg, 83 μmol, 0.05 equiv) and the reaction was allowed to proceed at 85 °C for 24 h. The reaction mixture was diluted with 15 mL water and 15 mL ethyl acetate and extracted three times with 15 mL ethyl acetate. The organic phases were combined, washed once with 50 mL saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by silica gel column chromatography (PE:EA = 5:1-1:2) to give compound 2-1 . MS [ESI, M+1]: 620.4.

Step B: To a solution of 2-1 (590 mg, 952 μmol, 1.00 equiv) in dichloromethane (10 mL) was added N -chlorosuccinimide (152.55 mg, 1.14 mmol, 1.2 equiv). The reaction mixture was allowed to react at 25 °C for 2 h. The reaction mixture was then washed twice with 10 mL of saturated aqueous sodium sulfite solution and the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to afford 2-2 . MS [ESI, M+1]: 654.3.

Step C: To a solution of 2-2 (584 mg, 892.70 μmol, 1.00 equiv.) in dichloromethane (7 mL) was added 7 mL of trifluoroacetic acid. The reaction mixture was allowed to react at 25 °C for 1 h and concentrated to give the crude product. The crude product was separated by preparative HPLC (water (0.05% aqueous ammonia, v/v)/acetonitrile) to give 2 . MS [ESI, M+1]: 554.3.

¹ H NMR (400 MHz, CDCl ₃ )8.54 (br s, 1H), 7.62-7.56 (m, 2H), 7.49-7.48 (m, 2H), 7.35 (br d, J = 7.2 Hz, 1H), 7.27-7.22 (m, 2H), 7.19-7.13 (m, 3H), 6.75-6.71 (m, 1H), 6, 40-6.30 (m, 2H), 4.18 (br s, 2H), 3.34 (br d, J = 3.6 Hz, 2H), 2.96 (br s, 3H), 2.91 (br s, 3H), 2.85 (br s, 2H), 2.71-2.42 (m, 2H ), 0.92 (br t, J= 6.8 Hz, 3H).

Example 3

Step A: Under nitrogen atmosphere, to a solution of compound 1-10 (500 mg, 775.67 μmol, 1.00 equiv), iodobenzene (205.71 mg, 1.01 mmol, 112.41 μL, 1.3 equiv), potassium hydroxide (261.12 mg, 4.65 mmol, 6 equiv) in 2-methyltetrahydrofuran (10 mL) and water (3 mL) was added bis(triphenylphosphine)palladium(II) dichloride (54.44 mg, 77.57 μmol, 0.1 equiv). The reaction mixture was purged with nitrogen three times and the reaction was allowed to proceed at 80 °C for 12 h under nitrogen atmosphere. The reaction mixture was diluted with 20 mL of water and separated, the aqueous phase was extracted three times with 50 mL of ethyl acetate. The organic phases were combined, washed once with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by preparative HPLC (water (0.1% formic acid, v/v)/acetonitrile) to give compound 3-1 . MS [ESI, M+1]: 595.3.

Step B: Under nitrogen atmosphere, to a solution of 3-1 (80 mg, 134.51 μmol, 1 equiv) in dichloromethane (5 mL) was added N -chlorosuccinimide (17.96 mg, 134.51 μmol, 1 equiv). The reaction mixture was allowed to react at 20 °C for 1 h. The reaction was quenched with 2 mL of saturated aqueous sodium sulfite solution and the reaction mixture was separated. The aqueous phase was extracted three times with 20 mL of ethyl acetate and the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to give 3-2 . MS [ESI, M+1]: 629.3.

Step C: Under nitrogen atmosphere, to a solution of compound 3-2 (90 mg, 143.04 μmol, 1 equiv.) in dichloromethane (5 mL) was added 1 mL of trifluoroacetic acid. The reaction mixture was allowed to react at 20 °C for 10 min and concentrated to give the crude product. The crude product was sequentially separated by preparative HPLC using water (0.1% trifluoroacetic acid, v/v)/acetonitrile and water (0.05% aqueous ammonia, v/v)/acetonitrile to give compound 3. MS [ESI, M+1]: 529.1.

¹ H NMR(400 MHz, METHANOL-d₄) δ = 7.67 (d,J = 2.4 Hz, 1H), 7.57-7.51 (m, 1H), 7.36 (d, J = 8.0 Hz, 1H), 7.31-7.09 (m, 8H), 6.77 (td, J = 5.6, 15.2 Hz, 1H), 6.59-6.50 (m, 2H), 4.25 (br t, J = 4.8 Hz, 2H), 3.43 (br dd, J = 2.0, 4.0 Hz, 2H), 3.07 (d, J= 2.0 Hz, 3H), 2.96 (d,J = 0.8 Hz, 3H), 2.94-2.88 (m, 2H), 2.55 (q, J= 7.6 Hz, 2H), 0.98 (t,J= 7.2 Hz, 3H).

The pH of compound 3 was adjusted to 3 with 1 M hydrochloric acid and the solvent was removed under reduced pressure to give the monohydrochloride of compound 3 .

^1H NMR (400 MHz, DMSO- _d6 ) 11.57 (s, 1H), 9.43 (br s, 2H), 7.67 (d, J = 1.6 Hz, 1H), 7.50 (d, J = 7.6 Hz, 1H), 7.40 (d, J = 8.0 Hz, 1H), 7.34-7.28 (m, 2H), 7.27-7.11 (m, 6H), 6.83 (d, J = 15.2 Hz, 1H), 6.67-6.53 ( m, 2H), 4.39 (t, J = 5.2 Hz, 2H), 3.77 (br d, J = 5.2 Hz, 2H), 3.28-3.19 (m, 2H), 3.03 (s, 3H), 2.87 (s, 3H), 2.48-2.42 (m, 2H), 0.90 (t, J = 7.2 Hz, 3H).

Example 4

Step A: Under nitrogen atmosphere, to a solution of compound 1-10 (500 mg, 775.67 μmol, 1.00 equiv), 1-bromo-3-methoxybenzene (188.60 mg, 1.01 mmol, 127.43 μL, 1.3 equiv), potassium hydroxide (261.12 mg, 4.65 mmol, 6 equiv) in 2-methyltetrahydrofuran (10 mL) and water (3 mL) was added bis(triphenylphosphine)palladium(II) dichloride (54.44 mg, 77.57 μmol, 0.1 equiv). The reaction mixture was purged with nitrogen three times and the reaction was allowed to proceed at 80 °C for 12 h under nitrogen atmosphere. The reaction mixture was diluted with 20 mL of water and separated, the aqueous phase was extracted three times with 50 mL of ethyl acetate. The organic phases were combined, washed once with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by preparative HPLC (water (0.1% formic acid, v/v)/acetonitrile) to give compound 4-1 . MS [ESI, M+1]: 625.3.

In Step B and Step C, referring to Step B and Step C in the synthesis of compound 3 , compound 4-2 (MS[ESI, M+1]: 625.3) and crude product compound 4 were obtained, respectively. The crude product was sequentially separated by preparative HPLC using water (0.1% trifluoroacetic acid, v/v)/acetonitrile system and water (0.05% aqueous ammonia, v/v)/acetonitrile system to give compound 4 . MS [ESI, M+1]: 559.1.

¹ H NMR(400 MHz, DMSO-d₆) δ = 11.45 (br s, 1H), 7.70-7.60 (m, 1H), 7.50 (br d, J = 7.8 Hz, 1H), 7.39 (br d, J = 7.8 Hz, 1H), 7.34-7.06 (m, 5H), 6.83-6.72 (m, 2H), 6.66-6.54 (m, 2H), 6, 53-6.44 (m, 1H), 4.16 (br t, J = 5.2 Hz, 2H), 3.69 (s, 3H), 3.30 (br d, J = 4.4 Hz, 2H), 2.98 (s, 3H), 2.84 (s, 3H), 2.80-2.73 (m, 2H), 2.45 (br d, J = 7.2 Hz, 2H), 2.06 (br d, J = 13.2 Hz, 1H), 0.91 (br t, J= 7.2 Hz, 3H).

The crude product was separated by preparative HPLC (water (0.05% hydrochloric acid, v/v)/acetonitrile system) to afford the monohydrochloride salt of compound 4. MS [ESI, M+1]: 559.3.

^1H NMR (400 MHz, DMSO- _d6 ) δ = 11.59 (s, 1H), 9.54 (br s, 2H), 7.73-7.66 (m, 1H), 7.49 ( d, J = 7.6 Hz, 1H), 7.40 (d, J = 8.0 Hz, 1H), 7.32-7.28 (m, 1H), 7.26 (br d, J = 2.4 Hz, 1H), 7.23-7.17 (m, 2H), 7.16-7.09 (m, 1H), 6.88-6.80 (m, 1H), 6.80-6.72 (m, 2H), 6.66 (d, J = 8.4 Hz, 1H), 6.63-6.55 (m, 1H), 4.51-4.31 (m, 2H), 3.78 (br d, J = 5.6 Hz, 2H), 3.69 (s, 3H ), 3.25 (br s, 2H), 3.02 (s, 3H), 2.85 (s, 3H), 2.45 (br d, J = 7.2 Hz, 2H), 0.96-0.84 (m, 3H)

Example 5

Step A: A solution of 5-1 (5 g, 42.32 mmol, 1 equiv) in N , N -dimethylformamide (50 mL) was cooled to 0 °C and sodium hydride (2.54 g, 63.49 mmol, 60% pure, 1.5 equiv ) was added portionwise. The reaction mixture was stirred at 0 °C for 0.5 h, 5-2 (8.22 g, 46.56 mmol, 5.96 mL, 1.1 equiv ) was added, the mixture was warmed to 20 °C and the reaction was allowed to proceed for 1 h before 50 mL of saturated aqueous ammonium chloride solution was added. The separated aqueous phase was extracted three times with 50 mL ethyl acetate and the organic phases were combined, washed twice with 50 mL saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give compound 5-3 . MS [ESI, M+1]: 259.1.

Step B: Under a nitrogen atmosphere at -70 °C, lithium diisopropylamide (2 M, 6.39 mL, 1.1 equiv) was added dropwise and slowly to a solution of 5-3 (3 g, 11.61 mmol, 1 equiv) in tetrahydrofuran (20 mL). The reaction mixture was stirred at -70 °C for 30 min and iodine (4.42 g, 17.42 mmol, 3.51 mL, 1.5 equiv) was added. The reaction mixture was warmed to 25 °C and allowed to react for 2 h before 20 mL of saturated aqueous ammonium chloride and 20 mL of saturated aqueous sodium sulfite were added. The aqueous phase was separated and washed three times with 50 ml of ethyl acetate, and the organic phases were combined, washed twice with 50 ml of saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1-3:1, v/v) to give compound 5-4 . MS [ESI, M+1]: 384.9.

Step C. To a solution of compound 5-4 (2 g, 5.21 mmol, 1 equiv ) in N , N -dimethylacetamide (10 mL) was added cesium carbonate (8 (3.39 g, 10.41 mmol, 2 equiv ), copper(I) iodide (49.57 mg, 260.29 μmol, 0.05 equiv ) , palladium acetate (58.44 mg, 260.29 μmol, 0.05 equiv ), and 1,1'-bis(diphenylphosphino)ferrocene (144.30 mg, 260.29 μmol, 0.05 equiv ). Then, compound 5-5 (1.31 g, 10.41 mmol, 2 equiv . ). The reaction mixture was allowed to react at 80°C for 12 h, 20 ml of ethyl acetate and 20 ml of water were added, filtered and separated. The aqueous phase was extracted three times with 20 ml of ethyl acetate and the organic phases were combined and washed once with 20 ml of saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by silica gel column chromatography (petroleum ether:ethyl acetate = 1:0-5:1, v/v) to give compound 5-6 . MS [ESI, M+1]: 311.1.

Stage D. To 10 ml of the compound solution5-6 (220 mg, 708.83 µmol, 1 equiv.) sodium hydroxide (4 M, 708.83 µl, 4 equiv) was added. The reaction mixture was allowed to react at 60°C for 2 h, 10 ml of water was added and then extracted three times with 50 ml of ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give the compound5-7. MS [ESI, M+1]: 171.2.

Step E: To a solution of 5-7 (110 mg, 646.26 μmol, 1 equiv ) in 2-methyltetrahydrofuran (10 mL) were added 5-8 (164.11 mg, 646.26 μmol, 1 equiv ) and tetrakis(triphenylphosphine)platinum(0) (16.08 mg, 12.93 μmol, 0.02 equiv ). The reaction mixture was allowed to react at 85 °C for 12 h under nitrogen atmosphere and then cooled to room temperature to afford 5-9 for direct use in the next reaction without purification.

Step F. Compound 5-10 (215.03 mg, 452.38 μmol, 0.7 equiv ), cesium carbonate (421.13 mg, 1.29 mmol, 2 equiv ), and bis(triphenylphosphine)palladium(II) dichloride (22.68 mg, 32.31 μmol, 0.05 equiv) were added to a solution of compound 5-9 (274.11 mg, 646.26 μmol, 1 equiv ) in 2-methyltetrahydrofuran (10 mL) at room temperature. The reaction system was purged with nitrogen three times before water (2.5 mL) was added. The reaction mixture was allowed to react at 30 °C for 12 h under nitrogen atmosphere to give compound 5-11 .

Step G. Compound 5-12 (131.84 mg, 646.26 μmol, 72.04 μL, 1 equiv ), aqueous potassium hydroxide solution (4 M, 1.13 mL, 7 equiv ) , and bis(triphenylphosphine)palladium(II) dichloride (22.68 mg, 32.31 μmol, 0.05 equiv ) were added to a solution of compound 5-11 (417.22 mg, 646.26 μmol, 1 equiv ) in 2-methyltetrahydrofuran (10 mL). The reaction system was allowed to react at 85 °C for 12 h under nitrogen atmosphere, then the system was cooled to room temperature, and 20 mL of water and 20 mL of ethyl acetate were added. The aqueous phase was extracted three times with 20 mL ethyl acetate and the organic phases were combined and washed once with 20 mL saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by reverse phase column chromatography (water (0.1% formic acid, v/v)/acetonitrile) to give compound 5-13 . MS [ESI, M+1]: 596.3.

Stage H. To the solution of the compound5-13(25 mg, 41.97 µmol, 1 equiv..) in dichloromethane (5 ml) was addedN-chlorosuccinimide (6.72 mg, 50.36 µmol, 1.2 equiv.). The reaction mixture was allowed to react at 25°C for 12 h. The reaction was quenched with 10 mL of saturated aqueous sodium sulfite solution. The aqueous phase was separated and extracted three times with 20 mL of dichloromethane, and the organic phases were combined and washed twice with 20 mL of saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give the compound5-14. MS [ESI, M+1]: 630.3.

Step I. To a solution of compound 5-14 (20 mg, 31.74 μmol, 1 equiv ) in dichloromethane (2 mL) was added 0.1 mL of trifluoroacetic acid. The reaction mixture was allowed to react at 25 °C for 1 h and concentrated to give the crude product. The crude product was separated by preparative HPLC (water (10 mM ammonium bicarbonate)/acetonitrile 45-75%) to give compound 5. MS [ESI, M+1]: 530.3.

¹ H NMR(400 MHz, METHANOL-d₄) δ = 8.28 (dd,J= 1.6, 4.8 Hz, 1H), 8.01 (dd, J= 1.6, 8.0 Hz, 1H), 7.70 (d, J= 2.0 Hz, 1H), 7.33-7.20 (m, 7H), 6.78 (td, J= 5.6, 15.2 Hz, 1H), 6.60-6.53 (m, 2H), 4.26 (t, J= 5.4 Hz, 2H), 3.44 (dd,J= 1.5, 5.6 Hz, 2H), 3.09 (s, 3H), 2.98 (s, 3H), 2.96-2.88 (m, 2H), 2.59-2, 50 (m, 2H), 0.98 (t, J= 7.2 Hz, 3H).

Example 6

Step A. Compound 6-1 (1.15 g, 2.49 mmol, 1 equiv), cesium carbonate (1.62 g, 4.99 mmol, 2 equiv) and bis(triphenylphosphine)palladium(II) dichloride (87.49 mg, 124.65 μmol, 0.05 equiv) were added to a solution of compound 1-8 (1.50 g, 3.54 mmol, 1.42 equiv) in 2-methyltetrahydrofuran (10 mL) at room temperature. The reaction system was purged with nitrogen three times before water (1 mL) was added. The reaction mixture was allowed to react at 30 °C for 12 h under nitrogen atmosphere to give compound 6-2 for direct use in the next reaction without purification.

Step B. 1-Bromo-3-methoxybenzene (558.81 mg, 2.99 mmol, 377.57 μL, 1.2 equiv), aqueous potassium hydroxide solution (4 M, 4.36 mL, 7 equiv) and bis(triphenylphosphine)palladium(II) dichloride (87.38 mg, 124.49 μmol, 0.05 equiv) were added to a solution of compound 6-2 (1.07 g, 1.66 mmol, 1.00 equiv) in 2-methyltetrahydrofuran (10 mL). The reaction system was purged with nitrogen three times and the reaction was carried out at 85 °C for 12 h under nitrogen atmosphere. After cooling to room temperature, 30 mL of water was added to the reaction mixture. The aqueous phase was extracted three times with 50 mL ethyl acetate, and the organic phases were combined and washed twice with 50 mL saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by silica gel column chromatography (petroleum ether:ethyl acetate = 1:0-4:5, v/v) to give compound 6-4 . MS [ESI, M+1]: 611.3.

Step C: To a solution of 6-4 (1.2 g, 1.96 mmol, 1 equiv) in dichloromethane (20 mL) was added N -chlorosuccinimide (314.84 mg, 2.36 mmol, 1.2 equiv). The reaction mixture was allowed to react at 25 °C for 12 h and then washed with 5 mL of saturated aqueous sodium sulfite solution. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give 6-5 . MS [ESI, M+1]: 645.2.

Step D: To a solution of compound 6-5 (450 mg, 697.47 μmol, 1 equiv.) in dichloromethane (5 mL) was added 5 mL of trifluoroacetic acid. The reaction mixture was allowed to react at 25 °C for 1 h under nitrogen atmosphere and concentrated to give the crude product. The crude product was separated by preparative HPLC (water (0.05% hydrochloric acid, v/v)/acetonitrile) to give compound 6. MS [ESI, M+1]: 545.2.

^1H NMR (400 MHz, DMSO- _d6 ) δ = 11.93-11.15 (m, 1H), 9.43 (br s, 2H), 8.24 (br d, J = 4.8 Hz , 1H), 7.77-7.62 (m, 1H), 7.48 (d, J = 7.6 Hz, 1H), 7.39 (d, J = 8.4 Hz, 1H), 7.33-7.26 (m, 1H), 7.25 (d, J = 3.2 Hz, 1H), 7. 26-7.22 (m, 1H), 7.22-7.17 (m, 2H), 7.15-7.08 (m, 1H), 6.84-6.78 (m, 1H), 6.76 (br d, J = 2.8 Hz, 1H), 6.19 (d, J = 15.4 Hz, 1H), 4.47-4.34 (m, 2H), 3.73 ( br d, J = 5.6 Hz, 2H), 3.69 (s, 3H), 3.22 (br s, 2H), 2.64 (d, J = 4.8 Hz, 3H), 2.47-2.40 (m, 2H), 1.12-0.72 (m, 3H).

Example 7

Step A. To a solution of compound 7-1 (1.2 g, 4.54 mmol, 1 equiv.) in dichloromethane (10 mL) and N , N -dimethylformamide (1 mL) was added sodium carbonate (2.41 g, 22.72 mmol, 5 equiv.). The reaction system was allowed to react at 25 °C for 15 min, cooled to 0 °C, acryloyl chloride (1.23 g, 13.63 mmol, 1.11 mL, 3 equiv.) was added, and the reaction was allowed to react at 25 °C for 11 h and 45 min. The system was added 10 mL of water, and extraction was performed three times with 20 mL of dichloromethane-methanol (10/1, v/v). The organic phases were combined, washed once with 10 mL of brine, dried over sodium sulfate, filtered and concentrated to give the crude product as compound 7-2 . MS [ESI, M+1]: 318.9.

Step B. Pd( _PPh3 ) _2Cl2 (73.01 mg, 104.01 μmol, 0.05 equiv) and _Cs2CO3 (1.36 g, 4.16 mmol _, 2 equiv) were added _to a solution of compounds 1-8 (1.25 g, 2.95 mmol, 1.42 equiv) and 7-2 (661.75 mg, 2.08 mmol, 1 equiv) in 2-methyltetrahydrofuran (5 mL). The reaction system was purged with nitrogen three times, 0.2 mL of water was added and the reaction was carried out at 30 °C for 12 h to afford compound 7-3 for direct use in the next reaction without purification. MS [ESI, M+1]: 488.3.

Step C: An aqueous solution of potassium hydroxide (4 M, 4.36 mL, 7 equiv.) and bis(triphenylphosphine)palladium(II) dichloride (72.72 mg, 103.61 μmol, 0.05 equiv.) was added to a solution of compound 7-3 (1.01 g, 2.07 mmol, 1 equiv.) and 2-chloro-4-fluoroiodobenzene (637.69 mg, 2.49 mmol, 1.2 equiv.) in 2-methyltetrahydrofuran (5 mL). The reaction system was purged with nitrogen three times and the reaction was carried out at 85 °C for 12 h under nitrogen atmosphere. The reaction system was added 30 mL of water and extracted three times with 50 mL of ethyl acetate. The organic phases were combined and washed twice with 50 mL of saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by silica gel column chromatography (petroleum ether:ethyl acetate = 1:0-2:3, v/v) to give compound 7-4 (360 mg, 734.74 μmol). MS [ESI, M+1]: 490.2.

Step D. To a solution of compound 7-4 (160 mg, 326.55 μmol, 1 equiv.) in dichloromethane (10 mL) was added N -chlorosuccinimide (52.33 mg, 391.86 μmol, 1.2 equiv.). The reaction system was purged with nitrogen three times, reacted at 25 °C for 12 h, and diluted with 10 mL of dichloromethane. The organic phase was washed twice with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude product. The crude product was separated by preparative HPLC (water (0.225% formic acid)/acetonitrile, v/v) to give compound 7 . MS [ESI, M+1]: 524.2.

^1H NMR (400 MHz, DMSO- _d6 ) δ = 11.47 (s, 1H), 8.24 (br t, J = 5.3 Hz, 1H), 7.72 (d, J = 2, 3 Hz, 1H), 7.68-7.54 (m, 1H), 7.52-7.34 (m, 3H), 7.26-7.11 (m, 4H), 6.61 (d, J = 8.7 Hz, 1H), 6.20 (dd, J = 10.0, 17.1 Hz, 1H), 6 .06 (dd, J = 2.3, 17.1 Hz, 1H), 5.58-5.54 (m, 1H), 4.16 (t, J = 5.6 Hz, 2H), 3.47-3.36 (m, 2H), 2.48-2.39 (m, 2H), 0.90 (t, J = 7.5 Hz, 3H).

Example 8

Step A. A solution of indolin-2-one 8-1 (10 g, 75.10 mmol, 1 equiv.) and triethylamine (22.80 g, 225.31 mmol, 31.36 mL, 3 equiv.) in dichloromethane (100 mL) was purged with nitrogen three times, cooled to -70 °C, and trifluoromethanesulfonic anhydride (46.62 g, 165.23 mmol, 27.26 mL, 2.2 equiv.) was added dropwise while maintaining the temperature below -60 °C. After complete addition, the reaction system was allowed to react dropwise at -60 °C for 2 h. 1 M hydrochloric acid was added dropwise and slowly to adjust the pH to 1 while maintaining the temperature below 30 °C. The reaction system was washed twice with 100 ml of 0.5 M hydrochloric acid solution and once with 100 ml of saturated saline solution. The organic phase was dried over anhydrous sodium sulfate and concentrated to give the crude product, which was compound 8-2 .

¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 7.86 (d, J = 8.4 Hz, 1 H), 7.54 (d, J = 8.8 Hz, 1 H), 7.35- 7.40 (m, 2 H), 6.61 (s, 1 H).

Step B. To a solution of compound 8-2 (15 g, 37.76 mmol, 1 equiv.) in triethylamine (150 mL) were added Pd( _PPh3 ) ₄ (4.36 g, 3.78 mmol, 0.1 equiv.) and copper(I) iodide (719.09 mg, 3.78 mmol, 0.1 equiv.). The system was purged with nitrogen three times and trimethylsilylacetylene (18.54 g, 188.80 mmol, 26.15 mL, 5 equiv.) The reaction system was allowed to react under nitrogen atmosphere at 65°C for 12 h. The reaction system was quenched with 100 mL of water and extracted three times with 150 mL of ethyl acetate. The organic phases were combined, washed twice with 100 ml of saturated brine and dried over sodium sulfate. The reaction mixture was filtered and concentrated to give the crude product. The crude product was separated by silica gel column chromatography (petroleum ether:ethyl acetate = 1:0 - 10:1, v/v) to give compound 8-3 .

¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 7.97 (d, J = 8.8 Hz, 1 H), 7.57 (d, J = 8.0 Hz, 1 H), 7.37- 7.44 (m, 2 H), 7.07 (s, 1 H), 0.31 (s, 9 H).

Step C: Potassium carbonate (6.72 g, 48.64 mmol, 2 equiv) was added to a solution of compound 8-3 (8.4 g, 24.32 mmol, 1 equiv) in methanol (50 mL), and the reaction system was allowed to react at 70 °C for 0.5 h. After cooling to room temperature, the reaction was quenched with 100 mL of water, and the reaction system was extracted three times with 150 mL of dichloromethane. The organic phases were combined, washed twice with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude product, compound 8-4 .

¹ H NMR (400 MHz, CDCl3) ppm 8.19 (s, 1 H), 7.51 (d, J = 8.4 Hz, 1 H), 7.24 (d, J = 7.6 Hz, 1 H), 7.17-7.19 (m, 1 H), 7.06-7.08 (m, 1 H), 6.75 (s, 1 H), 3.24 (s, 1 H).

Step D. _Pd2 (dba) ₃ (648.67 mg, 708.37 μmol, 0.05 equiv ), bis[(2-diphenylphosphino)phenyl]ether (DPEphos, 1.53 g, 2.83 mmol, 0.2 equiv), and DABCO (3.18 g, 28.33 mmol, 3.12 mL, 2 equiv) were added to a dry flask. The reaction system was purged with nitrogen three times and compound 8-4 (2 g, 14.17 mmol, 1 equiv), 1,1,1-trifluoroiodoethane (2.97 g, 14.17 mmol, 1.39 mL, 1 equiv), and toluene (20 mL) were added sequentially under nitrogen atmosphere. The reaction mixture was allowed to react at 80°C for 12 h, filtered and washed with 50 ml of ethyl acetate. The organic phase was concentrated to give the crude product. The crude product was separated by silica gel column chromatography (petroleum ether:ethyl acetate = 1:0 - 5/1, v/v) to give compound 8-5 . MS [ESI, M+1]: 224.1.

Step E: Tetrakis(triphenylphosphine)platinum(0) (55.75 mg, 44.80 μmol, 0.02 equiv) was added to a solution of 8-5 (0.5 g, 2.24 mmol, 1 equiv) and bis(pinacolato)diboron (568.87 mg, 2.24 mmol, 1 equiv) in 2-methyltetrahydrofuran (10 mL). The reaction mixture was allowed to react at 85 °C for 12 h under nitrogen atmosphere and then cooled to room temperature to afford 8-6 for direct use in the next reaction without purification.

Step F. Compound 1-9 (532.97 mg, 1.12 mmol, 0.50 equiv), Pd( _PPh3 ) _2Cl2 (78.70 mg, 112.13 μmol, 0.05 equiv) and cesium carbonate (1.46 g, _4.49 mmol, 2 equiv) were added to a solution of compound 8-6 (1.07 g, 2.24 mmol, 1 equiv) in 2-methyltetrahydrofuran (10 mL). The reaction system was purged with nitrogen three times, 2.5 mL of water was added and the reaction was carried out at 30 °C for 12 h to give compound 8-7 for direct use in the next reaction without purification.

Step G. Compound 8-8 (649.89 mg, 2.70 mmol, 1.2 equiv), aqueous potassium hydroxide solution (4 M, 3.93 mL, 7 equiv) and Pd( _PPh3 ) _2Cl2 (78.87 mg, 112.37 μmol, 0.05 equiv) _were added to a solution of compound 8-7 (1.57 g, 2.25 mmol, 1 equiv) in 2-methyltetrahydrofuran (10 mL). The reaction system was purged with nitrogen three times and the reaction was carried out at 85 °C for 12 h. After cooling to room temperature, the reaction system was diluted with 10 mL of water and 10 mL of ethyl acetate. The aqueous phase was extracted 3 times with 10 mL of ethyl acetate. The organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by silica gel column chromatography to give compound 8-9 . MS [ESI, M+1]: 686.2.

Step H: The reaction was carried out in a solution of compound 8-9 (500 mg, 729.20 μmol, 1 equiv) and N -chlorosuccinimide (116.84 mg, 875.04 μmol, 1.2 equiv) in dichloromethane (5 mL) at 20 °C for 12 h. The reaction was quenched with 10 mL of water and the aqueous phase was extracted 3 times with 20 mL of dichloromethane. The organic phases were combined, washed twice with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product, compound 8-10 . MS [ESI, M+1]: 720.4.

Step I: Trifluoroacetic acid (71.25 mg, 624.89 μmol, 46.27 μL, 1 equiv) was added to a solution of compound 8-10 (450 mg, 624.89 μmol, 1 equiv) in dichloromethane (5 mL). The reaction mixture was allowed to react at 20 °C for 1 h and concentrated to give the crude product. The crude product was sequentially separated by preparative HPLC (water (0.025% formic acid, v/v)/acetonitrile system), thin layer chromatography (petroleum ether/ethyl acetate = 0/1, v/v) and preparative HPLC (water (0.025% formic acid, v/v)/acetonitrile system) to afford compound 8. MS [ESI, M+1]: 620.2.

^1H NMR (400 MHz, DMSO -d6 ) δ ppm 11.79 (s, 1H), 8.50-8.60 (m, 2H) _, 8.23 (s, 1H), 7.85 (d, J = 2.4 Hz, 1 H), 7.56 (d, J = 7.6 Hz, 1 H), 7.34-7.47 (m, 2 H), 7.12-7.29 (m, 2 H), 6.70 (d, J = 8.4 Hz, 1 H), 6.55-6 .63 (m, 1 H), 6.46-6.53 (m, 1 H), 4.18 (t, J = 5.6 Hz, 2 H), 3.50-3.63 (m, 2 H), 3.31-3.32 (m, 2 H), 2.97 (s, 3 H), 2.83-2.87 (m, 3 H), 2.77 (t, J = 5.6 Hz , 2H).

Example 9

Step A. 2-Chloro-4-fluoroiodobenzene (692.40 mg, 2.70 mmol _, 1.2 equiv), aqueous potassium hydroxide solution (4 M, 3.93 mL, 7 equiv) and Pd( _PPh3 ) _2Cl2 (78.87 mg, 112.37 μmol, 0.05 equiv) were added to a solution of compound 8-7 (1.57 g, 2.25 mmol, 1 equiv) in 2-methyltetrahydrofuran (10 mL). The reaction system was purged with nitrogen three times and the reaction was carried out at 85 °C for 12 h. After cooling to room temperature, the reaction system was diluted with 10 mL of water and 10 mL of ethyl acetate. The aqueous phase was extracted 3 times with 10 mL of ethyl acetate. The organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by silica gel column chromatography to give compound 9-1 . MS [ESI, M+1]: 701.2.

Step B. The reaction was carried out in a solution of compound 9-1 (200 mg, 285.25 μmol, 1 equiv.) and N -chlorosuccinimide (45.71 mg, 342.30 μmol, 1.2 equiv.) in dichloromethane (10 mL) at 20 °C for 12 h. The reaction was quenched with 10 mL of water and the aqueous phase was extracted 3 times with 20 mL of dichloromethane. The organic phases were combined, washed twice with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product, compound 9-2 . MS [ESI, M+1]: 735.1.

Step C. Trifluoroacetic acid (3.08 g, 27.01 mmol, 2 mL, 132.47 equiv) was added to a solution of compound 9-2 (150 mg, 203.92 μmol, 1 equiv) in dichloromethane (2 mL). The reaction mixture was allowed to react at 20 °C for 1 h and concentrated to give the crude product. The crude product was separated twice by preparative HPLC (water (0.025% formic acid, v/v)/acetonitrile) to give compound 9. MS [ESI, M+1]: 635.2.

^1H NMR (EW16419-73-P1A, 400 MHz, DMSO - _d6 ) δ ppm 11.66 (s, 1 H), 7.79 (d, J = 2.00 Hz, 1 H), 7.53 (d, J = 7.6 Hz, 1 H), 7.42-7.49 (m, 3 H), 7.21-7.33 (m, 3 H), 7.13-7.19 (m, 1 H), 6.55-6.67 (m, 2 H), 6.46-6.53 ( m, 1 H), 4.16 (t, J = 5.6 Hz, 2 H), 3.41-3.62 (m, 2 H), 3.30-3.31 (m, 2H), 2.97 (s, 3H), 2.83 (s, 3 H), 2.77 (t, J = 5.6 Hz, 2 H).

Example 10

Step A. o -Iodobenzonitrile (104.22 mg, 572.59 μmol, 1 equiv), aqueous potassium hydroxide solution (4 M, 715.74 μl, 5 equiv) and Pd( _PPh3 ) _2Cl2 (20.10 mg, 28.63 μmol, 0.05 equiv) _were added to a solution of compound 8-7 (400 mg, 572.59 μmol, 1 equiv) in 2-methyltetrahydrofuran (10 ml) and water (2.5 ml). The reaction system was purged with nitrogen three times and the reaction was carried out at 85 °C for 12 h. After cooling to room temperature, the reaction system was diluted with 10 ml of water and 10 ml of ethyl acetate. The aqueous phase was extracted 3 times with 10 mL ethyl acetate. The organic phases were combined, washed with 20 mL saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by thin layer chromatography on silica gel to give compound 10-1 . MS [ESI, M+1]: 674.2.

Step B. The reaction was carried out in a solution of compound 10-1 (35 mg, 51.95 μmol, 1 equiv.) and N -chlorosuccinimide (8.32 mg, 62.34 μmol, 1.2 equiv.) in dichloromethane (5 mL) at 20 °C for 12 h. The reaction was quenched with 10 mL of saturated aqueous sodium sulfite solution and the aqueous phase was extracted 3 times with 20 mL of dichloromethane. The organic phases were combined, washed twice with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product, compound 10-2 . MS [ESI, M+1]: 708.2.

Step C: To a solution of compound 10-2 (30 mg, 42.36 μmol, 1 equiv.) in dichloromethane (3 mL) was added trifluoroacetic acid (770.00 mg, 6.75 mmol, 0.5 mL, 159.41 equiv.). The reaction mixture was allowed to react at 20 °C for 1 h and concentrated to give the crude product. The crude product was separated by preparative HPLC (water (0.025% formic acid, v/v)/acetonitrile system) and preparative HPLC (water (0.05% aqueous ammonia, v/v)/acetonitrile system) to give compound 10. MS [ESI, M+1]: 608.3.

^1H NMR (EW16419-216-P1A, 400 MHz, DMSO- _d6 ) δ = 11.59 (s, 1H), 7.81-7.68 (m, 4H), 7.60-7.42 ( m, 3H), 7.32-7.12 (m, 3H), 6.66-6.43 (m, 3H), 4.21-4.11 (m, 2H), 3.47-3.43 (m, 2H), 3.48-3.41 (m, 2H), 2.96 (s , 3H), 2.89 (s, 3H), 2.76 (t, J = 6.0 Hz, 2H).

Example 11

Step A: To a solution of compound 11-1 (100 mg, 154.52 μmol, 1 equiv.) in dichloromethane (1 mL) was added 1 mL of trifluoroacetic acid. The reaction mixture was allowed to react at 25 °C for 1 h under nitrogen atmosphere and concentrated to give the crude product. The crude product was separated by preparative HPLC (water (0.225% formic acid, v/v)/acetonitrile) to give compound 11 . MS [ESI, M+1]: 547.3.

¹ H NMR (400 MHz, _DMSO-d6 ) δ ppm 10.77 (s, 1 H), 8.90 (br s, 2 H), 7.74 (d, J = 2.45 Hz, 1 H ), 7.57 (d, J = 7.46 Hz, 1 H), 7.26-7.41 (m, 4 H), 7.18 (m, 1 H), 7.08 (t, J = 7.23 Hz, 1 H), 7.01 (t, J = 7.04 Hz, 1 H), 6.81 (d, J = 15.16 Hz, 1 H), 6.67 (d, J = 8.68 Hz, 1 H), 6.52-6.60 (m, 2 H), 4.37 (t, J = 5.07 Hz, 2 H), 3.81 (br d, J = 4.40 Hz, 2 H), 3.04 (s, 3 H), 2.88 (s, 3 H), 2.59-2, 75 (m, 2 H), 2.31-2.44 (m, 2 H), 1.01 (t, J = 7.52 Hz, 3 H).

Example 12

Step A: To a solution of compound 11-1 (100 mg, 154.52 μmol, 1 equiv.) in dichloromethane (3 mL) was added N -bromosuccinimide (33.00 mg, 185.42 μmol, 1.2 equiv.). The reaction system was purged with nitrogen three times and the reaction was allowed to proceed at 25 °C for 2 h. To the reaction mixture was added 10 mL of dichloromethane. The organic phase was washed twice with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product, compound 12-1 . MS [ESI, M+1]: 804.9.

Step B: To a solution of compound 12-1 (140.00 mg, 173.92 μmol, 1 equiv.) in dichloromethane (1 mL) was added 1 mL of trifluoroacetic acid. The reaction mixture was allowed to react at 25 °C for 1 h under nitrogen atmosphere and concentrated to give the crude product. The crude product was separated by preparative HPLC (water (0.1% trifluoroacetic acid, v/v)/acetonitrile) to give compound 12 . MS [ESI, M+1]: 627.1.

^1H NMR (400 MHz, _DMSO-d6 ) δ ppm 11.70 (s, 1 H), 8.91 (br s, 2 H), 7.78 (d, J = 2.02 Hz, 1 H ), 7.31-7.38 (m, 1H), 7.35-7.28 (m, 2H), 7.25-7.18 (m, 2H), 7.12-7.27 (m, 2 H), 7.19 (d, J = 13.28 Hz, 1 H), 6.79 (d, J = 15.18 Hz, 1 H), 6.68 (d, J = 8.54 Hz, 1 H), 6.54 (m, 1 H), 4.35 (br t, J = 4.96 Hz, 2 H), 3.97 (br s, 2 H), 3.79 (br d, J = 5.14 Hz, 2 H), 3.02 (s, 3 H), 2.87 (s, 3 H ), 2.36-2.47 (m, 2 H), 0.90 (t, J = 7.56 Hz, 3 H).

Example 13

Step A: To a solution of compound 11-1 (200 mg, 309.03 μmol, 1 equiv.) in dichloromethane (10 mL) was added N -iodosuccinimide (83.43 mg, 370.84 μmol, 1.2 equiv.). The reaction system was purged with nitrogen three times and allowed to react at 25 °C for 2 h. The reaction was quenched with 5 mL of saturated aqueous sodium sulfite solution. To the reaction mixture was added 10 mL of dichloromethane. The organic phase was washed twice with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product, compound 13-1 . MS [ESI, M+1]: 773.3.

Step B: To a solution of compound 13-1 (200 mg, 258.71 μmol, 1 equiv.) in dichloromethane (3 mL) was added 3 mL of trifluoroacetic acid. The reaction mixture was allowed to react at 25 °C for 1 h under nitrogen atmosphere and concentrated to give the crude product. The crude product was separated by preparative HPLC (water (0.1% trifluoroacetic acid, v/v)/acetonitrile) to give compound 13 . MS [ESI, M+1]: 673.1.

¹ H NMR (400 MHz, _DMSO-d6 ) δ ppm 11.79 (s, 1 H), 8.86 (br s, 2 H), 7.81 (m, 1 H) 7.47 (d, J = 8.84 Hz, 1 H), 7.39-7.42 (m, 1 H), 7.30-7.37 (m, 1 H), 7.27-7.29 (m, 2 H), 7.17-7.26 (m, 1 H), 7.11-7.15 (m, 1 H), 6 .88 (d, J =15.18 Hz, 1 H), 6.78 (d, J = 15.16 Hz, 1 H), 6.68 (d, J =8.66 Hz, 1 H), 6.54 (m, 1 H), 4.36 (t, J = 5.02 Hz, 2 H), 3.72-3.98 (m, 2 H), 3.19-3.44 (m, 2 H ), 3.02 (s, 3 H), 2.87 (s, 3 H), 2.30-2.48 (m, 2 H), 0.90 (t, J = 7.52 Hz, 3 H).

Example 14

Step A. 1-Bromo-2-chlorobenzene (317.20 mg, 1.66 mmol, 193.42 μL, 1.2 equiv), aqueous potassium hydroxide solution (4 M, 2.42 mL, 7 equiv) and Pd( _PPh3 ) _2Cl2 (48.45 _mg , 69.03 μmol, 0.05 equiv) were added to a solution of compound 1-10 (0.89 g, 1.38 mmol, 1 equiv) in 2-methyltetrahydrofuran (10 mL). The reaction system was purged with nitrogen three times and the reaction was carried out at 85 °C for 12 h. After cooling to room temperature, the reaction system was diluted with 30 mL of water. The aqueous phase was extracted 3 times with 50 mL of ethyl acetate. The organic phases were combined, washed twice with 50 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by silica gel column chromatography to give compound 14-1 . MS [ESI, M+1]: 629.3.

Step B: To a solution of compound 14-1 (170 mg, 270.19 μmol, 1 equiv.) in dichloromethane (10 mL) was added N -chlorosuccinimide (43.30 mg, 324.23 μmol, 1.2 equiv.). The reaction system was purged with nitrogen three times and allowed to react at 25 °C for 2 h. The reaction was quenched with 5 mL of saturated aqueous sodium sulfite solution. To the reaction mixture was added 10 mL of dichloromethane. The organic phase was washed twice with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product, compound 14-2 . MS [ESI, M+1]: 663.4.

Step C: To a solution of compound 14-2 (200 mg, 301.37 μmol, 1 equiv.) in dichloromethane (3 mL) was added 3 mL of trifluoroacetic acid. The reaction mixture was allowed to react at 25 °C for 1 h under nitrogen atmosphere and concentrated to give the crude product. The crude product was separated by preparative HPLC (water (0.1% trifluoroacetic acid, v/v)/acetonitrile) to give compound 14 . MS [ESI, M+1]: 563.2.

¹ H NMR (400 MHz, DMSO - _d6 ) δ ppm 11.54 (s, 1 H), 8.88 (br s, 2 H), 8.75 (m, 1 H), 7.81-8 .53 (m, 1 H), 7.77 (m, 2 H), 7.40-7.65 (m, 4 H), 7.22-7.35 (m, 1 H), 7.12-7.21 (m, 1 H), 6.78 (d, J = 15.28 Hz, 1 H), 6.65 (d , J = 8.40 Hz, 1 H), 6.54 (m, 1 H), 4.35 (t, J = 5.04 Hz, 2 H), 3.58 (br s, 2 H), 3.27 (br s, 2 H), 3.02 (s, 3 H), 2.87 (s, 3 H), 2.39-2.48 (m, 2 H), 0.90 (t, J = 7.52 Hz, 3 H).

Example 15

Step A. 1-Bromo-2,4-dichlorobenzene (239.70 mg, _{1.06 mmol, 1.2 equiv), aqueous potassium hydroxide solution (4 M, 1.55 mL, 7 equiv) and Pd(PPh3)2Cl2 ( 31.03} mg, 44.21 μmol, 0.05 equiv) were added to a solution of compound 1-8 (0.57 g, 884.26 μmol, 1 equiv) in 2-methyltetrahydrofuran (10 mL). The reaction system was purged with nitrogen three times and the reaction was carried out at 85 °C for 12 h. After cooling to room temperature, the reaction system was diluted with 10 mL of water and 10 mL of ethyl acetate. The aqueous phase was extracted 3 times with 10 mL of ethyl acetate. The organic phases were combined, washed 3 times with 10 mL of brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by reverse phase HPLC (under 0.1% trifluoroacetic acid conditions) to give compound 15-1 . MS [ESI, M+1]: 663.3.

Step B. The reaction was carried out in a solution of compound 15-1 (125 mg, 188.36 μmol, 1 equiv.) and N -chlorosuccinimide (30.18 mg, 226.03 μmol, 1.2 equiv.) in dichloromethane (10 mL) at 20 °C for 1 h. The reaction was quenched with 10 mL of water and the aqueous phase was extracted 3 times with 20 mL of dichloromethane. The organic phases were combined, washed twice with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product, compound 15-2 . MS [ESI, M+1]: 697.2.

Step C: To a solution of compound 15-2 (150 mg, 214.88 μmol, 1 equiv.) in dichloromethane (2 mL) was added trifluoroacetic acid (24.50 mg, 214.88 μmol, 15.91 μL, 1 equiv.). The reaction mixture was allowed to react at 20 °C for 1 h and concentrated to give the crude product. The crude product was separated by preparative HPLC (water (0.025% formic acid, v/v)/acetonitrile) to give compound 15. MS [ESI, M+3]: 599.3.

¹ H NMR (400 MHz, DMSO - _d6 ) δ ppm 11.52 (s, 1 H), 8.85 (br s, 2 H), 7.77 (dd, J = 2.4, 0.8 Hz, 1 H), 7.63 (d, J = 2.0 Hz, 1 H), 7.51 (d, J = 8.0 Hz, 1 H), 7.39-7.46 (m, 2 H), 7.31-7.37 (m, 2 H), 7.13-7.24 (m, 2 H), 6, 79 (d, J =15.2 Hz, 1 H), 6.69 (dd, J = 8.4, 0.61 Hz, 1 H), 6.50-6.58 (m, 1 H), 4.34-4.39 (m, 2 H), 3.80 (br d, J = 4.8 Hz, 2 H), 3.28 (br s, 2 H), 3.03 (s, 3 H), 2.87 ( s, 3 H), 2.42-2.47 (m, 2 H), 0.90 (t, J = 7.6 Hz, 3 H).

Example 16

Step A. 4-Bromo-2-methylthiophene (187.88 mg, _1.06 mmol, 1.2 equiv), aqueous potassium hydroxide solution (4 M, 1.55 mL, 7 equiv) and Pd( _PPh3 ) _2Cl2 (31.03 mg, 44.21 μmol, 0.05 equiv) were added to a solution of compound 1-8 (0.57 g, 884.26 μmol, 1 equiv) in 2-methyltetrahydrofuran (10 mL). The reaction system was purged with nitrogen three times and the reaction was carried out at 85 °C for 12 h. After cooling to room temperature, the reaction system was diluted with 10 mL of water and 10 mL of ethyl acetate. The aqueous phase was extracted 3 times with 10 mL of ethyl acetate. The organic phases were combined, washed three times with 10 mL of brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by reverse phase HPLC (water (0.1% trifluoroacetic acid, v/v)/acetonitrile) to give compound 16-1 . MS [ESI, M+1]: 615.4.

Step B. The reaction was carried out in a solution of compound 16-1 (65 mg, 105.73 μmol, 1 equiv) and N -chlorosuccinimide (16.94 mg, 126.87 μmol, 1.2 equiv) in dichloromethane (10 mL) at 20 °C for 12 h. The reaction was quenched with 10 mL of water and the aqueous phase was extracted 3 times with 20 mL of dichloromethane. The organic phases were combined, washed twice with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product, compound 16-2 . MS [ESI, M+1]: 649.2.

Step C: To a solution of compound 16-2 (100 mg, 154.03 μmol, 1 equiv.) in dichloromethane (2 mL) was added trifluoroacetic acid (17.56 mg, 154.03 μmol, 11.40 μL, 1 equiv.). The reaction mixture was allowed to react at 20 °C for 1 h and concentrated to give the crude product. The crude product was separated by preparative HPLC (water (0.025% formic acid, v/v)/acetonitrile) to give compound 16 (10.13 mg, 18.01 μmol). MS [ESI, M+1]: 549.4.

^1H NMR (EW16419-130-P1A, 400 MHz, DMSO - _d6 ) δ ppm 11.42 (s, 1 H), 7.72 (dd, J = 2.4, 0.67 Hz, 1 H) , 7.48 (d, J = 7.6 Hz, 1 H), 7.34-7.39 (m, 1 H), 7.23-7.28 (m, 1 H), 7.09-7.20 (m, 2 H), 7.04 (d, J = 1.6 Hz, 1 H ), 6.57-6.67 (m, 2 H), 6.50-6.56 (m, 2 H), 4.22 (t, J = 5.6 Hz, 2 H), 3.32-3.36 (m.5 H), 2.99 (s, 2 H), 2.84 (s, 3 H), 2.35-2.43 (m, 5 H), 0.93-0.99 (m, 3 H) .

Example 17

Step A. 1-Bromo-2,4-difluorobenzene (409.57 mg, _2.12 mmol, 1.2 equiv), aqueous potassium hydroxide solution (4 M, 3.09 mL, 7 equiv), and Pd( _PPh3 ) _2Cl2 (62.07 mg, 88.43 μmol, 0.05 equiv) were added to a solution of compound 1-8 (1.14 g, 1.77 mmol, 1 equiv) in 2-methyltetrahydrofuran (10 mL) and water (2 mL). The reaction system was purged with nitrogen three times and the reaction was carried out at 85 °C for 12 h. After cooling to room temperature, the reaction system was diluted with 10 mL of water and 10 mL of ethyl acetate. The aqueous phase was extracted 3 times with 10 mL of ethyl acetate. The organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by reversed-phase HPLC (water (0.1% trifluoroacetic acid, v/v)/acetonitrile) to give compound 17-1 . MS [ESI, M+1]: 631.4.

Step B. The reaction was carried out in a solution of compound 17-1 (287 mg, 455.03 μmol, 1 equiv.) and N -chlorosuccinimide (72.91 mg, 546.04 μmol, 1.2 equiv.) in dichloromethane (5 mL) at 25 °C for 1 h. The reaction was quenched with 10 mL of water and the aqueous phase was extracted 3 times with 20 mL of dichloromethane. The organic phases were combined, washed twice with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product, compound 17-2 . MS [ESI, M+1]: 665.2.

Step C. Trifluoroacetic acid (1.54 g, 13.51 mmol, 1 mL, 29.95 equiv) was added to a solution of compound 17-2 (300 mg, 451.01 μmol, 1 equiv) in dichloromethane (5 mL). The reaction mixture was allowed to react at 25 °C for 0.5 h and concentrated to give the crude product. The crude product was separated by preparative HPLC (water (0.025% formic acid, v/v)/acetonitrile) to give compound 17 . MS [ESI, M+1]: 565.2.

¹H NMR (EW16419-174-P1, 400 MHz, DMSO-d ₆) δ ppm 11.50 (s, 1 H), 8.19 (s, 1 H), 7.68 (s, 1 H), 7.50 (d, J= 7.82 Hz, 1 H), 7.28-7.41 (m, 2 H), 7.04-7.24 (m, 5 H), 6.45-6.65 (m, 3 H ), 4.16 (t, J= 5.75 Hz, 2 H), 3.32 (br d, J= 4.40 Hz, 2 H), 2.98 (s, 3 H), 2.83 (s, 3 H), 2.78 (t, J= 5.75 Hz, 2 H), 2.43 (q,J= 7.30 Hz, 2 H), 0.90 (t,J= 7.46 Hz, 3 H).

Example 18

Step A. 1-Bromo-3-methylbenzene (504.13 mg, 2.95 mmol, 357.54 μL, 1 equiv), aqueous potassium hydroxide solution (4 _{M, 20.63 mmol, 5.16 mL, 7 equiv) and Pd(PPh3)2Cl2 ( 103.44} mg, 147.38 μmol, 0.05 equiv) were added to a solution of compound 1-8 (1.9 g, 2.95 mmol, 1 equiv) in 2-methyltetrahydrofuran (5 mL). The reaction system was purged with nitrogen three times and the reaction was carried out at 85 °C for 12 h. After cooling to room temperature, the reaction system was diluted with 20 mL of water and 20 mL of ethyl acetate. The aqueous phase was extracted 3 times with 10 mL of ethyl acetate. The organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by reverse phase HPLC (0.1% trifluoroacetic acid) to give compound 18-1 . MS [ESI, M+1]: 609.3.

Step B: To a solution of 18-1 (271 mg, 445.16 μmol, 1 equiv) in dichloromethane (5 mL) was added N -chlorosuccinimide (71.33 mg, 534.19 μmol, 1.2 equiv). The reaction mixture was allowed to react at 25 °C for 1 h. The reaction was quenched with 10 mL of water and the aqueous phase was extracted 3 times with 20 mL of dichloromethane. The organic phases were combined, washed twice with 20 mL of brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product 18-2 . MS [ESI, M+1]: 643.4.

Step C: To a solution of compound 18-2 (250 mg, 388.67 μmol, 1 equiv.) in dichloromethane (5 mL) was added trifluoroacetic acid (770.00 mg, 6.75 mmol, 0.5 mL, 17.37 equiv.). The reaction mixture was allowed to react at 25 °C for 1 h and concentrated to give the crude product. The crude product was separated by preparative HPLC (water (0.025% formic acid, v/v)/acetonitrile) to give compound 18 . MS [ESI, M+1]: 543.3.

^1H NMR (400 MHz, DMSO- _d6 ) δ = 11.54 (s, 1H), 9.42 (br s, 2H), 7.67 (d, J = 2.0 Hz, 1H), 7 .49 (d, J = 7.6 Hz, 1H), 7.40 (d, J = 8.0 Hz, 1H), 7.28-7.01 (m, 6H), 6.95 (d, J = 7.6 Hz, 1H), 6.83 (d, J = 15.2 Hz, 1H), 6.69-6 .54 (m, 2H), 4.40 (t, J = 5.2 Hz, 2H), 3.79-3.75 (m, 2H), 3.24 (br s, 2H), 3.03 (s, 3H), 2.87 (s, 3H), 2.44 (q, J = 7.2 Hz, 2H), 2.27 (s, 3H), 0.90 (t, J = 7.6 Hz, 3H).

Example 19

Step A. A solution of 2,6-difluoro-3-iodopyridine 19-1 (10 g, 41.50 mmol, 1 equiv.) in N,N -dimethylformamide (80 mL) was cooled to 0 °C and sodium hydride (2.49 g, 62.25 mmol, 60% purity, 1.5 equiv.) was added portionwise while maintaining the temperature below 0 °C. After complete addition, the reaction system was allowed to react at 0 °C for 0.5 h. A solution of N -Boc-ethanolamine (6.69 g, 41.50 mmol, 6.43 mL, 1 equiv.) in N,N -dimethylformamide (20 mL) was added dropwise to the reaction system while maintaining the temperature below 0 °C. After completion of the above dropwise addition, the reaction system was reacted at 0°C for 1 hour. 50 ml of water was added to the reaction system for quenching, and extraction was performed 3 times with 50 ml of ethyl acetate. The organic phases were combined, washed twice with 50 ml of saturated brine, dried over sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was separated by silica gel column chromatography (petroleum ether:ethyl acetate = 1:0-5:1, v/v) to give compound 19-2 .

¹ H NMR (400 MHz, CHLOROFORM-d) δ = 7.85 (t, J = 8.4 Hz, 1H), 6.45-6.31 (m, 1H), 4.30-4.13 (m, 2H), 3.44 (br d, J = 4.8 Hz, 2H), 1.37 (s, 9H).

Step B: To a solution of compound 19-2 (2.04 g, 5.34 mmol, 1 equiv.) in ethanol (20 mL) was added methanolic hydrochloric acid (4 M, 2 mL, 1.50 equiv.). The reaction system was allowed to react at 45 °C for 2 h and concentrated under reduced pressure to give compound 19-3 . MS [ESI, M+1]: 283.4.

Step C. A solution of N , N -dimethylbromocrotonamide (615.03 mg, 3.20 mmol, 0.6 equiv.) in N , N -dimethylformamide (1 mL) was added dropwise to a solution of compound 19-3 (1.7 g, 5.34 mmol, 1 equiv., HCl) and N , N -diisopropylethylamine (2.07 g, 16.01 mmol, 2.79 mL, 3 equiv.) in N , N -dimethylformamide (5 mL). The reaction system was allowed to proceed at 25 °C for 12 h. A solution of (Boc) _2O (1.40 g, 6.40 mmol, 1.47 mL, 1.2 equiv.) in dichloromethane (2 mL) was added dropwise to the reaction system while maintaining the temperature below 0 °C. The reaction system was allowed to react at 25 °C for 2 hours. The reaction system was diluted with 20 mL of water and 20 mL of ethyl acetate and separated. The aqueous phase was extracted three times with 10 mL of ethyl acetate. The organic phases were combined, washed twice with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was separated by silica gel column chromatography (petroleum ether:ethyl acetate = 1:0-0:1, v/v) to give compound 19-4 . MS [ESI, M+1]: 494.2.

Step D. Compound 19-4 (440.66 mg, 893.28 μmol, 0.6 equiv), Pd( _PPh3 ) _2Cl2 ( _52.25 mg, 74.44 μmol, 0.05 equiv) and cesium carbonate (970.16 mg, 2.98 mmol, 2 equiv) were added to a solution of compound 1-8 (0.63 g, 1.49 mmol, 1 equiv) in 2-methyltetrahydrofuran (5 mL). The reaction system was purged with nitrogen three times, 1 mL of water was added and the reaction was carried out at 30 °C for 12 h to give compound 19-5 for direct use in the next reaction without purification. MS [ESI, M+1]: 663.4.

Step E. 1-Bromo-3-methylbenzene (278.68 mg, 1.49 mmol, 188.30 μL, 1 equiv), aqueous potassium hydroxide solution (4 M, 2.61 mL, 7 equiv) and Pd( _PPh3 ) _2Cl2 (52.25 _mg , 74.50 μmol, 0.05 equiv) were added to a solution of compound 19-5 (986.47 mg, 1.49 mmol, 1 equiv) in 2-methyltetrahydrofuran (20 mL). The reaction system was purged with nitrogen three times and the reaction was carried out at 85 °C for 12 h. After cooling to room temperature, the reaction system was diluted with 10 mL of water and 10 mL of ethyl acetate. The aqueous phase was extracted 3 times with 10 mL of ethyl acetate. The organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by reverse phase chromatography (0.1% trifluoroacetic acid) to give compound 19-6 . MS [ESI, M+1]: 643.4.

Step F: To a solution of compound 19-6 (0.1 g, 155.58 μmol, 1 equiv.) in dichloromethane (5 mL) was added N -chlorosuccinimide (24.93 mg, 186.70 μmol, 1.2 equiv.). The reaction system was allowed to proceed at 25 °C for 1 h. The reaction was quenched with 10 mL of saturated aqueous sodium sulfite solution and the aqueous phase was extracted 3 times with 20 mL of dichloromethane. The organic phases were combined, washed twice with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by preparative HPLC (water (0.025% formic acid, v/v)/acetonitrile system) to afford compound 19-7 . MS [ESI, M+1]: 677.4.

Step G: To a solution of compound 19-7 (20 mg, 29.53 μmol, 1 equiv.) in dichloromethane (3 mL) was added trifluoroacetic acid (770.00 mg, 6.75 mmol, 0.5 mL, 228.66 equiv.). The reaction mixture was allowed to react at 25 °C for 1 h and concentrated to give the crude product. The crude product was separated by preparative HPLC (water (0.05% hydrochloric acid, v/v)/acetonitrile) to give compound 19 . MS [ESI, M+1]: 577.3.

^1H NMR (400 MHz, METHANOL-d4) δ = 7.44 (dd, J = 8.4, 9.2 Hz, 1H), 7.37 (d, J = 8.0 Hz, 1H), 7 .26 (d, J = 8.0 Hz, 1H), 7.10-6.98 (m, 3H), 6.76-6.62 (m, 4H), 6.58-6.46 (m, 2H), 4.39-4.32 (m, 2H), 3.77 (d, J = 6.4 Hz, 2H), 3.61 (s, 3H), 3.33-3.27 (m, 2H), 3.00 (s, 3H), 2.88 (s, 3H), 2.46 (q, J = 7.6 Hz, 2H), 0.89 (t, J = 7.6 Hz, 3H).

Example 20

Step A: To a solution of 4-1 (964 mg, 1.54 mmol, 1 equiv.) in dichloromethane (20 mL) was added N -bromosuccinimide (329.55 mg, 1.85 mmol, 1.2 equiv.). The reaction system was purged with nitrogen three times and allowed to react at 25 °C for 1 h. The reaction was quenched with 10 mL of saturated aqueous sodium sulfite solution and the aqueous phase was extracted 3 times with 20 mL of dichloromethane. The organic phases were combined, washed twice with 20 mL of saturated brine, dried over anhydrous sodium sulfate and concentrated to give 20-1 . MS [ESI, M+1]: 703.3.

Step B. Compound 20-2 (1.07 g, 4.26 mmol, 1.19 mL, 3 equiv), cesium carbonate (1.39 g, 4.26 mmol, 3 equiv) and Pd(dppf) _Cl2.CH2Cl2 (116.06 mg, 142.11 μmol, 0.1 equiv) were _added to a solution of compound 20-1 (1 g, 1.42 mmol, 1 equiv) in dioxane (10 mL). The reaction _system was purged with nitrogen three times and the reaction was carried out at 110 °C for 3 h. After cooling to room temperature, 10 mL of water and 10 mL of ethyl acetate were added to the reaction system for quenching and it was separated. The aqueous phase was extracted 3 times with 10 mL of ethyl acetate. The organic phases were combined, washed with 10 mL of brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by reverse phase column chromatography (water (0.1% trifluoroacetic acid, v/v)/acetonitrile) to give compound 20-3 . MS [ESI, M+1]: 639.6.

Step C: To a solution of compound 20-3 (650 mg, 1.02 mmol, 1 equiv.) in dichloromethane (10 mL) was added trifluoroacetic acid (3.08 g, 27.01 mmol, 2.00 mL, 26.55 equiv.). The reaction mixture was allowed to react at 25 °C for 0.5 h and concentrated to give the crude product. The crude product was separated by preparative HPLC (water (0.05% hydrochloric acid, v/v)/acetonitrile) to give compound 20. MS [ESI, M+1]: 539.3.

^1H NMR (400 MHz, DMSO- _d6 ) δ = 10.80 (s, 1H), 9.27-9.03 (m, 2H), 7.67 (d, J = 2.0 Hz, 1H ), 7.48 (d, J = 7.6 Hz, 1H), 7.35-7.15 (m, 3H), 7.10-6.95 (m, 2H), 6.87-6.74 (m, 4H), 6.69-6.49 (m, 2H), 4.37 (br t, J = 5, 2 Hz, 2H), 3.78 (br d, J = 6.0 Hz, 2H), 3.70 (s, 3H), 3.25 (br s, 2H), 3.03(s, 3H), 2.87 (s, 3H), 2.45 (br d, J = 7.6 Hz, 2H), 2.24-2.18 (m, 3H), 0.87 (t, J = 7.4 Hz, 3H).

Example 21

Step A: A solution of compound 21-1, which is 4-iodophenol (10 g, 45.45 mmol, 1 equiv), N -Boc-ethanolamine (8.79 g, 54.54 mmol, 8.45 mL, 1.2 equiv), and triphenylphosphine (17.88 g, 68.18 mmol, 1.5 equiv) in tetrahydrofuran (80 mL) was cooled to 0 °C and a solution of diethyl azodicarboxylate (11.87 g, 68.18 mmol, 12.39 mL, 1.5 equiv) in tetrahydrofuran (10 mL) was added dropwise while maintaining the temperature below 0 °C. After complete addition, the reaction system was allowed to react at 25 °C for 12 hours. To the reaction system were added 50 ml of water and 50 ml of ethyl acetate for quenching, and extraction was performed 3 times with 50 ml of ethyl acetate. The organic phases were combined, washed with 50 ml of saturated brine, dried over sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was separated by silica gel column chromatography (petroleum ether:ethyl acetate = 1:0 - 5:1, v/v) to give compound 21-2 .

Step B. To a solution of compound 21-2 (14 g, 38.55 mmol, 1 equiv.) in methanol (100 mL) was added methanolic hydrochloric acid (4 M, 20 mL, 2.08 equiv.). The reaction system was allowed to react at 45 °C for 1 h and concentrated under reduced pressure to give compound 21-3 . MS [ESI, M+1]: 264.0.

Step C. A solution of N , N -dimethylbromocrotonamide (4.62 g, 24.04 mmol, 0.6 equiv.) in N , N -dimethylformamide (10 mL) was added dropwise to a solution of compound 21-3 (12 g, 40.06 mmol, 1 equiv., HCl) and N , N -diisopropylethylamine (15.53 g, 120.19 mmol, 20.93 mL, 3 equiv.) in N , N -dimethylformamide (80 mL). The reaction system was allowed to proceed at 25 °C for 12 h. To the reaction system was added dropwise a solution of (Boc) _2O (10.49 g, 48.07 mmol, 11.04 ml, 1.2 equiv.) in N , N -dimethylformamide (10 ml) at 0°C, and the reaction was carried out at 25°C for 12 hours. After the reaction system reached room temperature, 100 ml of water was added thereto for quenching, and separation was carried out. The aqueous phase was extracted 3 times with 100 ml of ethyl acetate. The organic phases were combined, washed twice with 100 ml of saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. The crude product was separated by silica gel column chromatography (petroleum ether:ethyl acetate=1:0-0:1, v/v) to give compound 21-4 . MS [ESI, M+1]: 475.1.

Step D. Compound 21-4 (2.52 g, 5.32 mmol, 0.6 equiv), Pd( _PPh3 ) _2Cl2 ( _311.01 mg, 443.09 μmol, 0.05 equiv) and cesium carbonate (5.77 g, 17.72 mmol, 2 equiv) were added to a solution of compound 1-8 (3.75 g, 8.86 mmol, 1 equiv) in 2-methyltetrahydrofuran (10 mL). The reaction system was purged with nitrogen three times, added 5 mL of water and allowed to react at 30 °C for 12 h to give compound 21-5 for direct use in the next reaction without purification.

Step E. 3-Iodoanisole (1.66 g, 8.86 mmol, 1.12 mL, 1 equiv), aqueous potassium hydroxide solution (4 M, 15.51 mL, 7 equiv), and Pd(dppf) _Cl2 (324.01 mg, 443.00 μmol, 0.05 equiv) were added to a solution of compound 21-5 (5.7 g, 8.86 mmol, 1 equiv) in 2-methyltetrahydrofuran (20 mL). The reaction system was purged with nitrogen three times and the reaction was carried out at 85 °C for 12 h. After cooling to room temperature, the reaction system was diluted with 20 mL of water and 20 mL of ethyl acetate. The aqueous phase was extracted 3 times with 20 mL of ethyl acetate. The organic phases were combined, washed with 20 mL of brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by reverse phase chromatography (0.1% trifluoroacetic acid) to give compound 21-6 . MS [ESI, M+1]: 624.4.

Step F: To a solution of 21-6 (1.51 g, 2.42 mmol, 1 equiv) in dichloromethane (20 mL) was added N -chlorosuccinimide (387.90 mg, 2.90 mmol, 1.2 equiv). The reaction mixture was allowed to react at 25 °C for 1 h. The reaction was quenched with 10 mL of saturated aqueous sodium sulfite solution and the aqueous phase was extracted 3 times with 20 mL of dichloromethane. The organic phases were combined, washed twice with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give 21-7 . MS [ESI, M+1]: 658.3.

Step G: To a solution of 21-7 (1.5 g, 2.28 mmol, 1 equiv) in dichloromethane (20 mL) was added trifluoroacetic acid (7.70 g, 67.53 mmol, 5 mL, 29.63 equiv). The reaction mixture was allowed to react at 25 °C for 0.5 h and concentrated to give the crude product. The crude product was separated by preparative HPLC (water (0.05% aqueous ammonia, v/v)/acetonitrile) to give 21 . MS [ESI, M+1]: 558.1.

^1H NMR (400 MHz, DMSO- _d6 ) δ = 11.39 (s, 1H), 7.48 (d, J = 7.6 Hz, 1H), 7.37 (d, J = 8.0 Hz, 1H), 7.21-7.09 (m, 3H), 6.87-6.70 (m, 5H), 6.69-6.56 (m, 3H), 6.54-6.46 (m, 1H), 3.92-3.85 (m, 2H), 3.66 (s, 3H), 3. 33-3.29 (m, 2H), 2.98 (s, 3H), 2.84 (s, 3H), 2.78 (br t, J = 5.6 Hz, 2H), 2.45-2.37 (m, 2H), 0.92-0.85 (m, 3H).

Examples 22 and 23

Step A: Trans -2-pentenoic acid (5 g, 49.94 mmol, 5.05 mL, 1 equiv) was dissolved in carbon tetrachloride (50 mL) and N -bromosuccinimide (11.56 g, 64.92 mmol, 1.3 equiv) was added. The reaction system was allowed to react at 80 °C for 12 h under nitrogen atmosphere, filtered and concentrated under reduced pressure to give the crude product as compound 22-1 . MS [ESI, M+1]: 179.1.

Step B. To compound 22-1 (3 g, 16.76 mmol, 1 equiv) was added 8 mL of thionyl chloride. The reaction system was allowed to react at 80°C for 12 h and concentrated under reduced pressure to give compound 22-2 .

Step C: To a solution of dimethylamine (1.33 g, 16.26 mmol, 1.49 mL, 1 equiv, hydrochloride) in dichloromethane (30 mL) was added sodium carbonate (3.45 g, 32.51 mmol, 2 equiv). To the reaction system was added dropwise a solution of compound 22-2 (3.21 g, 16.26 mmol, 1 equiv) in dichloromethane (12 mL) at 0 °C, and reacted at 25 °C for 2 h. The reaction system was diluted with 20 mL of water and 20 mL of dichloromethane and separated. The organic phase was washed twice with 30 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product as compound 22-3 . MS [ESI, M+1]: 206.1.

Step D. Compound 22-4 (6.83 g, 25.88 mmol, 1 equiv) was dissolved in DMF (100 mL) and diisopropylethylamine (6.69 g, 51.76 mmol, 9.02 mL, 2 equiv) was added. Compound 22-3 (3.2 g, 15.53 mmol, 0.6 equiv) was dissolved in DMF (50 mL), and then the resulting product was added to the reaction system. The reaction mixture was allowed to react at 25 °C for 12 h to give compound 22-5 for direct use in the next reaction without purification. MS [ESI, M+1]: 390.0.

Step E: To compound 22-5 (10.07 g, 25.87 mmol, 1 equiv) was added (Boc) _2O (6.78 g, 31.05 mmol, 7.13 mL, 1.2 equiv). The reaction mixture was allowed to react at 25 °C for 2 h under nitrogen atmosphere, 100 mL of water was added and extracted three times with 200 mL of ethyl acetate. The organic phases were combined, washed twice with 200 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by column chromatography to give compound 22-6 . MS [ESI, M+1]: 490.1.

Step F. Compound 22-6 (399.06 mg, 815.49 μmol, 0.6 equiv), water (5 mL), cesium carbonate (885.68 mg, 2.72 mmol, 2 equiv), and bis(triphenylphosphine)palladium(II) dichloride (47.70 mg, 67.96 μmol, 0.05 equiv) were added to a solution of compound 1-8 (575.14 mg, 1.36 mmol, 1 equiv) in 2-methyltetrahydrofuran (20 mL). The reaction system was purged with nitrogen three times and the reaction was carried out at 30 °C for 12 h under nitrogen atmosphere to give compound 22-7 .

Step G. m -Bromoanisole (305.05 mg, 1.63 mmol, 206.12 μL, 1.2 equiv), aqueous potassium hydroxide solution (4 M, 2.38 mL, 7 equiv) and bis(triphenylphosphine)palladium(II) dichloride (47.70 mg, 67.96 μmol, 0.05 equiv) were added to a solution of compound 22-7 (895.19 mg, 1.36 mmol, 1 equiv) in 2-methyltetrahydrofuran (20 mL). The reaction system was purged with nitrogen three times and the reaction was carried out at 85 °C for 12 h under nitrogen atmosphere. After cooling to room temperature, 20 mL of water were added to the reaction mixture. The aqueous phase was extracted three times with 30 mL ethyl acetate and the organic phases were combined and washed three times with 50 mL saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by reverse phase column chromatography (water (0.1% formic acid, v/v)/acetonitrile) to give compound 22-8 . MS [ESI, M+1]: 639.2.

Step H. The reaction was carried out in a solution of compound 22-8 (290 mg, 453.98 μmol, 1 equiv.) and N -chlorosuccinimide (72.75 mg, 544.78 μmol, 1.2 equiv.) in dichloromethane (5 mL) at 20 °C for 1 h. The reaction was quenched with 10 mL of saturated aqueous sodium sulfite solution. The aqueous phase was separated and extracted three times with 20 mL of dichloromethane and the organic phases were combined and washed twice with 20 mL of saturated brine. The organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to give compound 22-8' . MS [ESI, M+1]: 673.3.

Step I. To a solution of compound 22-8' (203 mg, 301.53 μmol, 1 equiv.) in dichloromethane (8 mL) was added 1 mL of trifluoroacetic acid. The reaction mixture was allowed to react at 20 °C for 1 h and concentrated to give the crude product. The crude product was separated by preparative HPLC (water (0.05% hydrochloric acid, v/v)/acetonitrile) to give compound 22-9 . MS [ESI, M+1]: 573.2.

Step J. Compound 22-9 (150 mg, 256.78 μmol, 1 equiv, HCl) was resolved by SFC (chiral IG (water (0.1% ammonia solution, v/v)/methanol)) to afford compound 22 or 23 , Rt = 2.290, MS [ESI, M+1]: 573.2; or to afford compound 23 or 22 , Rt = 2.474, MS [ESI, M+1]: 573.2.

¹ H NMR (400 MHz, CHLOROFORM-d) δ = 8.58 (s, 1H), 7.69-7.63 (m, 2H), 7.31-7.28 (m, 1H), 7, 24-7.12 (m, 5H), 6.77-6.71 (m, 3H), 6.63-6.58 (m,1H), 6.43 (d, J = 8.4 Hz, 1H), 6.33 (d, J = 15.2 Hz, 1H), 4.28-4.21 (m, 2H) , 3.72 (s, 3H), 3.37 (t, J = 6.8 Hz, 1H), 3.04 (s, 3H), 2.98 (s, 3H), 2.92-2.81 (m, 2H), 2.59-2.54 (m, 2H), 1.18 (d, J = 6.4 Hz, 3H), 0.99 (t, J = 8.0 Hz, 3H) .

¹ H NMR (400 MHz, CHLOROFORM-d) δ = 9.42 (s, 1H), 7.67-7.57 (m, 2H), 7.32-7.28 (m, 1H), 7, 24-7.11 (m, 5H), 6.76-6.71 (m, 3H), 6.57-6.36 (m, 3H), 4.35 (s, 2H), 3.89 (s, 1H), 3.71 (s, 3H), 3.23-3.06 (m, 2H), 2.94 ( d, J = 2, Hz, 6H), 2.59-2.53 (m, 2H), 1.34 (d, J = 6.0 Hz, 3H), 0.98 (t, J = 7.2 Hz, 3H).

Examples 24 and 25

Step A. Compound 22-6 (3.48 g, 7.11 mmol, 0.9 equiv), Pd( _PPh3 ) _2Cl2 (277.42 mg, 395.25 μmol, 0.05 equiv) and cesium carbonate (5.15 g, 15.81 mmol, 2 equiv) were added to a solution of _compound 1-8 (4.75 g, 11.23 mmol, 1.42 equiv) in 2-methyltetrahydrofuran (30 mL). The reaction system was purged with nitrogen three times, 6 mL of water was added and the reaction was carried out at 30 °C for 12 h to give compound 24-7 for direct use in the next reaction without purification.

Step B. Iodobenzene (1.94 g, 9.49 mmol, 1.06 mL, 1.2 equiv), aqueous potassium hydroxide solution (4 M, 13.84 mL, 7 equiv), and Pd( _PPh3 ) _2Cl2 (277.61 mg, 395.52 μmol, 0.05 equiv) were _added to a solution of compound 24-7 (5.21 g, 7.91 mmol, 1 equiv) in 2-methyltetrahydrofuran (20 mL). The reaction system was purged with nitrogen three times and the reaction was carried out at 85 °C for 12 h. After cooling the reaction system to room temperature, 30 mL of water was added to it and it was extracted three times with 50 mL of ethyl acetate. The organic phases were combined, washed twice with 50 mL of brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by reverse phase chromatography (0.1% formic acid) to give compound 24-8 . MS [ESI, M+1]: 609.3.

Step C: To a solution of compound 24-8 (2.1 g, 3.45 mmol, 1 equiv.) in dichloromethane (25 mL) was added N -chlorosuccinimide (552.76 mg, 4.14 mmol, 1.2 equiv.). The reaction system was allowed to proceed at 25 °C for 12 h. The reaction was quenched with 15 mL of saturated aqueous sodium sulfite solution. To the reaction mixture was added 30 mL of dichloromethane. The organic phase was washed twice with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give compound 24-8' . MS [ESI, M+1]: 643.3.

Step D: To a solution of 24-8' (1.9 g, 2.95 mmol, 1 equiv.) in dichloromethane (10 mL) was added 10 mL of trifluoroacetic acid. The reaction mixture was allowed to react at 25 °C for 1 h and concentrated to give the crude product. The crude product was separated by preparative HPLC (water (0.05% hydrochloric acid, v/v)/acetonitrile) to give 24-9 . MS [ESI, M+1]: 543.3.

Step E. Compound 24-9 (150 mg, 256.78 μmol, 1 equiv, HCl) was resolved by SFC (chiral IG (water (0.1% ammonia solution, v/v)/ethanol)) to afford compound 24 or 25 , Rt=2.148, MS [ESI, M+1]: 543.3; or to afford compound 25 or 24 , Rt=2.352, MS [ESI, M+1]: 543.3.

^1H NMR (400 MHz, DMSO- _d6 ) δ = 11.46 (s, 1H), 7.61 (d, J = 2.4 Hz, 1H), 7.49 (d, J = 7.6 Hz, 1H), 7.38 (d, J = 8.0 Hz, 1H), 7.29 (d, J = 7.6 Hz, 2H), 7.23 (d, J = 7.2 Hz, 1H), 7.21-7.16 (m, 3H), 7.15-7.09 (m, 2H), 6.55 (d, J = 8.8 Hz, 1H), 6.43-6.39 (m, 2H), 4.15-4.08 (m, 2H), 3.30-3.25 (m, 1H), 2.96 (s, 3H), 2.82 (s, 3H), 2.74-2.62 (m, 2H), 2.45 (br d, J = 7.6 Hz, 2H) , 1.06 (d, J = 6.8 Hz, 3H), 0.89 (t, J = 7.6 Hz, 3H).

^1H NMR (400 MHz, DMSO- _d6 ) δ = 11.61 (s, 1H), 9.51-9.18 (m, 2H), 7.64 (d, J = 2.8 Hz, 1H ), 7.39 (br d, J = 8.0 Hz, 4H), 7.29 (br s, 2H), 7.18-7.18 (m, 2H), 6.78 (d, J = 15.2 Hz, 1H), 6.61 (d, J = 8.8 Hz, 1H), 6.55-6 .47 (m, 1H), 4.34 (br s, 2H), 4.06-3.98 (m, 1H), 3.16 (br s, 2H), 2.99(s, 3H), 2.84 (s, 3H), 2.44 (br d, J = 7.6 Hz, 2H), 1.35 (d, J = 6.8 Hz, 3H), 0.88 (t , J = 7.6 Hz, 3H).

Example 26

Step A. Bis(triphenylphosphine)palladium(II) dichloride (116.81 mg, 166.42 μmol, 0.05 equiv ) and cesium carbonate (2.17 g, 6.66 mmol, 2 equiv ) were added sequentially to a solution of compounds 1-8 (2 g, 4.73 mmol, 1.42 equiv ) (in 2-methyltetrahydrofuran, theoretical value) and 26-1 (1.59 g, 3.33 mmol, 1 equiv ) in 2-methyltetrahydrofuran (15 mL). The reaction system was purged with nitrogen three times, 1 ml of water was added and the reaction was carried out at 30°C for 10 hours under nitrogen atmosphere and then cooled to room temperature to give compound 26-2 for direct use in the next reaction without purification. MS [ESI, M+1]: 646.4.

Step B. An aqueous solution of potassium hydroxide (4 M, 5.83 mL, 7 equiv ) and bis(triphenylphosphine)palladium(II) dichloride (116.88 mg, 166.51 μmol, 0.05 equiv ) was added to a solution of compound 26-2 (2.15 g, 3.33 mmol, 1 equiv ) and iodobenzene (815.28 mg, 4.00 mmol, 445.51 μL, 1.2 equiv ) in 2-methyltetrahydrofuran (10 mL). The reaction system was purged with nitrogen three times and the reaction was carried out at 85 °C for 12 h under nitrogen atmosphere. After cooling the reaction mixture to room temperature, 25 mL of water was added and extracted three times with 30 mL of ethyl acetate. The organic phases were combined and washed twice with 30 mL of saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by reverse phase column chromatography (trifluoroacetic acid system) to give compound 26-3 . MS [ESI, M+1]: 568.2.

Step C. To a solution of compound 26-3 (1.21 g, 2.14 mmol, 1 equiv ) in dichloromethane (15 mL) was added N -chlorosuccinimide (342.67 mg, 2.57 mmol, 1.2 equiv ). The reaction system was purged with nitrogen three times and the reaction was allowed to proceed at 25 °C for 2 h. The reaction mixture was diluted with 10 mL of dichloromethane and the organic phase was washed twice with 10 mL of brine. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was separated by preparative HPLC (water (10 mM ammonium bicarbonate)-acetonitrile) to give compound 26-4 . MS [ESI, M+1]: 602.2.

Step D. N , N -Diisopropylethylamine (85.86 mg, 664.32 μmol, 115.71 μL, 2 equiv ) and 2-(7-azabenzotriazol-1-yl) -N , N , N ', N' -tetramethyluronium hexafluorophosphate (151.56 mg, 398.59 μmol, 1.2 equiv ) were added to a solution of compound 26-4 (200 mg, 332.16 μmol, 1 equiv ) and azetidine trifluoroacetate (31.08 mg, 332.16 μmol, 36.73 μL, 1 equiv , hydrochloride) in N , N -dimethylformamide (10 mL). The reaction system was purged with nitrogen three times and the reaction was carried out at 25°C for 2 h. After cooling the reaction mixture to room temperature, 10 ml of water was added thereto and extraction was performed three times with 10 ml of ethyl acetate. The organic phases were combined and washed twice with 10 ml of saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product, which is compound 26-5 . MS [ESI, M+1]: 641.3.

Step E: To a solution of 26-5 (236 mg, 368.06 μmol, 1 equiv.) in dichloromethane (5 mL) was added trifluoroacetic acid (9.54 g, 83.66 mmol, 6.19 mL, 227.31 equiv.). The reaction mixture was allowed to react at 25 °C for 1 h under nitrogen atmosphere and concentrated to give the crude product. The crude product was separated twice by preparative HPLC (water (0.05% aqueous ammonia, v/v)-acetonitrile; water (10 mM ammonium bicarbonate)-acetonitrile) to give 26. MS [ESI, M+1]: 541.3.

¹ H NMR (400 MHz, METHANOL-d ₄ ) δ = 7.71 (d, J = 1.9 Hz, 1H), 7.54 (d, J = 7.8 Hz, 1H), 7.39-7.10 (m, 9H), 6.72 (td, J = 6.4, 15.4 Hz, 1H), 6.64-6.59 (m, 1H), 6.35-6.26 (m, 1H), 4.42-4.38 (m, 2H), 4.30 (t, J = 7.7 Hz, 2H), 4.07 (t, J = 7.8 Hz, 2H), 3.72 (dd, J = 1.3, 6.4 Hz, 2H), 3.25-3.22 (m, 2H), 2.56 (q, J = 7.4 Hz, 2H), 2.33 (td, J = 7.8, 15.6 Hz, 2H), 0.98 (t, J = 7.5 Hz, 3H).

Example 27

Stage A. To the solution of the compound27-1(6.84 g, 25.90 mmol, 1 equiv..) V N,N-dimethylformamide (50 ml) was addedN,N-diisopropylethylamine (6.70 g, 51.80 mmol, 9.02 ml, 2 equiv.). Ethyl bromocrotonate solution (3 g, 15.54 mmol, 2.14 mL, 0.6 equiv) was slowly and dropwise added to the reaction system..) V N,N-dimethylformamide (15 ml). The reaction system was allowed to react at 25°C for 22 h and then cooled to 0°C before adding (Boc)₂O (5.65 g, 25.90 mmol, 5.95 ml, 1 eq.). The reaction system was allowed to react at 25°C for 2 hours, 20 ml of water was added, and extraction was performed three times with 25 ml of ethyl acetate. The organic phases were combined, washed twice with 50 ml of saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was separated by reverse-phase column chromatography (water (0.1% trifluoroacetic acid, v/v)-acetonitrile) to give the compound26-1. MS [ESI, M+1]: 477.1.

Step B: To a solution of compound 27-3 (200 mg, 1.06 mmol, 1 equiv ) in dichloromethane (5.2 mL) was added trifluoroacetic acid (8.01 g, 70.23 mmol, 5.20 mL, 66.45 equiv ). The reaction mixture was allowed to react at 25 °C for 1 h and concentrated to give the crude product, compound 27-4 .

^1H NMR (EW19918-27-P1, 400 MHz, DMSO- _d6 ) δ = 11.32-10.91 (m, 1H), 9.24-8.78 (m, 1H), 4.61 ( d, J = 4.9 Hz, 1H), 4.49 (d, J = 4.9 Hz, 1H), 4.09-3.99 (m, 1H), 3.87-3.78 (m, 1H), 3.23-3.05 (m, 1H).

Step C. Bis(triphenylphosphine)palladium(II) dichloride (73.01 mg, 104.01 μmol, 0.05 equiv. . ) and cesium carbonate (1.36 g, 4.16 mmol, 2 equiv.) were added to the solution of compounds1-8(1.25 g, 2.95 mmol, 1.42 equiv..) (in 2-methyltetrahydrofuran, theoretical value) and26-1 (990.84 mg, 2.08 mmol, 1 eq..) in 2-methyltetrahydrofuran (15 ml). The reaction system was purged with nitrogen three times and 1 ml of water was added. The reaction mixture was allowed to react at 30°C for 8 hours under nitrogen atmosphere and then cooled to room temperature to obtain the compound27-5for direct use in the next reaction without purification. MS [ESI, M+1]: 646.4.

Step D. An aqueous solution of potassium hydroxide (4 M, 3.63 mL, 7 equiv ) and bis(triphenylphosphine)palladium(II) dichloride (72.84 mg, 103.78 μmol, 0.05 equiv) was added to a solution of compound 27-5 (1.34 g, 2.08 mmol, 1 equiv) and iodobenzene (508.13 mg, 2.49 mmol, 277.67 μL, 1.2 equiv ) in 2-methyltetrahydrofuran (8 mL). The reaction system was purged with nitrogen three times and the reaction was carried out at 85 °C for 12 h under nitrogen atmosphere. After cooling the reaction mixture to room temperature, 25 mL of water was added to it and extracted three times with 30 mL of ethyl acetate. The organic phases were combined and washed twice with 30 mL of saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The desired compound was confirmed by TLC chromatography (PE:EA = 5:1). The crude product was separated by column chromatography to give compound 27-6 . MS [ESI, M+1]: 596.3.

Stage E. To the solution of the compound27-6 (574 mg, 963.53 µmol, 1 equiv.) in dichloromethane (15 ml) was addedN-chlorosuccinimide (154.39 mg, 1.16 mmol, 1.2 equiv.). The reaction system was purged with nitrogen three times and allowed to react at 25°C for 2 hours. The reaction mixture was extracted and diluted with 10 mL of dichloromethane, and the organic phases were combined and washed twice with 20 mL of brine. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude product, which was compound27-7. MS [ESI, M+1]: 630.2.

Step F: To a solution of compound 27-7 (400 mg, 634.75 μmol, 1 equiv ) and lithium hydroxide monohydrate (266.36 mg, 6.35 mmol, 10 equiv ) in methanol (30 mL) was added water (10.00 g, 555.08 mmol, 10 mL, 874.49 equiv ). The reaction mixture was allowed to react at 25 °C for 6 h. The pH of the reaction mixture was adjusted to 7 with hydrochloric acid (3 M), after which 5 mL of water was added. The aqueous phase was extracted three times with 10 mL ethyl acetate and the organic phases were combined and washed twice with 10 mL saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product as compound 27-8 . MS [ESI, M+1]: 602.2.

Step G. N , N -Diisopropylethylamine (30.05 mg, 232.51 μmol, 40.50 μL, 2 equiv ) and 2-(7-azabenzotriazol-1-yl) -N , N , N ', N' -tetramethyluronium hexafluorophosphate (53.04 mg, 139.51 μmol, 1.2 equiv) were added to a solution of compounds 27-8 (70 mg, 116.26 μmol, 1 equiv ) and 27-4 (23.62 mg, 116.26 μmol, 1 equiv , trifluoroacetate) in N , N -dimethylformamide (3 mL). The reaction system was purged with nitrogen three times and the reaction was carried out at 25 °C for 4 h under nitrogen atmosphere. After cooling the reaction mixture to room temperature, 10 ml of water was added to it and extraction was performed three times with 10 ml of ethyl acetate. The organic phases were combined and washed twice with 10 ml of saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product, which is compound 27-9 . MS [ESI, M+1]: 673.3.

Step H: To a solution of compound 27-9 (120 mg, 178.25 μmol, 1 equiv ) in dichloromethane (3 mL) was added 3 mL of trifluoroacetic acid. The reaction mixture was allowed to react at 25 °C for 1 h under nitrogen atmosphere and concentrated to give the crude product. The crude product was separated twice by preparative HPLC (water (0.05% hydrochloric acid)-acetonitrile; water (0.225% formic acid v/v)-acetonitrile) to give compound 27. MS [ESI, M+1]: 573.1.

^1H NMR (400 MHz, DMSO- _d6 ) δ = 11.49 (s, 1H), 7.64 (d, J = 2.4 Hz, 1H), 7.50 (d, J = 7.6 Hz, 1H), 7.39 (d, J = 8.0 Hz, 1H), 7.35-7.27 (m, 2H), 7.26-7.11 (m, 6H), 6.64-6.55 (m, 2H), 6.16 (d, J = 15.6 Hz, 1H), 4.62 (d , J = 5.6 Hz, 1H), 4.50 (d, J = 5.6 Hz, 1H), 4.27-4.17 (m, 3H), 3.98-3.89 (m, 2H), 3.71-3.63 (m, 2H), 3.49-3.46 (m, 2H), 3.01-2.87 (m, 2H), 2.47-2.43 ( m, 2H), 0.90 (t, J = 7.2 Hz, 3H).

Experimental Example 1. Inhibition of MCF-7 Cell Proliferation

Experimental materials

EMEM medium was purchased from Wisent, and fetal bovine serum was purchased from Biosera. CellTiter-Glo reagent was from Promega. MCF-7 cells were purchased from the Cell Bank of the Type Culture Collection Committee of the Chinese Academy of Sciences. Nivo5 Multimarker Analyzer (PerkinElmer).

Way

MCF-7 cells were seeded in a white 384-well plate at a density of 600 cells/45 ml suspension/well. The plate was incubated in a CO ₂ incubator overnight.

On the day of treatment, the day 0 plate coated under the same conditions the day before was removed, centrifuged to remove the medium, 25 microliters of Promega CellTiter-Glo reagent were added to each well, and incubated at room temperature for 10 minutes to stabilize the luminescent signal. Readings were made using a PerkinElmer Nivo Multi-Marker Analyzer as a baseline at 0% inhibition.

The test compounds were serially diluted using a multichannel pipette to a concentration corresponding to the fifth decimal dilution, i.e. from 2 millimolar to 0.1 nanomolar, and the experiments were carried out in duplicate. 47.5 microliters of medium were added to the intermediate plate, 2.5 microliters of the serially diluted compounds were transferred to the corresponding wells of the intermediate plate and, after mixing, 5 microliters of the mixture per well were transferred to the plate with cells. The plate was incubated in a CO ₂ incubator for 6 days.

After 6 days of co-incubation with the compounds, the plate was centrifuged to remove the medium, 25 microliters of Promega CellTiter-Glo reagent were added to each well, and the plate was incubated at room temperature for 10 minutes to stabilize the luminescent signal. Readings were made using a PerkinElmer Nivo Multi-Marker Analyzer.

Data Analysis

The raw data were converted to the degree of inhibition using the formula: % inhibition = ((RFU _Comp. - AVER(RFU _{Neg. Contr.} ))/((AVER(RFU _{Day 0} ) - AVER(RFU _{Neg. Contr.} )) × 100%. A fitting curve was fitted and the IC ₅₀ was calculated.

The experimental results are shown in Table 1.

Table 1. Experimental results in vitro on inhibition of MCF-7 cell proliferation

Experimental example 2. Evaluation of DMPK properties

(1) Analysis of metabolic stability in liver microsomes

Objective: To determine the metabolic stability of test compounds in liver microsomes of humans, CD-1 mice and SD rats (Corning Co., Ltd., Miaotong Biological Technology Co., Ltd., Miaotong Biological Technology Co., Ltd.).

Methods: First, eight 96-well plates were prepared and named T0, T5, T10, T20, T30, T60, NCF60 and BLANK; except for the BLANK (in which the buffer was added at 10 μl/well), the compound solution was added to the plates at 10 μl/well. The prepared microsomes were added to 7 plates (80 μl/well), except for the T0 plate. The NCF60 plate was added with 10 μl/well buffer and incubated in a water bath at 37°C, and timing was started.

The prepared working solution of NADPH as a cofactor was divided into aliquots, which were added to a 96-well plate with shallow wells serving as loading sites. The solution was added to each plate using a 96-channel pipette at a rate of 10 μl/well and incubated in a water bath at 37°C to initiate the reaction.

At each time point, termination solution (cold acetonitrile containing 100 ng/mL tolbutamide and 100 ng/mL labetalol as internal standard) was added at 300 μL/well to stop the reaction, and the mixture was mixed well; the mixture was centrifuged at 4000 rpm for 20 min to precipitate the protein; the supernatant was transferred at 100 μL/well using a 96-channel pipette into a new 96-well plate containing HPLC-grade water at 300 μL/well, and the mixture was mixed well. The mixture was analyzed by LC/MS/MS. The results are shown in Table 2.

2) Analysis of inhibition of cytochrome P450 isoenzymes

The aim of the experiment was to determine the inhibitory effect of the test compound on the activity of cytochrome P450 isoenzymes of human liver microsomes (CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4; Corning Co., Ltd.).

Experimental procedures: First, the test compound (10 mM) was gradiently diluted to prepare working solutions (100× final concentration) with the following concentrations: 5, 1.5, 0.5, 0.15, 0.05, 0.015, and 0.005 mM, and simultaneously working solutions of positive control inhibitors of P450 isoenzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4) and their specific substrate mixtures (5 in 1) were prepared; human liver microsomes frozen in a refrigerator at -80°C were thawed on ice, and after all samples were thawed, human liver microsomes were diluted with PB to obtain a working solution of a specific concentration (0.253 mg/mL); 20 μl of the substrate mixture was added to the reaction plate (20 μl of PB was added to the well with the blank solution), and 158 μl of the human liver microsome working solution was also added to the reaction plate, which was then placed on ice until use; then 2 μl of each concentration of the test compound (N = 1) and the specific inhibitor (N = 2) were added to the corresponding well, and the corresponding organic solvent was added to the no inhibitor group (test compound or inhibitor positive control) as a control sample (the control sample of the test compound was 1:1 mixture of DMSO:MeOH; the positive control sample was 1:9 mixture of DMSO:MeOH); after preincubation in a water bath at 37°C for 10 min. 20 μL of coenzyme (NADPH) solution was added to the reaction plate and incubated in a water bath at 37°C for 10 min, 400 μL of cold acetonitrile solution (200 ng/mL tolbutamide and labetalol were internal standards) was added to terminate the reaction, and the reaction plate was placed on a shaker and shaken for 10 min; after centrifugation at 4000 rpm for 20 min, 200 μL of the supernatant was collected and added to 100 μL of water to dilute the sample, and finally the plate was sealed, rocked, vortexed uniformly, and detected by LC/MS/MS. The results are shown in Table 2.

(3) MDR1-MDCK bidirectional permeability analysis

In this study, the MDR1-MDCKII cell line was used as an in vitro model with permission from the Piet Borst Laboratory at the Netherlands Cancer Institute. It is a canine Madin-Darby kidney cell line transfected with the human multidrug resistance gene (MDR1). Its cells can stably express the efflux transporter P-gp and are thus suitable for screening P-gp substrates or inhibitors and predicting the permeability of compounds across barriers with strong efflux manifestations in the duodenum, blood-brain barrier, hepatocyte nucleus, nephrons, etc. This study involved the use of MDR1-MDCK II cells to investigate the bidirectional permeability of test compounds using the MDR1-MDCK II cell model.

Methods: The standard conditions are as follows:

- test concentration: 2 μM (DMSO ≤ 1%);

- number of repetitions: n = 2;

- direction: bidirectional transport, which involves two directions: A→B and B→A;

- incubation time: one time point, 2.5 hours;

- transfer buffer: HBSS buffer containing 10 mM Hepes, pH 7.4;

Incubation conditions: 37°C, 5% CO ₂ .

After incubation, the test solutions were collected at the dosing end and the receiving end and immediately mixed with cold acetonitrile solution containing the internal standard. LC/MS/MS was used to analyze the concentration of test compounds in all samples (including the initial dosing solution, samples from the dosing end, and samples from the receiving end). The apparent permeability coefficient, efflux coefficient, and other parameters were calculated. The results are presented in Table 2.

Table 2. Results of the conducted assessment of DMPK properties in vitro

(4) Pharmacokinetic study in mice

Objective: To determine the drug concentration in mouse blood plasma at different time points after intravenous and oral administration of the test compounds using LC/MS/MS, using female Balb/c mice as experimental animals. To study the pharmacokinetic properties of the test compounds using mice as an example and to evaluate the pharmacokinetic characteristics.

Experimental methodology

Experimental animals: 4 healthy female Balb/c mice were divided into 2 groups according to the principle of similarity in weight, 2 into group IV and 2 into group PO. The animals were purchased from Shanghai Lingchang Biotechnology Co., Ltd.

Receiving a medicine

Group IV: Suitable amounts of the compounds were weighed accordingly, prepared into 2 mg/ml solutions, then mixed to obtain a 0.5 mg/ml solution, which was stirred and sonicated until clear. The carrier medium was 15% HP-b-CD.

Group PO: An appropriate amount of the solution for group IV was taken and diluted to 0.4 mg/ml with 15% HP-b-CD.

Administration: After overnight fasting, the IV group was administered intravenously, the dose of the test compound was 1 mg/kg; the PO group was administered intragastrically, the dose of the test compound was 2 mg/kg.

Methods: In group IV, 30 μL of blood were collected from the saphenous vein of female Balb/c mice at 0.0833, 0.25, 0.5, 1, 2, 4, 8, and 24 hours after dosing and placed in a commercially available blood collection tube with EDTA-K2 anticoagulant added in advance. In group PO, 30 μL of blood were collected from the saphenous vein of female Balb/c mice at 0.25, 0.5, 1, 2, 4, 8, 12, and 24 hours after dosing and placed in a commercially available blood collection tube with EDTA-K2 anticoagulant added in advance. Blood samples were centrifuged (3200 g, 4°C, 10 min) to obtain plasma samples, which were transferred to pre-chilled centrifuge tubes, chilled on dry ice, and stored in an ultra-low temperature freezer at -60°C or below until LC-MS/MS analysis. Animals were allowed free access to food 4 h after administration. The LC/MS/MS method was used to determine the test compounds in mouse plasma after intravenous and oral administration. The linear range for this method was 2.00-2000 nM. The results are presented in Table 3.

(5) Pharmacokinetic study in rats

The aim of the experiment: to determine the concentration of the drug in rat blood plasma at different time points after intravenous and oral administration of the test compounds using SD female rats as experimental animals using LC/MS/MS. To study the pharmacokinetic properties of the test compounds using rats as an example and to evaluate the pharmacokinetic characteristics.

Experimental methodology

Experimental animals: 4 healthy female SD rats were divided into 2 groups according to the principle of similarity in weight, 2 into group IV and 2 into group PO. The animals used in this study were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

Receiving a medicine

Group IV: Suitable amounts of the compounds were weighed accordingly, prepared into 2 mg/ml solutions, then mixed to obtain a 0.5 mg/ml solution, which was stirred and sonicated until clear. The carrier medium was 15% HP-b-CD.

Group PO: An appropriate amount of the solution for group IV was taken and diluted to 0.4 mg/ml with 15% HP-b-CD.

Administration: After overnight fasting, the IV group was administered intravenously, the dose of the test compound was 1 mg/kg; the PO group was administered intragastrically, the dose of the test compound was 2 mg/kg.

Methods: In Group IV, 200 μL of blood were collected from the jugular vein of female SD rats at 0.0833, 0.25, 0.5, 1, 2, 4, 8, and 24 hours after dosing and placed in a commercially available blood collection tube with EDTA-K2 anticoagulant added in advance. In Group PO, 200 μL of blood were collected from the jugular vein of female SD rats at 0.25, 0.5, 1, 2, 4, 8, 12, and 24 hours after dosing and placed in a commercially available blood collection tube with EDTA-K2 anticoagulant added in advance. Blood samples were centrifuged (3200 g, 4°C, 10 min) to obtain plasma samples, which were transferred to pre-chilled centrifuge tubes, chilled on dry ice, and stored in an ultra-low temperature freezer at -60°C or below until LC-MS/MS analysis. Animals were allowed free access to food 4 h after administration. The LC/MS/MS method was used to determine the test compounds in mouse plasma after intravenous and oral administration. The linear range for this method was 2.00-2000 nM. The results are presented in Table 3.

The results of PK properties in vivo are presented in Table 3.

Table 3. Results of the conducted assessment of PK properties in vivo

Experimental example 3. Evaluation of effectiveness in vivo

This study aimed to evaluate the antitumor efficacy of the compounds according to the present application in MCF-7 cells of breast cancer xenograft in BALB/c nude mice (provided by Beijing Vital River Laboratory Animal Technology Co., Ltd., with the number of experimental animals in each experimental group equal to 7).

Three days before xenograft injection, mice were subcutaneously inoculated with 0.36 mg 60-day sustained-release estrogen tablets into the left scapula. At logarithmic growth phase, cells were harvested and counted. Cell density was adjusted to 10 × 10 ⁷ cells/mL, an equal volume of Matrigel was added, and mixed for inoculation. Each mouse was subcutaneously inoculated with xenograft using 0.2 mL of MCF-7 tumor cell suspension (10 × 10 ⁶ ) into the right scapula. On day 14 after tumor xenograft injection, mice were divided into groups and treated with drugs once daily at an average tumor volume of 200 mm ³ and body weight of 22.0-23.0 g. After group division, tumor volume and body weight were measured twice a week. The tumor growth rate (T/C) and tumor growth inhibition rate (TGI) were calculated from the last tumor measurement, namely, day 27 after division into groups, and the antitumor efficacy of the compound was evaluated by TGI (%) or the relative tumor cell proliferation rate T/C (%). TGI (%) represents the tumor growth inhibition rate. TGI (%) = [(1-(average tumor volume at the end of treatment in the treatment group - average tumor volume at the beginning of treatment in the treatment group))/(average tumor volume at the end of treatment in the vehicle control group - average tumor volume at the beginning of treatment in the vehicle control group)] × 100%, relative tumor cell proliferation rate T/C (%) = T _RTV /C _RTV × 100% (T _RTV : average RTV in the treatment group; C _RTV : average RTV in the negative control group). The relative tumor volume (RTV) was calculated based on the tumor measurement results. The formula is: RTV = V _t /V ₀ , where V ₀ is the tumor volume measured at division and administration (i.e., D ₀ ) and V _t is the tumor volume at the specific measurement, with T _RTV and C _RTV data measured on the same day. The results are as follows.

Table 4. Analysis of antitumor efficacy

a. Mean ± SEM.

Experimental Example 4. Suppression of Wet Uterine Weight Increase in Immature Rats

This study was designed to evaluate the uterine growth inhibition effect of the compounds of the present application in 18-21 day-old female immature rats (provided by Beijing Vital River Laboratory Animal Technology Co., Ltd., with the number of test animals in each experimental group being 5). In this study, 18 day-old female immature rats were orally administered the compound of the present application at a dose of 10 mg/kg and 0.1 mg/kg estradiol for three consecutive days, the control group was orally administered estradiol at a dose of 0.1 mg/kg for three consecutive days, and the blank group was not administered any drug except the corresponding vehicle. Three days after the administration, the rats were sacrificed and the uterus of the rats was weighed to observe the effect of the test compounds on the uterine growth inhibition effect in the rats. The inhibition rate = 100 * [(VEHICLE _EE - CONT.)/(VEHICLE _EE - VEHICLE)], where "VEHICLE _EE " is the wet uterine weight of the control group (at an oral dose of estradiol of 0.1 mg/kg); "CONT." is the wet uterine weight of the treatment group; "VEHICLE" is the wet uterine weight of rats in the blank group. The results are as follows.

Claims (57)

1. A compound of formula (II), its isomer or a pharmaceutically acceptable salt thereof, , where X is NH; Y is selected from the group consisting of N and CR ₇ ; Y ₁ is CH and Y ₂ is N; or Y ₁ is N and Y ₂ is CH or CF; or Y ₁ is CH and Y ₂ is CH; L represents or ; ring A is selected from the group consisting of phenyl, thienyl and pyridinyl; R ₁ is selected from the group consisting of H, halogen and C _1-6 alkyl; R ₂ is C _1-3 alkyl optionally substituted with 1, 2 or 3 R _b ; R ₃ is selected from the group consisting of H and C _1-3 alkyl; R ₁₃ is H or ; R ₄ is selected from the group consisting of COOH, , , -NHCH ₃ and -N(CH ₃ ) ₂ ; R ₅ is H; each of R ₆ , R ₇ , R ₈ and R ₉ independently represents H; each of R ₁₀ , R ₁₁ and R ₁₂ is independently selected from the group consisting of H, halogen, CN, C _1-3 alkyl and -OC _1-3 alkyl; and R _b is selected from the group consisting of H, F, Cl, Br and I. 2. The compound, its isomer or its pharmaceutically acceptable salt according to claim 1, wherein X is NH; Y is selected from the group consisting of CR ₇ ; Y ₁ is CH and Y ₂ is N or Y ₁ is N and Y ₂ is CH; L is ; and R ₁₃ represents . 3. A compound, its isomer or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein R _b is selected from the group consisting of F, Cl, Br and I. 4. A compound, its isomer or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, wherein ring A is selected from phenyl, And . 5. A compound, its isomer or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, wherein R ₁ is selected from the group consisting of H, F, Cl, Br, I and C _1-3 alkyl. 6. The compound, its isomer or its pharmaceutically acceptable salt according to claim 5, wherein R ₁ is selected from the group consisting of H, F, Cl, Br, I, Me and Et. 7. The compound, its isomer or its pharmaceutically acceptable salt according to claim 5, wherein R ₁ is selected from the group consisting of H, Cl, Br, I and Me. 8. A compound, its isomer or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, wherein R ₂ is selected from the group consisting of Me, Et, CF ₃ , CH ₂ CF ₃ , CHF ₂ and CH ₂ F. 9. The compound, its isomer or its pharmaceutically acceptable salt according to claim 8, wherein R ₂ is selected from the group consisting of ethyl and CH ₂ CF ₃ . 10. A compound, its isomer or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, wherein R ₃ is selected from the group consisting of H and methyl. 11. The compound, its isomer or its pharmaceutically acceptable salt according to claim 10, wherein R ₃ is H. 12. A compound, its isomer or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 11, wherein R ₄ is selected from the group consisting of , , -NHCH ₃ and -N(CH ₃ ) ₂ . 13. A compound, isomer or pharmaceutically acceptable salt thereof according to any one of claims 1 to 12, wherein each of R ₁₀ , R ₁₁ and R ₁₂ is independently selected from H, F, Cl, Br, I, CN, C _1-3 alkyl and -OC _1-3 alkyl. 14. The compound, its isomer or its pharmaceutically acceptable salt according to claim 13, wherein each of R ₁₀ , R ₁₁ and R ₁₂ is independently selected from H, F, Cl, Br, I, CN, Me, Et and OMe. 15. A compound, its isomer or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 14, wherein each of R ₁₀ , R ₁₁ and R ₁₂ is independently selected from H, F, Cl, Br, I, CN, Me and OMe. 16. A compound, its isomer or its pharmaceutically acceptable salt according to any one of claims 1 to 15, wherein the structural unit selected from And . 17. A compound, its isomer or its pharmaceutically acceptable salt according to claim 16, wherein the structural unit selected from And . 18. A compound, its isomer or its pharmaceutically acceptable salt according to any one of claims 1 to 17, wherein the structural unit selected from the group consisting of , , And . 19. A compound, its isomer or its pharmaceutically acceptable salt according to claim 18, wherein the structural unit selected from the group consisting of , , , , And . 20. A compound, its isomer or its pharmaceutically acceptable salt according to claim 19, wherein the structural unit selected from the group consisting of , , , , , , , , And . 21. A compound, its isomer or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 20, wherein R ₁₃ is H, , , or . 22. A compound, its isomer or a pharmaceutically acceptable salt thereof according to claim 21, wherein R ₁₃ is H, , or . 23. A compound, its isomer or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 22, wherein the structural unit selected from the group consisting of , , , And . 24. The compound, its isomer or its pharmaceutically acceptable salt according to claim 23, wherein the structural unit selected from the group consisting of , , And . 25. A compound, its isomer or its pharmaceutically acceptable salt according to any one of claims 1 to 24, wherein the compound, its isomer or its pharmaceutically acceptable salt is selected from , , , And , where R ₁ , R ₂ , R ₃ , R ₅ , R ₆ , R ₇ , R ₈ , R ₉ , R ₁₀ , R ₁₁ , R ₁₂ and R ₁₃ are as defined in any of paragraphs 1-24, R ₄₁ is H, R ₄₂ is selected from H and Me and R _d is selected from H and CH ₂ F. 26. A compound of the following formulas, its isomer or a pharmaceutically acceptable salt thereof: 27. A compound, its isomer or a pharmaceutically acceptable salt thereof according to claim 26, wherein the compound is selected from: 28. A pharmaceutical composition as an estrogen receptor antagonist, containing a therapeutically effective amount of a compound, its isomer or its pharmaceutically acceptable salt according to any one of claims 1 to 27 as an active ingredient and a pharmaceutically acceptable carrier. 29. Use of a compound, its isomer or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 27 or a pharmaceutical composition according to claim 28 in the preparation of a medicinal product for the treatment of estrogen receptor-positive breast cancer.