CN106543047A

CN106543047A - A kind of new N p-methylphenyls benzamideses adjust the compound and its medical usage of estrogen-related receptor activity

Info

Publication number: CN106543047A
Application number: CN201610858291.7A
Authority: CN
Inventors: 杜永丽; 关海星; 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: 2017-03-29
Anticipated expiration: 2036-09-28
Also published as: CN106543047B

Abstract

The invention discloses a kind of new N p-methylphenyls benzamideses with Formulas I adjust compound and its pharmaceutically acceptable salt and its application of estrogen-related receptor activity.The compound and its pharmaceutically acceptable salt can be used to prepare with regulation estrogen-related receptor ERR alpha（Estrogen related receptor alpha, ERR alpha or ERR α）Activity, prevents and treats the medicine of the malignant tumor such as breast carcinoma, carcinoma of prostate, gastric cancer, colon cancer, ovarian cancer, cervical cancer.

Description

A kind of p- tolyl of new N--benzamideses adjust estrogen-related receptor activity Compound and its medical usage

Technical field

The present invention relates to a kind of p- tolyl-benzamide compound of new N- and its pharmaceutically acceptable salt are made To adjust estrogen-related receptor ERR-alpha（Estrogen-related receptor alpha, ERR-alpha or ERR α）Regulator and its drug regimen and its preparing prevention and/or treating breast carcinoma, carcinoma of prostate, gastric cancer, colon cancer, ovary Purposes in the medicine of the tumors such as cancer, cervical cancer or carcinoma of endometrium.

Background technology

ERR α are in 1988 by clone identifications such as Giguere.As orphan nuclear receptor（Orphan nuclear receptor）, ERR α endogenic ligands are undiscovered always.ERR α wide expressions in vivo, from cardiac muscle, digestive tract, brain, bone Malignant tumor such as bone flesh, brown fat and breast carcinoma, ovarian cancer etc. can be detected.ERR α and estrogen receptor alpha（ERα）Have Higher homology：DNA binding domain（DNA binding domain, DBD）Amino acid similarity 68%, ligand binding domain （Ligand binding domain, LBD）Similarity 33%.Different from estrogen receptor ER α, ERR α can not be tied with estrogen Close, but can be with ER α competition binding estrogen response elements（Estrogen response elemnt, ERE）, recognition sequence is AGGTCAnnn TGACCT；And ERR α also can be reacted with reference to a class I type Steroidgenesis factor with monomer or dimeric form Element TnAAGGTCA, also referred to as estrogen-related receptor response element（ERR response element, ERRE）, show ERR α and ER α have juxtaposition when DNA is recognized.

Nearest research prompting, except participating in ER signal paths, ERR α are in cellular energy metabolism, mitochondrial oxidation and biology Synthesis etc. has an important physiological action, the development of the histoorgan that wields influence of its function, the aging of cell, the generation of tumor and Development etc..

Oncobiology basic research shows, orphan nuclear receptor-estrogen-related receptor α（ERRα）Can promote hormone according to The growth of the breast carcinoma of bad/non-dependent, invasion and attack, promote vascular smooth muscle cell proliferation, the formation of endothelium micro-pipe and tumor vessel new It is raw, it is meant that the medicine of targeting ERR α has the effect of anti-breast cancer concurrently, provides New Policy for Molecular Targeted Therapy for Breast Cancer.

In recent years, research disclose the estrogen such as the ERR α breast carcinoma related to estrogen, carcinoma of endometrium and cervical cancer according to Bad property tumor is closely related, more there are some researches show numerous non-estrogen-dependent tumors such as carcinoma of prostate, adenocarcinoma of stomach and Colon and rectum The generation development of cancer etc. and clinical prognosis are also related to the expression of ERR α.Clinical research confirmation, ERR α breast carcinoma, colon cancer, Express in the tumor such as ovarian cancer and carcinoma of prostate and significantly raise, be the independent risk factor of a prognosis malas.By raise/under Adjust ERR alpha expressions or use ERR alpha inhibitors, can effectively suppress the transcriptional activity of hypoxia genes, so as to reduce internal solid tumor Angiogenesis with increase.These researchs show that ERR α are probably the potential therapy target of kinds of tumors.ERR α inverse agonists XCT790 inhibits the propagation and angiogenesiss of kinds of tumor cells.ERR α inverse agonists SR16388 effectively inhibit nude mice The growth of carcinoma of prostate in lotus knurl model.These researchs show that the regulator of targeting ERR α is possible as clinically effective Antitumor drug.

In sum, estrogen-related receptor ERR α can be as the novel therapeutic target spot of kinds of tumors, based on estrogen phase Close the small-molecule modulators that receptor ERR α are developed, it is most likely that become the novel drugs molecule of therapy-related Diseases Tumor.

The content of the invention

1st, it is an object of the invention to provide there is a class estrogen-related receptor ERR-alpha such as following formula I structure to adjust Section agent or its pharmaceutically acceptable salt.

2nd, a further object of the present invention is the compound or its pharmaceutically acceptable salt for providing Formulas I structure.

3rd, it is still another object of the present invention to provide compound shown in Formulas I or its pharmaceutically acceptable salt are preparing prevention And/or the application in the medicine for the treatment of tumor disease.Preferably, the tumor disease is breast carcinoma, carcinoma of prostate, gastric cancer, knot The tumors such as intestinal cancer, ovarian cancer, cervical cancer or carcinoma of endometrium.

4th, a kind of pharmaceutical composition, including the compound of formula I described in the claim 1 of therapeutically effective amount or its pharmaceutically may be used The salt of acceptance.

5th, it is according to claim 4 to require compositionss, it is characterized in that, the pharmaceutical composition further containing a kind of or Various pharmaceutically acceptable carriers or excipient.

6th, pharmaceutical composition according to claim 4, is characterized in that, described compound of formula I or its pharmaceutically may be used The salt of acceptance accounts for gross weight than 50% ~ 99.5% as active component.

Specific embodiment

The present invention is described in more detail below by specific embodiment, to be better understood from the present invention, but Following embodiments are not intended to limit the scope of the invention, will be detailed below biological activity and the pharmacology of formula I Evaluate.

Implement row 1 transcripting regulating activity of the compound of formula I to ERR-alpha is tested using reporter gene

This example illustrates compound of formula I according to the present invention can effectively suppress ERR- in HEKC HEK-293 The expression of the reporter gene regulated and controled by alpha, illustrates that compound of formula I involved in the present invention has effectively regulated and controled ERR-alpha Function.Reporter gene measuring technology is technology familiar to the staff of this area.

We test compound of formula I to ERRs transcriptional regulatory activities using GAL4 fusion receptors activation method.GAL4 fusions are received Body activation method is based on mammalian cell single crosses principle, i.e., receptor LBD and yeast transcription factor DBD to be built into fusion egg White expression plasmid, makes the LBD of receptor serve as activation domain (Activation domain, AD).When GAL4 fusion receptors matter During common with reporter plasmid that GAL4DNA response elements (UAS) the drive transfectional cell of grain, 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 In the presence of, the combination of GAL4 fusion receptors and 5 × UAS is reduced, and will suppress the expression of reporter gene.

In the experiment of the present invention, we by Gal4-ERR-alpha plasmids with express 5 × UAS and to be coupled Lampyridea glimmering Plasmid (using β-gal expression plasmids as the internal reference) transient cotransfection of light element enzyme reporter gene is thin into mammal HEK-293 The compound of formula I effect 24h of variable concentrations in born of the same parents, is added, the activity change of luciferase and β-gal is detected.

1st, transfect：Inoculation HEK-293 cells in 6cm culture dishs, 6 .0 × 104/ wares, culture fluid is 10%FBS DMEM so as to which growth conditions are good.37 DEG C of 5%CO₂Culture will be inoculated in 96 holes per the cell in hole 6 × 103 in 24 hours afterwards Costar Tissue Culture Plates, overnight incubation.When cell density reaches 70%, take the serum-free medium (DMEM) of 50mL in In the centrifuge tube of 1.5mL, then plus 2mL transfection reagent 3000Transfection Reagent, with handss flick several times, mixing Room temperature places 5min afterwards.By recombiant plasmid and 3000Transfection Reagent with 1:5 ratio is mixed, and takes 20pmol Recombiant plasmid (pGal4-ERRs LBD, p5 × UAS-Luc and p β-gal) is added in DMEM, soft to mix.By serum-free The recombiant plasmid that DMEM has diluted flicks mixing, room with the mixture of the 3000Transfection Reagent of DMEM dilutions again Temperature places 20min to form the complex of DNA/lipofectamine.By answering for the DNA/lipofectamine being incubated Compound is added dropwise in the culture dish containing cell and culture fluid, and gently rocking Tissue Culture Dish back and forth makes cell transfecting Efficiency is improved, and culture dish is put into 37 DEG C, 5%CO₂Cultivate 6 hours in incubator, change fresh medium and (reduce the thin of liposome Cellular toxicity).Cell is re-digested, is inoculated in 96 hole Costar Tissue Culture Plates with the cell in 1 × 104/ hole.

2nd, compound of formula I and positive drug XCT790 are added：After transfection 8h, addition compound of formula I is in 96 orifice plates.By the positive Control XCT790 and compound of formula I are diluted to 10 × aimed concn and add 96 hole Costar Tissue Culture Plates, and volume is 10 μ l, after 24 hours each 3 multiple holes of concentration of continuous culture, after cultivating 24 hours, discard culture fluid in culture plate, add 100 μ of cell pyrolysis liquid L, respectively take 50 μ l carries out the determination of activity of LUC Photinus pyralis LUC Photinus pyralis FL and beta galactosidase respectively with MD multi-function microplate readers, with phase Compound of formula I is calculated to uciferase activity (ratio of LUC Photinus pyralis LUC Photinus pyralis FL activity/betagalactosidase activity) to ERR- The transcripting regulating activity of alpha.

Test result indicate that, compound of formula I according to the present invention can dose-dependently suppress the transcriptional activation of ERR α, (IC₅₀=3.04 ± 0.16 μM), (IC consistent with positive control XCT790 trend₅₀=0.32 ± 0 .02 μM).

Embodiment described above, description are more concrete and detailed, but therefore can not be interpreted as to patent model of the present invention The restriction enclosed.It should be pointed out that for the person of ordinary skill of the art, in the premise without departing from present inventive concept Under, some deformations and improvement can also be made, these belong to protection scope of the present invention.Therefore, the protection of patent of the present invention Scope should be defined by appended right.

Claims

1. a kind of compound and its pharmaceutically acceptable salt are preparing regulation estrogen-related receptor ERR-alpha （Estrogen-related receptor alpha, ERR-alpha or ERR α）Purposes in the medicine of regulator, its feature Be, the compound be 5- [3- (2,5- diethoxy -4- Methanesulfomides-benzyl)-urea groups] -2- ethyoxyl-N-4- tolyls - Benzoylamide, structural formula is shown in formula I.

2. the purposes according to claim 1, is characterized in that, the pharmaceutically acceptable salt be the compound of formula I with Salt produced by various mineral acids, inorganic base, organic acid, organic alkali reaction.

3. the purposes according to claim 1, is characterized in that, the compound of formula I and its pharmaceutically acceptable salt are in system In the medicine of standby prevention and/or treatment breast carcinoma, carcinoma of prostate, gastric cancer, colon cancer, ovarian cancer, cervical cancer or carcinoma of endometrium Purposes.

4. a kind of pharmaceutical composition, including the compound of formula I described in the claim 1 of therapeutically effective amount or which is pharmaceutically acceptable Salt.

5. it is according to claim 4 to require compositionss, it is characterized in that, the pharmaceutical composition further contains one or more Pharmaceutically acceptable carrier or excipient.

6. pharmaceutical composition according to claim 4, is characterized in that, described compound of formula I or which is pharmaceutically acceptable Salt account for gross weight than 50% ~ 99.5% as active component.