CN106539783B

CN106539783B - A kind of 3- bromophenyl ureas adjusts the active compound of estrogen-related receptor and its medical usage

Info

Publication number: CN106539783B
Application number: CN201610858261.6A
Authority: CN
Inventors: 杜永丽; 李蕊; 凌浩; 李群益
Original assignee: Qilu University of Technology
Current assignee: Qilu University of Technology
Priority date: 2016-09-28
Filing date: 2016-09-28
Publication date: 2019-08-20
Anticipated expiration: 2036-09-28
Also published as: CN106539783A

Abstract

The invention discloses a kind of, and the novel 3- bromophenyl ureas with Formulas I adjusts the active compound of estrogen-related receptor and its pharmaceutically acceptable salt and its application.The compound and its pharmaceutically acceptable salt, which can be used for preparing having, adjusts estrogen-related receptor ERR-alpha(Estrogen-related receptor alpha, ERR-alpha or ERR α) activity, prevent and treat the drug of the malignant tumours such as breast cancer, prostate cancer, gastric cancer, colon cancer, oophoroma, cervical carcinoma.

Description

A kind of 3- bromophenyl ureas adjusts the active compound of estrogen-related receptor and its doctor Learn purposes

Technical field

The present invention relates to a kind of 3- bromophenyl carbamide compounds and its pharmaceutically acceptable salt as adjusting estrogen phase Close receptor ERR-alpha(Estrogen-related receptor alpha, ERR-alpha or ERR α) regulator and its medicine Object combines and its in preparation prevention and/or treatment breast cancer, prostate cancer, gastric cancer, colon cancer, oophoroma, cervical carcinoma or uterus Purposes in the drug of the tumours such as endometrial carcinomas.

Background technique

ERR α is in 1988 by clone identifications such as Giguere.As orphan nuclear receptor (Orphan nuclear Receptor), ERR α endogenic ligand is undiscovered always.ERR α wide expression in vivo, from cardiac muscle, alimentary canal, brain, bone Malignant tumours such as bone flesh, brown fat and breast cancer, oophoroma etc. can detect.ERR α has with estrogen receptor alpha (ER α) Higher homology: DNA binding domain (DNA binding domain, DBD) amino acid similarity 68%, ligand binding domain (Ligand binding domain, LBD) similitude 33%.Different from estrogen receptor ER α, ERR α cannot be with estrogen knot It closes, but can identify that sequence is with ER α competitive binding estrogen response element (Estrogen response elemnt, ERE) AGGTCAnnn TGACCT；And ERR α also can combine one kind I type Steroidgenesis factor to react in the form of monomer or dimer Element TnAAGGTCA, also referred to as estrogen-related receptor response element (ERR response element, ERRE), show ERR α and ER α have juxtaposition when identifying DNA.

Nearest research prompt, in addition to participating in ER signal path, ERR α is in cellular energy metabolism, mitochondrial oxidation and biology Synthesis etc. has an important physiological action, the development for the histoorgan that wields influence of function, the aging of cell, the generation of tumour and Development etc..

Oncobiology basic research show orphan nuclear receptor-estrogen-related receptor α (ERR α) can promote hormone according to Growth, the invasion of bad/non-dependent breast cancer promote vascular smooth muscle cell proliferation, the formation of endothelium micro-pipe and tumor vessel new It is raw, it is meant that the drug of targeting ERR α has both the effect of anti-breast cancer, provides new strategy for Molecular Targeted Therapy for Breast Cancer.

In recent years, research disclose the estrogen such as relevant to the estrogen breast cancer of ERR α, carcinoma of endometrium and cervical carcinoma according to Rely property tumour closely related, more some researches show that numerous non-estrogen dependent tumors such as prostate cancer, sdenocarcinoma of stomach and Colon and rectums The occurrence and development of cancer etc. and clinical prognosis are also related to the expression of ERR α.Clinical research confirmation, ERR α breast cancer, colon cancer, Significant up-regulation is expressed in the tumours such as oophoroma and prostate cancer, is the independent risk factor an of prognosis mala.By up-regulation/under It adjusts ERR alpha expression or uses ERR alpha inhibitor, can effectively inhibit the transcriptional activity of hypoxia genes, to reduce internal solid tumor Angiogenesis and growth.These are studies have shown that ERR α may be the potential therapy target of kinds of tumors.ERR α inverse agonist XCT790 inhibits the proliferation and angiogenesis of kinds of tumor cells.ERR α inverse agonist SR16388 effectively inhibits nude mice The growth of prostate cancer in lotus knurl model.These are studies have shown that target the regulator of ERR α possible as clinically effective Anti-tumor drug.

In conclusion estrogen-related receptor ERR α can be used as the novel therapeutic target spot of kinds of tumors, it is based on estrogen phase Close the small-molecule modulators that receptor ERR α developed, it is most likely that the novel drugs molecule as treatment related disease tumour.

Summary of the invention

1, the purpose of the present invention is to provide a kind of estrogen-related receptor ERR-alpha tune having such as following formula I structure Save agent or its pharmaceutically acceptable salt；

2, another object of the present invention is to provide the compound of Formulas I structure or its pharmaceutically acceptable salt；

3, a further object of the present invention is to provide compounds shown in Formulas I or its pharmaceutically acceptable salt prevents in preparation And/or the application in the drug for the treatment of tumor disease.Preferably, the tumor disease is breast cancer, prostate cancer, gastric cancer, knot The tumours such as intestinal cancer, oophoroma, cervical carcinoma or carcinoma of endometrium.

Specific embodiment:

The present invention is described in more detail below by specific embodiment, to better understand the present invention, but Following embodiments are not intended to limit the scope of the invention.

It will be detailed below the bioactivity and pharmacological evaluation of formula Compound I.

Implementation column 1:

Using reporter gene test compound of formula I to the transcripting regulating activity of ERR-alpha

This example illustrates compound of formula I of the present invention can effectively inhibit in human embryonic kidney cells HEK-293 The expression for the reporter gene that ERR-alpha is regulated and controled illustrates that compound of formula I according to the present invention has effectively regulated and controled ERR- The function of alpha.Reporter gene measuring technology is technology known to the staff of this field.

We are using GAL4 fusion receptors activation method test compound of formula I to ERRs transcriptional regulatory activity.GAL4 fusion by Body activation method is that receptor LBD is built into yeast transcription factor DBD and is merged egg based on mammalian cell single crosses principle White expression plasmid makes the LBD of receptor serve as activation domain (Activation domain, AD).When GAL4 fusion receptors matter When grain and the reporter plasmid of GAL4DNA response element (UAS) driving transfect cell jointly, in the effect of receptor stimulating agent Under, GAL4 fusion receptors are incorporated on 5 × UAS, drive the expression of reporter gene.And in receptor inverse agonists or antagonist Under the action of, the combination of GAL4 fusion receptors and 5 × UAS are reduced, and will inhibit the expression of reporter gene.

In experiment of the invention, we are by Gal4-ERR-alpha plasmid and 5 × UAS of expression and to be coupled firefly glimmering It is thin that plasmid (using β-gal expression plasmid as internal reference) transient cotransfection of light element enzyme reporter gene enters mammal HEK-293 In born of the same parents, the compound of formula I effect that various concentration is added for 24 hours, detects the activity change of luciferase and β-gal.

1, transfect: inoculation HEK-293 cell is in 6cm culture dish, the ware of 6 .0 × 104/, culture solution 10%FBS DMEM keeps its growth conditions good.37 DEG C of 5%CO₂After culture 24 hours, by the cell inoculation in every hole 6 × 103 in 96 holes Costar tissue culture plate, overnight incubation.When cell density reaches 70%, take the serum-free medium (DMEM) of 50mL in Then plus the transfection reagent 3000Transfection Reagent of 2mL it in the centrifuge tube of 1.5mL, is flicked several times with hand, mixing It is placed at room temperature for 5min later.It is mixed in recombinant plasmid and 3000Transfection Reagent with the ratio of 1:5, takes 20pmol Recombinant plasmid (pGal4-ERRs LBD, p5 × UAS-Luc and p β-gal) is added in DMEM, soft to mix.By serum-free Mixture of the recombinant plasmid that DMEM has diluted again with the diluted 3000Transfection Reagent of DMEM flicks mixing, room Temperature places 20min to form the compound of DNA/lipofectamine.By answering for the DNA/lipofectamine being incubated for It closes object to be added dropwise in the culture dish containing cell and culture solution, gently rocking Tissue Culture Dish back and forth makes cell transfecting Efficiency improves, and culture dish is put into 37 DEG C, 5%CO₂It is cultivated 6 hours in incubator, changes fresh medium and (reduce liposome Cytotoxicity).Cell is re-digested, with the cell inoculation in 1 × 104/ hole in 96 hole Costar tissue culture plates.

2, compound of formula I and positive drug XCT790 is added: after transfection 8h, compound of formula I is added into 96 orifice plates.It will be positive Control XCT790 and compound of formula I are diluted to 10 × aimed concn and 96 hole Costar tissue culture plates are added, and volume is 10 μ l, after 24 hours each 3 multiple holes of concentration of continuous culture discard culture solution in culture plate after culture 24 hours, and 100 μ of cell pyrolysis liquid is added L takes 50 μ l to carry out the determination of activity of firefly luciferase and beta galactosidase with MD multi-function microplate reader respectively, respectively with phase Compound of formula I is calculated to ERR- to uciferase activity (firefly luciferase activity/betagalactosidase activity ratio) The transcripting regulating activity of alpha.

The experimental results showed that compound of formula I of the present invention can dose-dependently inhibit the transcriptional activation of ERR α, (IC₅₀=0.64 ± 0.08 μM), (IC consistent with positive control XCT790 trend₅₀.02 μM of=0 .32 ± 0).

Embodiment described above, description is more specific and detailed, and but it cannot be understood as to the invention patent model The limitation enclosed.It should be pointed out that for those of ordinary skill in the art, in the premise for not departing from present inventive concept Under, various modifications and improvements can be made, and these are all within the scope of protection of the present invention.Therefore, the protection of the invention patent Range should be subject to appended right.

Claims

1. a kind of compound and its pharmaceutically acceptable salt adjust estrogen-related receptor ERR-alpha in preparation Purposes in (Estrogen-related receptor alpha, ERR-alpha or ERR α) inverse agonist, feature It is that the compound is N- { 4- [3- (the bromo- phenyl of 3-)-methyl] -2,5- diethoxy-phenyl }-ethyl sulfonamide, and structural formula is such as Shown in Formulas I:

。

2. the purposes according to claim 1, characterized in that the pharmaceutically acceptable salt be the compound of formula I with Various inorganic acids, inorganic base, organic acid, organic base react generated salt.