CN111233661B

CN111233661B - Compound for targeted ubiquitination degradation of ERR alpha protein and medicinal composition and application thereof

Info

Publication number: CN111233661B
Application number: CN201811445971.1A
Authority: CN
Inventors: 丁克; 彭丽洁; 张振声; 任小梅; 张章; 雷冲; 李姗
Original assignee: Jinan University
Current assignee: Jinan University
Priority date: 2018-11-29
Filing date: 2018-11-29
Publication date: 2023-03-31
Anticipated expiration: 2038-11-29
Also published as: CN111233661A

Abstract

The invention provides a compound with a structure shown in a formula (I), which has the effects of inhibiting ERR alpha protein activity and degrading ERR alpha protein activity, has stronger subtype selectivity, and can also effectively inhibit triple negative breast cancer MDA-MB-231 cell migration. Therefore, the invention can be used for diseases related to the abnormal expression of ERR alpha protein, such as various cancers.

Description

Compound for targeted ubiquitination degradation of ERR alpha protein and medicinal composition and application thereof

Technical Field

The invention relates to the field of pharmaceutical chemistry, in particular to a compound for targeted ubiquitination degradation of ERR alpha protein, a pharmaceutical composition thereof and application thereof.

Background

The breast cancer is a tumor with strong heterogeneity, and researchers have clearer and systematic cognition on the molecular complexity of the breast cancer along with the development and common application of technologies such as genomics, epigenomics, transcriptomics and proteomics. Despite the complex mechanisms, breast cancer is clinically classified and treated mostly by the expression degree of Estrogen Receptor (ER), progesterone receptor (PgR) and human epidermal growth factor receptor 2 (human epidermal growth factor receptor-2, herr 2/ErbB 2). Wherein, clinically effective hormone treatment means including aromatase inhibitor, selective estrogen regulator and the like can be adopted for ER +, pgR + or/and HER2+ type breast cancer. Especially for HER2 high-expression breast cancer, the chemical drugs Lapatinib, the biological drugs pertuzumab and trastuzumab which are on the market have become the treatment first choice. However, about 15-20% of breast cancer cases worldwide are currently diagnosed with triple-negative breast cancer (TNBC, i.e., ER-, pgR-, HER 2-), because TNBC is not suitable for endocrine therapy and drug therapy for HER2 targets, and is highly malignant and has poor prognosis. Therefore, there is a great clinical need to develop a strategy with different mechanisms of action to treat TNBC.

Cell metabolic reprogramming is an important marker of cancer cells and plays a crucial role in the occurrence and development of tumors. With the intensive research on the energy metabolism of tumor cells, targeted 'metabolic reprogramming' becomes a new direction for the research on anti-tumor drugs.

Estrogen-related receptors (ERRs) are members of the nuclear receptor superfamily, which are known for their homology to Estrogen Receptors (ERs). Including three types ERR alpha, ERR beta and ERR gamma. ERR β is associated with early development, and ERR α and ERR γ are considered as new potential targets for metabolic disorders. Where ERR α is the first most discovered and studied member of this subgroup. The derivative is widely expressed in tissues with vigorous energy metabolism, such as heart, kidney, skeletal muscle, gastrointestinal tract and the like, is directly combined with peroxisome proliferator-activated receptor coactivators (peroxisome proliferator-activated receptor-gamma coactivator 1, PGC-1) to regulate the expression levels of regulatory factors in the processes of tricarboxylic acid cycle, glycolysis and the like, such as PGC-1, middle-chain acyl-coenzyme A dehydrogenase (MCAD), pyruvate dehydrogenase kinase (PDK 4) and the like, thereby playing an important role in maintaining metabolic energy homeostasis. As a major metabolic regulator, the ERR (α)/PGC-1 (β) transcriptional axis mediates metabolic adaptation in various types of cancer cells. For example, alterations in metabolism driven by ERR α can significantly enhance proliferation, migration, angiogenesis, metastasis and drug resistance of human breast cancer cells. Research shows that the inhibition of ERR alpha can obviously inhibit the development of breast cancer. Furthermore, ERR α overexpression is often detected in Her2+ and triple negative breast tumors, and the clinical prognosis of breast cancer patients is closely related. Therefore, ERR alpha is a potential new target for developing anti-tumor drugs, and has potential application value in the aspects of enhancing the research on the target and developing compounds related to the target in personalized treatment of cancers.

Traditional drug development, including the development of ERR α small molecule drugs described above, has focused primarily on directly regulating protein or enzyme activity. The development and use of modulators, particularly inhibitors, of protein activity has been the mainstay of drug development. Small molecule inhibitors exert their protein activity modulating effects by binding to the active site, and belong to the "occupancy-driven" model. However, small molecule inhibitors have several disadvantages: firstly, the long-term use of small molecule drugs can generate drug resistance; secondly, in order to achieve the required effect, the small molecular compound needs to keep higher concentration in the cell, thereby causing off-target and generating adverse reaction; meanwhile, many potential target proteins do not have pockets capable of being directly combined by small molecule inhibitors, such as non-enzyme proteins like transcription factors, and the development of the small molecule inhibitors is difficult.

Ubiquitin-mediated protein degradation is the predominant negative regulatory mode of intracellular proteins. The ubiquitin-proteasome system (UPS) is responsible for cleaning unwanted or harmful proteins from cells, is an intracellular "cleaner" responsible for cleaning defective proteins from cells. Proteolytic targeting chimeras (protein-targeting chimeras) are essentially hybrid bifunctional small molecule compounds with a mode of action different from the "occupancy-driven" mode of traditional small molecule inhibitors, and PROTACs can specifically degrade target proteins by using the ubiquitin-proteasome pathway. One end of the molecule is combined with target protein, the other end is combined with E3 ubiquitin ligase of ubiquitin-protease system, ubiquitination target protein is marked on the target protein, and finally the ubiquitination target protein is hydrolyzed by proteasome. The process belongs to an "event driven" mode. The PROTACS molecules in "event-driven" mode have multiple cycles to remove multiple stoichiometric ratios of the target protein compared to traditional small molecule inhibitors; after degradation, the target protein can only restore the protein function through resynthesis; the effect can be achieved by binding any pocket without occupying an active pocket.

The first batch of bifunctional molecules of PROTACs was reported in the desheies group 2001, the initial technology was the short peptide-based PROTAC technology, and the targets for successful degradation included MetAP2, androgen receptor, etc. The related reported PROTAC are polypeptide compounds, and the compounds have the defects of large molecular weight, poor cell permeability, poor activity and the like, so the development of the technology is limited. In recent years, the discovery of E3 ubiquitin ligase specific small molecule ligands such as CRBN, VHL, cIAP and the like makes the PROTACs technology have great breakthrough. The first small molecule PROTAC was reported by the Crew topic group in 2008: PROTAC _ AR. The PROTAC _ AR connects the MDM2E3 ubiquitin ligase small molecule inhibitor nutlin with the nonsteroidal androgen receptor small molecule ligand through a polyethylene glycol linker, thereby successfully realizing AR degradation in the HeLa cells of the human cervical carcinoma. In the following years, researchers of various research institutions respectively develop CRABP-II _ PROTAC based on a claP1E3 ubiquitin ligase ligand, BRD4_ PROTAC and Bcr-Abl _ PROTAC based on a CRBN E3 ubiquitin ligase ligand, ERR alpha _ PROTAC and BRD4_ PROTAC based on a VHL ubiquitin ligase ligand, and successfully realize the effective degradation of related target proteins.

Therefore, inverse agonists with stronger activity to ERR α proteins were synthesized by design, and the proteolytic targeting chimeras (PROTACs) technique was applied to ERR α proteins. Can provide important reference for ERR alpha as a drug development target, the treatment of triple negative breast cancer and the development and application feasibility of PROTAC technology.

Disclosure of Invention

In view of the above problems, it is an object of the present invention to provide a novel compound for targeted ubiquitination degradation of ERR α protein, or a pharmaceutically acceptable salt, solvate, isomer, metabolite or prodrug thereof.

Based on the above purpose, the invention provides the following technical scheme:

a compound having the structure of formula (i) or a pharmaceutically acceptable salt or stereoisomer or prodrug molecule thereof:

/>

wherein, R1 is selected from:

1)H；

2) A halogen;

3)C _1～ C ₃ an alkyl group;

4)C ₃ ～C ₆ a cycloalkyl group;

5)C ₁ ～C ₃ an alkoxy group;

6) A fluoromethyl group;

7) A cyano group;

l is optionally selected from:

1)–NH–(CH ₂ ) _m –；

2)–NH–((CH ₂ ) ₂ –O) _n –(CH ₂ ) ₂ –；

3)–NH–(CH ₂ ) _u –O–(CH ₂ ) _v –

wherein m is 0,1, 2, 3, 4, 5, 6, 7 or 8;

n is 0,1, 2, 3, 4, 5 or 6;

u is 0,1, 2, 3, 4, 5, 6, 7 or 8;

v is 0,1, 2, 3, 4, 5, 6, 7 or 8;

4) L is absent

Y is optionally selected from:

1)–CH ₂ –,–C(＝O)–；

2) Y is absent;

b is optionally selected from:

1)H；

2)OR ₂ (ii) a R2 is optionally selected from: h, C ₁ ～C ₃ Alkyl radical, C ₃ ～C ₆ A cycloalkyl group;

3)

r3 is optionally selected from: h, C ₁ ～C ₃ Alkyl radical, C ₃ ～C ₆ A cycloalkyl group.

Another object of the present invention is to provide the use of a compound having the structure of formula (i) or a pharmaceutically acceptable salt thereof or a stereoisomer thereof or a prodrug molecule thereof, specifically as follows:

the use of a compound having the structure of formula (i) or a pharmaceutically acceptable salt or stereoisomer thereof, or a prodrug molecule thereof, in the preparation of an ERR α protein inhibitor:

wherein R1 is selected from:

1)H；

2) Halogen;

3)C _1～ C ₃ an alkyl group;

4)C ₃ ～C ₆ a cycloalkyl group;

5)C ₁ ～C ₃ an alkoxy group;

6) A fluoromethyl group;

7) A cyano group;

l is optionally selected from:

1)–NH–(CH ₂ ) _m –；

2)–NH–((CH ₂ ) ₂ –O) _n –(CH ₂ ) ₂ –；

3)–NH–(CH ₂ ) _u –O–(CH ₂ ) _v –

wherein m is 0,1, 2, 3, 4, 5, 6, 7 or 8;

n is 0,1, 2, 3, 4, 5 or 6;

u is 0,1, 2, 3, 4, 5, 6, 7 or 8;

v is 0,1, 2, 3, 4, 5, 6, 7 or 8;

4) L is absent;

y is optionally selected from:

1)–CH ₂ –,–C(＝O)–；

2) Y is absent;

b is optionally selected from:

1)H；

3)

In some of these embodiments, R ₁ Is optionally selected from: halogen, fluoromethyl, cyano.

In some of these embodiments, L is selected from:

1)–NH–(CH ₂ ) _m –；

2)–NH–((CH2) ₂ –O) _n –(CH ₂ ) ₂ –；

3)–NH–(CH ₂ ) _u –O–(CH ₂ ) _v –

wherein m is 0,1, 2, 3, 4, 5, 6, 7 or 8;

n is 0,1, 2, 3, 4, 5 or 6;

u is 0,1, 2, 3, 4, 5, 6, 7 or 8;

v is 0,1, 2, 3, 4, 5, 6, 7 or 8;

in some of these embodiments, Y is selected from: -CH2-, -C (= O) -.

In some embodiments, B is selected from: OR (OR) ₂ Or

R ₂ Is optionally selected from: h, C ₁ ～C ₃ Alkyl radical, C ₃ ～C ₆ A cycloalkyl group; r is ₃ Is optionally selected from: h, C ₁ ～C ₃ Alkyl radical, C ₃ ～C ₆ A cycloalkyl group.

In some of these embodiments, the compound has the structure of formula (ii):

l is optionally selected from:

1)–NH–(CH ₂ ) _m –；

2)–NH–((CH ₂ ) ₂ –O) _n –(CH ₂ ) ₂ –；

3)–NH–(CH ₂ ) _u –O–(CH ₂ ) _v –

wherein m is 0,1, 2, 3, 4, 5, 6, 7 or 8;

n is 0,1, 2, 3, 4, 5 or 6;

u is 0,1, 2, 3, 4, 5, 6, 7 or 8;

v is 0,1, 2, 3, 4, 5, 6, 7 or 8;

4) L is absent;

y is optionally selected from:

1)–CH ₂ –,–C(＝O)–；

2) Y is absent;

b is optionally selected from:

1)H；

3)

In some of these embodiments, the compound has the structure of formula (iii):

l is optionally selected from:

1)–NH–(CH ₂ ) _m –；

2)–NH–((CH ₂ ) ₂ –O) _n –(CH ₂ ) ₂ –；

3)–NH–(CH ₂ ) _u –O–(CH ₂ ) _v –

wherein m is 0,1, 2, 3, 4, 5, 6, 7 or 8;

n is 0,1, 2, 3, 4, 5 or 6;

u is 0,1, 2, 3, 4, 5, 6, 7 or 8;

v is 0,1, 2, 3, 4, 5, 6, 7 or 8;

In some of these embodiments, the compound is selected from:

(Z) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-fluoroacrylic acid;

(Z) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2- (trifluoromethyl) acrylic acid;

(E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanoacrylic acid;

(Z) -methyl 3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-fluoroacrylate;

(Z) -methyl 3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2- (trifluoromethyl) acrylate;

(E) -methyl 3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanoacrylate;

(Z) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-fluoroacrylamide;

(Z) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2- (trifluoromethyl) acrylamide;

(E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanoacrylamide;

(Z) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -N-hydroxy-2- (trifluoromethyl) acrylamide;

(Z) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-fluoro-N-hydroxyacrylamide;

(E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyano-N-hydroxyacrylamide;

(E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyano-N- (2-methoxyethyl) acrylamide;

(E) -5- (3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanoacrylamido) pentanoic acid;

(E) -3- (2- (3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanoacrylamido) ethoxy) propanoic acid;

(2s, 4r) -1- ((S) -2- (3- ((E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanoacrylamido) propionylamino) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide;

(2s, 4r) -1- ((S) -2- (4- ((E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanoacrylamide) butyrylamino) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide;

(2s, 4r) -1- ((S) -2- (6- ((E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanopropionylamino) hexanamide) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide;

(2s, 4r) -1- ((S) -2- (3- ((E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanopropionylamino) propionylamino) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide;

(2s, 4r) -1- ((S, E) -16- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2- (tert-butyl) -15-cyano-4, 14-dioxo-7, 10-dioxa-3, 13-diazahexadec-15 enoyl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide;

(2S, 4R) -1- ((S, E) -19- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2- (tert-butyl) -18-cyano-4, 17 dioxo-7, 10, 13-trioxa-3, 16-diaza-nona-18 enoyl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide;

(2s, 4r) -1- ((S) -2- (((2- ((E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanopropionylamino) ethoxy) methyl) amino) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide;

(2s, 4r) -1- ((S) -2- ((4- ((E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanopropionylamino) butyl) amino) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide;

(2s, 4r) -1- ((S) -2- (7- ((E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanopropionylamino) heptanamido) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide;

(2S, 4R) -1- ((S) -2- (8- ((E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanopropionylamino) octan) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide.

In some of these embodiments, the compound is selected from:

(2S, 4R) -1- ((S, E) -16- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2- (tert-butyl) -15-cyano-4, 14-dioxo-7, 10-dioxa-3, 13-diaza-hexadeca-15 enoyl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide;

The application of the compound or the pharmaceutically acceptable salt or the stereoisomer or the prodrug molecule thereof in preparing the medicine for preventing or treating the diseases related to the abnormal expression of the ERR alpha protein activity.

In some of these embodiments, the diseases associated with aberrant expression of ERR α protein activity include: tumor, hyperglycemia, diabetes, obesity, hyperlipidemia, hypercholesterolemia, hyperlipoproteinemia, hypertriglyceridemia, hypertension, hyperinsulinemia, hyperuricemia, parkinson disease, and Alzheimer disease.

The compound or the pharmaceutically acceptable salt or the stereoisomer or the prodrug molecule thereof can be applied to the preparation of the drugs for treating or preventing tumors or preventing the postoperative recurrence of the tumors.

In some of these embodiments, the tumor is: non-small cell lung cancer, malignant melanoma, prostate cancer, renal cancer, bladder cancer, ovarian cancer, colon cancer, rectal cancer, breast cancer, cervical cancer, lung cancer, larynx cancer, nasopharyngeal carcinoma, pancreatic cancer, or multiple myeloma, B lymphoma, and leukemia.

The invention also aims to provide an ERR alpha protein inhibitor, which has the following specific technical scheme:

an ERR alpha protein inhibitor contains the compound or the pharmaceutically acceptable salt or stereoisomer or prodrug molecules thereof as an active ingredient.

The invention also aims to provide a specific technical scheme of the medicine for treating or preventing the tumor or preventing the postoperative recurrence of the tumor, which comprises the following steps:

the active ingredient of the medicine for treating or preventing tumor or preventing postoperative recurrence of tumor contains the compound or pharmaceutically acceptable salt or stereoisomer or prodrug molecule thereof.

Based on the technical scheme, the invention has the following beneficial effects:

the compound provided by the invention has the activities of inhibiting ERR alpha protein and degrading ERR alpha protein, has stronger subtype selectivity, and can also effectively inhibit the MDA-MB-231 cell migration of triple negative breast cancer. Therefore, the invention can be used for diseases related to the abnormal expression of ERR alpha protein, such as various cancers.

The compound can effectively inhibit ERR alpha protein and has the function of degrading ERR alpha target protein. The protein degradation mechanism is that one end of the molecule is combined with ERR alpha target protein, the other end is combined with E3 ubiquitin ligase of ubiquitin-protease system, ubiquitination target protein, and finally, degradation of ERR alpha target protein is successfully realized through proteasome. The compound obviously inhibits the migration of tumor cells, has good subtype selection effect, and has no obvious inhibition effect or degradation effect on ERR beta and ERR gamma. The compound provides important references for research and development of ERR alpha as a drug development target, treatment of triple negative breast cancer and development and application feasibility of PROTAC technology.

Drawings

FIG. 1 shows the result of measurement of ERR α protein level in MDA-MB-231 cells;

FIG. 2 shows the results of experiments in which compounds inhibited MDA-MB-231 cell migration.

Detailed Description

In the compounds of the invention, when any variable (e.g. R) ₁ 、R ₂ Etc.) occur more than one time in any constituent, then the definition of each occurrence is independent of the definition of each other occurrence. Also, combinations of substituents and variables are permissible only if such combinations result in stable compounds. The line drawn from a substituent into the ring system indicates that the indicated bond can be attached to any ring atom that can be substituted. If the ring system is polycyclic, it means that such a bond is only attached to any suitable carbon atom of the adjacent ring. It is understood that one of ordinary skill in the art can select a compound of the inventionThe substituents and substitution patterns of (a) provide compounds that are chemically stable and can be readily synthesized by techniques in the art and methods set forth below from readily available starting materials. If a substituent is itself substituted with more than one group, it is understood that these groups may be on the same carbon atom or on different carbon atoms, so long as the structure is stable. The phrase "optionally substituted with one or more substituents" is considered equivalent to the phrase "optionally substituted with at least one substituent" and preferred embodiments in this case will have from 0 to 3 substituents.

The terms "alkyl" and "alkylene" as used herein are intended to include both branched and straight chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms. For example, "C _1-5 Alkyl radical "middle" C _1-5 The definition of "includes groups having 1, 2, 3, 4, or 5 carbon atoms in a straight or branched chain arrangement. For example, "C _1-5 Alkyl "specifically includes methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, pentyl. The term "cycloalkyl" refers to a monocyclic saturated aliphatic hydrocarbon group having the specified number of carbon atoms. For example, "cycloalkyl" includes cyclopropyl, methyl-cyclopropyl, 2-dimethyl-cyclobutyl, 2-ethyl-cyclopentyl, cyclohexyl, and the like.

The term "heterocycle" or "heterocyclyl" as used herein refers to a 5-to 7-membered aromatic or nonaromatic heterocycle containing 1 to 4 heteroatoms selected from O, N and S and includes bicyclic groups. "Heterocyclyl" thus includes the above-mentioned heteroaryl groups, as well as the dihydro and tetrahydro analogs thereof. Further examples of "heterocyclyl" include, but are not limited to: imidazolyl, indolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, oxetanyl, pyranyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, quinoxalinyl, tetrazolyl, thiadiazolyl, thiazolyl, thienyl, triazolyl, l, 4-dioxanyl, pyrrolidinyl, dihydroimidazolyl, dihydroisoxazolyl, dihydroisothiazolyl, dihydrooxadiazolyl, dihydrooxazolyl, dihydropyrazinyl, dihydropyrazolyl, dihydropyridinyl, dihydropyrimidyl, dihydropyrrolyl, dihydrotetrazolyl, dihydrothiadiazolyl, dihydrothiazolyl, dihydrothienyl, dihydrotriazolyl, dihydroazetidinyl, tetrahydrofuryl, and tetrahydrothienyl, and N-oxides thereof. Attachment of the heterocyclic substituent may be through a carbon atom or through a heteroatom.

As understood by those skilled in the art, "halogen" as used herein is meant to include fluorine, chlorine, bromine and iodine.

Unless otherwise defined, alkyl, cycloalkyl, aryl, and heterocyclyl substituents may be unsubstituted or substituted. For example, (C) _1-6 ) Alkyl groups may be substituted with one, two or three substituents selected from OH, halogen, alkoxy, dialkylamino or heterocyclyl, e.g., morpholinyl, piperidinyl, and the like.

The invention includes the free forms of the compounds of formula (i) and also pharmaceutically acceptable salts and stereoisomers thereof. Some specific exemplary compounds herein are protonated salts of amine-based compounds. The term "free form" refers to the amine compound in a non-salt form. Included within the pharmaceutically acceptable salts are not only exemplary salts of the particular compounds described herein, but are all typical pharmaceutically acceptable salts of the free forms of the compounds of formula (I). The free form of a particular salt of the compound may be isolated using techniques known in the art. For example, the free form can be regenerated by treating the salt with a dilute aqueous solution of a suitable base, such as a dilute aqueous NaOH solution, a dilute aqueous potassium carbonate solution, dilute aqueous ammonia, and a dilute aqueous sodium bicarbonate solution. The free forms differ somewhat from their respective salt forms in certain physical properties, such as solubility in polar solvents, but for the purposes of the invention such acid and base salts are otherwise pharmaceutically equivalent to their respective free forms.

Pharmaceutically acceptable salts of the invention can be synthesized from compounds of the invention containing a basic or acidic moiety by conventional chemical methods. In general, salts of basic compounds are prepared by ion exchange chromatography or by reaction of the free base with a stoichiometric amount or excess of an inorganic or organic acid in the form of the desired salt in an appropriate solvent or combination of solvents. Similarly, salts of acidic compounds are formed by reaction with a suitable inorganic or organic base.

Thus, pharmaceutically acceptable salts of the compounds of the present invention include the conventional non-toxic salts of the compounds of the present invention formed by the reaction of a basic compound of the present invention and an inorganic or organic acid. For example, conventional non-toxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like, as well as those prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxy-benzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, trifluoroacetic and the like.

If the compounds of the invention are acidic, suitable "pharmaceutically acceptable salts" refer to salts prepared from pharmaceutically acceptable non-toxic bases including inorganic and organic bases, salts derived from inorganic bases including aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic, manganous, potassium, sodium, zinc, and the like. Particularly preferred are ammonium, calcium, magnesium, potassium and sodium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases including salts of primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins such as arginine, betaine, caffeine, choline, N, N' -dibenzylethylenediamine, diethylamine, 2-dimethylaminoethanol, aminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, histidine, hydroxycobalamin, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine-piperidine, polyamine resins, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine and the like.

Berg et al, "Pharmaceutical Salts" J.pharm.Sci.1977:66:1-19 describe in more detail the preparation of the pharmaceutically acceptable salts described above and other typical pharmaceutically acceptable salts.

In addition to standard methods known in the literature or exemplified in experimental procedures, the compounds of the invention can be prepared using reactions as shown in the following schemes. The following illustrative schemes are therefore for illustrative purposes and are not limited to the compounds listed or any particular substituents.

Metabolites of the compounds referred to herein and pharmaceutically acceptable salts thereof, as well as prodrugs that are convertible in vivo into the structures of the compounds referred to herein and pharmaceutically acceptable salts thereof, are also encompassed by the claims herein.

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

Example 1: (Z) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-fluoroacrylic acid

Step 1: synthesis of 4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxybenzaldehyde

Compound 26 (3.07g, 10mmol) and vanillin (1.52g, 10mmol) were dissolved in DMF (10 ml), and K was added ₂ CO ₃ (2g, 14mmol), and stirred at 80 ℃ for 3 hours. After cooling to room temperature, the resulting mixture was extracted with ethyl acetate and water. The organic layer was separated, washed with brine, and Na ₂ SO ₄ And (5) drying. After the solvent was dried under reduced pressure, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 7) to give product 27 (3.21g, 8.50mmol, 85%) as a white solid: ¹ H NMR(400MHz,CDCl ₃ )δ9.87(s,1H),7.97(d,J＝8.4Hz,2H),7.85(d,J＝8.5Hz,1H),7.48(d,J＝1.7Hz,1H),7.42(dd,J＝8.2,1.8Hz,1H),6.93(d,J＝8.2Hz,1H),5.47(s,2H),3.99(s,3H).MS(ESI)m/z 379[M+H] ⁺ 。

step 2: synthesis of ethyl (Z) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-fluoroacrylate

Compound 27 (3.78g, 10mmol) was dissolved in anhydrous THF (20 ml) under an argon atmosphere. n-BuLi (10mmol, 6.3ml) was added dropwise at 0 ℃. After stirring was continued for 30 minutes, 10ml of a THF solution of ethyl 2- (diethoxyphosphoryl) acetate (10mmol, 2.24g) was added dropwise. After stirring for 1.5 hours, a saturated aqueous solution of ammonium chloride was added to quench the reaction, and the resulting mixture was extracted with ethyl acetate and water. The organic layer was separated, washed with brine, na ₂ SO ₄ And (5) drying. After spin-drying the solvent under reduced pressure, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 7) to give the product 28 (3.0 g,6.43mmol, 64%) as a white solid: ¹ H NMR(400MHz,DMSO-d ₆ )δ8.16(d,J＝8.1Hz,1H),8.09(s,1H),8.02(d,J＝8.2Hz,1H),7.41(d,J＝1.8Hz,1H),7.15(s,1H),7.14–7.08(m,1H),7.01(d,J＝8.5Hz,1H),5.37(s,2H),4.22(q,J＝7.1Hz,2H),3.79(s,3H),1.18(t,J＝7.1Hz,3H).MS(ESI)m/z 467[M+H] ⁺ 。

and 3, step 3: synthesis of (Z) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-fluoroacrylic acid

Compound 28 (2g, 4.3mmol) was dissolved in methanol (8 ml), naOH (520mg, 13mmol) was added, and then (8 ml) water was added. After stirring overnight at room temperature, water was added and a 4N hydrochloric acid solution was added dropwise to precipitate, which was filtered and washed with water to give the desired product 1 (1.7g, 3.9mmol, 91%): ¹ H NMR(400MHz,CDCl ₃ )δ7.97(d,J＝8.2Hz,1H),7.94(s,1H),7.83(d,J＝8.1Hz,1H),7.42(d,J＝1.8Hz,1H),7.05(dd,J＝8.4,1.7Hz,1H),6.97(d,J＝24.0Hz,1H),6.78(d,J＝8.4Hz,1H),5.42(s,2H),3.92(s,3H).MS(ESI)m/z 439[M+H] ⁺ 。

example 2: (Z) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2- (trifluoromethyl) acrylic acid

The procedure differs from that of example 1 in that, directly after the synthesis of compound 27, the following reaction is carried out to prepare (Z) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2- (trifluoromethyl) acrylic acid:

compound 27 (3.78g, 10mmol), 3-trifluoropropionic acid (10mmol, 0.88ml) was dissolved in anhydrous THF (20 ml) under argon protection. Titanium tetrachloride (169mol, 1.8ml) was added dropwise at 0 ℃. Stirring was continued for 30 minutes and triethylamine (5.6 ml, 40mmol) was added dropwise. After stirring overnight at room temperature, the mixture was extracted with ethyl acetate and water. The organic layer was separated, washed with brine and Na ₂ SO ₄ And (5) drying. After spin-drying the solvent under reduced pressure, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 1) to give product 2 (3.1g, 6.3mmol,63% yield) as a yellow solid: ¹ H NMR(400MHz,CDCl ₃ )δ8.14(s,1H),8.01–7.93(m,2H),7.85(d,J＝8.1Hz,1H),7.12–7.01(m,2H),6.85(d,J＝8.3Hz,1H),5.44(s,2H),3.94(s,3H).MS(ESI)m/z 487[M-H] ⁺ 。

example 3: (E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy-3-methoxyphenyl-2-cyanoacrylic acid

The procedure differs from that of example 1 in that, directly after the synthesis of compound 27, the following reaction is carried out:

compound 27 (2.9g, 7.7mmol) and 2-cyanoacetic acid (977.9mg, 11.4mmol) were dissolved in acetonitrile (20 ml), followed by addition of piperidine (1.5 ml) and stirring at 80 ℃ for 3 hours. After cooling to room temperature, the resulting mixture was treated with water. Hydrochloric acid (2N) was then added to precipitate a solid. The mixture was extracted with ethyl acetate and water. The organic layer was separated, washed with brine and Na ₂ SO ₄ And (5) drying. The solvent was dried by spinning under reduced pressure, and the product was purified by silica gel column chromatography (petroleum ether/ethyl acetate)Ester = 1) to give the product as a pale yellow solid 3 (2.6 g,5.8mmol, 75.6%): ¹ H NMR(400MHz,DMSO-d ₆ )δ13.79(s,1H),8.27(s,1H),8.18(d,J＝8.1Hz,1H),8.11(s,1H),8.02(d,J＝8.1Hz,1H),7.81(d,J＝2.0Hz,1H),7.70(dd,J＝8.6,1.9Hz,1H),7.26(d,J＝8.6Hz,1H),5.46(s,2H),3.83(s,3H).MS(ESI)m/z[M+H] ⁺ :446.

example 4: (Z) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-fluoroacrylic acid methyl ester

Compound 1 (193mg, 0.44mmol) prepared in example 1 was dissolved in anhydrous DMF (3 ml). Adding K ₂ CO ₃ (121.6 mg, 0.88mmol) and CH ₃ I (41. Mu.L, 0.66 mmol). After stirring at room temperature for 1 hour, ethyl acetate and saturated NaHCO were added ₃ The resulting mixture was extracted. The organic layer was separated, washed with brine, na ₂ SO ₄ And (5) drying. After filtration and evaporation, the residue was purified by silica gel column chromatography (PE: EA = 2) to give 4 (190mg, 0.42mmol, 95%) as a white solid: ¹ H NMR(400MHz,CDCl ₃ )δ7.98(d,J＝8.2Hz,1H),7.94(s,1H),7.83(d,J＝8.2Hz,1H),7.44(d,J＝2.0Hz,1H),7.02(dd,J＝8.4,1.7Hz,1H),6.84(d,J＝24.3Hz,1H),6.78(d,J＝8.4Hz,1H),5.42(s,2H),3.94(s,3H),3.84(s,3H).MS(ESI)m/z 453[M+H] ⁺ 。

example 5: (Z) -methyl 3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2- (trifluoromethyl) acrylate

The synthesis method is substantially the same as in example 4, except that the starting material used in this example is compound 2 prepared in example 2.

¹ H NMR(300MHz,CDCl ₃ )δ8.01–7.93(m,3H),7.85(d,J＝8.1Hz,1H),7.06–6.98(m,2H),6.83(d,J＝8.3Hz,1H),5.43(s,2H),3.93(s,3H),3.89(s,3H).MS(ESI)m/z 503[M+H] ⁺ 。

Example 6: (E) -methyl 3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanoacrylate

The synthesis method was substantially the same as in example 4, except that the starting material used in this example was compound 3 prepared in example 3.

¹ H NMR(400MHz,DMSO-d ₆ )δ8.35(s,1H),8.18(d,J＝7.7Hz,1H),8.12(s,1H),8.02(d,J＝7.8Hz,1H),7.84(s,1H),7.75(d,J＝8.2Hz,1H),7.28(d,J＝8.4Hz,1H),5.47(s,2H),3.84(d,J＝6.5Hz,6H).MS(ESI)m/z 482[M+Na] ⁺ .

Example 7: (Z) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-fluoroacrylamide

Compound 1 (96mg, 0.22mmol) prepared in example 1 was dissolved in DCM (2 mL), oxalyl chloride (0.5 mL) and one drop of DMF were added, stirred for 5h and the solvent was spin-dried under reduced pressure. The crude product after spin drying was dissolved in DCM (2 mL) and 28% aqueous ammonia (1 mL) was added at 0 ℃. After stirring at room temperature for 2 hours, the mixture was washed with DCM and H ₂ O extraction, separation of the organic layer, washing with brine, na ₂ SO ₄ And (5) drying. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 3) after spin-drying the solvent under reduced pressure to give product 7 (52mg, 0.12mmol, 55%) as a white solid: ¹ H NMR(400MHz,CDCl ₃ )δ7.97(d,J＝8.2Hz,1H),7.93(s,1H),7.83(dd,J＝9.0,5.1Hz,2H),7.08(dd,J＝8.4,1.9Hz,1H),6.72(t,J＝18.9Hz,2H),5.94(d,J＝221.9Hz,2H),5.42(s,2H),3.95(s,3H).MS(ESI)m/z 438[M+H] ⁺ .

example 8: (Z) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2- (trifluoromethyl) acrylamide

The synthesis method was substantially the same as in example 7, except that the starting material used in this example was compound 2 prepared in example 2.

¹ H NMR(400MHz,CDCl ₃ )δ7.98(t,J＝3.9Hz,2H),7.95(s,1H),7.85(d,J＝8.1Hz,1H),7.03(s,1H),7.00(d,J＝8.4Hz,1H),6.83(d,J＝8.3Hz,1H),5.90(s,2H),5.42(s,2H),3.92(s,3H).MS(ESI)m/z 488[M+H] ⁺ .

Example 9: (E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanoacrylamide

The synthesis method is substantially the same as in example 7, except that the starting material used in this example is compound 3 prepared in example 3.

¹ H NMR(400MHz,DMSO-d ₆ )δ8.18(d,J＝8.2Hz,1H),8.12(d,J＝7.4Hz,2H),8.03(d,J＝8.1Hz,1H),7.88-7.64(m,3H),7.57(dd,J＝8.5,1.9Hz,1H),7.24(d,J＝8.5Hz,1H),5.44(s,2H),3.84(s,3H).MS(ESI)m/z 445[M+H] ⁺ .

Example 10: (Z) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -N-hydroxy-2- (trifluoromethyl) acrylamide

Step 1: synthesis of (Z) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -N- ((tetrahydro-2H-pyran-2-yl) oxy) -2- (trifluoromethyl) acrylamide

Compound 2 prepared in example 2 (215mg, 0.44mmol) and O- (tetrahydro-2H-pyran-2-yl) hydroxylamine (51.5mg, 0.44mmol) were dissolved in anhydrous DMF (3 ml), followed by the addition of EDCI (168.7mg, 0.88mmol) and HOBT (119mg, 0.88mmol). After stirring overnight, the mixture was extracted with ethyl acetate and water. The organic layer was separated, washed with brine, na ₂ SO ₄ And (5) drying. After the solvent was dried under reduced pressure, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 6) to give the product 29 (111.6 mg,0.19mmol, 43%) as a pale yellow oil: ¹ H NMR(400MHz,CDCl ₃ )δ8.12(s,1H),7.98(d,J＝8.2Hz,1H),7.93(s,1H),7.83(d,J＝8.2Hz,1H),7.37(d,J＝1.8Hz,1H),6.97(dd,J＝8.3,1.9Hz,1H),6.78(d,J＝8.3Hz,1H),5.41(s,2H),5.38(dd,J＝4.8,2.5Hz,1H),3.96(s,3H),3.68–3.60(m,1H),1.94–1.81(m,2H),1.79–1.55(m,6H).MS(ESI)m/z588[M+H] ⁺ .

and 2, step: synthesis of (Z) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -N-hydroxy-2- (trifluoromethyl) acrylamide

Compound 29 (100mg, 0.17mmol) was dissolved in methanol (4 mL), and concentrated hydrochloric acid (4 mL) was added dropwise. After stirring overnight, water was added to precipitate, which was filtered and washed with water to give the product 10 as a white solid (80.5mg, 0.16mmol, 94%): ¹ H NMR(400MHz,CDCl ₃ )δ8.06(s,1H),7.98(d,J＝8.2Hz,1H),7.94(s,1H),7.83(d,J＝8.1Hz,1H),7.28(d,J＝1.8Hz,1H),6.98(dd,J＝8.3,1.8Hz,1H),6.80(d,J＝8.3Hz,1H),5.42(s,2H),3.95(s,3H).MS(ESI)m/z 504[M+H] ⁺ .

example 11: (Z) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-fluoro-N-hydroxyacrylamide

The synthesis was as in example 10, except that in example 10, the starting material used in step 1 was compound 1 prepared in example 1.

¹ H NMR(400MHz,DMSO)δ11.38(s,1H),9.31(d,J＝1.6Hz,1H),8.16(d,J＝8.2Hz,1H),8.09(s,1H),8.02(d,J＝8.2Hz,1H),7.57(d,J＝1.9Hz,1H),7.11(dd,J＝8.4,1.8Hz,1H),6.99(d,J＝8.4Hz,1H),6.71(d,J＝27.5Hz,1H),5.35(s,2H),3.77(s,3H).MS(ESI)m/z 454[M+H] ⁺ .

Example 12: (E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyano-N-hydroxyacrylamide

The synthesis was as in example 10, except that in example 10, the starting material used in step 1 was compound 3 prepared in example 3.

¹ H NMR(400MHz,DMSO)δ11.24(s,1H),9.30(s,1H),8.18(d,J＝8.3Hz,1H),8.11(s,1H),8.03(d,J＝11.2Hz,2H),7.72(s,1H),7.58(d,J＝8.5Hz,1H),7.23(d,J＝8.5Hz,1H),5.44(s,2H),3.83(s,3H).MS(ESI)m/z 461[M+H] ⁺ .

Example 13: (E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyano-N- (2-methoxyethyl) acrylamide

2-Methoxyethyl-1-amine (39.8mg, 0.53mmol), HATU (216.7mg, 0.57mmol) and DIPEA (0.22ml, 1.3mmol) were added to a solution of intermediate compound 3 (200mg, 0.44mmol) in DMF (3 mL). After stirring at room temperature for 1 hour, the resulting mixture was extracted with ethyl acetate and saturated NaHCO ₃ And (4) extracting. The organic layer was separated, washed with brine, na ₂ SO ₄ And (5) drying. The solvent was spin-dried under reduced pressure, and the mixture was purified by silica gel column chromatography (petroleum ether)Ethyl acetate =3: 1) The residue was purified to give white solid 13 (183mg, 0.36mmol,82% yield): ¹ H NMR(400MHz,DMSO-d ₆ )δ8.36(s,1H),8.18(d,J＝7.9Hz,1H),8.12(d,J＝6.2Hz,2H),8.03(d,J＝8.1Hz,1H),7.72(s,1H),7.59(d,J＝8.4Hz,1H),7.24(d,J＝8.4Hz,1H),5.44(s,2H),3.84(s,3H),3.43(d,J＝4.6Hz,2H),3.38(d,J＝5.2Hz,2H),3.27(s,3H).HRMS(ESI ⁺ ):calculated for C ₂₃ H ₂₀ F ₆ N ₂ O ₄ [M+H]+:503.1400,found 503.1383.

example 14: (E) -5- (3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanoacrylamido) pentanoic acid

Tert-butyl 5-aminovalerate (91.8mg, 0.53mmol), HATU (216.7mg, 0.57mmol) and DIPEA (0.22ml, 1.3mmol) were added to a solution of intermediate 3 (200mg, 0.44mmol) in DMF (3 mL). After stirring at room temperature for 1 hour, the resulting mixture was taken up with ethyl acetate and saturated NaHCO ₃ And (4) extracting. The organic layer was separated, washed with brine, na ₂ SO ₄ And (5) drying. After the solvent was dried under reduced pressure, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 2). TFA (1 ml) was added to a solution of the tert-butyl ester intermediate in DCM (2 ml). After stirring for 1h, the solvent was dried under reduced pressure, toluene (3X 3 mL) was added and residual TFA was evaporated to give 14 as a yellow solid (155mg, 0.28mmol, 93%): ¹ H NMR(400MHz,DMSO-d ₆ )δ12.00(s,1H),8.37(t,J＝5.7Hz,1H),8.18(d,J＝8.2Hz,1H),8.11(d,J＝3.9Hz,2H),8.03(d,J＝8.1Hz,1H),7.71(d,J＝2.0Hz,1H),7.58(dd,J＝8.6,2.0Hz,1H),7.24(d,J＝8.6Hz,1H),5.44(s,2H),3.84(s,3H),3.21(d,J＝5.7Hz,2H),2.24(t,J＝6.8Hz,2H),1.57-1.42(m,4H).HRMS(ESI ⁺ ):calculated for C ₂₅ H ₂₂ F ₆ N ₂ O ₅ [M+H] ⁺ :545.1506,found 545.1495.

example 15: (E) -3- (2- (3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanoacrylamido) ethoxy) propanoic acid

The synthesis method is as in example 14, wherein the difference from example 14 is that the raw materials used in this example are compound 3 and tert-butyl 3- (2-aminoethoxy) propionate (compound 47 in example 19).

¹ H NMR(400MHz,DMSO-d ₆ )δ12.17(s,1H),8.34(s,1H),8.18(s,1H),8.12(s,2H),8.03(s,1H),7.72(s,1H),7.58(s,1H),5.42(d,J＝23.2Hz,2H),3.84(s,2H),3.62(d,J＝6.0Hz,2H),3.48(s,2H),3.36(s,2H),3.32(s,3H).HRMS(ESI ⁺ ):calculated for Chemical Formula:C ₂₅ H ₂₂ F ₆ N ₂ O ₆ [M+H] ⁺ :561.1455,found 561.1444.

Example 16: (2S, 4R) -1- ((S) -2- (3- ((E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanoacrylamido) propionylamino) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide

Step 1: preparation of tert-butyl 3- (((benzyloxy) carbonyl) amino) propionate

3- (((benzyloxy) carbonyl) amino) propionic acid (848.3mg, 3.8mmol), DMAP (92.8mg, 0.76mmol) and DCC (862.5mg, 4.2mmol) were dissolved in DCM (5 mL) and t-butanol (0.7mL, 7.6 mmol) was added. Stir at room temperature for 3 hours. The solvent was spin dried under reduced pressure to give a residue. Column chromatography (petroleum ether/ethyl acetate = 6) of the residue afforded 33 (949.8mg, 3.4mmol,89.0% yield) as a colorless oil: 7.40-7.28 (m,5H),5.29(s,1H),5.09(s,2H),3.42(dd,J＝12.1,6.1Hz,2H),2.45(t,J＝6.0Hz,2H),1.44(s,9H).MS(ESI)m/z 280[M+H] ⁺ .

step 2: preparation of tert-butyl 3-Aminopropionate

A10% palladium on carbon catalyst (90 mg) was added to a solution of tert-butyl ester compound 33 (642.5mg, 2.3mmol,1 eq) in ethanol (6 mL). The hydrogen was replaced and the reaction system was stirred at 45 ℃ for 24 hours. The reaction mixture was filtered through a pad of celite, washed with ethyl acetate and the filtrate evaporated to give 34 (159.7 mg,1.1mmol,48% yield) as a colourless oil: ¹ H NMR(400MHz,CDCl ₃ )δ2.92(t,J＝6.2Hz,2H),2.36(t,J＝6.2Hz,2H),1.44(s,9H).MS(ESI)m/z 146[M+H] ⁺ .

and step 3: preparation of tert-butyl (E) -3- (3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanoacrylamido-propionate

Compound 34 (76.6mg, 0.53mmol), HATU (216.6mg, 0.57mmol) and DIPEA (0.24ml, 1.3 mmol) were added to a solution of intermediate compound 3 (200mg, 0.44mmol) in dry DMF (3 mL). After stirring at room temperature for 1 hour, the resulting mixture was taken up with ethyl acetate and saturated NaHCO ₃ And (4) extracting. The organic layer was separated, washed with brine, na ₂ SO ₄ And (5) drying. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 2): ¹ H NMR(400MHz,CDCl ₃ )δ8.21(s,1H),7.95(d,J＝5.7Hz,2H),7.85(d,J＝8.3Hz,1H),7.74(d,J＝2.0Hz,1H),7.39(dd,J＝8.5,2.1Hz,1H),6.94(t,J＝5.8Hz,1H),6.87(d,J＝8.4Hz,1H),5.46(s,2H),3.98(s,3H),3.66(dd,J＝12.1,6.0Hz,2H),2.55(t,J＝6.1Hz,2H),1.49(s,9H).MS(ESI)m/z 573[M+H] ⁺ .

and 4, step 4: preparation of 4- (4-methylthiazol-5-yl) benzonitrile

4-bromobenzylnitrile (7.0g, 38.5mmol), pd (OAc) was measured ₂ (43.2mg, 0.19mmol) and KOAc (5.67g, 57.8mmol) were placed in a two-necked flask, and DMAc (40 mL) and 4-methylthiazole (5.3mL, 57.8mmol) were added under argon atmosphere, and the mixture was heated at 150 ℃ overnight. After cooling, the solvent DMAC was spin-dried, extracted with water and ethyl acetate, the organic layer was separated and washed with saturated brine, na ₂ SO ₄ After drying and spin-drying the solvent under reduced pressure, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 5) to give 38 (5.9g, 29.5mmol,77% yield) as a white solid: ¹ H NMR(400MHz,CDCl ₃ )δ8.76(s,1H),7.7-7.69(m,2H),7.60-7.53(m,2H),2.57(s,3H).MS(ESI)m/z 201[M+H] ⁺ .

and 5: preparation of (4- (4-methylthiazol-5-yl) phenyl) methanamine

Compound 38 (5.0 g, 25mmol) was dissolved in methanol (125 ml), coCl2 (4.5 g, 35mmol) was added under ice-bath conditions and NaBH was added portionwise ₄ (3.8g, 100mmol). After stirring at room temperature for 90 minutes, the reaction was quenched with ammonia and water, and the reaction was filtered through celite, the celite was washed with DCM: meOH: TEA =10: ¹ H NMR(400MHz,CD ₃ OD_SPE)δ8.87(s,1H),7.45(s,4H),3.85(s,2H),3.31(s,1H).MS(ESI)m/z 205[M+H] ⁺ .

and 6: preparation of (2S,4R) -4-hydroxy-2- ((4- (4-methylthiazol-5-yl) benzyl) carbamoyl) pyrrolidine-1-carboxylic acid tert-butyl ester

HATU (5.3g, 14mmol), DIPEA (4.6ml, 28mmol) and (2S, 4R) -1- (tert-butoxycarbonyl) -4-hydroxypyrrolidine-2-carboxylic acid (2.15g, 9.3mmol) were added to a solution of compound 39 (1.9g, 9.3mmol) in DMF (15 ml). After stirring at 25 ℃ for 1 hour, DMF was spin-dried under reduced pressure and taken up with ethyl acetate and saturated NaHCO ₃ And (4) extracting with an aqueous solution. The organic layer was separated, washed with brine and Na ₂ SO ₄ And (5) drying. The solvent was dried under reduced pressure and purified by silica gel column chromatography (MeOH: CH) ₂ Cl ₂ = 4)] ⁺ .

And 7: preparation of tert-butyl ((S) -1- ((2S, 4R) -4-hydroxy-2- ((4- (4-methylthiazol-5-yl) benzyl) carbamoyl) pyrrolidin-1-yl) -3,3 dimethyl-1-oxobutan-2-yl) carbamic acid tert-butyl ester

Trifluoroacetic acid (10 ml) was added to a solution of compound 40 (2.6 g, 6.2mmol) in DCM (10 ml). After stirring for 1 hour, the solvent was removed by rotary drying under reduced pressure, toluene was added and the residual trifluoroacetic acid was removed by evaporation. The crude product was used in the next step without further purification. HATU (3.5g, 9.3mmol), DIPEA (5.1ml, 31mmol) and (S) -2- ((tert-butoxycarbonyl) amino) -3, 3-dimethylbutyric acid (1.43g, 6.2 mmol) were added to a solution of the crude product obtained above in DMF (10 ml). After stirring for 1 hour at 25 ℃, the DMF was spin-dried, washed with ethyl acetate and saturated NaHCO ₃ And (4) extracting with an aqueous solution. The organic layer was separated, washed with brine, na ₂ SO ₄ And (5) drying. The solvent was dried under reduced pressure and purified by silica gel column chromatography (MeOH: CH) ₂ Cl ₂ = 4)] ⁺ .

And 8: preparation of (2S, 4R) -1- ((S) -2-amino-3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide

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Trifluoroacetic acid (10 ml) was added to a solution of compound 41 (2.4 g,4.5 mmol) in DCM (10 ml). After stirring for 1 hour, the solvent was removed by rotary drying under reduced pressure, toluene was added and the residual trifluoroacetic acid was removed by evaporation. Extract with saturated aqueous sodium bicarbonate with DCM: meOH =10 ₂ SO ₄ And (5) drying. The solvent was dried by spinning under reduced pressure and the mixture was purified by silica gel column chromatography (MeOH: CH) ₂ Cl ₂ = 1): ¹ H NMR(400MHz,CDCl ₃ )δ8.66(s,1H),7.63(s,1H),7.33(s,4H),4.76(t,J＝7.6Hz,1H),4.47(d,J＝18.4Hz,2H),4.32(d,J＝11.9Hz,1H),3.77(d,J＝11.0Hz,1H),3.60(dd,J＝11.1,3.6Hz,1H),3.34(d,J＝36.7Hz,1H),2.89(dd,J＝14.5,7.2Hz,1H),2.50(s,3H),2.36(s,1H),2.20–2.09(m,1H),0.94(s,9H).MS(ESI)m/z 431[M+H] ⁺ .

and step 9: preparation of (2S, 4R) -1- ((S) -2- (3- ((E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanoacrylamido) propionylamino) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide.

Trifluoroacetic acid (1 ml) was added to a solution of compound 35 (100mg, 0.17mmol) in DCM (2 ml). After stirring for 1 hour, the solvent was removed by rotary drying under reduced pressure, toluene (3X 3 mL) was added and the residual trifluoroacetic acid was removed by evaporation and the crude product was used in the next step without further purification. HATU (83.6 mg, 0.22mmol), DIPEA (85. Mu.L, 0.51 mmol) and 42 (86.1mg, 0.2mmol) were added to a DMF solution of the crude product obtained above. After stirring at 25 ℃ for 1 hour, the mixture was stirred with ethyl acetate and saturated NaHCO ₃ Extracting with saturated water solution. Separation of organic matterLayer, washed with brine and Na ₂ SO ₄ And (5) drying. The solvent was dried under reduced pressure and purified by silica gel column chromatography (MeOH: CH) ₂ Cl ₂ = 4): ¹ H NMR(400MHz,CDCl ₃ )δ8.66(s,1H),8.17(s,1H),7.94(d,J＝8.7Hz,2H),7.84(d,J＝8.1Hz,1H),7.66(d,J＝1.9Hz,1H),7.40-.7.28(m,6H),7.23(s,1H),6.84(d,J＝8.5Hz,1H),6.53(t,J＝9.9Hz,1H),5.42(s,2H),4.72(t,J＝8.1Hz,1H),4.58(dd,J＝16.4,7.9Hz,3H),4.29(dd,J＝15.1,5.2Hz,1H),4.08(d,J＝11.0Hz,1H),3.94(s,3H),3.75(dd,J＝12.8,6.4Hz,1H),3.67-.3.51(m,2H),3.38(s,1H),2.58-.2.45(m,6H),2.16(dd,J＝13.5,8.0Hz,1H),2.05(d,J＝7.8Hz,1H),0.93(s,9H).HRMS(ESI ⁺ ):calculated for C ₄₅ H ₄₆ F ₆ N ₆ O ₇ S[M+H] ⁺ :929.3126,found 929.3132.

example 17: (2S, 4R) -1- ((S) -2- (4- ((E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanoacrylamide) butyrylamino) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide

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The synthesis was as in example 16.

¹ H NMR(400MHz,CDCl ₃ )δ8.67(s,1H),8.21(s,1H),7.95(d,J＝6.7Hz,2H),7.85(d,J＝8.5Hz,1H),7.68(d,J＝1.9Hz,1H),7.42-.7.31(m,6H),7.28(s,1H),6.87(d,J＝8.5Hz,1H),6.57(d,J＝8.5Hz,1H),5.45(s,2H),4.74(t,J＝8.0Hz,1H),4.62-.48(m,3H),4.32(dd,J＝14.9,5.2Hz,1H),4.17(d,J＝11.4Hz,1H),3.97(s,3H),3.61(dd,J＝11.3,3.4Hz,1H),3.54-.3.35(m,3H),2.59-2.48(m,4H),2.35(dd,J＝11.7,6.0Hz,2H),2.15(dd,J＝13.4,8.1Hz,1H),1.97-.1.85(m,2H),0.95(s,9H).HRMS(ESI ⁺ ):calculated for C ₄₆ H ₄₈ F ₆ N ₆ O ₇ S[M+H] ⁺ :943.3282,found 943.3287.

Example 18: (2S, 4R) -1- ((S) -2- (6- ((E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanopropionylamino) hexanamide) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide

The synthesis was as in example 16.

¹ H NMR(400MHz,CDCl ₃ )δ8.67(s,1H),8.19(s,1H),7.96(s,2H),7.85(d,J＝7.7Hz,1H),7.69(s,1H),7.35(s,6H),6.86(d,J＝8.3Hz,1H),6.48(s,1H),6.17(d,J＝7.6Hz,1H),5.45(s,2H),4.73(t,J＝7.7Hz,1H),4.56(dd,J＝21.1,7.0Hz,3H),4.33(dd,J＝14.9,4.6Hz,1H),4.11(d,J＝9.9Hz,1H),3.97(s,3H),3.59(d,J＝9.5Hz,1H),3.39(d,J＝5.8Hz,3H),2.50(s,3H),2.19(dd,J＝22.4,6.1Hz,3H),1.70-1.54(m,4H),1.40(dd,J＝26.9,12.1Hz,3H),0.93(s,9H).HRMS(ESI ⁺ ):calculated for C ₄₈ H ₅₂ F ₆ N ₆ O ₇ S[M+H] ⁺ :971.3595,found 971.3595.

Example 19: (2S, 4R) -1- ((S) -2- (3- ((E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanopropionylamino) propionylamino) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide

Step 1: synthesis of tert-butyl 3- (2-hydroxyethoxy) propionate

To a solution of anhydrous ethylene glycol (10g, 161.1mmol) in anhydrous THF (40 mL) was added metallic sodium (62.0 mg, 2.70mmol), and the mixture was stirred at room temperature for 2 hours. Tert-butyl acrylate (6.90, 53.7 mmol) was added and stirred overnight. After quenching with water, the solvent THF was dried under reduced pressure and extracted with ethyl acetate andand (4) water extraction. The organic layer was separated, washed with brine, and Na ₂ SO ₄ And (5) drying. After filtration and evaporation, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 2) to give 44 (5.0 g,26.3mmol,49% yield) as a colorless oil: ¹ H NMR(400MHz,CDCl ₃ )δ3.77-.3.67(m,4H),3.61-.3.53(m,2H),2.54-.2.43(m,3H),1.45(s,9H).MS(ESI)m/z 191[M+H] ⁺ .

step 2: synthesis of tert-butyl 3- (2- (toluenesulfonyloxy) ethoxy) propionate

To a solution of p-toluenesulfonyl chloride (3.23g, 17mmol) in CH at-10 deg.C ₂ Cl ₂ (5 mL) the solution was added dropwise to 44 (2.5g, 13.1mmol), NEt ₃ (2.4mL, 17mmol) and DMAP (402.8mg, 3.3mmol) in anhydrous CH ₂ Cl ₂ (5 mL) in solution. The reaction mixture was stirred at room temperature for 8 hours. The resulting mixture was taken up in saturated NaHCO ₃ Extracted with DCM. The organic layer was separated, washed with brine and Na ₂ SO ₄ And (5) drying. After drying DCM under reduced pressure, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 7) to give 45 (3.9g, 11.3mmol, 86%) as a colorless oil: ¹ H NMR(400MHz,CDCl ₃ )δ7.81(d,J＝8.3Hz,2H),7.36(d,J＝8.0Hz,2H),4.15(dd,J＝5.4,4.2Hz,2H),3.65(dd,J＝8.1,4.3Hz,4H),2.46(s,3H),2.43(t,J＝6.4Hz,2H),1.45(s,9H).MS(ESI)m/z 345[M+H] ⁺ .

and 3, step 3: synthesis of tert-butyl 3- (2-azidoethoxy) propionate

Adding NaN ₃ (2.8g, 43mmol) was added to a solution of 45 (3.0g, 8.6mmol) in DMF (8 ml). The reaction mixture was heated at 100 ℃ for 3 hours. After cooling to room temperature, the mixture was extracted with ethyl acetate and water. The organic layer was separated, washed with saturated brine and Na ₂ SO ₄ And (5) drying. After spin-drying the solvent under reduced pressure, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 8) to give a colorless oil 46 (1.7g, 7.9mmol, 92%): ¹ H NMR(400MHz,CDCl ₃ )δ3.72(t,J＝6.4Hz,2H),3.65-.3.58(m,2H),3.35(t,J＝5.0Hz,2H),2.51(t,J＝6.4Hz,2H),1.45(s,9H).MS(ESI)m/z 216[M+H] ⁺ .

and 4, step 4: synthesis of tert-butyl 3- (2-aminoethoxy) propionate

Mixing PPh ₃ (1.84g, 7.0mmol) and water (3 mL) were added to a solution of 46 (1.0g, 4.7mmol) in THF (18 mL) and stirred at room temperature overnight. The solvent was evaporated under reduced pressure and purified by silica gel column chromatography (2% MeOH/CH) ₂ Cl ₂ To 10% MeOH ₂ Cl ₂ ) The residue was purified to give 47 as a colorless oil (525mg, 2.8mmol, 60%): ¹ HNMR(400MHz,CDCl ₃ )δ3.68(t,J＝6.4Hz,2H),3.46(t,J＝5.2Hz,2H),2.83(t,J＝5.2Hz,2H),2.48(t,J＝6.4Hz,2H),1.44(s,11H).MS(ESI)m/z 190[M+H] ⁺ .

and 5: synthesis of tert-butyl (E) -3- (2- (3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanoacrylamido (ethoxy) propionate

Compound 47 (107.7 mg, 0.53mmol), HATU (216.6 mg, 0.57mmol) and DIPEA (0.24ml, 1.3 mmol) were added to a solution of intermediate compound 3 (200mg, 0.44mmol) in dry DMF (3 mL). After stirring at room temperature for 1 hour, the resulting mixture was extracted with ethyl acetate and saturated NaHCO ₃ And (4) solution extraction. The organic layer was separated, washed with brine and Na ₂ SO ₄ And (5) drying. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 2) after spin-drying the solvent under reduced pressure to give 48 (185mg, 0.30mmol,68% yield) as a pale yellow solid: ¹ H NMR(400MHz,CDCl ₃ )δ8.21(s,1H),7.95(d,J＝5.0Hz,2H),7.85(d,J＝8.2Hz,1H),7.73(d,J＝2.0Hz,1H),7.40(dd,J＝8.5,2.0Hz,1H),6.87(d,J＝8.4Hz,1H),6.76(s,1H),5.46(s,2H),3.98(s,3H),3.72(t,J＝6.3Hz,2H),3.61(d,J＝2.5Hz,4H),2.52(t,J＝6.3Hz,2H),1.46(s,9H).MS(ESI)m/z 617[M+H] ⁺ .

and 6: synthesis of (2S, 4R) -1- ((S) -2- (3- ((E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanopropionylamino) propionylamino) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide

Trifluoroacetic acid (1 ml) was added to a solution of compound 48 (100mg, 0.16mmol) in DCM (2 ml). After stirring for 1 hour, the solvent was spin dried under reduced pressure, toluene (3X 3 mL) was added and the residual trifluoroacetic acid was removed by evaporation. The crude product was used in the next step without further purification. HATU (61mg, 0.16mmol), DIPEA (85. Mu.L, 0.51 mmol) and 42 (70mg, 0.16mmol) were added to a solution of the above crude product in DMF (2 ml) and stirred at 25 ℃ for 1 hour. With ethyl acetate and saturated NaHCO ₃ The resulting mixture was solution extracted. The organic layer was separated, washed with brine, na ₂ SO ₄ And (5) drying. The solvent was dried by spinning under reduced pressure and then purified by silica gel column chromatography (MeOH: CH) ₂ Cl ₂ = 4): ¹ H NMR(400MHz,CDCl ₃ )δ8.66(s,1H),8.20(s,1H),7.95(d,J＝7.0Hz,2H),7.85(d,J＝8.2Hz,1H),7.67(d,J＝2.0Hz,1H),7.40-7.32(m,6H),7.20(t,J＝5.1Hz,1H),7.04(d,J＝8.7Hz,1H),6.87(d,J＝8.5Hz,1H),5.44(s,2H),4.77(t,J＝8.0Hz,1H),4.59(q,J＝6.6Hz,2H),4.51(s,1H),4.30(dd,J＝15.0,5.1Hz,1H),4.10(d,J＝11.3Hz,1H),3.95(s,3H),3.74(td,J＝9.9,4.0Hz,2H),3.67-3.54(m,5H),3.13(s,1H),2.56-2.45(m,6H),2.13(dd,J＝13.5,8.1Hz,1H),0.94(s,9H).HRMS(ESI ⁺ ):calculated for C ₄₇ H ₅₀ F ₆ N ₆ O ₈ S[M+H] ⁺ :973.3388,found 973.3346.

example 20: (2S, 4R) -1- ((S, E) -16- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2- (tert-butyl) -15-cyano-4, 14-dioxo-7, 10-dioxa-3, 13-diaza-hexadeca-15 enoyl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide

Synthesis procedure as in example 19

¹ H NMR(400MHz,CDCl ₃ )δ8.66(s,1H),8.19(s,1H),7.95(d,J＝7.1Hz,2H),7.84(d,J＝8.2Hz,1H),7.69(d,J＝1.8Hz,1H),7.46-7.29(m,6H),7.06(d,J＝7.6Hz,2H),6.86(d,J＝8.5Hz,1H),5.44(s,2H),4.72(t,J＝8.0Hz,1H),4.63-4.46(m,3H),4.31(dd,J＝15.0,5.2Hz,1H),4.11(d,J＝10.8Hz,1H),3.96(s,3H),3.72(d,J＝6.8Hz,2H),3.67-3.27(m,10H),2.58-2.38(m,6H),2.12(dd,J＝13.5,8.1Hz,1H),0.94(s,9H).HRMS(ESI ⁺ ):calculated for C ₄₉ H ₅₄ F ₆ N ₆ O ₉ S[M+H] ⁺ :1017.3650,found 1017.3635.

Example 21: (2S, 4R) -1- ((S, E) -19- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2- (tert-butyl) -18-cyano-4, 17 dioxo-7, 10, 13-trioxa-3, 16-diaza-nona-18 enoyl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide

Synthesis procedure as in example 19

¹ H NMR(400MHz,CDCl ₃ )δ8.66(s,1H),8.21(s,1H),7.95(d,J＝4.6Hz,2H),7.85(d,J＝8.1Hz,1H),7.70(d,J＝1.8Hz,1H),7.44-7.30(m,6H),7.05-6.94(m,2H),6.87(d,J＝8.5Hz,1H),5.45(s,2H),4.73(t,J＝8.0Hz,1H),4.61-4.44(m,3H),4.33(dd,J＝15.0,5.2Hz,1H),4.13(d,J＝11.5Hz,1H),3.97(s,3H),3.71(t,J＝6.9Hz,2H),3.68-3.52(m,14H),3.47(s,1H),2.57-2.42(m,6H),2.13(dd,J＝13.4,8.2Hz,1H),0.93(s,9H).HRMS(ESI ⁺ ):calculated for C ₅₁ H ₅₈ F ₆ N ₆ O ₁₀ S[M+H] ⁺ :1061.3912,found 1061.3887.

Example 22: (2S, 4R) -1- ((S) -2- (((2- ((E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanopropionylamino) ethoxy) methyl) amino) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide

Step 1: synthesis of tert-butyl 2- (2- (benzyloxy) ethoxy) acetate

Potassium tert-butoxide (2.24g, 20mmol) was added to a solution of compound 49 (3.00g, 20mmol) in anhydrous tert-butanol (24 mL), and stirred at room temperature for 30 minutes. Tert-butyl bromoacetate (3.90g, 20mmol) was added at 10 ℃ by injection and stirred at room temperature for 16 hours. With ethyl acetate and H ₂ And (4) extracting. The organic layer was separated, washed with brine, na ₂ SO ₄ And (5) drying. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 6) after spin-drying the solvent under reduced pressure to give colorless oil 51 (2.60g, 9.76mmol, 49%): ¹ H NMR(400MHz,CDCl ₃ )δ7.39–7.22(m,5H),4.58(s,2H),4.04(s,2H),3.74(dd,J＝5.7,3.4Hz,2H),3.67(dd,J＝5.9,3.4Hz,2H),1.47(s,9H).MS(ESI)m/z 267[M+H] ⁺ .

and 2, step: synthesis of tert-butyl (2-hydroxyethoxy) acetate

10% Palladium on carbon catalyst (200 mg) was added to a solution of intermediate 51 (2.40g, 9.01mmol) in ethanol (6 mL). The hydrogen was replaced and the reaction system was stirred at 45 ℃ for 24 hours. The reaction mixture was filtered through a pad of celite, washing with ethyl acetate. Filtrate is extracted with ethyl acetate and H ₂ And (4) extracting. The combined organic layers were washed with brine and Na ₂ SO ₄ And (5) drying. After filtration and evaporation, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 1) to give colorless oil 52 (680 mg,3.86mmol, 43%): ¹ H NMR(400MHz,CDCl ₃ )δ7.39–7.22(m,5H),4.58(s,2H),4.04(s,2H),3.74(dd,J＝5.7,3.4Hz,2H),3.67(dd,J＝5.9,3.4Hz,2H),1.47(s,9H).

and 3, step 3: synthesis of (2s, 4r) -1- ((S) -2- (((2- ((E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanopropionylamino) ethoxy) methyl) amino) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide compound intermediate 52 the synthetic method for final product 22 was as in steps 2-6 of example 19.

¹ H NMR(400MHz,CDCl ₃ )δ8.65(s,1H),8.22(s,1H),7.94(d,J＝6.8Hz,2H),7.84(d,J＝8.2Hz,1H),7.68(s,1H),7.46–7.29(m,6H),7.15(d,J＝8.6Hz,1H),7.07(s,1H),6.86(d,J＝8.4Hz,1H),5.44(s,2H),4.72(t,J＝7.8Hz,1H),4.53(dd,J＝16.0,7.9Hz,3H),4.31(dd,J＝15.0,5.4Hz,1H),4.04(t,J＝12.9Hz,2H),3.94(s,3H),3.77-3.69(m,1H),3.62(dd,J＝15.8,4.0Hz,4H),2.49(s,4H),2.11(dd,J＝13.5,8.1Hz,1H),1.99(s,2H),0.94(s,9H).HRMS(ESI ⁺ ):calculated for C ₄₆ H ₄₈ F ₆ N ₆ O ₈ S[M+H] ⁺ :959.3231,found 959.3239.

Example 23: (2S, 4R) -1- ((S) -2- ((4- ((E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanopropionylamino) butyl) amino) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide

Step 1:

tert-butyl (5- (((S) -1- ((2S, 4R) -4-hydroxy-2- ((4- (4-methylthiazol-5-yl) benzyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) -5-oxopentyl) carbamic acid tert-butyl ester

HATU (133mg, 0.35mmol), DIPEA (0.20ml, 1.2mmol) and 53 (100mg, 0.23mmol) were added to a solution of 5- ((tert-butoxycarbonyl) amino) pentanoic acid (61mg, 0.28mmol) in DMF (2 ml). After stirring at room temperature for 1 hour, the resulting mixture was extracted with ethyl acetate and saturated NaHCO ₃ And (4) extracting with an aqueous solution. The organic layer was separated, washed with brine, na ₂ SO ₄ And (5) drying. The solvent was dried under reduced pressure and the residue was purified by silica gel column chromatography (DCM/MeOH = 20) to give 54 (89mg, 0.14mmol,61% yield) as a white solid: ¹ H NMR(400MHz,CDCl ₃ )δ8.68(s,1H),7.40-7.32(m,5H),6.33(d,J＝8.3Hz,1H),4.72(t,J＝7.9Hz,2H),4.62-4.49(m,3H),4.33(dd,J＝15.0,5.2Hz,1H),4.10(d,J＝9.9Hz,1H),3.60(d,J＝8.5Hz,1H),3.08(d,J＝6.0Hz,2H),2.51(s,4H),2.26-2.11(m,3H),1.59(d,J＝15.0Hz,2H),1.48-1.39(m,12H),0.93(s,9H).MS(ESI)m/z 602[M+H] ⁺ .

step 2:

(2S, 4R) -1- ((S) -2- ((4- ((E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanopropionylamino) butyl) amino) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide

TFA (1 ml) was added to a solution of compound 54 (80mg, 0.13mmol) in DCM (2 ml). After stirring for 1 hour, the solvent was spin dried under reduced pressure, toluene (3X 3 mL) was added and residual TFA was removed by evaporation. The crude product was used in the next step without further purification. HATU (76mg, 0.20mmol), DIPEA (110. Mu.L, 0.65 mmol) and intermediate compound 42 (73mg, 0.16mmol) were added to a solution of the above crude product in DMF (2 ml). Stirred at 25 ℃ for 1 hour, and then saturated NaHCO with ethyl acetate ₃ The resulting mixture was extracted with an aqueous solution. The organic layer was separated, washed with brine, na ₂ SO ₄ And (5) drying. The solvent was dried by spinning under reduced pressure and then purified by silica gel column chromatography (MeOH: CH) ₂ Cl ₂ = 4) purification of the residue to give 23 (81mg, 0.085mmol, 65%) white solid: ¹ H NMR(400MHz,CDCl ₃ )δ8.69(s,1H),8.22(s,1H),7.97(d,J＝6.8Hz,2H),7.87(d,J＝8.2Hz,1H),7.71(d,J＝2.0Hz,1H),7.41-7.31(m,6H),6.91-6.80(m,2H),6.31(d,J＝5.8Hz,1H),5.45(d,J＝14.3Hz,2H),4.74(t,J＝8.0Hz,1H),4.62-4.53(m,3H),4.35(dd,J＝15.0,5.3Hz,1H),4.15(d,J＝11.2Hz,1H),3.98(s,3H),3.64(dd,J＝11.3,3.3Hz,1H),3.57-3.30(m,3H),2.52(s,4H),2.29(dt,J＝15.0,7.5Hz,2H),2.17(dd,J＝13.4,8.0Hz,1H),1.70(dd,J＝13.4,6.7Hz,2H),1.66-1.56(m,2H),0.96(s,9H).HRMS(ESI ⁺ ):calculated for C ₄₇ H ₅₀ F ₆ N ₆ O ₇ S[M+H] ⁺ :957.3439,found 957.3435.

example 24: (2S, 4R) -1- ((S) -2- (7- ((E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanopropionylamino) heptanamido) -3, 3-dimethylbutanoyl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide

The synthesis was as in example 23

¹ H NMR(400MHz,CDCl ₃ )δ8.67(s,1H),8.17(s,1H),7.95(d,J＝5.9Hz,2H),7.85(d,J＝8.2Hz,1H),7.69(d,J＝1.5Hz,1H),7.41-7.31(m,6H),6.86(d,J＝8.4Hz,1H),6.44(t,J＝5.4Hz,1H),6.18(d,J＝8.7Hz,1H),5.45(s,2H),4.73(t,J＝8.0Hz,1H),4.60-4.50(m,3H),4.34(dd,J＝15.0,5.2Hz,1H),4.12(d,J＝11.4Hz,1H),3.97(s,3H),3.60(dd,J＝11.3,3.1Hz,1H),3.49-3.34(m,3H),2.50(s,4H),2.27-2.11(m,3H),1.60(dd,J＝13.9,7.0Hz,4H),1.33(d,J＝4.6Hz,4H),0.93(s,9H).HRMS(ESI ⁺ ):calculated for C ₄₉ H ₅₄ F ₆ N ₆ O ₇ S[M+H] ⁺ :985.3752,found 985.3723.

Example 25: (2S, 4R) -1- ((S) -2- (8- ((E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanopropionylamino) octan) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide

Synthesis procedure as in example 23

¹ H NMR(400MHz,CDCl ₃ )δ8.67(s,1H),8.20(s,1H),7.95(d,J＝6.8Hz,2H),7.85(d,J＝8.2Hz,1H),7.70(d,J＝2.0Hz,1H),7.40-7.31(m,6H),6.86(d,J＝8.5Hz,1H),6.41(t,J＝5.7Hz,1H),6.17(d,J＝8.8Hz,1H),5.45(s,2H),4.72(t,J＝8.0Hz,1H),4.60-4.50(m,3H),4.33(dd,J＝15.0,5.2Hz,1H),4.11(d,J＝10.9Hz,1H),3.97(s,3H),3.60(dd,J＝11.4,3.5Hz,1H),3.39(dd,J＝13.4,6.7Hz,3H),2.57-2.46(m,4H),2.24-2.09(m,3H),1.58(dd,J＝14.1,7.2Hz,4H),1.32(d,J＝3.4Hz,6H),0.93(s,9H).HRMS(ESI ⁺ ):calculated for C ₅₀ H ₅₆ F ₆ N ₆ O ₇ S[M+H] ⁺ :999.3908,found 999.3888.

Example 26: binding Capacity test with ERRa

Binding Capacity of Compounds described in examples 1-25 to ERRa Using the TR-FRET assay, a commercial screening kit (LanthaScreen) ^TM Estrogen Related Receptor alpha TR-FRET Coactator Assay, invitrogen PV 4663) which adopts the principle of fluorescence resonance energy transfer, and the specific method comprises the following steps: the compounds were prepared in 5 concentration-gradient (2X) solutions using Buffer, and 10. Mu.l of each concentration solution was taken in a 384-well blackboard (Thermo, # 267461), and 5. Mu.l of ERR alpha-LBD (20nM, 4X) solution was added to each well, followed by 5. Mu.l of a mixed solution comprising fluo-resein-PGC 1 α (2. Mu.M, 4X) and Tb anti-GST antibody (2nM, 4X); the reaction was left at room temperature for 1 hour away from light and absorbance at 495nm and 520nm was measured.

And making a positive correlation with the concentration of the compound, making a curve of the correlation with the concentration, and calculating an IC50 value.

Compound numbers and corresponding TR-FRET activity results are listed in table 1.

TABLE 1 binding of Compounds to ERR alpha

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In conclusion, in the TR-FRET detection, when the R1 site is halogen fluorine, the activity of the corresponding compounds is relatively poor. When the R1 site is trifluoromethyl, the corresponding acid, amide and hydroxylamine have certain activity, but the methyl ester does not. When R is ₁ In the case of cyano, the series of compounds are more active in inhibiting the binding of ERR alpha to PGC-1.

Example 27: effect of Compounds on ERR α expression in Breast cancer MDA-MB-231 cells

Cell lines: breast cancer MDA-MB-231 cell line was purchased from the american standard biological collection (ATCC).

Detection was performed using a conventional Western Blot (immunoblotting), as follows. The MDA-MB-231 cells are planted on a 12-well plate according to a certain quantity, after being adhered to the wall in an incubator and cultured overnight, a compound with a certain concentration is added for acting for 4 hours, and lysis is carried out by using a lysis solution to collect samples. Then, an appropriate amount of the sample was subjected to SDS-PAGE, after which proteins were transferred to a nitrocellulose membrane using a semidry electrotransfer system, the nitrocellulose membrane was blocked in a blocking solution (5% skim milk powder diluted in TBS containing 0.1% Tween 20) at room temperature for 2h, and then the membrane was incubated overnight at 4 ℃ in primary antibody solutions (1. Washed three times with TBS containing 0.1-vol Tween 20, 15min each time. The membrane was placed in a secondary antibody solution (horseradish peroxidase-labeled goat anti-rabbit IgG,1, 1000 diluted in TBS containing 0.1% Tween 20) for a reaction at room temperature for 1h. After washing the membrane three times as above, the membrane was developed with ECL plus reagent and photographed by Amersham Imager 600 system.

TABLE 2 ERR alpha protein levels in MDA-MB-231 cells

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From table 2 (calculated from fig. 1), it can be seen that the ERR α protein levels in MDA-MB-231 cells were significantly degraded in the above compounds, examples 16-25, of which example 23 was the strongest, and the ERR α protein degradation efficiency was 39%, 83%, and 96% at 10nM, 30nM, and 100nM, respectively.

Example 28: in-vitro migration inhibition effect of compound on breast cancer cell MDA-MB-231

The method comprises the following steps: the Tanswell cell was purchased from Corning, USA, and has a pore size of 8 μm. Prior to use, the same amount of MDA-MB-231 cells were added to the upper chamber in 200. Mu.l of serum-free PRMI-1640 medium containing different compound concentrations, to the lower chamber in 800. Mu.l of medium containing 10% fetal bovine serum, and the chambers were placed at 37 ℃ and 5% CO ₂ After 24h incubation in the incubator, the chamber was removed, the cells were fixed at the bottom of the chamber by immersing the cells in crystal violet for 20 minutes, the cells which did not pass through the pores in the upper layer of the chamber were gently wiped off with a cotton swab, and then washed with clean water. The number of migrated cells in 6 different fields in each chamber was counted by observing and photographing under a 200-fold microscope, and the average value was calculated.

As can be seen from FIG. 2, of the above compounds, the compound of example 23 significantly inhibited MDA-MB-231 cell migration, with inhibition rates of about 30% and 50% at 1.0. Mu.M and 6.0. Mu.M, respectively.

The technical features of the above-mentioned embodiments can be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the following embodiments are not described, however, as long as there is no contradiction between the combinations of the technical features, the combinations should be considered as the scope of the present description.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is specific and detailed, but not to be understood as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

Claims

1. A compound having the structure of formula (i) or a pharmaceutically acceptable salt or stereoisomer or prodrug molecule thereof:

wherein R1 is selected from:

a cyano group;

l is optionally selected from:

1)–NH–(CH ₂ ) _m –；

2)–NH–((CH ₂ ) ₂ –O) _n –(CH ₂ ) ₂ –；

3)–NH–(CH ₂ ) _u –O–(CH ₂ ) _v –

wherein m is 0,1, 2, 3, 4, 5, 6, 7 or 8;

n is 0,1, 2, 3, 4, 5 or 6;

u is 0,1, 2, 3, 4, 5, 6, 7 or 8;

v is 0,1, 2, 3, 4, 5, 6, 7 or 8;

4) L is absent;

y is optionally selected from:

1)–C(＝O)–；

2) Y is absent;

b is optionally selected from:

1)H；

2)OR ₂ ；R ₂ is optionally selected from: h, C ₁ ～C ₃ Alkyl radical, C ₃ ～C ₆ A cycloalkyl group;

3)

R ₃ is optionally selected from: h, C ₁ ～C ₃ Alkyl radical, C ₃ ～C ₆ A cycloalkyl group.

2. The use of a compound having the structure of formula (i) or a pharmaceutically acceptable salt or stereoisomer thereof, or a prodrug molecule thereof, in the preparation of an ERR α protein inhibitor:

wherein R is ₁ Selected from:

a cyano group;

l is optionally selected from:

1)–NH–(CH ₂ ) _m –；

2)–NH–((CH ₂ ) ₂ –O) _n –(CH ₂ ) ₂ –；

3)–NH–(CH ₂ ) _u –O–(CH ₂ ) _v –

wherein m is 0,1, 2, 3, 4, 5, 6, 7 or 8;

n is 0,1, 2, 3, 4, 5 or 6;

u is 0,1, 2, 3, 4, 5, 6, 7 or 8;

v is 0,1, 2, 3, 4, 5, 6, 7 or 8;

4) L is absent;

y is optionally selected from:

1)–C(＝O)–；

2) Y is absent;

b is optionally selected from:

1)H；

3)

3. The use of a compound according to claim 2, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, wherein L is selected from any of:

1)–NH–(CH ₂ ) _m –；

2)–NH–((CH2) ₂ –O) _n –(CH ₂ ) ₂ –；

3)–NH–(CH ₂ ) _u –O–(CH ₂ ) _v –

wherein m is 0,1, 2, 3, 4, 5, 6, 7 or 8;

n is 0,1, 2, 3, 4, 5 or 6;

u is 0,1, 2, 3, 4, 5, 6, 7 or 8;

v is 0,1, 2, 3, 4, 5, 6, 7 or 8.

4. The use of a compound according to claim 2, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, wherein Y is selected from: -C (= O) -.

5. The compound of claim 2, or a pharmaceutically acceptable salt or stereoisomer or prodrug molecule thereof, wherein B isSelected from: OR (OR) ₂ Or

6. The use of a compound according to claim 5, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, wherein the compound has the structure of formula (iii):

7. the use of a compound according to claim 2, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, wherein the compound is selected from the group consisting of:

(E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanoacrylamide; (E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyano-N-hydroxyacrylamide;

(E) -3- (2- (3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanoacrylamido) ethoxy) propionic acid;

8. The use of a compound according to claim 7, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, wherein the compound is selected from the group consisting of:

(2s, 4r) -1- ((S) -2- (7- ((E) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2-cyanopropionylamino) heptanamido) -3, 3-dimethylbutanoyl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide;

9. Use of a compound or a pharmaceutically acceptable salt or stereoisomer thereof or a prodrug molecule thereof for the manufacture of a medicament for the prevention or treatment of a disease associated with aberrant expression of ERR α protein activity, wherein the compound or pharmaceutically acceptable salt or stereoisomer thereof or prodrug molecule thereof is as defined in any one of claims 1 to 8.

10. The use according to claim 9, wherein the diseases associated with the abnormal expression of ERR α protein activity comprise: tumors, hyperglycemia, diabetes, obesity, hyperlipidemia, hypercholesterolemia, hyperlipoproteinemia, hypertriglyceridemia, hypertension, hyperinsulinemia, hyperuricemia, parkinson's disease, and Alzheimer's disease.

11. Use of a compound or a pharmaceutically acceptable salt or a stereoisomer or a prodrug molecule thereof for the manufacture of a medicament for the treatment or prevention of a tumour or for the prevention of post-operative recurrence of a tumour, characterised in that the compound or a pharmaceutically acceptable salt or a stereoisomer or a prodrug molecule thereof is as defined in any one of claims 1 to 8.

12. The use of claim 11, wherein the tumor is: non-small cell lung cancer, malignant melanoma, prostate cancer, renal cancer, bladder cancer, ovarian cancer, colon cancer, rectal cancer, breast cancer, cervical cancer, lung cancer, larynx cancer, nasopharyngeal carcinoma, pancreatic cancer or multiple myeloma, B lymphoma, and leukemia.

13. An ERR alpha protein inhibitor, characterized in that the active ingredient contains the compound or the pharmaceutically acceptable salt or the stereoisomer or the prodrug molecule thereof according to claim 1, or the compound or the pharmaceutically acceptable salt or the stereoisomer or the prodrug molecule thereof according to any one of claims 2 to 8.

14. A medicament for treating or preventing tumors or preventing postoperative recurrence of tumors, wherein the active ingredient comprises the compound of claim 1 or a pharmaceutically acceptable salt or stereoisomer thereof or a prodrug molecule thereof, or the compound of any one of claims 2 to 8 or a pharmaceutically acceptable salt or stereoisomer thereof or a prodrug molecule thereof.