WO2018130123A1

WO2018130123A1 - Pentacyclic compound as selective estrogen receptor down-regulator and use thereof

Info

Publication number: WO2018130123A1
Application number: PCT/CN2018/071699
Authority: WO
Inventors: 王勇; 赵立文; 张小猛; 刘奇; 徐岩; 雷时海; 王小伟; 张雁; 庞司林
Original assignee: 南京圣和药业股份有限公司
Priority date: 2017-01-11
Filing date: 2018-01-08
Publication date: 2018-07-19
Also published as: CN108299420A; TW201825489A; CN108299420B

Abstract

The present invention falls within the field of medical chemistry, and relates to a pentacyclic compound as a selective estrogen receptor down-regulator and the use thereof. In particular, provided in the present invention are a compound as shown in formula I or an isomer, a pharmaceutically acceptable salt, a solvate, a crystal or a prodrug thereof, the preparation method therefor and pharmaceutical compositions containing the compounds and the use of the compounds or compositions for treating and/or preventing estrogen receptor-associated diseases. The compound of the present invention has an even better anti-tumour activity, a longer dosing interval and fewer side effects, and seems to be very promising as a therapeutic agent against breast cancer.

Description

Pentacyclic compound as a selective estrogen receptor down-regulation agent and application thereof

Technical field

The invention belongs to the field of medical chemistry, and particularly relates to a compound as a selective estrogen receptor down-regulator (SERD) or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal, an electronic isostere or a prodrug And their preparation and pharmaceutical compositions containing these compounds and the use of these compounds or compositions for the treatment and/or prevention of estrogen receptor-related diseases.

Background technique

The Estrogen Receptor (ER) is a ligand-activated transcriptional regulatory protein that mediates the induction of multiple biological effects through interaction with endogenous estrogens. Endogenous estrogens include 17β-estradiol and estrone. ER has been found to have two isoforms: ER-α and ER-β, which are encoded by two different genes located on human chromosomes 6 and 14, respectively, and ER-α is widely expressed in various tissues, ER- The expression of β is limited to the female reproductive system and tissues such as brain and bone. Both contain 6 domains and 4 functional regions, and the N-terminal A/B functional region has a non-ligand-dependent transcriptional activation domain AF-1 with constitutive activity through basal transcription factors. Co-activators and other transcription factors act to activate transcription of the target gene, in addition to multiple phosphorylation sites. The DNA binding domain (DBD) consisting of the C domain can specifically bind to the target DNA and contain a nuclear localization signal, and also has a dimerization interface, which plays an important role in the dimerization of the receptor. The D domain is the hinge region responsible for the connection of the DBD and the ligand binding domain (LBD). The C-terminal E domain constitutes LBD, which determines the specific binding of ER to ligands such as estrogen, has a ligand-dependent transcriptional activation domain AF-2, and LBD also has a strong dimerization interface, in the absence of In the case of the body, it still functions, and it is a key part of the receptor's dimerization. The LBD consists of 12 alpha helices and one beta fold, forming a three-layer anti-parallel sandwich structure in which H5, H6, H9 and H10 form the intermediate layer, and H1, H2, H3, H4 and H7, H8, H11 form the outer layers respectively. Two layers, wherein H12 contains AF-2 with a hydrophobic surface facing the ligand binding pocket and a hydrophilic surface facing outward. The LBD of the ER is wedge-shaped, and the H12 is in the groove of the ligand binding pocket and encloses the ligand binding pocket. When ER binds to an agonist, the conformation of H12 becomes more stable, suitable for the binding of coactivators, which in turn activates transcription of the target gene. When ER binds to the antagonist, the position of H12 changes and occupies the binding site of the coactivator, which in turn produces estrogen antagonism.

The research on drugs targeting estrogen receptors has been carried out for many years, and some successes have been made in the clinic. In particular, it has recently been found that selective estrogen receptor down-regulation has a stronger anti-estrogen effect, such as interfering with estrogen receptors. Stability and cause its degradation. However, there is still a need to develop more estrogen receptor down-regulators, especially selective estrogen receptor down-regulations, which make the drugs more excellent, such as better efficacy, less side effects, longer dosing intervals. Etc., thus better for preventing or treating estrogen receptor-related diseases.

Summary of the invention

It is an object of the present invention to provide a compound of the formula I having a selective estrogen receptor down-regulating activity or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal, an isostere or a former medicine,

Another object of the present invention is to provide a process for the preparation of a compound of the formula I according to the invention, or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal, an isostere or a prodrug.

A further object of the present invention is to provide a combination of a compound of the formula I according to the invention or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal, an isostere or a prodrug and a pharmaceutically acceptable carrier And a composition comprising a compound of formula I of the invention, or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal, an isostere or a prodrug, and another drug or drugs.

A further object of the present invention is to provide a compound of the formula I according to the invention, or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal, an isostere or a prodrug for the treatment and/or prophylaxis of an estrogen receptor A method of related diseases, and a compound of the formula I according to the invention, or an isomer, pharmaceutically acceptable salt, solvate, crystal, electron isostere or prodrug thereof, for use in the treatment and/or prevention of a female The use of drugs for hormone receptor-related diseases.

In view of the above purposes, the present invention provides the following technical solutions:

In a first aspect, the present invention provides a compound of the formula I, or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal, an isostere or a prodrug,

among them,

Each of R ¹ and R ^{2 is} independently selected from the group consisting of hydrogen, halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxy, cyano, amino, a monoalkylamino group, an alkyl acylamino group, an alkyl acyl group, an amino acyl group, an alkylamino acyl group, a dialkylamino group, and a cycloalkyl group;

R ³ and R ⁹ are each independently selected from the group consisting of hydrogen, alkyl acyl, amino acyl, alkyl amino acyl, alkyl, haloalkyl, hydroxyalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, heteroaryl. Base and aryl group;

R ⁴ , R ⁵ , R ⁶ and R ⁷ are each independently selected from the group consisting of hydrogen, halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxy, a cyano group, an amino group, a monoalkylamino group, an alkyl acylamino group, an alkyl acyl group, an amino acyl group, an alkylamino acyl group, and a dialkylamino group; or R ⁴ and R ⁵ together with the carbon atom to which they are attached form a carbonyl group, a ring An alkyl group; or R ⁶ , R ⁷ together with the carbon atom to which they are attached form a carbonyl group, a cycloalkyl group; or R ⁴ , R ⁶ together with the carbon atom to which they are attached form a cycloalkyl group;

Each R ^{8 is} independently selected from the group consisting of hydrogen, halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxy, cyano, amino, monoalkyl An amino group, an alkyl acylamino group, an alkyl acyl group, an amino acyl group, an alkylamino acyl group, a bisalkylamino group, a cycloalkyl group, and a boronic acid;

X, Y, Z, W are each independently selected from N and C(R ¹⁰ ), wherein R ^{10 is} selected from the group consisting of hydrogen, halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxy Alkoxy, nitro, carboxy, cyano, amino, monoalkylamino, alkyl acylamino, alkyl acyl, amino acyl, alkylamino acyl, bisalkylamino and cycloalkyl;

Chemical bond

Independently

or

And

m, n, and o are independently 1, 2, 3, or 4, respectively.

In some specific embodiments, the compound of the present invention is a compound of Formula I or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal, an isostere or a prodrug thereof, wherein: X, Y, Z and W are both C (R ¹⁰ ).

In other specific embodiments, the compound of the present invention is a compound of Formula I or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal, an isostere or a prodrug thereof, wherein: X, Y One of Z, W and N is N, and the other three are C (R ¹⁰ ).

In other specific embodiments, the compound of the present invention is a compound of Formula I or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal, an isostere or a prodrug thereof, wherein: X, Y Two of Z, W, and N are N, and the other two are C (R ¹⁰ ).

In other specific embodiments, the compound of the present invention is a compound of Formula I or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal, an isostere or a prodrug thereof, wherein: X, Y Three of Z, W, and N are N, and the other one is C (R ¹⁰ ).

In some specific embodiments, the compound of Formula I of the present invention is a compound of Formula Ia below, or an isomer thereof, a pharmaceutically acceptable salt, solvate, crystal, or isostere or prodrug thereof,

Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , X, R ¹⁰ , m, n, o, a chemical bond

As defined in Formula I.

In other specific embodiments, the compound of Formula I of the present invention is a compound of Formula Ib, or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal, an isostere or a prodrug,

Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , Z, R ¹⁰ , m, n, o, a chemical bond

As defined in Formula I.

In other specific embodiments, the compound of Formula I of the present invention is a compound of Formula Ic, or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal, an isostere or a prodrug,

Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , Y, X, R ¹⁰ , m, n, o, chemical bond

As defined in Formula I.

In other specific embodiments, the compound of Formula I of the present invention is a compound of Formula Id, or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal, an isostere or a prodrug,

Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , W, Z, R ¹⁰ , m, n, o, a chemical bond

As defined in Formula I.

In other specific embodiments, the compound of Formula I of the present invention is a compound of Formula Ie below, or an isomer, pharmaceutically acceptable salt, solvate, crystal, ortho-form or prodrug thereof,

Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , Y, Z, R ¹⁰ , m, n, o, chemical bond

As defined in Formula I.

In some preferred embodiments, the compound of the present invention is a compound of the formula I, Ia, Ib, Ic, Id or Ie or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal, an isostere Or a prodrug, where:

Each of R ¹ and R ^{2 is} independently selected from the group consisting of hydrogen, fluorine, chlorine, bromine, iodine, hydroxyl, C _1-6 alkyl, halo C _1-6 alkyl, hydroxy C _1-6 alkyl, C _{1- 6} alkoxy, halo C _1-6 alkoxy, hydroxy C _1-6 alkoxy, nitro, carboxy, cyano, amino, mono C _1-6 alkylamino, C _1-6 alkyl acyl An amino group, a C _1-6 alkyl acyl group, an amino acyl group, a C _1-6 alkylamino acyl group, a bis C _1-6 alkylamino group, and a C _3-10 cycloalkyl group;

Further preferably, each of R ¹ and R ^{2 is} independently selected from the group consisting of hydrogen, fluorine, chlorine, bromine, iodine, hydroxyl, C _1-3 alkyl, halogenated C _1-3 alkyl, hydroxy C _1-3 alkyl , C _1-3 alkoxy, halo C _1-3 alkoxy, hydroxy C _1-3 alkoxy, nitro, carboxy, cyano, amino, mono C _1-3 alkylamino, C _{1- a 3} -alkylacylamino group, a C _1-3 alkyl acyl group, an amino acyl group, a C _1-3 alkylamino acyl group, a bis C _1-3 alkylamino group, and a C _3-6 cycloalkyl group;

Still more preferably, each of R ¹ and R ^{2 is} independently selected from the group consisting of hydrogen, fluorine, chlorine, bromine, iodine, hydroxyl, methyl, ethyl, propyl, isopropyl, trifluoromethyl, trifluoroethyl , hydroxymethyl, hydroxyethyl, hydroxypropyl, 2-hydroxypropyl, methoxy, ethoxy, propoxy, isopropoxy, nitro, carboxyl, cyano, amino, methylamino, B Base amino, propylamino, isopropylamino, dimethylamino, diethylamino, methylethylamino, dipropylamino, methylpropylamino, ethylpropylamino, methylacylamino, ethylacyl Amino, vinyl acylamino, methyl acyl, ethyl acyl, vinyl acyl, amino acyl, methyl amino acyl, ethyl amino acyl, cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

R ³ and R ⁹ are each independently selected from hydrogen, C _1-10 alkyl acyl, amino acyl, C _1-10 alkylamino acyl, C _1-10 alkyl, halo C _1-10 alkyl, hydroxy C _1-10 alkyl, C _2-10 alkenyl, C _2-10 alkynyl, C _3-10 cycloalkyl, C _3-10 heterocyclyl, C _6-18 aryl and C _5-18 heteroaryl The group may be one or more of halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxy, cyano, amino, monoalkane a base amino group, a dialkylamino group, a cycloalkyl group, a heterocyclic group, an aryl group and a heteroaryl group;

Further preferably, R ³ and R ⁹ are each independently selected from the group consisting of hydrogen, C _1-6 alkyl acyl, amino acyl, C _1-6 alkylamino acyl, C _1-6 alkyl, halo C _1-6 alkane , hydroxy C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-8 cycloalkyl, C _3-8 heterocyclyl, C _6-12 aryl and C _{5- 12} heteroaryl group, the group may be substituted with one or more halogen, hydroxy, C _1-6 alkyl, halo C _1-6 alkyl, hydroxy C _1-6 alkyl, C _1-6 alkoxy, Halogenated C _1-6 alkoxy, hydroxy C _1-6 alkoxy, nitro, carboxy, cyano, amino, mono C _1-6 alkylamino, bis C _1-6 alkylamino, C _{3 a -10} cycloalkyl group, a C _3-10 heterocyclic group, a C _6-18 aryl group, and a C _5-18 heteroaryl group;

Still more preferably, R ³ and R ⁹ are each independently selected from the group consisting of hydrogen, formyl, acetyl, propionyl, aminoacyl, methylaminoacyl, ethylaminoacyl, propylaminoacyl, methyl, ethyl, Propyl, isopropyl, butyl, isobutyl, pentyl, isopentyl, neopentyl, hexyl, trifluoromethyl, trifluoroethyl, 2,2-difluoropropyl, 2-fluoro- 2-methylpropyl, (S)-3-fluoro-2-methylpropyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, 2-hydroxypropyl, vinyl, propenyl, butenyl, 3-methyl-2-butenyl, C _2-6 alkynyl, C _3-6 cycloalkyl, C _3-6 heterocyclyl, C _6-10 aryl and C _5-10 heteroaryl, The group may be one or more halogen, hydroxy, C _1-3 alkyl, halo C _1-3 alkyl, hydroxy C _1-3 alkyl, C _1-3 alkoxy, halogen C _{1- alkoxy,} a C _1-3 alkoxy group, hydroxy, nitro, carboxy, cyano, amino, mono C _1-3 alkyl group, a di C _1-3 alkyl amino, C _3-6 cycloalkyl, C _3-6 heterocyclyl, C _6-10 aryl and C _5-10 heteroaryl are substituted.

R ⁴ , R ⁵ , R ⁶ and R ⁷ are each independently selected from the group consisting of hydrogen, fluorine, chlorine, bromine, iodine, hydroxyl, C _1-6 alkyl, halogenated C _1-6 alkyl, hydroxy C _1-6 alkane , C _1-6 alkoxy, halo C _1-6 alkoxy, hydroxy C _1-6 alkoxy, nitro, carboxy, cyano, amino, mono C _1-6 alkylamino, C _{1 a -6} alkyl acylamino group, a C _1-6 alkyl acyl group, an amino acyl group, a C _1-6 alkylamino acyl group, and a bis C _1-6 alkylamino group;

Further preferably, R ⁴ , R ⁵ , R ⁶ , R ⁷ are each independently selected from the group consisting of hydrogen, fluorine, chlorine, bromine, iodine, hydroxyl, C _1-3 alkyl, halogenated C _1-3 alkyl, hydroxy C _1-3 alkyl, C _1-3 alkoxy, halo C _1-3 alkoxy, hydroxy C _1-3 alkoxy, nitro, carboxy, cyano, amino, mono C _1-3 alkyl An amino group, a C _1-3 alkyl acylamino group, a C _1-3 alkyl acyl group, an amino acyl group, a C _1-3 alkylamino acyl group, and a bis C _1-3 alkylamino group;

Still more preferably, R ⁴ , R ⁵ , R ⁶ , R ⁷ are each independently selected from the group consisting of hydrogen, fluorine, chlorine, bromine, iodine, hydroxyl, methyl, ethyl, propyl, isopropyl, monofluoromethyl. , difluoromethyl, trifluoromethyl, trifluoroethyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, 2-hydroxypropyl, methoxy, ethoxy, propoxy, isopropoxy , nitro, carboxyl, cyano, amino, methylamino, ethylamino, propylamino, isopropylamino, dimethylamino, diethylamino, methylethylamino, dipropylamino, methylpropylamino , ethyl propylamino, methyl acylamino, ethyl acylamino, vinyl acylamino, methyl acyl, ethyl acyl, vinyl acyl, amino acyl, methyl amino acyl, ethyl amino acyl and propyl amino Acyl group.

R ⁴ , R ⁵ together with the carbon atom to which they are attached form a carbonyl group or a C _3-10 cycloalkyl group;

Further preferably, R ⁴ , R ⁵ together with the carbon atom to which they are attached form a carbonyl group, a C _3-6 cycloalkyl group;

Still more preferably, R ⁴ , R ⁵ together with the carbon atom to which they are attached form a carbonyl group, a cyclopropyl group, a cyclobutyl group, a cyclopentyl group and a cyclohexyl group.

In some preferred embodiments, the compound of the invention is a compound of formula I or an isomer thereof, a pharmaceutically acceptable salt, solvate, crystal, ortho isomer or prodrug thereof, wherein:

R ⁶ and R ⁷ together with the carbon atom to which they are attached form a carbonyl group, a C _3-10 cycloalkyl group;

Further preferably, R ⁶ and R ⁷ together with the carbon atom to which they are attached form a carbonyl group, a C _3-6 cycloalkyl group;

Still more preferably, R ⁶ , R ⁷ together with the carbon atom to which they are attached form a carbonyl group, a cyclopropyl group, a cyclobutyl group, a cyclopentyl group and a cyclohexyl group.

R ⁴ , R ⁶ together with the carbon atom to which they are attached form a C _3-10 cycloalkyl group;

Further preferably, R ⁴ , R ⁶ together with the carbon atom to which they are attached form a C _3-6 cycloalkyl group;

Still more preferably, R ⁴ , R ⁶ together with the carbon atom to which they are attached form a cyclopropyl group, a cyclobutyl group, a cyclopentyl group and a cyclohexyl group.

Each R ^{8 is} independently selected from the group consisting of hydrogen, halogen, hydroxy, C _1-6 alkyl, halo C _1-6 alkyl, hydroxy C _1-6 alkyl, C _1-6 alkoxy, halo C _{1 -6} alkoxy, hydroxy C _1-6 alkoxy, nitro, carboxy, cyano, amino, mono C _1-6 alkylamino, C _1-6 alkyl acylamino, C _1-6 alkyl acyl , aminoacyl group, C _1-6 alkylamino acyl group, bis C _1-6 alkylamino group, C _3-10 cycloalkyl group and boric acid;

Further preferably, each R ^{8 is} independently selected from the group consisting of hydrogen, halogen, hydroxy, C _1-3 alkyl, halo C _1-3 alkyl, hydroxy C _1-3 alkyl, C _1-3 alkoxy, Halogen C _1-3 alkoxy, hydroxy C _1-3 alkoxy, nitro, carboxy, cyano, amino, mono C _1-3 alkylamino, C _1-3 alkyl acylamino, C _{1- a 3-} alkyl acyl group, an amino acyl group, a C _1-3 alkylamino acyl group, a bis C _1-3 alkylamino group, a C _3-6 cycloalkyl group, and a boronic acid;

Still more preferably, each R ^{8 is} independently selected from the group consisting of hydrogen, fluorine, chlorine, bromine, iodine, hydroxyl, methyl, ethyl, propyl, isopropyl, trifluoromethyl, trifluoroethyl, hydroxymethyl Base, hydroxyethyl, hydroxypropyl, 2-hydroxypropyl, methoxy, ethoxy, propoxy, isopropoxy, nitro, carboxyl, cyano, amino, methylamino, ethylamino, Propylamino, isopropylamino, dimethylamino, diethylamino, methylethylamino, dipropylamino, methylpropylamino, ethylpropylamino, methylacylamino, ethylacylamino, ethylene Alkylamino, methylacyl, ethylacyl, vinylacyl, aminoacyl, methylaminoacyl, ethylaminoacyl, propylaminoacyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and boric acid .

R ^{10 is} selected from the group consisting of hydrogen, halogen, hydroxy, C _1-6 alkyl, halo C _1-6 alkyl, hydroxy C _1-6 alkyl, C _1-6 alkoxy, halo C _1-6 alkoxy a group, a hydroxy C _1-6 alkoxy group, a nitro group, a carboxyl group, a cyano group, an amino group, a mono C _1-6 alkylamino group, a C _1-6 alkyl acylamino group, a C _1-6 alkyl acyl group, an amino acyl group, a C _1-6 alkylamino acyl group, a bis C _1-6 alkylamino group, and a C _3-10 cycloalkyl group;

Further preferably, R ^{10 is} selected from the group consisting of hydrogen, halogen, hydroxy, C _1-3 alkyl, halogenated C _1-3 alkyl, hydroxy C _1-3 alkyl, C _1-3 alkoxy, halogenated C _{1 -3} alkoxy, hydroxy C _1-3 alkoxy, nitro, carboxy, cyano, amino, mono C _1-3 alkylamino, C _1-3 alkyl acylamino, C _1-3 alkyl acyl An aminoacyl group, a C _1-3 alkylamino acyl group, a bis C _1-3 alkylamino group, and a C _3-6 cycloalkyl group;

Still more preferably, R ^{10 is} selected from the group consisting of hydrogen, fluorine, chlorine, bromine, iodine, hydroxyl, methyl, ethyl, propyl, isopropyl, trifluoromethyl, trifluoroethyl, hydroxymethyl, hydroxyethyl Base, hydroxypropyl, 2-hydroxypropyl, methoxy, ethoxy, propoxy, isopropoxy, nitro, carboxyl, cyano, amino, methylamino, ethylamino, propylamino, Isopropylamino, dimethylamino, diethylamino, methylethylamino, dipropylamino, methylpropylamino, ethylpropylamino, methylacylamino, ethylacylamino, vinylacylamino, Methyl acyl, ethyl acyl, vinyl acyl, amino acyl, methyl amino acyl, ethyl amino acyl, propyl amino acyl, cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

In some specific embodiments, a compound of Formula I, Ia, Ib, Ic, Id or Ie according to the invention, or an isomer, pharmaceutically acceptable salt, solvate, crystal, or isoster thereof or a prodrug wherein each of R ¹ , R ² , R ^{8 is} independently selected from the group consisting of fluorine, chlorine, methyl, ethyl, propyl, methoxy, ethoxy, propoxy, fluoromethoxy, Fluoromethoxy, trifluoromethoxy, fluoroethoxy, difluoroethoxy, R ^{3 is} selected from

R ⁴ is a methyl group, and R ⁵ , R ⁶ , R ⁷ and R ⁹ are H.

The present invention provides the following specific compounds:

Or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal, an isostere or a prodrug.

In another aspect, the invention provides a process for the preparation of a compound of the formula of the invention, comprising:

1) a compound of formula 1 and reacting with trifluoromethanesulfonic anhydride to produce a compound of formula 2;

2) a compound of formula 2 is reacted with a compound of formula 3 to produce a compound of formula 4;

3) a compound of formula 5 is reacted with a compound of formula 6 to produce a compound of formula 7;

4) reacting a compound of formula 4 with a compound of formula 7 to produce a compound of formula 8;

5) A compound of formula 8 is obtained by hydrolysis of a compound of formula 8.

Wherein the compound of Formula 1, the compound of Formula 3, the compound of Formula 5 or the compound of Formula 6 can be obtained by commercially available or conventionally known methods known in the art, R ¹ , R ² , R ³ , R ⁴ And R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , X, m, n and o have the meanings in the formula I; M is an alkyl group.

In a third aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal, an isostere or a prodrug.

In some embodiments, the present invention provides a pharmaceutical composition comprising a compound of the present invention, or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal, an isostere or a prodrug, further comprising a selected from the group consisting of One or more agents in the composition group: SERD, SERM, tyrosinase inhibitor, EGFR inhibitor, VEGFR inhibitor, Bcr-Abl inhibitor, c-kit inhibitor, c-Met inhibitor, Raf Inhibitor, MEK inhibitor, histone deacetylase inhibitor, VEGF antibody, EGF antibody, HIV protein kinase inhibitor, HMG-CoA reductase inhibitor, and the like.

In some embodiments, the invention provides a compound of the invention, or an isomer, pharmaceutically acceptable salt, solvate, crystal, or isoster thereof or prodrug thereof, and a compound comprising the same, or an isomer thereof, Pharmaceutical compositions of pharmaceutically acceptable salts, solvates, crystals, electron isosteres or prodrugs for use in the treatment and/or prevention of estrogen receptor related diseases.

The compound of the present invention or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal, an isostere or a prodrug thereof may be mixed with a pharmaceutically acceptable carrier, diluent or excipient to prepare a pharmaceutical preparation. To be suitable for oral or parenteral administration. Methods of administration include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, and oral routes. The formulation may be administered by any route, for example by infusion or bolus injection, by a route of absorption through the epithelium or skin mucosa (e.g., oral mucosa or rectum, etc.). Administration can be systemic or topical. Examples of the orally administered preparations include solid or liquid dosage forms, specifically, tablets, pills, granules, powders, capsules, syrups, emulsions, suspensions and the like. The formulations may be prepared by methods known in the art and comprise carriers, diluents or excipients conventionally employed in the field of pharmaceutical formulations.

In a fourth aspect, the present invention provides a compound of the formula I of the present invention, or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal, an isostere or a prodrug thereof, or a pharmaceutical composition comprising the same Use of a medicament for the treatment and/or prevention of an estrogen receptor-related disease, wherein the estrogen receptor-associated disease or condition includes, but is not limited to, a cancer associated with ER-α dysfunction (eg, bone cancer) , breast cancer, colorectal cancer, endometrial cancer, prostate cancer, ovarian cancer, and uterine cancer, etc.), leiomyomas (such as uterine leiomyoma, etc.), central nervous system (CNS) defects (such as alcoholism, migraine) Etc., cardiovascular system defects (such as aortic aneurysm, myocardial infarction susceptibility, aortic valve sclerosis, cardiovascular disease, coronary artery disease, hypertension, etc.), blood system defects (such as deep vein thrombosis, etc.), immunity and Inflammatory diseases (such as Graves' disease, arthritis, multiple sclerosis, cirrhosis, etc.), susceptibility to infection (such as hepatitis B, chronic liver disease, etc.), metabolic defects (such as bone density, cholestasis, hypospadias) Obesity, osteoarthritis, osteopenia, osteoporosis, etc.), neurological deficits (eg Alzheimer's disease, Parkinson's disease, migraine, dizziness, etc.), mental disorders (eg anorexia nervosa, attention deficit) With hyperactivity disorder (ADHD), dementia, major depressive disorder, mental illness, etc.) and reproductive defects (such as abnormal menarche age, endometriosis, infertility, etc.). In some embodiments, the invention relates to a method of treating an estrogen receptor-related disorder comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula I or an isomer thereof, pharmaceutically acceptable a salt, solvate, crystallization, electron isostere or prodrug, or a pharmaceutical composition comprising the same, wherein the estrogen receptor related diseases include, but are not limited to, cancer associated with ER dysfunction (eg, bone cancer) , breast cancer, colorectal cancer, endometrial cancer, prostate cancer, ovarian cancer, and uterine cancer, etc.), leiomyomas (such as uterine leiomyoma, etc.), central nervous system (CNS) defects (such as alcoholism, migraine) Etc., cardiovascular system defects (such as aortic aneurysm, myocardial infarction susceptibility, aortic valve sclerosis, cardiovascular disease, coronary artery disease, hypertension, etc.), blood system defects (such as deep vein thrombosis, etc.), immunity and Inflammatory diseases (such as Graves' disease, arthritis, multiple sclerosis, cirrhosis, etc.), susceptibility to infection (such as hepatitis B, chronic liver disease, etc.), metabolic defects (such as bone density, cholestasis, Hypospadias, obesity, osteoarthritis, osteopenia, osteoporosis, etc., neurological deficits (eg Alzheimer's disease, Parkinson's disease, migraine, dizziness, etc.), mental disorders (eg anorexia nervosa) , attention deficit hyperactivity disorder (ADHD), dementia, major depressive disorder, mental illness, etc.) and reproductive defects (such as abnormal menarche age, endometriosis, infertility, etc.).

The compound of the present invention has more excellent antitumor activity, a longer dosing interval, and less side effects.

Terminology

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

"Hydrogen", "carbon", "oxygen" in the compounds of the invention include all isotopes thereof. Isotopes are understood to include those atoms having the same number of atoms but having different mass numbers. For example, isotopes of hydrogen include lanthanum and cerium, carbon isotopes include ¹³ C and ¹⁴ C, and oxygen isotopes include ¹⁶ O and ¹⁸ O.

The "halogen" of the present invention means fluorine, chlorine, bromine or iodine. "Halo" as used in the present invention means substituted by fluorine, chlorine, bromine or iodine.

The "alkyl group" of the present invention means a linear or branched saturated aliphatic hydrocarbon group, preferably a linear or branched group having 1 to 6 carbon atoms, further preferably a linear or branched having 1 to 3 carbon atoms. Chain groups, non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethyl Propyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, and the like. The alkyl group can be substituted or unsubstituted, and when substituted, the substituent can be at any point of attachment that can be used.

The "alkylene group" of the present invention means a group in which an alkyl group is formally removed from a hydrogen atom, such as a methylene group (-CH ₂ -), an ethylene group (-CH ₂ -CH ₂ -), a propylene group (-CH ₂ -CH ₂ -CH ₂ -, -CH(CH ₃ )CH ₂ -), etc., and a "C _1-10 alkyl group" as used herein means a C _1-10 alkane. The group is formally removed from the group remaining by a hydrogen atom, and the " _C1-6 alkyl group" means a group in which a _C1-6 alkyl group is formally removed from a hydrogen atom. The alkylene group can be substituted or unsubstituted, and when substituted, the substituent can be at any point of attachment that can be used.

The "haloalkyl group" of the present invention means an alkyl group substituted with at least one halogen.

The "hydroxyalkyl group" of the present invention means an alkyl group substituted with at least one hydroxyl group.

The "alkoxy group" of the present invention means an -O-alkyl group. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, propoxy, n-propoxy, isopropoxy, isobutoxy, sec-butoxy, and the like. The alkoxy group can be optionally substituted or unsubstituted, and when substituted, the substituent can be at any point of attachment that can be used.

The "cycloalkyl group" of the present invention means a cyclic saturated hydrocarbon group. Suitable cycloalkyl groups may be substituted or unsubstituted monocyclic, bicyclic or tricyclic saturated hydrocarbon groups having 3 to 10 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl.

The "heterocyclyl" of the present invention means 3- to 10-- having a ring carbon atom and 1 to 4 ring hetero atoms each of which is independently selected from the group consisting of nitrogen, oxygen, sulfur, boron, phosphorus, and silicon. A group of a non-aromatic ring system (the "C _3-10 membered heterocyclic group" of the present invention has the same meaning as the "3-10 membered heterocyclic group"). In a heterocyclic group containing one or more nitrogen atoms, the point of attachment may be a carbon or nitrogen atom as long as the valency permits. The heterocyclyl group may alternatively be a monocyclic ("monocyclic heterocyclic") or a fused, bridged or spiro ring system (eg, a bicyclic system ("bicyclic heterocyclyl")) And it can be saturated or can be partially unsaturated. The heterocyclic bicyclic ring system may include one or more heteroatoms in one or both rings. "Heterocyclyl" also includes ring systems wherein a heterocycle, as defined above, is fused to one or more carbocyclyl groups (wherein the point of attachment is on a carbocyclic group or on a heterocyclic ring), or a ring system The heterocyclic ring, as defined above, is fused to one or more aryl or heteroaryl groups (wherein the point of attachment is on the heterocyclic ring), and in such cases, the number of ring members continues to be referred to as hetero The number of ring members in the ring system. Unless otherwise specified, each instance of a heterocyclic group is independently optionally substituted, that is, unsubstituted ("unsubstituted heterocyclic") or substituted with one or more substituents ("substituted hetero Ring base"). In certain embodiments, the heterocyclyl group is an unsubstituted 3-10 membered heterocyclyl. In certain embodiments, the heterocyclyl group is a substituted 3-10 membered heterocyclyl. Fused to the C ₆ aryl ring (also referred to as 5,6-bicyclic heterocycle) Exemplary 5-membered heterocyclyl groups include, but are not limited to, indolinyl, iso indolinyl, two Hydrobenzofuranyl, dihydrobenzothienyl, benzoxazolinone, and the like. Exemplary 6-membered heterocyclyl groups (also referred to herein as 6,6-bicyclic heterocycles) fused to an aryl ring include, but are not limited to, tetrahydroquinolyl, tetrahydroisoquinolinyl, and the like. .

The "aryl" of the present invention means an aromatic system which may comprise a monocyclic or fused polycyclic ring, preferably a monocyclic or fused bicyclic aromatic system containing from 6 to 18 carbon atoms, preferably from about 6 to about 12 carbon atoms. Suitable aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, tetrahydronaphthyl, anthracenyl, indanyl. The aryl group can be optionally substituted or unsubstituted, and when substituted, the substituent can be at any available point of attachment.

The "heteroaryl" of the present invention means an aryl group having at least one carbon atom replaced by a hetero atom, and the hetero atom is O, S, N. Suitable heteroaryl groups include, but are not limited to, imidazolyl, benzimidazolyl, imidazopyridyl, quinazolinone, pyrrolyl, imidazolidinyl, furyl, thienyl, pyrazolyl, oxazolyl, thiazole Base, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl and the like. Wherein the "heteroaryl group" of the present invention is preferably composed of 5 to 18 atoms (abbreviated as "5-18 membered heteroaryl group" or "C _5-18 membered heteroaryl group" in the present invention), further preferably A heteroaryl group having 5-12 atoms and at least one atom being a hetero atom. Suitable five- to twelve-membered heteroaryl groups include, but are not limited to, pyrimidinyl, pyridyl, pyrazinyl, pyridazinyl, pyrimidopyrazolyl, pyrimidoimidazolyl, and the like. The heteroaryl group can be optionally substituted or unsubstituted, and when substituted, the substituent can be at any point of attachment that can be used.

The "isomer" of the present invention is a compound having the same molecular formula but differing in nature or on the bond sequence of its atom or in the spatial arrangement of its atoms. Stereoisomers are isomers whose atoms are spatially distinct. Stereoisomers that are not mirror images of each other are diastereomers and that the stereoisomers that are non-overlapping mirror images of each other are enantiomers. When a compound has an asymmetric center, for example, it is bonded to four different groups, a pair of enantiomers is possible. Enantiomers are characterized by the absolute configuration of their asymmetric centers and are described by Rahn and S-sequence rules of Cahn and Prelog, or by methods of rotating the plane of polarized light by molecules and designated as right-handed or left-handed ( That is, as (+) or (-)-isomers, respectively. The chiral compound can exist as a single enantiomer or a mixture thereof. An equal proportion of mixtures comprising enantiomers is referred to as a "racemic mixture".

The "pharmaceutically acceptable salts" of the present invention refer to salts of the compounds of the present invention which are safe and effective for use in mammals and which have the desired biological activity.

The "solvate" of the present invention refers in a conventional sense to a complex formed by a combination of a solute (such as an active compound, a salt of an active compound) and a solvent (such as water). Solvent refers to a solvent known or readily determinable by those skilled in the art. In the case of water, the solvate is generally referred to as a hydrate such as a hemihydrate, a monohydrate, a dihydrate, a trihydrate or a substituted amount thereof.

The in vivo action of a compound of formula (I) can be exploited, in part, by one or more metabolites formed in the human or animal body following administration of a compound of formula (I). As described above, the in vivo action of the compound of formula (I) can also be exerted via metabolism of the precursor compound ("prodrug"). The "prodrug" of the present invention refers to a compound which is converted into a compound of the formula (I) by a reaction with an enzyme, gastric acid or the like under physiological conditions in a living body, that is, a compound converted by oxidation, reduction, hydrolysis or the like of an enzyme. A compound of the compound of (I) and/or a compound which is converted into a compound of the formula (I) by a hydrolysis reaction such as gastric acid or the like. Suitable pharmaceutically acceptable prodrugs of the compound of formula I having a carboxy group are, for example, cleavable esters thereof in vivo. An in vivo cleavable ester of a compound of formula I comprising a carboxyl group is, for example, a pharmaceutically acceptable ester which is cleaved in the human or animal body to produce the parent acid. Suitable pharmaceutically acceptable esters for carboxyl groups include alkyl esters such as methyl esters, ethyl esters and tert-butyl esters, alkoxymethyl esters such as methoxymethyl esters; alkanoyloxy groups a base such as pivaloyloxy ester; 3-decyl ester; a cycloalkylcarbonyloxyalkyl ester such as cyclopentylcarbonyloxymethyl ester and 1-cyclohexylcarbonyloxyethyl ester; -Oxo-1,3-dioxolyl methyl ester, such as 5-methyl-2-oxo-1,3-dioxol-4-ylmethyl ester And alkoxycarbonyloxyalkyl esters such as methoxycarbonyloxymethyl ester and 1-methoxycarbonyloxyethyl ester. Suitable pharmaceutically acceptable prodrugs of the compound of formula I having a carboxy group are, for example, in vivo cleavable amides such as N-alkyl amides and N,N-dialkyl amides such as N-methyl amide, N Ethyl amide, N-propyl amide, N,N-dimethyl amide, N-ethyl-N-methyl amide or N,N-diethyl amide.

The bioisostere of the present invention (or simply "e-isostere") is used to define that one or more of the atoms (or radicals) have been similarly spatially and/or electronically characterized by those atoms to which they are substituted. A generally recognized term in the art of a drug analog substituted with an atom (or a group of atoms), such as an isostere of a carboxyl group of the present invention, may be an aminoacyl group, in particular, the isostere of the -COOH of the present invention may Is -CH ₂ NH ₂ .

The "crystallization" of the present invention means that the internal structure is a solid formed by regularly repeating constituent atoms (or a group thereof) in three dimensions, and is different from an amorphous solid having an internal structure not having such a regularity.

A "pharmaceutical composition" according to the invention is meant to include any one of the compounds described herein, including the corresponding isomers, prodrugs, solvates, pharmaceutically acceptable salts or chemically protected forms thereof, and one or A mixture of a plurality of pharmaceutically acceptable carriers. The purpose of a pharmaceutical composition is to facilitate the administration of a compound to an organism. The compositions are typically used in the manufacture of a medicament for the treatment and/or prevention of a disease mediated by one or more kinases.

The "pharmaceutically acceptable carrier" of the present invention means a carrier which does not cause significant irritation to an organism and does not interfere with the biological activity and properties of the administered compound, and includes all solvents, diluents or other excipients, dispersing agents, Surfactant isotonicity agent, thickener or emulsifier, preservative, solid binder, lubricant, and the like. Unless any conventional carrier medium is incompatible with the compounds of the invention. Some examples of pharmaceutically acceptable carriers include, but are not limited to, sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethylcellulose, And cellulose and cellulose acetate; malt, gelatin and the like.

"Excipient" as used herein refers to an inert substance that is added to a pharmaceutical composition to further facilitate administration of the compound. Excipients can include calcium carbonate, calcium phosphate, various sugars and various types of starch, cellulose derivatives, gelatin, vegetable oils, polyethylene glycols.

The "use in a medicament for treating and/or preventing an estrogen receptor-associated disease" of the present invention means that an estrogen receptor-related disease can be improved, and growth, development and/or inhibition of an estrogen receptor-related disease can be suppressed. Or transferring, or reducing the risk of an estrogen receptor-related disease, primarily administering a therapeutically and/or prophylactically effective amount of a compound of the invention to a human or animal in need thereof to inhibit, slow or reverse the estrogen receptor in the subject The growth, development, or spread of a body-related disease that improves or reduces the risk of estrogen receptor-related diseases, including diseases associated with estrogen receptor alpha and Diseases associated with estrogen receptor beta, such as cancers associated with estrogen receptor dysfunction (eg, bone cancer, breast cancer, colorectal cancer, endometrial cancer, prostate cancer, ovarian cancer, and uterine cancer), smoothing Fibroids (eg uterine leiomyoma, etc.), central nervous system (CNS) defects (eg alcoholism, migraine, etc.), cardiovascular system defects (eg aortic aneurysm, myocardial infarction susceptibility, aorta Plaque sclerosis, cardiovascular disease, coronary artery disease, hypertension, etc.), blood system defects (such as deep vein thrombosis, etc.), immune and inflammatory diseases (such as Graves' disease, arthritis, multiple sclerosis, cirrhosis, etc.) Susceptibility to infection (such as hepatitis B, chronic liver disease, etc.), metabolic defects (such as bone density, cholestasis, hypospadias, obesity, osteoarthritis, osteopenia, osteoporosis, etc.), neurological defects ( For example, Alzheimer's disease, Parkinson's disease, migraine, dizziness, etc.), mental disorders (such as anorexia nervosa, attention deficit hyperactivity disorder (ADHD), dementia,

Serious depressive disorder, mental illness, etc.) and reproductive defects (such as abnormal menarche age, endometriosis, infertility, etc.).

detailed description

The following representative examples are intended to better illustrate the invention and are not intended to limit the scope of the invention. The materials used in the following examples are commercially available unless otherwise specified.

Example 1: 3',5'-Difluoro-4'-((1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9 -tetrahydro-1H-pyrido[3,4-b]indol-1-yl)-[1,1'-biphenyl]-4-carboxylic acid

Step a: Synthesis of 2-fluoro-2-methylpropan-1-ol

In a 500 mL three-necked flask, methyl 2-fluoro-2-methylpropionate (10 g, 83.3 mmol) was added, dissolved in 200 mL of anhydrous tetrahydrofuran, and the reactor was cooled to about -10 ° C in a low temperature reactor, and lithium hydride was slowly added in portions. Aluminum (3.8 g, 100 mmol) was maintained at this temperature for 1 h. After the reaction was completed, 3.8 mL of water, 3.8 mL of 15% NaOH solution and 7.6 mL of water were added dropwise, and the mixture was stirred for 15 min and then filtered. The filter cake was washed with a small amount of THF. ESI-Ms m/z: 93.1 [M+H] ⁺ .

Step b: Synthesis of 2-fluoro-2-methylpropyl trifluoromethanesulfonate

In a 250 mL reaction flask, 2-fluoro-2-methylpropan-1-ol (2.9 g, 31.5 mmol), 2,6-lutidine (5.1 mL, 44.1 mmol) was dissolved in 30 mL of dichloromethane. After low temperature cooling to -10 °C. Trifluoromethanesulfonic anhydride (5.83 mL, 34.7 mmol) was dissolved in 20 ml of dichloromethane, and then added dropwise to the above reaction mixture, and the reaction was continued for 1 h. After the reaction, the reaction solution was washed with 2N hydrochloric acid (20 mL×2×), saturated aqueous sodium hydrogen carbonate (20 mL×2×) and saturated aqueous sodium chloride (20 mL×2×) The methylene chloride was removed by pressure to give the title compound. ESI-Ms m/z: 225.1 [M+H] ⁺ .

Step c: Synthesis of (S)-2-amino-3-(1H-indol-3-yl)propan-1-ol

In a 100 mL reaction flask, L-tryptophan methyl ester hydrochloride (2.0 g, 7.85 mmol), methanol (1.5 mL) was added sequentially, followed by triethylamine (1.5 mL), diethyl ether (50 mL). Stir at 10 ° C - 0 ° C for 1 h. After 1 h, the white solid formed by the reaction was removed by filtration, and the filtrate was collected and evaporated to give a colorless oil, and then 15 mL of methanol was added thereto, and sodium borohydride (356 mg, 9.42 mmol) was added to the mixture in an ice bath for 30 min. The reaction mixture was transferred to room temperature and stirred for several hours until the starting material disappeared. After completion of the reaction, the reaction was quenched with EtOAc (EtOAc)EtOAc. Used directly in the next step. ESI-Ms m/z: 191.2 [M+H] ⁺ .

Step d: Synthesis of (S)-benzyl-(1-hydroxy-3-(1H-indol-3-yl)propan-2-yl)carbamate

(S)-2-Amino-3-(1H-indol-3-yl)propan-1-ol (74 g, 389.5 mmol) was placed in a 2 L eggplant bottle, 600 mL water and 600 mL acetone were added and stirred at room temperature. Sodium carbonate (70 g, 662.1 mmol) was added, the mixture was cooled to 0 ° C, benzyl chloroformate (66 g, 389.5 mmol) was added dropwise, stirred at the same temperature for 30 min, and transferred to room temperature for 14 h. The mixture was adjusted to pH 2 with EtOAc (EtOAc)EtOAc. ESI-Ms m/z: 325.1 [M+H] ⁺ .

Step e: Synthesis of (S)-2-(((benzyloxy)carbonyl)amino)-3-(1H-indol-3-yl)propyl-4-methylbenzenesulfonate

(S)-Benzyl-(1-hydroxy-3-(1H-indol-3-yl)propan-2-yl)carbamate (110 g, 339.5 mmol), triethylamine (95 mL, 679.0 Methyl) and 1 L of dichloromethane were mixed in a 2 L eggplant bottle, stirred until dissolved, and cooled to -3 °C. A solution of p-toluenesulfonyl chloride (71.2 g, 373.5 mmol) in dichloromethane (100 mL) was added dropwise and the mixture was stirred at the same temperature for 1 h and then allowed to react at room temperature for 20 h. The organic layer was dried over anhydrous sodium sulfate (MgSO4). ESI-Ms m/z: 479.1 [M+H] ⁺ .

Step f: Synthesis of (R)-1-(1H-indol-3-yl)propan-2-amine

(S)-2-(((Benzyloxy)carbonyl)amino)-3-(1H-indol-3-yl)propyl-4-methylbenzenesulfonate (110 g, 632.2 mmol) was dissolved in In 1.2 L of ethanol, palladium hydroxide was added, and the mixture was stirred under a hydrogen atmosphere at room temperature for 2 days. ESI-Ms m/z: 175.1 [M+H] ⁺ .

Step g: Synthesis of (R)-N-(1-(1H-indol-3-yl)propan-2-yl)-2-fluoro-2-methylpropan-1-amine

In a 250 mL reaction flask, the product obtained in step b was added 2-fluoro-2-methylpropyltrifluoromethanesulfonate (2 g, 30 mmol), (R)-1-(1H-indol-3-yl)propene 2-Amine (4.3g, 25mmol) and diisopropylethylamine (8.7mL, 50mmol), dissolved in 50mL dioxane, reacted under argon at 90 ° C for 2h, stop the reaction, concentrate, and purify by column chromatography The title compound was obtained. ESI-Ms m/z: 249.1 [M+H] ⁺ .

Step h: Synthesis of 3',5'-difluoro-4'-formyl-[1,1'-biphenyl]-4-carboxylic acid methyl ester

(4-(Methoxycarbonyl)phenyl)boronic acid (1.97 g, 11 mmol), 4-bromo-2,6-difluorobenzaldehyde (2.21 g, 10 mmol), 1,3-bis(diphenylphosphine) Propane) nickel dichloride (54 mg, 0.1 mmol), potassium phosphate (8.48 g, 40 mmol) and 60 mL of dioxane were mixed in a 100 mL eggplant bottle, heated to 110 ° C under argon atmosphere, and reacted for 10 h at the same temperature. . The organic solvent was concentrated under reduced pressure. EtOAc (EtOAc)EtOAc. The residue was purified by silica gel column chromatography eluting ESI-Ms m/z: 277.1 [M+H] ⁺ .

Step i: 3',5'-difluoro-4'-((1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9- Synthesis of Methyl Tetrahydro-1H-pyrido[3,4-b]indol-1-yl)-[1,1'-biphenyl]-4-carboxylate

Methyl 3',5'-difluoro-4'-formyl-[1,1'-biphenyl]-4-carboxylate (0.66 g, 2.4 mmol), ice vinegar (1 mL), toluene (20 mL) And (R)-N-(1-(1H-indol-3-yl)propan-2-yl)-2-fluoro-2-methylpropan-1-amine (0.6 g, 2 mmol) was mixed in 50 mL of eggplant The flask was heated to 80 ° C under argon atmosphere and stirred overnight. It was diluted with 100 mL of ethyl acetate, washed with 50 mL of saturated sodium hydrogen sulfate and 50 mL of saturated sodium chloride. ESI-Ms m/z: 507.1 [M+H] ⁺ .

Step j: 3',5'-difluoro-4'-((1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9- Synthesis of tetrahydro-1H-pyrido[3,4-b]indol-1-yl)-[1,1'-biphenyl]-4-carboxylic acid

3',5'-Difluoro-4'-((1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro -1H-pyrido[3,4-b]indol-1-yl)-[1,1'-biphenyl]-4-carboxylic acid methyl ester (0.5 g, 1 mmol), 3 mL of methanol and 6 mL of tetrahydrofuran In a 50 mL eggplant bottle, stir, cool to 0 ° C, add sodium hydroxide solution (2 mL) dropwise, and react at room temperature for 2 h. The pH was adjusted to 6 with 2M EtOAc EtOAc (EtOAc)EtOAc.

^{1 H NMR (400MHz, DMSO-} d 6) δ: 13.04 (s, 1H), 10.65 (s, 1H), 8.03-8.01 (d, 2H), 7.91-7.89 (d, 2H), 7.52-7.49 (d , 2H), 7.44 - 7.42 (d, 1H), 7.21 - 7.19 (d, 1H), 7.02 - 7.01 (m, 2H), 5.28 (s, 1H), 3.33 - 3.32 (m, 1H), 2.94 - 2.85 (m, 2H), 2.63 - 2.61 (m, 1H), 2.52 - 2.40 (m, 1H), 1.22 - 1.01 (m, 9H). ESI-Ms m/z: 493.1 [M+H] ⁺ .

Example 2: 3'-Chloro-5'-fluoro-4'-((1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4, 9-tetrahydro-1H-pyrido[3,4-b]indol-1-yl)-[1,1'-biphenyl]-4-carboxylic acid

The preparation method was similar to the preparation method of Example 1, except that the starting material 4-bromo-2,6-difluorobenzaldehyde was replaced with 4-bromo-2-chloro-6-fluorobenzaldehyde to give the title compound. ^{1 H NMR (400MHz, DMSO-} d 6) δ: 13.02 (s, 1H), 10.64 (s, 1H), 8.01-7.97 (d, 2H), 7.80-7.76 (d, 2H), 7.42-7.39 (d , 3H), 7.20–7.17 (d, 1H), 7.02–6.93 (m, 2H), 5.26 (s, 1H), 3.48–3.45 (d, 1H), 2.88–2.79 (m, 2H), 2.58–2.53 (m, 1H), 2.42 - 2.36 (m, 1H), 1.23 - 1.01 (m, 9H). ESI-Ms m/z: 509.2 [M+H] ⁺ .

Example 3: 3',5'-Dichloro-4'-((1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9 -tetrahydro-1H-pyrido[3,4-b]indol-1-yl)-[1,1'-biphenyl]-4-carboxylic acid

The preparation method was similar to the preparation method of Example 1, except that the starting material 4-bromo-2,6-difluorobenzaldehyde was replaced with 4-bromo-2,6-dichlorobenzaldehyde to give the title compound. ^{1 H NMR (400MHz, DMSO-} d 6) δ: 13.00 (s, 1H), 10.65 (s, 1H), 8.00-7.98 (d, 2H), 7.38-7.45 (d, 2H), 7.30-7.35 (d , 3H), 7.20–7.18 (d, 1H), 7.02–6.93 (m, 2H), 5.25 (s, 1H), 3.48–3.45 (d, 1H), 2.88–2.79 (m, 2H), 2.58–2.53 (m, 1H), 2.42 - 2.36 (m, 1H), 1.22 - 1.01 (m, 9H). ESI-Ms m/z: 525.1 [M+H] ⁺ .

Example 4: 3',5'-Difluoro-4'-((1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9 -tetrahydro-1H-pyrido[3,4-b]indol-1-yl)-[1,1'-biphenyl]-3-carboxylic acid

The preparation method was similar to the preparation method of Example 1, except that the starting material (4-(methoxycarbonyl)phenyl)boronic acid was replaced by (3-(methoxycarbonyl)phenyl)boronic acid to give the title compound. ^{1 H NMR (400MHz, DMSO-} d 6) δ: 12.98 (s, 1H), 10.63 (s, 1H), 8.23-8.21 (d, 1H), 7.99-7.97 (m, 2H), 7.62-7.58 (m , 1H), 7.46 - 7.41 (d, 3H), 7.21 - 7.19 (d, 1H), 7.02 - 6.94 (d, 2H), 5.28 (s, 1H), 3.58 - 3.52 (m, 1H), 2.94 - 2.90 (m, 2H), 2.62 - 2.46 (m, 2H), 1.26 - 1.07 (m, 9H). ESI-Ms m/z: 493.1 [M+H] ⁺ .

Example 5: 3'-Fluoro-4'-((1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro- 1H-pyrido[3,4-b]indol-1-yl)-3,5'-dimethoxy-[1,1'-biphenyl]-4-carboxylic acid

The preparation method is similar to the preparation method of Example 1, except that the starting material (4-(methoxycarbonyl)phenyl)boronic acid is replaced by (3-methoxy-4-(methoxycarbonyl)phenyl)boronic acid. And the starting material 4-bromo-2,6-difluorobenzaldehyde was replaced with 4-bromo-2-methoxy-6-fluorobenzaldehyde to give the title compound. ^{1 H NMR (400MHz, DMSO-} d 6) δ: 12.70 (s, 1H), 10.41 (s, 1H), 8.24 (s, 1H), 7.68 (d, 1H), 7.40-7.37 (m, 2H), 7.24(s,1H), 7.18–7.16(m,1H),7.07(s,1H),6.97–6.93(m,2H),5.37(s,1H),3.95(s,3H),3.92(s, 3H), 3.60–3.57 (m, 1H), 3.10–2.87 (m, 2H), 2.59–2.55 (d, 1H), 2.51–2.42 (m, 1H), 1.18–1.03 (m, 9H). ESI- Ms m/z: 535.3 [M+H] ⁺ .

Example 6: 3'-Fluoro-4'-((1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro- 1H-pyrido[3,4-b]indol-1-yl)-5'-methoxy-[1,1'-biphenyl]-4-carboxylic acid

The preparation method is similar to the preparation method of Example 1, except that the starting material 4-bromo-2,6-difluorobenzaldehyde is replaced with 4-bromo-2-methoxy-6-fluorobenzaldehyde to obtain the title compound. . ^{1 H NMR (400MHz, DMSO-} d 6) δ: 13.02 (s, 1H), 10.40 (s, 1H), 7.95 (s, 1H), 7.66-7.64 (d, 1H), 7.36-7.34 (m, 3H ), 7.23 (s, 1H), 7.15 - 7.12 (m, 1H), 7.03 (s, 1H), 6.95 - 6.90 (m, 2H), 5.35 (s, 1H), 3.90 (s, 3H), 3.60 - 3.56 (m, 1H), 3.07–2.86 (m, 2H), 2.59–2.55 (d, 1H), 2.50–2.40 (m, 1H), 1.18–1.02 (m, 9H). ESI-Ms m/z: 505.3 [M+H] ⁺ .

Example 7: 2,3'-Difluoro-4'-((1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9- Tetrahydro-1H-pyrido[3,4-b]indol-1-yl)-5',6-dimethoxy-[1,1'-biphenyl]-4-carboxylic acid

The preparation method is similar to the preparation method of Example 1, except that the starting material (4-(methoxycarbonyl)phenyl)boronic acid is replaced by (2-fluoro-6-methoxy-4-(methoxycarbonyl). Phenyl)boronic acid, and the starting material 4-bromo-2,6-difluorobenzaldehyde was replaced with 4-bromo-2-methoxy-6-fluorobenzaldehyde to give the title compound. ^{1 H NMR (400MHz, DMSO-} d 6) δ: 13.00 (s, 1H), 10.40 (s, 1H), 7.67 (d, 1H), 7.40-7.37 (m, 2H), 7.25 (s, 1H), 7.17–7.15(m,1H),7.06(s,1H), 6.98–6.95(m,2H), 5.36(s,1H), 4.00(s,3H),3.96(s,3H),3.57–3.55 ( m,1H), 3.11–2.89 (m, 2H), 2.60–2.57 (d, 1H), 2.52–2.43 (m, 1H), 1.19–1.03 (m, 9H). ESI-Ms m/z: 553.3 [ M+H] ⁺ .

Example 8: 2-Chloro-3'-fluoro-4'-((1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9 -tetrahydro-1H-pyrido[3,4-b]indol-1-yl)-5'-methoxy-[1,1'-biphenyl]-4-carboxylic acid

The preparation method is similar to the preparation method of Example 1, except that the starting material (4-(methoxycarbonyl)phenyl)boronic acid is replaced by (2-chloro-4-(methoxycarbonyl)phenyl)boronic acid, and The title compound was obtained by substituting 4-bromo-2,6-difluorobenzaldehyde as 4-bromo-2-methoxy-6-fluorobenzaldehyde. ^{1 H NMR (400MHz, DMSO-} d 6) δ: 13.01 (s, 1H), 10.42 (s, 1H), 7.69 (d, 1H), 7.41-7.38 (m, 3H), 7.27 (s, 1H), 7.18–7.15(m,1H),7.08(s,1H), 7.00–6.96(m,2H),5.35(s,1H),3.96(s,3H),3.57–3.55(m,1H),3.11– 2.90 (m, 2H), 2.60 - 2.57 (d, 1H), 2.52 - 2.43 (m, 1H), 1.19 - 1.03 (m, 9H). ESI-Ms m/z: 539.1 [M+H] ⁺ .

Example 9: 3'-Fluoro-4'-((1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro- 1H-pyrido[3,4-b]indol-1-yl)-2,5'-dimethoxy-[1,1'-biphenyl]-4-carboxylic acid

The preparation method is similar to the preparation method of Example 1, except that the starting material (4-(methoxycarbonyl)phenyl)boronic acid is replaced by (2-methoxy-4-(methoxycarbonyl)phenyl)boronic acid. And the starting material 4-bromo-2,6-difluorobenzaldehyde was replaced with 4-bromo-2-methoxy-6-fluorobenzaldehyde to give the title compound. ^{1 H NMR (400MHz, DMSO-} d 6) δ: 12.72 (s, 1H), 10.42 (s, 1H), 7.68 (d, 1H), 7.41-7.37 (m, 3H), 7.26 (s, 1H), 7.19–7.16 (m, 1H), 7.09 (s, 1H), 7.01–6.95 (m, 2H), 5.36 (s, 1H), 4.00 (s, 3H), 3.96 (s, 3H), 3.57–3.55 ( m,1H), 3.11–2.89 (m, 2H), 2.60–2.57 (d, 1H), 2.52–2.43 (m, 1H), 1.19–1.03 (m, 9H). ESI-Ms m/z: 535.3 [ M+H] ⁺ .

Example 10: 3',5'-Difluoro-4'-((1R,3S)-2-(2-fluoro-2-methylpropyl)-3-(fluoromethyl)-2,3, 4,9-tetrahydro-1H-pyrido[3,4-b]indol-1-yl)-3-methoxy-[1,1'-biphenyl]-4-carboxylic acid

Step a: Synthesis of (S)-(1-fluoro-3-(1H-indol-3-yl)propan-2-yl)carbamate

(S)-2-(((Benzyloxy)carbonyl)amino)-3-(1H-indol-3-yl)propyl-4-methylbenzenesulfonate (4.0 g, 8.36 mmol) The reaction was heated at 85 ° C for 4 hours in a solution of tetrabutylammonium fluoride in tetrahydrofuran (1M, 12 mL). After completion of the reaction, the mixture was evaporated. EtOAcjjjjjjjjjjjjjjjj Compound. ESI-Ms m/z: 327.1 [M+H] ⁺ .

Step b: Synthesis of (S)-1-fluoro-3-(1H-indol-3-yl)propan-2-amine

The title compound was obtained in the same manner as in Example 1 Step 1 using benzyl (S)-(1-fluoro-3-(1H-indol-3-yl)propan-2-yl)carbamate as the starting material. ESI-Ms m/z: 193.1 [M+H] ⁺ .

Step c: Synthesis of (S)-2-fluoro-N-(1-fluoro-3-(1H-indol-3-yl)propan-2-yl)-2-methylpropan-1-amine

Starting from (S)-1-fluoro-3-(1H-indol-3-yl)propan-2-amine, the title compound was obtained from m. ESI-Ms m/z: 267.1 [M+H] ⁺ .

Step d: 3',5'-difluoro-4'-((1R,3S)-2-(2-fluoro-2-methylpropyl)-3-(fluoromethyl)-2,3,4 Synthesis of 9-tetrahydro-1H-pyrido[3,4-b]indol-1-yl)-3-methoxy-[1,1'-biphenyl]-4-carboxylic acid

The preparation method is similar to the preparation method of Example 1, except that the starting material (4-(methoxycarbonyl)phenyl)boronic acid is replaced by (3-methoxy-4-(methoxycarbonyl)phenyl)boronic acid. And replacing (R)-N-(1-(1H-indol-3-yl)propan-2-yl)-2-fluoro-2-methylpropan-1-amine with (S)-2- Fluorine-N-(1-fluoro-3-(1H-indol-3-yl)propan-2-yl)-2-methylpropan-1-amine gave the title compound. ^{1 H NMR (400MHz, DMSO-} d 6) δ: 12.72 (s, 1H), 10.74 (s, 1H), 7.67-7.65 (d, 1H), 7.61-7.59 (d, 2H), 7.53-7.51 (d , 1H), 7.40–7.37 (m, 2H), 7.21–6.19 (d, 1H), 7.04–6.96 (m, 2H), 5.09 (s, 1H), 4.89–4.84 (m, 1H), 4.77–4.72 (m, 1H), 4.66–4.62 (m, 1H), 3.92 (s, 3H), 3.60–3.57 (d, 1H), 2.85–2.73 (m, 2H), 2.53–2.43 (m, 1H), 1.18 –1.10(m,6H).ESI-Ms m/z: 541.2 [M+H] ⁺ .

Example 11: 3'-Fluoro-4'-((1R,3S)-2-(2-fluoro-2-methylpropyl)-3-(fluoromethyl)-2,3,4,9- Synthesis of tetrahydro-1H-pyrido[3,4-b]indol-1-yl)-3,5'-dimethoxy-[1,1'-biphenyl]-4-carboxylic acid

The preparation method is similar to the preparation method of Example 10 except that the starting material 4-bromo-2,6-difluorobenzaldehyde is replaced with 4-bromo-2-methoxy-6-fluorobenzaldehyde to obtain the title compound. . ^{1 H NMR (400MHz, DMSO-} d 6) δ: 12.75 (s, 1H), 10.43 (s, 1H), 7.67 (d, 1H), 7.41-7.36 (m, 3H), 7.26 (s, 1H), 7.20–7.16(m,1H), 7.10(s,1H), 7.01–6.95(m,2H), 5.35(s,1H), 4.31(s,3H), 4.25(d,2H),3.96(s, 3H), 3.58–3.55 (m, 1H), 3.11–2.89 (m, 2H), 2.60–2.57 (d, 1H), 2.52–2.43 (m, 1H), 1.19–1.03 (m, 6H). ESI- Ms m/z: 553.5 [M+H] ⁺ .

Example 12: 3',5'-Difluoro-4'-((1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9 Synthesis of tetrahydro-1H-pyrido[3,4-b]indol-1-yl)-3-methoxy-[1,1'-biphenyl]-4-carboxylic acid

The preparation method is similar to the preparation method of Example 1, except that the starting material (4-(methoxycarbonyl)phenyl)boronic acid is replaced by (3-methoxy-4-(methoxycarbonyl)phenyl)boronic acid. The title compound was obtained. ^{1 H NMR (400MHz, DMSO-} d 6) δ: 12.72 (s, 1H), 10.45 (s, 1H), 7.85-7.81 (m, 1H), 7.53-7.50 (m, 3H), 7.34-7.30 (m , 2H), 7.19-7.17 (m, 1H), 6.96–6.80 (m, 2H), 5.26 (s, 1H), 3.90 (s, 3H), 3.56–3.49 (m, 1H), 2.92–2.81 (m) , 2H), 2.60 - 2.44 (m, 2H), 1.25 - 1.06 (m, 9H). ESI-Ms m/z: 523.2 [M+H] ⁺ .

Example 13: 2,3',5'-trifluoro-4'-((1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4 Synthesis of 9-tetrahydro-1H-pyrido[3,4-b]indol-1-yl)-6-methoxy-[1,1'-biphenyl]-4-carboxylic acid

The preparation method is similar to the preparation method of Example 1, except that the starting material (4-(methoxycarbonyl)phenyl)boronic acid is replaced by (2-fluoro-6-methoxy-4-(methoxycarbonyl). Phenyl)boronic acid to give the title compound. ^{1 H NMR (400MHz, DMSO-} d 6) δ: 12.70 (s, 1H), 10.88 (s, 1H), 7.86-7.81 (m, 1H), 7.68-7.62 (m, 2H), 7.55-7.52 (d , 1H), 7.32 (s, 1H), 7.20-7.17 (m, 1H), 6.95 - 6.81 (m, 2H), 5.25 (s, 1H), 3.79 (s, 3H), 3.57 - 3.50 (m, 1H) ), 2.96 - 2.91 (m, 2H), 2.60 - 2.44 (m, 2H), 1.27 - 1.05 (m, 9H). ESI-Ms m/z: 541.2 [M+H] ⁺ .

Example 14: 2-Chloro-3',5'-difluoro-4'-((1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3 Synthesis of 4,9-tetrahydro-1H-pyrido[3,4-b]indol-1-yl)-[1,1'-biphenyl]-4-carboxylic acid

The preparation method is similar to the preparation method of Example 1, except that the starting material (4-(methoxycarbonyl)phenyl)boronic acid is replaced by (2-chloro-4-(methoxycarbonyl)phenyl)boronic acid. The title compound was obtained. ^{1 H NMR (400MHz, DMSO-} d 6) δ: 13.10 (s, 1H), 10.95 (s, 1H), 7.99-7.91 (m, 2H), 7.84-7.80 (m, 1H), 7.56-7.30 (m , 3H), 7.20-7.16 (m, 1H), 7.04–6.96 (d, 2H), 5.26 (s, 1H), 3.59–3.54 (m, 1H), 2.97–2.93 (m, 2H), 2.64–2.45 (m, 2H), 1.28 - 1.09 (m, 9H). ESI-Ms m/z: 527.1 [M+H] ⁺ .

Example 15: 3',5'-Difluoro-4'-((1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9 Synthesis of tetrahydro-1H-pyrido[3,4-b]indol-1-yl)-2-methoxy-[1,1'-biphenyl]-4-carboxylic acid

The preparation method is similar to the preparation method of Example 1, except that the starting material (4-(methoxycarbonyl)phenyl)boronic acid is replaced by (2-methoxy-4-(methoxycarbonyl)phenyl)boronic acid. The title compound was obtained. ^{1 H NMR (400MHz, DMSO-} d 6) δ: 12.75 (s, 1H), 10.68 (s, 1H), 8.00-7.96 (d, 1H), 7.88-7.76 (m, 3H), 7.53-7.50 (d , 1H), 7.34–7.32 (m, 2H), 7.00–6.91 (d, 2H), 5.27 (s, 1H), 3.79 (s, 3H), 3.50–3.46 (m, 1H), 2.88–2.75 (m) , 2H), 2.60 - 2.42 (m, 2H), 1.27 - 1.05 (m, 9H). ESI-Ms m/z: 523.2 [M+H] ⁺ .

Example 16: 6-(3,5-Difluoro-4-((1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9 -tetrahydro-1H-pyrido[3,4-b]indol-1-yl)phenyl)nicotinic acid

The preparation method is similar to the preparation method of Example 1, except that the starting material (4-(methoxycarbonyl)phenyl)boronic acid is replaced by 6-(4,4,5,5-tetramethyl-1,3, Methyl 2-dioxaborolan-2-yl)nicotinate, the title compound was obtained. ^{1 H NMR (400MHz, DMSO-} d 6) δ: 13.45 (s, 1H), 10.76 (s, 1H), 831 (s, 1H), 8.10-8.05 (d, 1H), 7.83-7.79 (d, 2H ), 7.48–7.41 (d, 2H), 7.25–7.20 (d, 1H), 7.05–6.97 (m, 2H), 5.28 (s, 1H), 3.50–3.47 (d, 1H), 2.88–2.79 (m) , 2H), 2.60–2.55 (m, 1H), 2.42–2.36 (m, 1H), 1.22–1.01 (m, 9H). ESI-Ms m/z: 494.1 [M+H] ⁺ .

Example 17: 3',5'-Dichloro-4'-((1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9 -tetrahydro-1H-pyrido[3,4-b]indol-1-yl)-3-methoxy-[1,1'-biphenyl]-4-carboxylic acid

The preparation method is similar to the preparation method of Example 1, except that the starting material (4-(methoxycarbonyl)phenyl)boronic acid is replaced by (3-methoxy-4-(methoxycarbonyl)phenyl)boronic acid. And the starting material 4-bromo-2,6-difluorobenzaldehyde was replaced with 4-bromo-2,6-dichlorobenzaldehyde to give the title compound. ^{1 H NMR (400MHz, DMSO-} d 6) δ: 12.71 (s, 1H), 10.38 (s, 1H), 8.02-8.01 (d, 1H), 7.75-7.74 (d, 1H), 7.73-7.71 (d , 1H), 7.42-7.41 (d, 2H), 7.39-7.38 (m, 1H), 7.18-7.16 (d, 1H), 7.00-6.91 (m, 2H), 5.64 (s, 1H), 3.94 (s , 3H), 3.76-3.73 (m, 1H), 3.13-3.09 (m, 1H), 3.03-2.95 (m, 1H), 2.67-2.64 (d, 1H), 2.34-2.23 (m, 1H), 1.19 -1.06 (m, 9H). ESI-Ms m/z: 555.2 [M+H] ⁺ .

Example 18: 3',5'-Difluoro-4'-((1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9 -tetrahydro-1H-pyrido[3,4-b]indol-1-yl)-[1,1'-biphenyl]-4-carboxamide

100 mg (197 mmol) of the product of step 1 of Example 1 3',5'-difluoro-4'-((1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl -2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indol-1-yl)-[1,1'-biphenyl]-4-carboxylic acid methyl ester, 20 mL of ammonia The methanol solution was mixed in a 50 mL eggplant bottle, heated to 65 ° C, and stirred overnight. Concentration and purification by silica gel column chromatography ^{1 H NMR (400MHz, DMSO-} d 6) δ: 10.6 (s, 1H), 8.06 (s, 1H), 7.98-7.96 (d, 2H), 7.87-7.85 (d, 2H), 7.52-7.42 (m , 4H), 7.21–7.19(d,1H), 7.03–9.64(m,2H), 5.28(s,1H), 3.56–3.52(m,1H),2.94–2.89(m,2H),2.63–2.41 (m, 2H), 1.26 - 1.07 (m, 9H). ESI-Ms m/z: 492.1 [M+H] ⁺ .

Example 19: 3'-Chloro-5'-fluoro-4'-((1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4, 9-tetrahydro-1H-pyrido[3,4-b]indol-1-yl)-3-methoxy-[1,1'-biphenyl]-4-carboxylic acid

The preparation method is similar to the preparation method of Example 1, except that the starting material (4-(methoxycarbonyl)phenyl)boronic acid is replaced by (3-methoxy-4-(methoxycarbonyl)phenyl)boronic acid. And the starting material 4-bromo-2,6-difluorobenzaldehyde was replaced with 4-bromo-2-chloro-6-fluorobenzaldehyde to give the title compound. ^{1 H NMR (400MHz, DMSO-} d 6) δ: 12.75 (s, 1H), 10.49 (s, 1H), 7.77 (s, 1H), 7.65-7.60 (m, 2H), 7.42-7.35 (m, 3H ), 7.19–7.17(d,1H), 7.00–6.92 (m, 2H), 5.38 (s, 1H), 3.91 (s, 3H), 3.70–3.66 (m, 1H), 3.08–2.93 (m, 2H) ), 2.64–2.60 (d, 1H), 2.38–2.27 (m, 1H), 1.16–1.05 (m, 9H). ESI-Ms m/z: 539.1 [M+H]

+.

Example 20: 6-(3-Fluoro-4-((1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro -1H-pyrido[3,4-b]indol-1-yl)-5-methoxyphenyl)nicotinic acid

The preparation method is similar to the preparation method of Example 1, except that the starting material (4-(methoxycarbonyl)phenyl)boronic acid is replaced with 5-(methoxycarbonyl)-2-pyridineboronic acid, and the starting material is 4- The bromine-2,6-difluorobenzaldehyde was replaced with 4-bromo-2-methoxy-6-fluorobenzaldehyde to give the title compound. ^{1 H NMR (400MHz, DMSO-} d 6) δ: 13.40 (s, 1H), 10.42 (s, 1H), 9.12 (s, 1H), 8.31-8.29 (d, 1H), 8.18-8.16 (d, 1H ), 7.71 (s, 1H), 7.43 - 7.38 (m, 1H), 7.16 - 7.15 (d, 2H), 6.97 - 6.93 (m, 2H), 5.38 (s, 1H), 3.97 (s, 3H), 3.60–3.34(m,1H), 2.97–2.84(m,2H), 2.67–2.29(m,2H),1.16–1.05(m,9H).ESI-Ms m/z:506.2[M+H] ⁺ .

Example 21: 2-Chloro-3',5'-difluoro-4'-((1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3 ,4,9-tetrahydro-1H-pyrido[3,4-b]indol-1-yl)-6-methoxy-[1,1'-biphenyl]-4-carboxylic acid

The preparation method is similar to the preparation method of Example 1, except that the starting material (4-(methoxycarbonyl)phenyl)boronic acid is replaced by (3-methoxy-6-chloro-4-(methoxycarbonyl). Phenyl)boronic acid to give the title compound. ^{1 H NMR (400MHz, DMSO-} d 6) δ: 13.41 (s, 1H), 10.77 (s, 1H), 7.65-7.64 (d, 1H), 7.56-7.55 (d, 1H), 7.41-7.40 (d , 1H), 7.24–7.22 (d, 1H), 7.02–6.98 (m, 4H), 5.3 (s, 1H), 3.79 (s, 3H), 3.52–3.51 (m, 1H), 2.91–2.85 (m) , 2H), 2.62–2.61 (d, 1H), 2.51–2.49 (m, 1H), 1.25–1.08 (m, 9H). ESI-Ms m/z: 557.2 [M+H] ⁺ .

Example 22: 2-(3,5-Difluoro-4-((1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9 -tetrahydro-1H-pyrido[3,4-b]indol-1-yl)phenyl)pyrimidine-5-carboxylic acid

The preparation method was similar to the preparation method of Example 1, except that the starting material (4-(methoxycarbonyl)phenyl)boronic acid was replaced with 5-(methoxycarbonyl)-2-pyrimidineboronic acid to give the title compound. ^{1 H NMR (400MHz, DMSO-} d 6) δ: 13.50 (s, 1H), 10.65 (s, 1H), 9.32 (s, 1H), 8.00-7.98 (m, 1H), 7.44-7.18 (m, 4H ), 7.08–6.88 (m, 2H), 5.30 (s, 1H), 3.55 (m, 1H), 2.96–2.91 (m, 2H), 2.63–2.59 (m, 1H), 2.41-2.37 (m, 1H) ), 1.30–1.25 (m, 9H). ESI-Ms m/z: 495.2 [M+H] ⁺ .

Example 23 1-(3,5-Difluoro-4-((1R,3R)-2-((S)-3-fluoro-2-methylpropyl)-3-methyl-2,3, 4,9-tetrahydro-1H-pyrido[3,4-b]indol-1-yl)phenyl)azetidin-3-carboxylic acid

Step a Synthesis of (S)-3-fluoro-2-methylpropionic acid methyl ester

Under a argon atmosphere, to a solution of methyl (R)-3-hydroxy-2-methylpropanoate (2.0 g, 16.93 mmol) dissolved in 20 mL of dichloromethane, N,N-diethyl- 1,1,2,3,3,3-hexafluoropropylamine. After the dropwise addition was completed, the reaction mixture was stirred at room temperature for one hour, then heated to reflux and stirred for 4 hours. The reaction mixture was again cooled to room temperature and stirred for 8 hours. After the reaction was completed, the reaction mixture was poured into EtOAc EtOAc. The title compound was obtained and used directly in the next reaction.

Step b Synthesis of (S)-3-fluoro-2-methylpropanol

In a 500 mL three-necked flask, methyl (S)-3-fluoro-2-methylpropanoate (2.3 g, 19.15 mmol) was added, dissolved in 100 mL of anhydrous tetrahydrofuran, and the reactor was cooled to about 0 ° C in a low temperature reactor. Lithium aluminum hydride (0.78 g, 20.53 mmol) was slowly added. After the addition was completed, the reaction mixture was stirred at 0 ° C and room temperature for 1 h each. After the reaction was completed, sodium sulfate decahydrate was added portionwise, and the mixture was stirred for 1 hr.

Step c Synthesis of (S)-3-fluoro-2-methylpropyltrifluoromethanesulfonate

In a 250 mL reaction flask, (S)-3-fluoro-2-methylpropanol (1.8 g, 19.54 mmol), 2,6-lutidine (2.8 mL, 23.7 mmol), with 20 mL dichloromethane After dissolution, it was cooled to -10 ° C. Trifluoromethanesulfonic anhydride (3.13 mL, 18.63 mmol) was dissolved in 10 mL of dichloromethane, and then added dropwise to the above reaction mixture, and the reaction was continued for 1 h. After the reaction was completed, the reaction mixture was washed with EtOAc (2×20 mL) Methane gave the title compound.

Synthesis of Step d(S)-N-((R)-1-(1H-indol-3-yl)propan-2-yl)-3-fluoro-2-methylpropyl-1-amine

In a 100 mL reaction flask, (S)-3-fluoro-2-methylpropyltrifluoromethanesulfonate (2.72 g, 12.10 mmol), (R)-1-(1H-indol-3-yl) was added. Propyl-2-amine (1.75 g, 10.04 mmol) and diisopropylethylamine (3.32 ml, 20.08 mmol), dissolved in 30 mL of dioxane,

The reaction was carried out at 90 ° C for 2 h under argon. The reaction was quenched, concentrated and purified by column chromatography ESI-Ms m/z: 249.1 [M+H] ⁺ .

Step e (1R,3R)-1-(4-bromo-2,6-difluorophenyl)-2-((S)-3-fluoro-2-methylpropyl)-3-methyl-2, Synthesis of 3,4,9-tetrahydro-1H-pyrido[3,4-b]indole

In a 50 mL single-mouth bottle, 4-bromo-2,6-difluorobenzaldehyde (0.535 g, 2.42 mmol) and (S)-N-((R)-1-(1H-indol-3-yl) were added. Prop-2-yl)-3-fluoro-2-methylpropyl-1-amine (0.50 g, 2.0 mmol), added 4 mL of acetic acid and 20 mL of toluene, reacted at 90 ° C for 3 h, concentrated, purified by column chromatography Compound 0.61 g. ESI-Ms m/z: 451.1 [M+H] ⁺ .

Step f 1-(3,5-Difluoro-4-((1R,3R)-2-((S)-3-fluoro-2-methylpropyl)-3-methyl-2,3,4 Synthesis of methyl 9-tetrahydro-1H-pyrido[3,4-b]indol-1-yl)phenyl)azetidin-3-carboxylate

In a microwave reactor, (1R,3R)-1-(4-bromo-2,6-difluorophenyl)-2-((S)-3-fluoro-2-methylpropyl)-3- Methyl-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole (910 mg, 2.02 mmol), azetidine-3-carboxylic acid methyl ester hydrochloride (367 mg) , 2.42 mmol), tris(dibenzylideneacetone)dipalladium (370 mg, 0.404 mmol), 2-dicyclohexylphosphino-2,4,6-triisopropylbiphenyl (391 mg, 0.82 mmol) and cesium carbonate (2.0 g, 6.06 mmol) was added to dioxane (20 mL). After the argon gas was sufficiently replaced, the mixture was heated to 100 ° C for microwave reaction. After the reaction was completed, the mother liquor was filtered and concentrated. This reaction product was used directly in the next step. ESI-Ms m/z: 486.3 [M+H] ⁺ .

Step g 1-(3,5-Difluoro-4-((1R,3R)-2-((S)-3-fluoro-2-methylpropyl)-3-methyl-2,3,4 Synthesis of 9-tetrahydro-1H-pyrido[3,4-b]indol-1-yl)phenyl)azetidin-3-carboxylic acid

In a 50 mL single-mouth bottle, 1-(3,5-difluoro-4-((1R,3R)-2-((S)-3-fluoro-2-methylpropyl)-3-methyl-) Methyl 2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indol-1-yl)phenyl)azetidin-3-carboxylate (0.62 g, 1.307 mmol) Dissolve with 10mL of tetrahydrofuran and 5mL of methanol, add 7.5M sodium hydroxide solution (3.0mL, 22.5mmol), react at room temperature for 3h, adjust the pH to 6.5 with 2N hydrochloric acid, extract with ethyl acetate, wash with saturated sodium chloride, anhydrous sulfuric acid The sodium was dried and concentrated to give the title compound. ^{1 H NMR (400MHz, d6-} DMSO): 12.53 (s, 1H), 10.50 (s, 1H), 7.40 (d, 1H), 7.23-7.19 (m, 1H), 7.02-6.94 (m, 2H), 6.08(d,2H), 5.02(s,1H), 4.44(d,1H), 4.34(d,1H),4.01(m,2H),3.97(m,2H),2.82(dd,1H), 2.50 (m, 3H), 2.21 (m, 1H), 2.04 (m, 1H), 1.93 (m, 1H), 1.06 (d, 3H), 0.77 (d, 3H). ESI-Ms m/z: 472.5 [ M+H] ⁺ .

Example 24 1-(4-((1R,3R)-2-(2,2-Difluoropropyl)-3-methyl-2,3,4,9-tetrahydro-1H-pyrido[3 ,4-b]indol-1-yl)-3,5-difluorophenyl)azetidin-3-carboxylic acid

Step a Synthesis of 2,2-difluoro-2-methylpropyl-1-ol

In a 250 mL three-necked flask, 2,2-difluoro-2-methylpropionic acid (5 g, 45.5 mmol) was added, dissolved in 100 mL of anhydrous tetrahydrofuran, and the reactor was cooled to about -10 ° C in a low temperature reactor. Lithium aluminum (2.1 g, 54.2 mmol) was maintained at this temperature for 1 h. After the reaction was completed, 2.1 mL of water, 2.1 mL of a 15% aqueous sodium hydroxide solution, and 4.2 mL of water were added dropwise, and the mixture was stirred for 15 min, then filtered, and the filter cake was washed with a small amount of THF. ESI-Ms m/z: 97.0 [M+H] ⁺ .

Synthesis of step b(R)-N-(1-(1H-indol-3-yl)propan-2-yl)-2,2-difluoropropyl-1-amine

The preparation method is the same as the preparation method of the step cd of Example 23, except that the raw material (S)-3-fluoro-2-methylpropanol is replaced with 2,2-difluoro-2-methylpropyl-1-ol. The title compound was obtained. ESI-Ms m/z: 253.0 [M+H] ⁺ .

Step c 1-(4-((1R,3R)-2-(2,2-Difluoropropyl)-3-methyl-2,3,4,9-tetrahydro-1H-pyrido[3, Synthesis of 4-b]non-1-yl)-3,5-difluorophenyl)azetidin-3-carboxylic acid

The preparation method was similar to the preparation method of the procedure of Example 23, except that the starting material (S)-N-((R)-1-(1H-indol-3-yl)propan-2-yl)-3- Replacement of fluoro-2-methylpropyl-1-amine with (R)-N-(1-(1H-indol-3-yl)propan-2-yl)-2,2-difluoropropyl-1 -Amine to give the title compound. ^{1 H NMR (500MHz, DMSO-} d 6) δ: 12.5 (s, 1H), 10.53 (s, 1H), 7.42-7.40 (d, 1H), 7.23-7.21 (d, 1H), 7.03-6.95 (m , 2H), 6.14-6.12 (d, 2H), 5.12 (s, 1H), 4.07-4.03 (m, 2H), 3.93-3.90 (m, 2H), 3.57-3.53 (m, 1H), 3.50-3.46 (m, 1H), 3.10-3.01 (m, 1H), 2.91-2.88 (m, 1H), 2.61-2.57 (m, 2H), 1.50-1.43 (t, 3H), 1.11-1.01 (d, 3H) .ESI-Ms m/z: 476.20 [M+H] ⁺ .

According to the synthesis method of Examples 1-24 of the present invention, the compounds of Examples 25-43 were synthesized using different commercially available starting materials, and the characterization parameters of these compounds are shown in Table 1:

Table 1:

Experimental Example 1 Evaluation of ER level activity of compounds based on cell level in vitro

Experimental material

The control compound is the chemical name (E)-3-(3,5-difluoro-4-((1R,3R)-2-)) disclosed in Example 1 of WO 2014/191726 (PCT/GB2014/051607). -fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indol-1-yl)phenyl)acrylic acid The compound (AZD9496) was prepared by the method described in WO 2014/191726 and identified by hydrogen spectroscopy and mass spectrometry.

Reagents: phosphate buffer (DPBS), trypan blue, PolarScreen ER Alpha competitor Assay, purchased from Invitrogen;

Fetal bovine serum (FBS), trypsin, DMEM, penicillin-streptomycin (Pen/Strep), purchased from GIBCO;

Dimethyl sulfoxide (DMSO), activated carbon, Formaldehyde solution, purchased from Sigma;

MCF-7 cells, purchased from AATCC;

Estrogen Receptor alpha (D8H8) Rabbit mAb, purchased from CST;

Goat anti-Rabbit IgG(H+L)Secondary Antibody,Alexa

488conjugate, purchased from Thermo Corporation;

Tween 20, purchased from EIA GRADE.

Instruments: Biosafety cabinet, CO ₂ incubator, purchased from Thermo Scientific;

Centrifuge, purchased from Eppendorf;

Cell counter, purchased from Invitrogen;

Inverted microscope, purchased from Olympus;

Multiflow, purchased from BioTeck;

Vortex mixer, purchased from IKA;

Envision, purchased from Perkin Elmer.

2. Experimental methods

2.1. Preparation of cell culture fluids and compounds

Preparation of serum-free FBS: Weigh 1g of activated carbon and 50mL fetal bovine serum at 4 ° C for 24h, then filter through 0.22μM filter, spare;

Preparation of cell culture medium: 50 mL of FBS, 5 mL of penicillin-streptomycin was added to 445 mL of DMEM, and mixed for use. Serum-free FBS was used in the preparation of serum-free cell culture media.

Compound Preparation: The compound of the present invention and the control compound prepared in the above examples, each compound was formulated into 100 mM with DMSO, and sequentially diluted to 10 nM, 3.33 nM, 1.11 nM, 0.37 nM, 0.123 nM, 0.041 nM, 0.014 nM, 0.0045. nM, 0.0015 nM, 0.0005 nM.

2.2. Inoculation of cells

The cell-discarded culture medium in the logarithmic growth phase of the T75 cell culture flask was washed once with 10 mL of DPBS. Add 2 mL of trypsin-digested cells, and place at 37 ° C for 2 minutes. Observe most of the cells in a round shape under microscope. Add 5 mL of serum-free cell culture solution to stop digestion. Pipette repeatedly and digest the cells to make cell suspension. Add 10 mL of cell culture solution, mix and count, dilute to 1500 cells/40 μL of cell suspension, and place the cells into a 384-well cell culture plate with a Multiflow instrument, 40 μL/well; equilibrate at room temperature for 20 min. Incubate in a 37 ° C cell culture incubator for 24 h.

2.3. Adding compounds

The compound was added to the cell culture plate with Acho instrument at a final concentration of 0.3% DMSO; it was centrifuged at 1000 rpm for 1 min at room temperature, and then cultured in a 37 ° C cell culture incubator for 24 h.

2.4. Immunofluorescence experiment

The cell culture medium was aspirated, the cells were washed once with PBS, and the cells were fixed with a final concentration of 3.7% paraformaldehyde solution (diluted in PBS) for 20 min at room temperature; the cells were washed twice with PBS and diluted with a final concentration of 0.5% Tween-20 (PBS). Penetrate for 1 h at room temperature; wash cells twice with PBS-T (0.05% Tween-20 in PBS), add ER antibody dilution (1:1000 in ER level assay, dilute with 1% milk in PBS-T) Incubate for 1.5 h at room temperature; wash cells three times with PBS-T, add a secondary antibody dilution (1:1000, diluted with 1% milk in PBS) 2 μg/mL of Hochest 33342, incubate for 40 min at room temperature; PBS-T wash The cells were washed 3 times and PBS was washed twice; Acumen read the ratio of ER positive signal value to nuclear signal value. The experimental results are shown in Table 2.

Table 2

"-" means untested

As can be seen from the above experimental results, the compounds of the present invention have a good inhibitory activity against ER levels based on cellular levels.

Experimental Example 2 Evaluation of in vitro cell viability of compounds

Experimental material

Test compound: The compound of the present invention and the control compound prepared in the above examples, each compound was formulated into 10 mM with DMSO, and then diluted 3 times to 100.00 nM, 33.33 nM, 11.11 nM, 3.70 nM, 1.23 nM, 0.41 nM, 0.14 nM, 0.045 nM, 0.015 nM.

The breast cancer cell line MCF-7 was purchased from Nanjing Kaiji Biotechnology Co., Ltd.

Reagents: MEM, FBS, Trypsin-EDTA, Penicillin-Streptomycin, purchased from GIBCO, USA;

Luminescent Cell Viability Assay Kit, purchased from Progema, USA; Paclitaxel, purchased from Sichuan Tai Chi Pharmaceutical Company.

2. Experimental methods

2.1. Cell seeding

Cultured expanded MCF-7 cells were trypsinized, resuspended in fresh medium and counted. The resuspended cells were adjusted to 2 × 10 ⁴ cells/mL, and 96-well cell culture plates were added, and 100 μL of each well was added to each well. Incubate for 24 h at 37 ° C, 5% CO ₂ .

2.2. Adding compounds

The compound was formulated into 200× working solution in DMSO, diluted to 2× working solution with the culture solution, and then transferred to 100 μL to the test well, and incubated at 37° C. under 5% CO ₂ for 96 h.

2.3. Fluorescence readings

Add 50 μL to the hole to be tested

Luminescent Cell Viability Assay buffer and shake gently. After 10 minutes, the fluorescence reading was read on the Envison and the cell survival rate (cell survival (%)) was calculated as the cell survival (%) = (Com-Min) / (Max-Min), where Max is For the control of the vehicle control group, Min is the reading of the cell-free control group, Com is the reading of the compound treatment group, and the data is processed by XLfit, and the IC _{50 is} fitted. The experimental results are shown in Table 3.

table 3

"-" means untested

As can be seen from the above experiments, the compounds of the present invention showed good inhibitory activity against MCF-7 breast cancer cells.

Experimental Example 3 In vivo pharmacokinetic evaluation of compounds

Test compound: The compound of the present invention and the control compound prepared in the above examples were prepared in an amount of 2 mg/kg for oral administration and 1 mg/kg for the test sample.

Balb/c mice were purchased from Beijing Weitong Lihua Experimental Animal Co., Ltd.

The mice were orally administered with 2 mg/kg, and after a single dose of 1 mg/kg intravenously, blood was collected from the orbital venous plexus at 2 min, 5 min, 15 min, 30 min, 1 h, 2 h, 6 h, 10 h, 24 h, and centrifuged for plasma treatment. The LC-MS/MS was used for the detection, and the measured blood concentration at each time point was plotted as a drug concentration-time curve, and the pharmacokinetic parameters were calculated. The experimental results are shown in Table 4.

Table 4

受试化合物Test compound	T _1/2(h) T _1/2 (h)	Cmax(ng/ml)Cmax(ng/ml)	AUC(hng/ml)AUC (hng/ml)	F(％)F (%)
AZD9496AZD9496	1.11.1	703.7703.7	1229.91229.9	45.545.5
实施例1Example 1	12.112.1	1277.71277.7	16073.716073.7	80.580.5
实施例5Example 5	7.67.6	1210.71210.7	13602.113602.1	79.079.0
实施例6Example 6	8.98.9	900.6900.6	12634.712634.7	67.567.5
实施例12Example 12	12.412.4	1148.21148.2	15699.915699.9	88.288.2
实施例15Example 15	6.66.6	617.0617.0	7626.27626.2	75.375.3
实施例16Example 16	6.76.7	439.2439.2	5737.05737.0	71.071.0
实施例18Example 18	4.44.4	140.3140.3	798.1798.1	24.324.3
实施例19Example 19	15.415.4	1519.51519.5	22055.422055.4	76.176.1
实施例20Example 20	4.34.3	403.1403.1	3563.23563.2	51.251.2
实施例23Example 23	1.31.3	647647	1948.91948.9	58.158.1
实施例24Example 24	4.34.3	410.2410.2	2413.52413.5	86.686.6

The above experimental results show that the half-life (T _1/2 ), peak concentration (Cmax), area under the curve (AUC) and bioavailability (F) of the compounds of the present invention are significantly superior to the control compound AZD9496. When the half-life of the compound is prolonged and the bioavailability is increased, the administration interval of the compound is prolonged, and the drug action time is prolonged, thereby improving the therapeutic effect of the compound and reducing the dose of the compound, thereby making the drug more effective and safer. Therefore, the compound of the present invention has more excellent antitumor activity in vivo, and the administration interval will be longer.

Experimental Example 4 Evaluation of the efficacy of compound in vivo MCF-7 subcutaneous xenograft model

Cell culture

MCF-7 breast cancer cells (purchased from KGI) were cultured in an MEM medium containing 10% fetal calf serum, 100 U/ml penicillin and 100 μg/ml streptomycin in a 37 ° C, 5% CO 2 incubator. The initial concentration of the cell culture was 1×10 ⁶ /mL, and the cells were subcultured every 3 to 4 days after the cells were over. Tumor cells in the logarithmic growth phase are used for inoculation of tumors in vivo.

2. Sustained release tablets and cell inoculation

Two to three days before cell inoculation, a beta-estradiol sustained release tablet (Estradiol-17β 60 days SE121 0.72 mg, purchased from Innovative Research of America) was inoculated into the left back of each mouse. One week after the inoculation, the animals were urinated three times a week, and if necessary, the animals were urinated daily.

A PBS containing 10 × 10 ⁶ cells was mixed with 100 μL of Matrigel (final volume 200 μL) to inoculate the right rear side of the mouse. 3. Tumor measurement and experimental indicators

Test compound: The compound of the present invention and the control compound prepared in the above examples. The compound of the present invention and the control compound were administered once daily (QD) for three weeks (3 W).

The experimental indicator is to investigate whether tumor growth is inhibited, delayed or cured. Tumor diameters were measured with vernier calipers three times a week. The calculation formula of tumor volume is: V = 0.5a × b ² , and a and b represent the long diameter and short diameter of the tumor, respectively.

The antitumor effect of the test compound was evaluated by relative tumor growth rate T/C (%) and tumor inhibition rate (%). The relative tumor growth rate T/C (%) was calculated as follows: T/C% = T _RTV / C _RTV × 100% (T _RTV : treatment group RTV; C _RTV : negative control group RTV). Relative tumor volume (RTV) was calculated according to the results of tumor measurement, and the formula was calculated as RTV=V _t /V ₀ , where V ₀ is the tumor volume measured at the time of group administration (ie, d ₀ ), and V _t is The tumor volume at one measurement, T _RTV and C _RTV took the same day data. Tumor inhibition rate (%) = (1-T/C) × 100%.

The experimental results of the antitumor efficacy evaluation of the test compound on the human breast cancer cell line MCF-7 subcutaneous xenograft model are shown in Table 5. The corresponding T/C (%) values of the compounds of the examples in the table are the respective examples. The T/C (%) mean of the tumor volume calculated on the 21st day after administration of the animals in each dose group of the compound.

table 5

The experimental results show that, unexpectedly, the compound of the present invention has a significantly better antitumor effect against breast cancer than the positive drug AZD9496. The compounds of Example 1, Example 5 and Example 24 of the present invention were significantly more effective than the same dose of AZD9496 at a dose of 0.6 mg/kg, and were more effective at a dose of 2 mg/kg than AZD9496 at a dose of 6 mg/kg. Drug effect. Some of the compounds of the present invention produced an effect equivalent to the positive drug AZD9496 (0.6 mg/kg VS 6 mg/kg) at a tenth dose.

While the invention has been described hereinabove, it will be understood by those skilled in the art that various modifications and changes of the invention may be made without departing from the spirit and scope of the invention. The scope of the invention is not limited to the details described above, but rather to the claims.

Claims

a compound of the formula I or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal or a prodrug,

among them,

Each of R 1 and R 2 is independently selected from the group consisting of hydrogen, halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxy, cyano, amino, a monoalkylamino group, an alkyl acylamino group, an alkyl acyl group, an amino acyl group, an alkylamino acyl group, a dialkylamino group, and a cycloalkyl group;

R 3 and R 9 are each independently selected from the group consisting of hydrogen, alkyl acyl, amino acyl, alkyl amino acyl, alkyl, haloalkyl, hydroxyalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, heteroaryl. Base and aryl group;

R 4 , R 5 , R 6 and R 7 are each independently selected from the group consisting of hydrogen, halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxy, a cyano group, an amino group, a monoalkylamino group, an alkyl acylamino group, an alkyl acyl group, an amino acyl group, an alkylamino acyl group, and a dialkylamino group; or R 4 and R 5 together with the carbon atom to which they are attached form a carbonyl group, a ring An alkyl group; or R 6 , R 7 together with the carbon atom to which they are attached form a carbonyl group, a cycloalkyl group; or R 4 , R 6 together with the carbon atom to which they are attached form a cycloalkyl group;

Each R 8 is independently selected from the group consisting of hydrogen, halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxy, cyano, amino, monoalkyl An amino group, an alkyl acylamino group, an alkyl acyl group, an amino acyl group, an alkylamino acyl group, a bisalkylamino group, a cycloalkyl group, and a boronic acid;

X, Y, Z, W are each independently selected from N and C(R 10 ), wherein R 10 is selected from the group consisting of hydrogen, halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxy Alkoxy, nitro, carboxy, cyano, amino, monoalkylamino, alkyl acylamino, alkyl acyl, amino acyl, alkylamino acyl, bisalkylamino and cycloalkyl;

Chemical bond
Independently
And

m, n, and o are independently 1, 2, 3, or 4, respectively.
The compound according to claim 1 or an isomer thereof, a pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein X, Y, Z and W are all C(R 10 ).
The compound according to claim 1 or an isomer thereof, a pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein each of R 1 and R 2 is independently selected from the group consisting of hydrogen, fluorine, chlorine, bromine and iodine. , hydroxy, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkoxy, hydroxy C 1-6 Alkoxy, nitro, carboxyl, cyano, amino, mono C 1-6 alkylamino, C 1-6 alkyl acylamino, C 1-6 alkyl acyl, amino acyl, C 1-6 alkylamino An acyl group, a di C 1-6 alkylamino group and a C 3-10 cycloalkyl group, and R 3 and R 9 are each independently selected from hydrogen, C 1-10 alkyl acyl group, amino acyl group, C 1-10 alkylamino acyl group. , C 1-10 alkyl, halo C 1-10 alkyl, hydroxy C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl, C 3-10 cycloalkyl, C 3- a 10 heterocyclic group, a C 6-18 aryl group and a C 5-18 heteroaryl group, which group may be one or more halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy , hydroxyalkoxy, nitro, carboxy, cyano, amino, monoalkylamino, dialkylamino, cycloalkyl, heterocyclyl, aryl and Heteroaryl substitution.
The compound according to any one of claims 1 to 3, or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein R 4 , R 5 , R 6 and R 7 are each independently Selected from hydrogen, fluorine, chlorine, bromine, iodine, hydroxyl, C 1-6 alkyl, halogenated C 1-6 alkyl, hydroxy C 1-6 alkyl, C 1-6 alkoxy, halogenated C 1 -6 alkoxy, hydroxy C 1-6 alkoxy, nitro, carboxy, cyano, amino, mono C 1-6 alkylamino, C 1-6 alkyl acylamino, C 1-6 alkyl acyl An aminoacyl group, a C 1-6 alkylamino acyl group and a bis C 1-6 alkylamino group.
A compound according to any one of claims 1 to 3, or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein R 4 and R 5 are taken together with the carbon atom to which they are attached a carbonyl group or a C 3-10 cycloalkyl group; or R 6 , R 7 together with the carbon atom to which they are attached form a carbonyl group or a C 3-10 cycloalkyl group; or R 4 , R 6 together with the carbon atom to which they are attached C 3-10 cycloalkyl.
The compound according to any one of claims 1 to 5, or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein each R 8 is independently selected from the group consisting of hydrogen, halogen, hydroxyl, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkoxy, hydroxy C 1-6 alkoxy , nitro, carboxyl, cyano, amino, mono C 1-6 alkylamino, C 1-6 alkyl acylamino, C 1-6 alkyl acyl, amino acyl, C 1-6 alkylamino acyl, double C 1-6 alkylamino, C 3-10 cycloalkyl and boric acid.
The compound according to claim 1 or an isomer thereof, a pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein R 10 is selected from the group consisting of hydrogen, halogen, hydroxy, C 1-6 alkyl, halogen C 1-6 alkyl, hydroxy C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkoxy, hydroxy C 1-6 alkoxy, nitro, carboxy, cyano, amino , mono C 1-6 alkylamino, C 1-6 alkyl acylamino, C 1-6 alkyl acyl, amino acyl, C 1-6 alkylamino acyl, bis C 1-6 alkylamino and C 3 -10 cycloalkyl.
A compound according to claim 1 or an isomer thereof, a pharmaceutically acceptable salt, solvate, crystal or prodrug, wherein the compound is a compound selected from the group consisting of:
A pharmaceutical composition comprising the compound of any one of claims 1 to 8, or an isomer thereof, a pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, and a pharmaceutically acceptable carrier.
The compound of any one of claims 1-8, or an isomer thereof, a pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, or the pharmaceutical composition of claim 9 for use in the treatment and/or Or the use of drugs to prevent estrogen receptor-related diseases.