Design,Synthesis,and Antitumor Activity Evaluation Of Pyrazinyl Bis-aryl Ureas

Bis-aryl urea compounds are a kind of small molecule kinase inhibitors and extremely important in medicinal chemistry for their biological activities.Sorafenib with urea as the typical representative of pharmacophore is listed drug which is mainly used for not surgical removal of the treatment of advanced cancer.Sorafenib is poor water-soluble and more side effects and produces a great pain to the patient.Because of these drawbacks of sorafenib,synthetic analogues to obtain good efficacy,of potential drug molecules have most significance by bioisosterism.Pyrazine derivatives are a kind of important pyrazine alkaloid.We make pyrazine as matrix by modifying the structure of pyrazine compounds,which have good activity in anti-tumor,antibacterial,enzyme inhibitors,antioxidation,and other functions.Combining pyrazine basis for maternal with benzene ring of Sorafenib to generate new compounds become a hot research topic in this paper.This paper consists of four chapters:The first chapter mainly introduced the synthetic methods of bis-aryl urea compounds and pyrazine with base advantage in pyrazine derivatives and other biological activities.The second chapter mainly discussed a serie of pyrazinyl bis-aryl urea were synthesized.2,3-dichloro pyrazine reaction with 3-amino benzyl amine,then the product was dissolved in dichloromethane reaction with isocyanate at room temperature to get target compounds.The structure of the target compounds were confirmed by NMR,HRMS.Have applied for patent protection.The third chapter mainly discussed antitumor biological activity of pyrazine bis-aryl urea compounds.The compound 5x were tested growth inhibit activity by the MTT method with four kinds of cancer cells in vitro which are HepG2,MGC-803,NCI-H460,T24,HL-7702.The results show that the synthesis of target compound 5x of T24 tumor cells with higher antitumor activity(IC50=1.25±0.06 ?M).The experiment of compound 5x induced T24 tumor cell necrosis,block T24 cells in G0/G1 phase,lead to ROS level decrease.As cells mitochondrial membrane injury which caused hypoxia of intercellular calcium ion concentration increase at the same time,mitochondrial membrane potential was decreased and necrosis occurs,which leads to the proliferation inhibition HepG2 liver cancer cell lines,human gastric cancer cell line MGC-803,human non-small cell lung cancer NCI-H460 cell lines,human T24 bladder cancer cell lines and normal liver cell line the HL-7702 cells.And then we use software data measured active best cell lines.The fourth chapter is mainly at the molecular level,through the WB experiment on the cycle,apoptosis,programmed necrosis,RAF/MEK/ERK pathway,autophagy,iron death related proteins were detected.We detected that when the compound 5x hatched the T24 cells,the results showed that the cycle related proteins demonstrated the blocking of T24 cells in G0/G1 phase.It is also proved that apoptosis is not caused by programmed necrosis.In the end of autophagy and iron death,it is confirmed that compound 5x inhibits the proliferation of bladder cancer cell line T24 by regulating the protein signaling pathway.It can effectively interfere the RAF/MEK/ERK pathway,and induce both autophagy and iron death in T24 cells to inhibit the proliferation of tumor cells.To sum up,in this study,we synthesized and obtained 25 Pyrazine urea urea compounds,all of them were new compounds,and their activity was screened,the optimum compound 5x was obtained,and the inhibition mechanism of 5x on the proliferation inhibition activity of T24 cells was investigated.The important significance of this study is to establish a good theoretical basis for further research and development of the new chemical entity with less toxic and side effects,and provide a new way for the treatment of bladder cancer.