阅读说明：本技术 一类含羟肟酸基团的二芳基乙烯类LSD1/HDACs双靶点抑制剂、其制备方法及应用 (Hydroximic acid group-containing diarylethene LSD1/HDACs double-target inhibitor, and preparation method and application thereof ) 是由 段迎超 靳林峰 关圆圆 袁航 文郁康 陈书慧 秦文平 张少杰 于 2020-06-09 设计创作，主要内容包括：本发明涉及一类含有羟肟酸基团的二芳基乙烯类LSD1/HDACs双靶点抑制剂、其制备方法及在制备抗肿瘤药物中的应用,属于药物化学技术领域。所述的化合物具有如下通式：<Image he="106" wi="211" file="111976DEST_PATH_IMAGE001.GIF" imgContent="drawing" imgFormat="GIF" orientation="portrait" inline="no"></Image>其中<Image he="46" wi="60" file="69478DEST_PATH_IMAGE002.GIF" imgContent="drawing" imgFormat="GIF" orientation="portrait" inline="no"></Image>优选3-位或4-位单取代；R<Sub>2</Sub>优选OH、OCH<Sub>3</Sub>、F、H；R<Sub>3</Sub>优选H、CH<Sub>2</Sub>OH；R<Sub>4</Sub>优选为H、CH<Sub>3</Sub>、F、OCH<Sub>3</Sub>；R<Sub>5</Sub>优选H、OH、CH<Sub>2</Sub>OH、CH<Sub>3</Sub>、F、OCH<Sub>3</Sub>；X为N或者CH；Y为N或者CH。本发明所述化合物对LSD1和HDAC1/6均具有较强的抑制活性,且对LSD1具有很好的选择性,多个化合物对人结肠癌HCT-116细胞株和人胃癌MGC-803细胞株的体外抗肿瘤活性优于上市药物SAHA。为LSD1/HDACs双靶点抑制剂类药物的研发提供了基础,可作为进一步开发的候选或者先导化合物用于开发抗肿瘤治疗药物。(The invention relates to a hydroximic acid group-containing diarylethene LSD1/HDACs double-target inhibitor, a preparation method thereof and application thereof in preparing antitumor drugs, belonging to the technical field of pharmaceutical chemistry. The compounds have the following general formula: wherein Preferably 3-or 4-monosubstituted; r 2 OH and OCH are preferred 3 、F、H；R 3 Preferably H, CH 2 OH；R 4 Preferably H, CH 3 、F、OCH 3 ；R 5 H, OH and CH are preferred 2 OH、CH 3 、F、OCH 3 (ii) a X is N or CH; y is N or CH. The compounds have stronger inhibitory activity to LSD1 and HDAC1/6, and have good selectivity to LSD1, and the in vitro anti-tumor activity of a plurality of compounds to human colon cancer HCT-116 cell strains and human gastric cancer MGC-803 cell strains is superior to that of the marketed medicine SAHA. Provides a foundation for the research and development of LSD1/HDACs double-target inhibitor medicines, and can be used as a candidate or lead compound for further development to develop anti-tumor therapeutic medicines.)

1. A diarylethene compound containing hydroximic acid group is characterized by having a structure shown in a general formula (I-5):

in the general formula (I-5), the substituentIs monosubstituted; r₂Is OH, OCH₃F, H; r₃Is H, CH₂Any one of OH; r₄Is H, CH₃、F、OCH₃Any one of the above; r₅Is H, OH, CH₂OH、CH₃、F、OCH₃Any one of the above; x is N or CH; y is N or CH.

2. A class of hydroxamic acid group containing diarylethene compounds as claimed in claim 1 wherein:3-or 4-mono-substitution is selected.

3. A class of diarylethenes containing hydroxamic acid groups according to claim 1 wherein one of the following compounds is selected:

4. a method for preparing a class of hydroximic acid group-containing diarylethene compounds as claimed in claim 1 or 2, which is carried out by the following steps:

(1) preparation of Compound I-3: stirring a compound I-1 and substituted benzyl diethyl phosphonate (a compound I-2) in N, N-Dimethylformamide (DMF) at room temperature in the presence of a strong basic compound for reaction, pouring a reaction system into ice water after the reaction is finished, performing suction filtration, washing, collecting a solid, and drying to obtain a compound I-3; wherein the strong alkaline compound is selected from one of potassium tert-butoxide, sodium methoxide and sodium hydride;

(2) preparation of Compound I-4: carrying out reflux stirring reaction on the compound I-3 and substituted phenylboronic acid or pyridine boric acid in toluene in the presence of an alkaline compound and a palladium catalyst to obtain a compound I-4; wherein the alkaline compound is selected from one of potassium carbonate, sodium bicarbonate, cesium carbonate and potassium phosphate, and the palladium catalyst is selected from one of tetrakis (triphenylphosphine) palladium, palladium acetate, bis (dibenzylideneacetone) palladium and palladium dichloride;

(3) preparation of Compound I-5: in dichloromethane solution, Compound I-4 and NH₂OK/NH₂And (3) reacting with an OH methanol solution, after the reaction is finished, concentrating the reaction system in vacuum, adding ethyl acetate and water into the concentrate for extraction, washing, and separating an organic phase by column chromatography to obtain a compound I-5.

5. Use of a class of hydroximic acid group containing diarylethenes according to any of claims 1 to 3 in the manufacture of a medicament, as active ingredients in the manufacture of a medicament of the LSD1/HDAC dual target inhibitor class.

6. The use of a class of diarylethenes containing hydroxamic acid groups as claimed in claim 5 in the manufacture of a medicament for use as active ingredients in the manufacture of anti-colon, lung and stomach cancer drugs.

Technical Field

The invention particularly relates to a diaryl ethylene LSD1/HDACs double-target inhibitor containing hydroximic acid groups, a preparation method thereof and application thereof in preparing anti-tumor drugs, belonging to the technical field of pharmaceutical chemistry.

Background

Histone lysine specific demethylase 1(LSD1) was first reported by professor Shi Yang of harvard university in 2004, and is a Flavin Adenine Dinucleotide (FAD) -dependent amino oxidase that specifically removes mono-and di-methyl groups of histone H3K 4. The discovery of LSD1 confirmed that there is a dynamic balance of methylation and demethylation of histones. In addition, LSD1 can remove the methyl group of non-histone such as p53, DNMT1, STAT3, E2F1, MYPT1, ERA and HIF-1, and further regulate the stability and activity of the downstream gene. A large number of researches show that the expression level of LSD1 in various tumors such as lymphoma, acute myelogenous leukemia, prostate cancer, lung cancer, gastric cancer, estrogen receptor negative breast cancer, oral cancer, colon cancer, synovial sarcoma, neuroblastoma and the like is remarkably increased, and the LSD1 is closely related to the poor prognosis of various malignant tumors such as lung cancer, acute myelogenous leukemia, colon cancer, breast cancer and the like. The RNA interference technology is used for reducing the expression quantity of the LSD1 or inhibiting the activity of the LSD1 by using a small molecule inhibitor, so that the LSD1 can inhibit the proliferation, the metastasis and the invasion of cancer cells, is one of hot target proteins for the research and development of the current antitumor drugs, and currently, a plurality of LSD1 small molecule inhibitors are in phase I and phase II clinical tests and are used for treating acute myelogenous leukemia and non-small cell lung cancer.

Histone Deacetylases (HDACs) are a class of epigenetic regulatory proteins responsible for removing acetyl groups from histone tails. In various malignant tumors such as leukemia, lymphoma, cervical cancer, colorectal cancer, breast cancer and the like, the expression and activity of the HDACs family members are obviously up-regulated, and the adverse prognosis of various malignant tumors such as leukemia, lymphoma, cervical cancer, colorectal cancer and the like is positively correlated with the high expression of HDACs. At present, 5 hdac inhibitors are approved by FDA and CFDA to be marketed for treating various tumors such as malignant lymphoma and myeloma, and a plurality of hdac inhibitor candidate drugs are in clinical trials.

Both LSD1 and HDACs inhibitor can re-express some genes silenced by external source in tumor cells to induce apoptosis, and there is a close cross-talk relationship between LSD1 and HDACs. LSD1 and HDAC1/2 are co-present in NuRD, CoREST, Sin3A multiple co-suppression complexes and are involved in regulating transcription of multiple genes. LSD1 activity is regulated by HDACs, and LSD1 activity is also inhibited upon inhibition of HDACs activity. In human breast cancer cells, HDAC5 can promote the stability and demethylation activity of LSD1 by up-regulating the expression of USP 28. In various cancers such as bladder cancer, breast cancer, lung cancer and the like, the reduction of the expression level of LSD1 or the inhibition of the activity of LSD1 can remarkably enhance the sensitivity of cancer cells to HDACs inhibitors. The simultaneous inhibition of the activity of LSD1 and HDACs by small molecule inhibitors has a synergistic antitumor effect. Therefore, a novel and high-activity LSD1/HDACs double-target inhibitor is found, and a synergistic anti-tumor effect of '1 +1> 2' is exerted by simultaneously inhibiting LSD1, HDACs and mutual cross-talk signal channel transduction, so that a novel high-efficiency anti-tumor lead compound is expected to be found, and the novel high-activity LSD1/HDACs double-target inhibitor has very important significance for researching the biological functions of LSD1 and HDACs and researching novel anti-tumor drugs. In order to find a novel LSD1/HDACs double-target inhibitor, a class of diarylethene compounds containing hydroximic acid groups is explored and synthesized, and the LSD1 and HDACs double-inhibition activity and in-vitro anti-tumor activity of the diarylethene compounds are verified as the starting points of the application, so that no reports of the synthesis, LSD1/HDACs inhibition activity and anti-tumor activity of the compounds are found at present.

Disclosure of Invention

From the above, it is an object of the present invention to provide a class of diarylethene compounds containing hydroximic acid groups, which provides the possibility of new drug screening.

It is another object of the present invention to provide a process for the preparation of such hydroxamic acid group containing diarylethene compounds.

The invention further aims to provide the application of the compound in preparing anti-tumor medicines by taking LSD1/HDACs as targets.

In order to achieve the above purpose, the structural general formula of the hydroximic acid group-containing diarylethene compound related by the invention is as follows:

in the general formula I-5, the substituent groupIs monosubstituted; r₂Is OH, OCH₃F, H; r₃Is H, CH₂Any one of OH; r₄Is H, CH₃、F、OCH₃Any one of the above; r₅Is H, OH, CH₂OH、CH₃、F、OCH₃Any one of the above; x is N or CH; y is N or CH.

Preferably:3-or 4-mono-substituted.

Preferably: in the general formula I-5,position of substituent group, R₂、R₃、R₄、R₅The substituents represented by X and Y are shown in the following table:

to achieve the second objective, the synthesis reaction scheme of the compound of the present invention is shown as the following formula:

synthetic routes to compounds of general formula I:

the preparation method of the compound I-3 comprises the steps of stirring the compound I-1 and substituted benzyl diethyl phosphonate (the compound I-2) in N, N-Dimethylformamide (DMF) at room temperature in the presence of a strong basic compound for reaction, pouring a reaction system into ice water after the reaction is finished, carrying out suction filtration, washing, collecting a solid, and drying to obtain the compound I-3. Wherein the strong alkaline compound is selected from one of potassium tert-butoxide, sodium methoxide and sodium hydride.

The preparation method of the compound I-4 comprises the step of carrying out reflux stirring reaction on the compound I-3 and various substituted phenylboronic acids or pyridine boric acids in toluene in the presence of an alkaline compound and a palladium catalyst to obtain the compound I-4. Wherein the alkaline compound is selected from one of potassium carbonate, sodium bicarbonate, cesium carbonate and potassium phosphate, and the palladium catalyst is selected from one of tetrakis (triphenylphosphine) palladium, palladium acetate, bis (dibenzylideneacetone) palladium and palladium dichloride.

The compound I-5 is prepared by dissolving the compound I-4 and NH in dichloromethane₂OK/NH₂Reacting OH with methanol solution, vacuum concentrating the reaction system after the reaction is finished, extracting the concentrate with ethyl acetate and water, washing, and separating the organic phase by column chromatographyTo obtain the compound I-5.

The invention has the advantages that: the synthesized hydroximic acid group-containing diarylethene compounds have stronger LSD1/HDACs double inhibitory activity and in-vitro anti-tumor activity. The compound reported by the invention has stronger inhibitory activity to LSD1 and HDAC1/6, and has good selectivity to LSD 1. Inhibition IC of LSD1, HDAC1 of several compounds₅₀Less than 1 μ M, and less than 50nM of inhibitory activity on HDAC 6. The LSD1/HDACs double-target preparation shows better in-vitro anti-tumor activity on human colon cancer HCT-116 cell strains and human gastric cancer MGC-803 cell strains, and the in-vitro anti-tumor activity of a plurality of compounds is better than that of a positive control marketed drug HDACs inhibitor SAHA. The compound represents a LSD1/HDACs double-target inhibitor with a brand-new structure, provides a foundation for the research and development of LSD1/HDACs double-target inhibitor medicines, and provides an effective tool for the biological function research of LSD1 and HDACs. Can be used as a candidate or lead compound for further development to develop anti-tumor, anti-virus, anti-AIDS and other disease treatment drugs, and the synthesis method is simple and is beneficial to popularization and application.

Detailed Description

The technical solution of the present invention will be described in detail below with reference to examples.