Study On The Antitumor Activity And Mechanism Of Action Of 2-phenylthiazole Derivatives

Objective:Tumors are major diseases that endangering human physical and mental health,and their incidence is increasing year by year.However,existing anti-tumor drugs have shortcomings such as toxicity and drug resistance,resulting in serious consequences.Therefore,the development of new anti-tumor drugs with advanced mechanisms of action and good therapeutic index has important application value in the field of tumor-related drug research and development.Compounds containing thiazole mother nucleus are widely used in the field of medicine and play an important role in anti-tumor drugs.Acetylcholinesterase（ACh E）also has several“non-classical”functions in non-neuronal cells,including regulation of cell proliferation,apoptosis,invasion,and etc.Studies have found changes in the expression and functional activity of ACh E in human tumors,suggesting that ACh E inhibitors（ACh EIs）may have antitumor activity.The research group designed and synthesized 27 2-phenylthiazole compounds with ACh E inhibition activity,and on this basis,10 thiazole compounds 2-（4-Chlorophenyl）thiazole-4 carboxylic acid derivatives and 2-[（4-trifluoromethyl）phenyl]thiazole-4 carboxylic acid derivatives were synthesized with 2-phenylthiazole as the parent nucleus.To determine whether the 37 2-phenylthiazole compounds previously synthesized for the purpose of anticholinesterase activity and the newly synthesized compounds have antitumor activity and preliminarily explore their antitumor mechanisms.Methods:This project uses 4-chlorothiobenzamide or 4-trifluoromethylthiobenzamide and ethyl3-bromopyruvate as the reaction raw material,with ethanol as the solvent,and reacts for 6 hours under reflux conditions at 80°C,and finally 2-（4-chlorophenyl）thiazole-4-carboxylate or 2-（4-trifluoromethylphenyl）thiazole-4-carboxylate ethyl ester was obtained.Then,using the compound as a raw material,the resulting ethyl carboxylate was then hydrolyzed with sodium hydroxide to form 2-（4-chlorophenyl）thiazole-4-carboxylic acid.Finally,it reacts with different alcohol compounds under the catalysis of concentrated sulfuric acid to form the final product.Antitumor cell activity screening was performed on 37 compounds with 2-phenylthiazole structure by CCK-8 method.The migration inhibition effect of preferred compounds on tumor cells was determined by cell invasion and scratch experiments,the inhibition of cell proliferation mechanism and the growth cycle of tumor cells were determined and analyzed by Annexin V-FITC/PI double staining method and PI single staining method,the differential genes were analyzed and verified by transcriptome sequencing results,and the anti-tumor mechanism of compounds was discussed by analyzing gene expression and signaling pathway changes in tumor cells after the action of the compound by transcriptome sequencing.Results:A total of 10 compounds of 2-phenyl thiazole backbone were obtained,and their structure was confirmed by ¹H NMR,¹³C NMR characterization,and HRMS.The research group conducted anti-tumor activity tests on 37 2-phenylthiazole derivatives by CCK-8 method.One compound,2-（4-（benzyloxy）phenyl）-N-（2-（piperidin-1-yl）ethyl）thiazole-4-carboxamide（A6）,was screened out with potent antitumor activity,especially against breast cancer MCF-7 cells.Further mechanistic studies have shown that A6 significantly inhibits migration and invasion.Flow cytometry results showed that A6 can induce apoptosis in MCF-7 cells,but has no significant effect on the cell cycle.Next,we use RNA-seq to further explore the underlying mechanisms by which A6 inhibits MCF-7 cells.The results showed that A6-treated MCF-7 cells caused a total of153 gene upregulation and 81 gene downregulation.GO and KEGG enrichment analysis showed A6 mainly disrupted sterol/cholesterol pathway,Ras signaling pathway,VEGF signaling pathway,and etc.Conclusion:In this project,one compound,2-（4-（benzyloxy）phenyl）-N-（2-（piperidin-1-yl）ethyl）thiazole-4-carboxamide（A6）,was screened out with potent antitumor activity.A6 can significantly inhibit migration and invasion and induce apoptosis,and interfere with gene expression and signaling pathways within tumor cells.This study has laid a certain foundation for the research and development of new anti-tumor drugs.