Design,Synthesis,and Antitumor Activity Of Bis-aryl Ureas Based On Lead Compound Sorafenib

Bis-aryl urea compounds is a kind of small molecule kinase inhibitors,sorafenib has certain curative effect on liver cancer,non-small cell lung cancer,and melanoma.They are mainly used for not surgical removal of the treatment of advanced kidney cancer,liver cancer and lung cancer.However,As sorafenib represented compound on the market,which it poor water-soluble and more side effects.Because of these drawbacks of sorafenib,synthetic analogues to obtain good efficacy,low toxicity of potential drug molecules have most significance.Quinazoline and pyrazine compounds has extensive biological activities in medicine,They were as many tumor target enzyme inhibitors and become the focus in the anti-tumor drugs.This paper consists of five chapters:Synthetic methods of bis-aryl urea compounds and their biological activity were summarized in the first chapter.The second chapter mainly discussed a serie of pyrazinyl bis-aryl urea were synthesized.2,3dichloro 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 using NMR,HRMS.It has applied for patent protection.The third chapter mainly discussed antitumor biological activity of pyrazine bis-aryl urea compounds.There other two series of compound 5 and compound 6 were tested growth inhibit activity by the MTT method with four kinds of cancer cells in vitro which are NCI-H460,Hela,A549 and MGC-803.The results show that the synthesis of target compound 6g of A549 tumor cells with higher antitumor activity(IC50= 5.21±0.16 μM).The experiment of compound 6g induced A549 tumor cell necrosis,block A549 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.The fourth chapter discussed antitumor mechanisms at the molecular level.The expression level of cycle-related proteins,which cyclin D1,CDK4 as the representative,were decreased.Apoptosis related proteins Bax,caspase-3 and caspase-8 up-regulated but Bcl-2 was decreased,.This result show compound 6g regulated the expression of proteins related to cell cycle and it also induced A549 apoptosis by the caspase pathway.But with the extension of incubation time,programmed necrosis pathway key proteins,such as RIP3,p-MLKL and caspase-8,expression level were increased,which fully shows that compound 6g is through cycle pathways and programmed necrosis pathway to realize multi-channel synergy eventually to suppress tumor growth.The fifth chapter mainly discussed quinazoline bis-aryl urea compounds which their antitumor biological activity.By screening experiment shows that compound 8g activity against tumor cells A549 cell toxicity is superior than positive control group sorafenib and gefitinib(IC50=8.74±0.66 μM).At the same time,experiments show that compound 8g induced A549 tumor cells apoptosis,blocked A549 cells in G0/G1 phase.ROS level of cell was increased,which making the mitochondrial membrane injury and causing hypoxia of intercellular calcium ion concentration increase at the same time,mitochondrial membrane potential decreased,release the upstream factor protein gene expression level of activation of caspases,eventually induce tumor cell apoptosis inhibiting tumor cell proliferation.In summary,as the leading compound of sorafenib,we synthesized two series of bis-aryl urea derivatives which were determined by MTT method to study the antitumor activity of these compounds.A preferred compound 6g was obtain.At the cellular and molecular level,the mechanism of 6g on the proliferation inhibition activity of A549 cells was studied.According to the similar method,the compound 8g exert its anti-tumor activity was deeply investigated.The significance of this study is to lay a good theoretical foundation for the further research and development of new chemical entities,which are better than sorafenib,and low toxicity,and provide a new idea for the treatment of non small cell lung cancer.