CN111233661A - Compound for targeted ubiquitination degradation of ERR α protein and medicinal composition and application thereof - Google Patents

Abstract

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

Description

Compound for targeted ubiquitination degradation of ERR α 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 α protein, and a pharmaceutical composition 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, HER2/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 have become the treatment first choice. However, about 15-20% of the breast cancer cases worldwide are currently diagnosed with triple-negative breast cancer (TNBC, i.e., ER-, PgR-, HER2-), because TNBC is not suitable for endocrine therapy and drug therapy against HER2 target, and has high malignancy and 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 of anti-tumor drugs.

The ERR β is related to early development, and ERR β and ERR γ are considered as new potential targets of metabolic disorders, wherein ERR α is the first most discovered and studied in this subgroup, which is widely expressed in tissues with vigorous energy metabolism such as heart, kidney, skeletal muscle, gastrointestinal tract, etc., and is directly associated with peroxisome proliferator-activated receptor co-activator (peroxisome proliferator-activated receptor-gamma coactivator 1, PGC-1), the expression levels of regulatory factors during regulation of tricarboxylic acid cycle and glycolysis, such as C-1, medium-chain acyl-coenzyme A dehydrogenase (PDD), pyruvate dehydrogenase kinase (MCAK 4), and the like, thereby the expression levels of regulatory factors during regulation of tricarboxylic acid cycle and glycolysis, such as the expression levels of C-1, medium-chain acyl-coenzyme A dehydrogenase (PDD), pyruvate dehydrogenase kinase (MCAK 4), and the like, are significantly correlated with the development of tumor metastasis-inhibiting genes, such as a primary tumor metastasis inhibiting factor, and inhibiting the development of breast cancer, breast cancer metastasis, breast cancer.

The small molecule inhibitors play a role in regulating the protein activity by combining active sites, belong to a mode of occupying drive, but have the defects that firstly, the long-term use of the small molecule drugs can generate drug resistance, secondly, in order to achieve the required effect, the small molecule compounds need to keep higher concentration in cells, thereby causing off-target and generating adverse reaction, and meanwhile, a plurality of potential target proteins do not have pockets which can be directly combined with the small molecule inhibitors, such as non-enzyme proteins such as transcription factors, and the difficulty in developing the small molecule inhibitors is higher.

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 (PROTACs) are essentially hybrid bifunctional small molecule compounds, with the mode of action different from the "occupancy driving" mode of traditional small molecule inhibitors, and the 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, target protein is ubiquitinated, target protein is marked with 'label', and finally ubiquitinated 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 desheies topic group reports the first batch of bifunctional molecules of PROTACs in 2001, the initial technology is a PROTAC technology based on short peptide fragments, targets for successful degradation comprise MetAP2, androgen receptors and the like, related reported PROTACs are polypeptide compounds, the defects of large molecular weight, poor cell permeability, poor activity and the like exist in the compounds, the development of the technology is limited, in recent years, E3 ubiquitin ligase specific small molecule ligands such as CRBN, VHL, cIAP and the like are found, the PROTAC technology is enabled to make a huge breakthrough, in 2008, Crew topic group reports the first small molecule PROTAC, PROTAC _ AR, which connects MDM2E3 ubiquitin ligase small molecule inhibitor nutlin with non-steroidal androgen receptor small molecule ligand through polyethylene glycol linker, the degradation of AR in human cervical carcinoma HeLa cells is successfully realized, and in the following years, researchers of various scientific research institutions respectively develop CRABBP-II _ TAC based on CLAP1E3 ligase ligand, BRE 3 ligase based on non-steroidal androgen receptor small molecule ligand nutlin, BRBcr 2 ligase based on CRAB ligand, BRD 2 ligase, BRTAC, and CRAC target protein linked with BRTAC, and BRTAC are successfully realized on the related protein.

Therefore, by designing and synthesizing an inverse agonist with stronger activity on ERR α protein and applying the proteolytic targeting chimeras (PROTACs) technology to ERR α protein, important references can be provided for ERR α as a drug development target, the treatment of triple negative breast cancer and the development and application feasibility of the PROTAC technology.

Disclosure of Invention

In view of the above problems, an object of the present invention is 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) halogen;

3)C_1～C₃an alkyl group;

4)C₃～C₆a cycloalkyl group;

5)C₁～C₃an alkoxy group;

6) a fluoromethyl group;

7) a cyano group;

l is optionally selected from:

1)–NH–(CH₂)_m–；

2)–NH–((CH₂)₂–O)_n–(CH₂)₂–；

3)–NH–(CH₂)_u–O–(CH₂)_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)–CH₂–,–C(＝O)–；

2) y is absent;

b is optionally selected from:

1)H；

2)OR₂(ii) a R2 is optionally selected from: h, C₁～C₃Alkyl radical, C₃～C₆A cycloalkyl group;

3)

r3 is optionally selected from: h, C₁～C₃Alkyl radical, C₃～C₆A 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 application of a compound with a structure shown as a formula (I) or a pharmaceutically acceptable salt or a stereoisomer or a prodrug molecule thereof in preparing an ERR α protein inhibitor:

wherein R1 is selected from:

1)H；

2) halogen;

3)C_1～C₃an alkyl group;

4)C₃～C₆a cycloalkyl group;

5)C₁～C₃an alkoxy group;

6) a fluoromethyl group;

7) a cyano group;

l is optionally selected from:

1)–NH–(CH₂)_m–；

2)–NH–((CH₂)₂–O)_n–(CH₂)₂–；

3)–NH–(CH₂)_u–O–(CH₂)_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)–CH₂–,–C(＝O)–；

2) y is absent;

b is optionally selected from:

1)H；

2)OR₂(ii) a R2 is optionally selected from: h, C₁～C₃Alkyl radical, C₃～C₆A cycloalkyl group;

3)

r3 is optionally selected from: h, C₁～C₃Alkyl radical, C₃～C₆A cycloalkyl group.

In some of these embodiments, R₁Is optionally selected from: halogen, fluoromethyl, cyano.

In some of these embodiments, L is selected from:

1)–NH–(CH₂)_m–；

2)–NH–((CH2)₂–O)_n–(CH₂)₂–；

3)–NH–(CH₂)_u–O–(CH₂)_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: -CH 2-, -C (═ O) -.

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

R₂Is optionally selected from: h, C₁～C₃Alkyl radical, C₃～C₆A cycloalkyl group; r₃Is optionally selected from: h, C₁～C₃Alkyl radical, C₃～C₆A cycloalkyl group.

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

l is optionally selected from:

1)–NH–(CH₂)_m–；

2)–NH–((CH₂)₂–O)_n–(CH₂)₂–；

3)–NH–(CH₂)_u–O–(CH₂)_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)–CH₂–,–C(＝O)–；

2) y is absent;

b is optionally selected from:

1)H；

2)OR₂(ii) a R2 is optionally selected from: h, C₁～C₃Alkyl radical, C₃～C₆A cycloalkyl group;

3)

r3 is optionally selected from: h, C₁～C₃Alkyl radical, C₃～C₆A cycloalkyl group.

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

l is optionally selected from:

1)–NH–(CH₂)_m–；

2)–NH–((CH₂)₂–O)_n–(CH₂)₂–；

3)–NH–(CH₂)_u–O–(CH₂)_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;

r3 is optionally selected from: h, C₁～C₃Alkyl radical, C₃～C₆A cycloalkyl group.

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) propionic 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-diaza-hexadeca-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-dimethylbutanoyl) -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) -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-diaza-hexadeca-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-dimethylbutanoyl) -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.

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 α protein activity.

In some of these embodiments, the disease associated with aberrant expression of ERR α protein activity includes tumors, hyperglycemia, diabetes, obesity, hyperlipidemia, hypercholesterolemia, hyperlipoproteinemia, hypertriglyceridemia, hypertension, hyperinsulinemia, hyperuricemia, Parkinson's disease, and Alzheimer's 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 α protein inhibitor, which has the following specific technical scheme:

an ERR α protein inhibitor contains the above compound or its pharmaceutically acceptable salt or stereoisomer or its prodrug molecule as 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 α protein and degrading ERR α protein, has stronger subtype selectivity, and can also effectively inhibit MDA-MB-231 cell migration of triple negative breast cancer, so that the compound can be used for diseases related to abnormal expression of ERR α protein, such as various cancers.

The compound can effectively inhibit ERR α protein and has the function of degrading ERR α target protein, the protein degradation mechanism is that one end of the molecule is combined with ERR α target protein, the other end is combined with E3 ubiquitin ligase of ubiquitin-protease system, target protein is ubiquitinated, and finally the degradation of ERR α target protein is successfully realized by proteasome.

Drawings

FIG. 1 shows the results of the 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、R₂Etc.) 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 to be understood that substituents and substitution patterns on the compounds of the present invention may be selected by one of ordinary skill in the art to provide compounds that are chemically stable and that can be readily synthesized by those skilled in the art and by the 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_1-5Alkyl radical "middle" C_1-5The definition of "includes groups having 1, 2, 3, 4, or 5 carbon atoms in a linear or branched arrangement. For example, "C_1-5Alkyl "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 analogues 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, tetrahydrofuranyl 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 pharmaceutically acceptable salts include not only the exemplary salts of the particular compounds described herein, but also 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 suitable inorganic or organic bases.

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 acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, and the like, as well as those prepared from organic acids such as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanilic acid, 2-acetoxy-benzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethane disulfonic acid, oxalic acid, isethionic acid, trifluoroacetic acid, 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' -dibenzylethylenediamine, diethylamine, 2-dimethylaminoethanol, aminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, histidine, hydroxycobalamin, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, piperdine, polyamine resins, procaine, purines, 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 and pharmaceutically acceptable salts thereof to which this application relates, and prodrugs that can be converted in vivo to the structures of the compounds and pharmaceutically acceptable salts thereof to which this application relates, are also included in the claims of this application.

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

Dissolving compound 26(3.07g, 10mmol) and vanillin (1.52g,10mmol) in DMF (10ml), adding K₂CO₃(2g, 14mmol) at 80 deg.CStirred 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 Na₂SO₄And (5) drying. After drying the solvent under reduced pressure, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 7: 1) to give product 27 as a white solid (3.21g, 8.50mmol, 85%):¹H NMR(400MHz,CDCl₃)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 dry THF (20ml) under argon. n-BuLi (10mmol,6.3mL) was added dropwise at 0 deg.C (1.6 mmol/mL). After stirring for a further 30 minutes, 10ml of ethyl 2- (diethoxyphosphoryl) acetate (10mmol,2.24g) in THF are 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, Na₂SO₄And (5) drying. After drying the solvent under reduced pressure, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 7: 1) to give product 28 as a white solid (3.0g, 6.43mmol, 64%):¹H NMR(400MHz,DMSO-d₆)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/z467[M+H]⁺。

and 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 (8ml), NaOH (520mg,13mmol) was added, and water (8ml) 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%):¹H NMR(400MHz,CDCl₃)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, after synthesis of compound 27, the following reaction was directly carried out to give (Z) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2- (trifluoromethyl) acrylic acid:

compound 27(3.78g, 10mmol), 3,3, 3-trifluoropropionic acid (10mmol,0.88ml) was dissolved in anhydrous THF (20ml) under argon. Titanium tetrachloride (16mmol,1.8ml) was added dropwise at 0 ℃. Stirring was continued for 30 min and triethylamine (5.6ml,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 Na₂SO₄And (5) drying. After spin-drying of the solvent under reduced pressure, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate ═ 1:1) to give product 2(3.1g,6.3mmol, 63% yield) as a yellow solid:¹H NMR(400MHz,CDCl₃)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 (20ml), followed by piperidine (1.5ml), which was stirred 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 the solid. The mixture was extracted with ethyl acetate and water. The organic layer was separated, washed with brine and Na₂SO₄And (5) drying. After spin-drying of the solvent under reduced pressure, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate ═ 1:1) to give the product as a light yellow solid 3(2.6g, 5.8mmol, 75.6%):¹H NMR(400MHz,DMSO-d₆)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 K₂CO₃(121.6mg, 0.88mmol) and CH₃I (41. mu.L, 0.66 mmol). After stirring at room temperature for 1 hour, ethyl acetate and saturated NaHCO were added₃The resulting mixture was extracted. The organic layer was separated, washed with brine, Na₂SO₄And (5) drying. After filtration and evaporation, the residue was purified by silica gel column chromatography (PE: EA ═ 2:1) to give 4(190mg, 0.42mmol, 95%) as a white solid:¹H NMR(400MHz,CDCl₃)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) -3- (4- ((2, 4-bis (trifluoromethyl) benzyl) oxy) -3-methoxyphenyl) -2- (trifluoromethyl) acrylic acid methyl ester

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

¹H NMR(300MHz,CDCl₃)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.

¹H NMR(400MHz,DMSO-d₆)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 (2mL), oxalyl chloride (0.5mL) 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 (2mL) and 28% aqueous ammonia (1mL) was added at 0 ℃. After stirring at room temperature for 2 hours, the mixture was taken up with DCM and H₂O extraction, separation of the organic layer, washing with brine and Na₂SO₄And (5) drying. The solvent was dried under reduced pressure and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 3:1) to give product 7(52mg, 0.12mmol, 55%) as a white solid:¹H NMR(400MHz,CDCl₃)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.

¹H NMR(400MHz,CDCl₃)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 was substantially the same as in example 7, except that the starting material used in this example was compound 3 prepared in example 3.

¹H NMR(400MHz,DMSO-d₆)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 as in example 2(215mg, 0.44mmol) and O- (tetrahydro-2H-pyran-2-yl) hydroxylamine (51.5mg,0.44mmol) were dissolved in anhydrous DMF (3ml) and EDCI (168.7mg,0.88mmol) and HOBT (119mg,0.88mmol) were added. After stirring overnight, the mixture was extracted with ethyl acetate and water. The organic layer was separated, washed with brine, Na₂SO₄And (5) drying. After drying the solvent under reduced pressure, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate ═ 6:1) to give the product 29(111.6mg, 0.19mmol, 43%) as a pale yellow oil:¹H NMR(400MHz,CDCl₃)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]⁺.

step 2: 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 (4mL) and concentrated hydrochloric acid (4mL) was added dropwise. After stirring overnight, water was added to precipitate, which was filtered and washed with water to give product 10 as a white solid (80.5mg,0.16mmol, 94%):¹H NMR(400MHz,CDCl₃)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.

¹H 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/z454[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.

¹H 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-methoxyethon-1-amine (39.8mg, 0.53mmol), HATU (216.7mg, 0.57mmol) and DIPEA (0.22ml, 1.3mmol) were added to intermediate compound 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 NaHCO₃And (4) extracting. The organic layer was separated, washed with brine, Na₂SO₄And (5) drying. After spin-drying the solvent under reduced pressure, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate ═ 3:1) to give 13(183mg, 0.36mmol, 82% yield) as a white solid:¹H NMR(400MHz,DMSO-d₆)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 C₂₃H₂₀F₆N₂O₄[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-aminopentanoate (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 NaHCO₃And (4) extracting. The organic layer was separated, washed with brine, Na₂SO₄And (5) drying. After the solvent was dried by spinning under reduced pressure, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate ═ 2:1) to give a yellow solid tert-butyl ester intermediate (178mg, 0.30mmol, 68%). TFA (1ml) was added to a solution of the above tert-butyl ester intermediate in DCM (2 ml). After stirring for 1h, the solvent was dried under reduced pressure, toluene (3X 3mL) was added and residual TFA was evaporated to give 14 as a yellow solid (155mg, 0.28mmol, 93%):¹H NMR(400MHz,DMSO-d₆)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⁺):calculatedfor C₂₅H₂₂F₆N₂O₅[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) propionic 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).

¹H NMR(400MHz,DMSO-d₆)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 ChemicalFormula:C₂₅H₂₂F₆N₂O₆[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 (5mL) and tert-butanol (0.7mL, 7.6mmol) was added. Stir at room temperature for 3 hours. Decompression is revolvedThe solvent was dried to give a residue. The residue was purified by column chromatography (petroleum ether/ethyl acetate ═ 6:1) to give 33(949.8mg, 3.4mmol, 89.0% yield) as a colourless 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 ] M/z]⁺.

Step 2: preparation of tert-butyl 3-aminopropionate

10% Palladium on carbon catalyst (90mg) 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(159.7mg, 1.1mmol, 48% yield) as a colourless oil:¹HNMR(400MHz,CDCl₃)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, 1.3mmol) 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 NaHCO₃And (4) extracting. The organic layer was separated, washed with brine, Na₂SO₄And (5) drying. The solvent was dried under reduced pressure and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate ═ 2:1) to give 35(170mg, 0.30mmol, 68% yield) as a pale yellow solid:¹H NMR(400MHz,CDCl₃)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-bromobenzonitrile (7.0g, 38.5mmol), Pd (OAc) was measured₂(43.2mg, 0.19mmol) and KOAc (5.67g, 57.8mmol) in a two-necked flask under argon, DMAc (40mL) and 4-methylthiazole (5.3mL, 57.8mmol) were added 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 brine, Na₂SO₄After drying and spin-drying of the solvent under reduced pressure, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate ═ 5: 1) to give 38(5.9g, 29.5mmol, 77% yield) as a white solid:¹H NMR(400MHz,CDCl₃)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) methylamine

Compound 38(5.0g,25mmol) was dissolved in methanol (125ml), CoCl2(4.5g,35mmol) was added under ice-bath conditions and NaBH was added portionwise₄(3.8g, 100 mmol). After stirring at room temperature for 90 minutes, the reaction was quenched with aqueous ammonia and water, the reaction mixture was filtered through celite, the celite was washed with DCM: MeOH: TEA ═ 10:1:0.1 solvent, the resulting filtrate was extracted with saturated aqueous sodium bicarbonate and DCM: MeOH ═ 10:1 solvent, the solvent was dried under reduced pressure, and the residue was purified by silica gel column chromatography (first ethyl acetate and then DCM: MeOH ═ 10:1) to give 39(1.9g,9.3mmol, 37% yield) as a yellow oil:¹HNMR(400MHz,CD₃OD_SPE)δ8.87(s,1H),7.45(s,4H),3.85(s,2H),3.31(s,1H).MS(ESI)m/z205[M+H]⁺.

step 6: preparation of tert-butyl (2S, 4R) -4-hydroxy-2- ((4- (4-methylthiazol-5-yl) benzyl) carbamoyl) pyrrolidine-1-carboxylate

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 NaHCO₃And (4) extracting with an aqueous solution. The organic layer was separated, washed with brine and Na₂SO₄And (5) drying. The solvent was dried under reduced pressure and purified by silica gel column chromatography (MeOH: CH)₂Cl₂Purify the residue to give 40(2.6g, 6.2mmol, 67%) as a white solid, ms (esi) M/z 418[ M + H ]]⁺.

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 (10ml) was added to a solution of compound 40(2.6g, 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-dimethylbutanoic acid (1.43g,6.2mmol) 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 and taken up with ethyl acetate and saturated NaHCO₃And (4) extracting with an aqueous solution. The organic layer was separated, washed with brine, Na₂SO₄And (5) drying. The solvent was dried under reduced pressure and purified by silica gel column chromatography (MeOH: CH)₂Cl₂Purification of the residue (4: 96) gave 41 as a colourless solid (2.4g, 4.5mmol, 73% yield) ms (esi) M/z 531[ M + H)]⁺.

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

Trifluoroacetic acid (10ml) was added to a solution of compound 41(2.4g, 4.5mmol) 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 and DCM: MeOH 10:1 solvent, separate the organic layer, wash with brine, and wash with Na₂SO₄And (5) drying. The solvent was dried under reduced pressure and purified by silica gel column chromatography (MeOH: CH)₂Cl₂Purify the residue 1:10) to give 42 as a white solid (1.2g, 2.8mmol, 62% yield):¹HNMR(400MHz,CDCl₃)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 (1ml) 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 3mL) 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 (83.6mg,0.22mmol), DIPEA (85. mu.L, 0.51mmol) 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 NaHCO₃And (4) extracting with saturated aqueous solution. The organic layer was separated, washed with brine and washed withNa₂SO₄And (5) drying. The solvent was dried under reduced pressure and purified by silica gel column chromatography (MeOH: CH)₂Cl₂4:96) to give 16(62mg, 0.067mmol, 39%) as a colorless solid:¹HNMR(400MHz,CDCl₃)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 C₄₅H₄₆F₆N₆O₇S[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

The synthesis was as in example 16.

¹H NMR(400MHz,CDCl₃)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 forC₄₆H₄₈F₆N₆O₇S[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.

¹H NMR(400MHz,CDCl₃)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 C₄₈H₅₂F₆N₆O₇S[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 (40mL) was added sodium metal (62.0mg, 2.70mmol) and stirred at room temperature for 2 hours. Tert-butyl acrylate (6.90,53.7mmol) was added and stirred overnight. After quenching with water, the solvent THF was spin-dried under reduced pressure and extracted with ethyl acetate and water. The organic layer was separated, washed with brine and Na₂SO₄And (5) drying. After filtration and evaporation, the mixture is purified by silica gel column chromatography (stone)Oil ether/ethyl acetate ═ 2:1) the residue was purified to give 44 as a colorless oil (5.0g, 26.3mmol, 49% yield):¹H NMR(400MHz,CDCl₃)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

P-toluenesulfonyl chloride (3.23g, 17mmol) in CH at-10 deg.C₂Cl₂(5mL) the solution was added dropwise to 44(2.5g, 13.1mmol), NEt₃(2.4mL, 17mmol) and DMAP (402.8mg, 3.3 mmol) in anhydrous CH₂Cl₂(5mL) in solution. The reaction mixture was stirred at room temperature for 8 hours. The resulting mixture was taken up in saturated NaHCO₃Extracted with DCM. The organic layer was separated, washed with brine and Na₂SO₄And (5) drying. DCM was dried under reduced pressure and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 7: 1) to give 45(3.9g, 11.3mmol, 86%) as a colourless oil:¹H NMR(400MHz,CDCl₃)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 step 3: synthesis of tert-butyl 3- (2-azidoethoxy) propionate

Adding NaN₃(2.8g, 43mmol) was added to a solution of 45(3.0g, 8.6mmol) 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 Na₂SO₄And (5) drying. The solvent was dried under reduced pressure and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 8:1) to give a colorless oil 46(1.7g, 7.9mmol, 92%):¹H NMR(400MHz,CDCl₃)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 PPh₃(1.84g, 7.0mmol) and water (3mL) were added to a solution of 46(1.0g, 4.7mmol) in THF (18mL) and stirred at room temperature overnight. The solvent was evaporated under reduced pressure and purified by silica gel column chromatography (2% MeOH/CH)₂Cl₂To 10% MeOH CH₂Cl₂) The residue was purified to give 47 as a colorless oil (525mg, 2.8mmol, 60%):¹HNMR(400MHz,CDCl₃)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(107.7mg, 0.53mmol), HATU (216.6mg, 0.57mmol) and DIPEA (0.24mL, 1.3mmol) 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 NaHCO₃And (4) extracting the solution. The organic layer was separated, washed with brine and Na₂SO₄And (5) drying. The solvent was dried under reduced pressure and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate ═ 2:1) to give 48(185mg, 0.30mmol, 68% yield) as a pale yellow solid:¹H NMR(400MHz,CDCl₃)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]⁺.

step 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 (1ml) 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 3mL) 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, 0.51mmol) and 42(70mg, 0.16mmol) were added to a solution of the above crude product in DMF (2ml) and stirred at 25 ℃ for 1 hour. With ethyl acetate and saturated NaHCO₃The resulting mixture was solution extracted. The organic layer was separated, washed with brine, Na₂SO₄And (5) drying. The solvent was dried under reduced pressure and then purified by silica gel column chromatography (MeOH: CH)₂Cl₂4:96) to give 19 as a white solid (72mg, 0.074mmol, 46%):¹H NMR(400MHz,CDCl₃)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 C₄₇H₅₀F₆N₆O₈S[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-diazahexadec-15 enoyl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide

Synthesis procedure as in example 19

¹H NMR(400MHz,CDCl₃)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 C₄₉H₅₄F₆N₆O₉S[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

¹H NMR(400MHz,CDCl₃)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 C₅₁H₅₈F₆N₆O₁₀S[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 dry tert-butanol (24mL) and stirred at room temperature for 30 minutes. Tert-butyl bromoacetate (3.90g, 20mmol) was added by injection at 10 ℃ and stirred at room temperature for 16 h. With ethyl acetate and H₂And (4) extracting. The organic layer was separated, washed with brine, Na₂SO₄And (5) drying. The solvent was dried under reduced pressure and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 6:1) to give 51(2.60g,9.76mmol, 49%) as a colorless oil:¹H NMR(400MHz,CDCl₃)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/z267[M+H]⁺.

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

10% Palladium on carbon catalyst (200mg) was added to a solution of intermediate 51(2.40g, 9.01mmol) in EtOH (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. The filtrate is extracted with ethyl acetate and H₂And (4) extracting. The combined organic layers were washed with brine and Na₂SO₄And (5) drying. After filtration and evaporation, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 1:1) to give 52 as a colorless oil (680mg, 3.86mmol, 43%):¹H NMR(400MHz,CDCl₃)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 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 synthesis of end product 22 is as for steps 2-6 in example 19.

¹H NMR(400MHz,CDCl₃)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 C₄₆H₄₈F₆N₆O₈S[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 taken up with ethyl acetate and saturated NaHCO₃And (4) extracting with an aqueous solution. The organic layer was separated, washed with brine, Na₂SO₄And (5) drying. The solvent was dried under reduced pressure and chromatographed over silica gel (DCM/MeOH ═ silica gel)20:1) purification of the residue to give 54 as a white solid (89mg, 0.14mmol, 61% yield):¹H NMR(400MHz,CDCl₃)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/z602[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 (1ml) 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 3mL) 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, 0.65mmol) and intermediate compound 42(73mg, 0.16mmol) were added to a solution of the above crude product in DMF (2 ml). Stirring at 25 deg.C for 1 hr, adding ethyl acetate and saturated NaHCO₃The resulting mixture was extracted with an aqueous solution. The organic layer was separated, washed with brine, Na₂SO₄And (5) drying. The solvent was dried under reduced pressure and then purified by silica gel column chromatography (MeOH: CH)₂Cl₂4:96) to give 23 as a white solid (81mg, 0.085mmol, 65%):¹H NMR(400MHz,CDCl₃)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 C₄₇H₅₀F₆N₆O₇S[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

Synthesis procedure as in example 23

¹H NMR(400MHz,CDCl₃)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 C₄₉H₅₄F₆N₆O₇S[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

¹H NMR(400MHz,CDCl₃)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⁺):calculatedfor C₅₀H₅₆F₆N₆O₇S[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)^TMEstrogen Related Receptor alpha TR-FRET Coactivator assay, Invitrogen PV4663) using the principle of fluorescence resonance energy transfer, the method comprises preparing 5 concentration gradient (2X) solutions of the compounds with Buffer, taking 10. mu.l of each concentration solution to 384-well blackboard (Thermo, #267461), adding 5. mu.l of ERR alpha-LBD (20nM, 4X) solution to each well, adding 5. mu.l of a mixed solution comprising fluoro-resein-PGC 1 α (2. mu.M, 4X) and Tb anti-GST antibody (20nM, 4X), placing at room temperature, and reacting in the dark for 1 hr to detect the absorptions at 495nM and 520 nM.

And (4) making a positive correlation with the concentration of the compound, making a concentration-related curve, and calculating an IC50 value.

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

Binding of the compounds of Table 1 to ERR α

In conclusion, in the TR-FRET detection, when the R1 site is halogen fluorine, the activity of the corresponding compound 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 is₁In the cyano group, the series of compounds have stronger activity of inhibiting the combination of ERR α and 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 taking a proper amount of sample to carry out SDS-PAGE electrophoresis, transferring the protein to a nitrocellulose membrane by using a semi-dry electrotransfer system after the electrophoresis is finished, sealing the nitrocellulose membrane in a sealing solution (5 percent of skimmed milk powder is diluted in TBS containing 0.1 percent of Tween 20) at room temperature for 2h, and then respectively putting the membranes in a primary antibody solution (1: 1000 is diluted in TBS containing 0.1 percent of Tween 20) to incubate at 4 ℃ overnight. Three washes with TBS containing 0.1% Tween 20, 15min each. The membrane was placed in a secondary antibody solution (horseradish peroxidase-labeled goat anti-rabbit IgG, 1: 1000 diluted in TBS with 0.1% Tween 20) and reacted for 1h at room temperature. 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 α protein levels in MDA-MB-231 cells

From table 2 (calculated from fig. 1), it can be seen that the above compounds, examples 16-25, significantly degraded ERR α protein levels in MDA-MB-231 cells, with example 23 being the strongest, and the efficiency of degrading ERR α protein at 10nM, 30nM, and 100nM was 39%, 83%, and 96%, 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 had a pore size of 8 μm. Prior to use, the same amount of MDA-MB-231 cells were placed in 200. mu.l of serum-free PRMI-1640 medium containing different compound concentrations in the upper chamber, 800. mu.l of medium containing 10% fetal bovine serum was placed in the lower chamber, and the chamber was placed at 37 ℃ and 5% CO₂After 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 more specific and detailed, but not construed 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 (16)

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:

1)H；

2) halogen;

3)C₁～C₃an alkyl group;

4)C₃～C₆a cycloalkyl group;

5)C₁～C₃an alkoxy group;

6) a fluoromethyl group;

7) a cyano group;

l is optionally selected from:

1)–NH–(CH₂)_m–；

2)–NH–((CH₂)₂–O)_n–(CH₂)₂–；

3)–NH–(CH₂)_u–O–(CH₂)_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)–CH₂–,–C(＝O)–；

2) y is absent;

b is optionally selected from:

1)H；

2)OR₂；R₂is optionally selected from: h, C₁～C₃Alkyl radical, C₃～C₆A cycloalkyl group;

3)

R₃is optionally selected from: h, C₁～C₃Alkyl radical, C₃～C₆A cycloalkyl group.

2. The application of a compound with a structure shown as a formula (I) or a pharmaceutically acceptable salt or a stereoisomer or a prodrug molecule thereof in preparing an ERR α protein inhibitor:

wherein R is₁Is optionally selected from:

1)H；

2) halogen;

3)C₁～C₃an alkyl group;

4)C₃～C₆a cycloalkyl group;

5)C₁～C₃an alkoxy group;

6) a fluoromethyl group;

7) a cyano group;

l is optionally selected from:

1)–NH–(CH₂)_m–；

2)–NH–((CH₂)₂–O)_n–(CH₂)₂–；

3)–NH–(CH₂)_u–O–(CH₂)_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)–CH₂–,–C(＝O)–；

2) y is absent;

b is optionally selected from:

1)H；

2)OR₂；R₂is optionally selected from: h, C₁～C₃Alkyl radical, C₃～C₆A cycloalkyl group;

3)

R₃is optionally selected from: h, C₁～C₃Alkyl radical, C₃～C₆A cycloalkyl group.

3. The use of a compound according to claim 2, or a pharmaceutically acceptable salt or stereoisomer or prodrug molecule thereof, wherein R is₁Is optionally selected from: halogen, fluoromethyl, cyano.

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 L is any selected from:

1)–NH–(CH₂)_m–；

2)–NH–((CH2)₂–O)_n–(CH₂)₂–；

3)–NH–(CH₂)_u–O–(CH₂)_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.

5. A compound according to claim 2 or a pharmaceutically acceptable salt thereofUse of a stereoisomer or prodrug molecule thereof, wherein Y is selected from the group consisting of: -CH₂–,–C(＝O)–。

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

R₂Is optionally selected from: h, C₁～C₃Alkyl radical, C₃～C₆A cycloalkyl group; r₃Is optionally selected from: h, C₁～C₃Alkyl radical, C₃～C₆A cycloalkyl group.

7. The use of a compound according to any one of claims 2 to 6, or a pharmaceutically acceptable salt or stereoisomer thereof, or a prodrug molecule thereof, wherein the compound has the structure of formula (II):

l is optionally selected from:

1)–NH–(CH₂)_m–；

2)–NH–((CH₂)₂–O)_n–(CH₂)₂–；

3)–NH–(CH₂)_u–O–(CH₂)_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)–CH₂–,–C(＝O)–；

2) y is absent;

b is optionally selected from:

1)H；

2)OR₂；R₂is optionally selected from: h, C₁～C₃Alkyl radical, C₃～C₆A cycloalkyl group;

3)

R₃is optionally selected from: h, C₁～C₃Alkyl radical, C₃～C₆A cycloalkyl group.

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 has the structure of formula (iii):

l is optionally selected from:

1)–NH–(CH₂)_m–；

2)–NH–((CH₂)₂–O)_n–(CH₂)₂–；

3)–NH–(CH₂)_u–O–(CH₂)_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;

r3 is optionally selected from: h, C₁～C₃Alkyl radical, C₃～C₆A cycloalkyl group.

9. 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:

(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) propionic 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-diaza-hexadeca-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-dimethylbutanoyl) -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.

10. Use of a compound according to claim 9, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, wherein the compound is selected from:

(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-diaza-hexadeca-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-dimethylbutanoyl) -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.

11. Use of a compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt or stereoisomer thereof, or a prodrug molecule thereof, in the preparation of a medicament for the prevention or treatment of a disease associated with aberrant expression of ERR α protein activity.

12. The use according to claim 11, wherein the diseases associated with the abnormal expression of ERR α protein activity include tumors, hyperglycemia, diabetes, obesity, hyperlipidemia, hypercholesterolemia, hyperlipoproteinemia, hypertriglyceridemia, hypertension, hyperinsulinemia, hyperuricemia, Parkinson's disease, and Alzheimer's disease.

13. Use of a compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt or stereoisomer thereof, or a prodrug molecule thereof, in the manufacture of a medicament for the treatment or prevention of a tumour or for the prevention of postoperative recurrence of a tumour.

14. The use of claim 13, 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.

15. An ERR α protein inhibitor, wherein the active ingredient comprises a compound as claimed in any one of claims 1 to 10 or a pharmaceutically acceptable salt or stereoisomer thereof or a prodrug molecule thereof.

16. A medicament for treating or preventing tumors or preventing postoperative recurrence of tumors, wherein the active ingredient comprises a compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt or stereoisomer thereof or a prodrug molecule thereof.

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