Design,Synthesis And Anti-tumor Activity Evaluation Of Novel Combretastatin Derivatives As Tubulin And HDAC Dual-targeting Inhibitors

Purpose:in order to obtain novel dual-targeting HDAC inhibitors,structure modification work was performed on Combretastatin to dually target histone deacetylase?HDAC?and tubulin.Methods:through checking a large number of literatures,a feasible synthetic route was designed;The target compounds were synthesized by reactions of condensation,substitution,and hydrolysis;Thin layer chromatography was used to monitor the reaction process and the purity of all intermediates and final products.Purification of intermediate and final products by extraction,recrystallization,filtration,distillation,column chromatography,etc.The structures of the target compounds were finally determined by ¹H NMR,¹³C NMR,and mass spectrums;The enzyme inhibition abilities of the synthesized compounds were tested by fluorescence assay;MTT assay was used to evaluate the anti-proliferative abilities of the compounds to different tumor cell lines;Then the selected representative compounds were tested to inhibit tubulin polymerization;The compound with the best anti-proliferative activity was selected to further test its effect on the regulation of tumor cell cycle,migration,and the effect on inducing apoptosis of tumor cells;at last,two molecular docking experiments were carried out.Results:20 final products were obtained.The structures of these 20 compounds were confirmed by ¹H NMR,¹³C NMR,mass spectrums.The results of enzyme inhibitory activity test showed that the20 compounds inhibited the enzyme activity with different degrees at the concentration of1?M,Among them,the inhibition ability of 8e was the strongest,and that of 4d was the second;Three cell lines,Hela,SGC-7901,A549,were selected to test the anti-proliferative abilities of the compounds with the hydroxamic acid as the zinc ion binding group,The results showed that 8a was the most potent compound against the proliferation of tumor cells.8e and 4d with the best enzyme activity and 8a with the best cell activity were screened to the inhibitory activity of microtubule polymerization.The results showed that8e had the best ability to resist the microtubule polymerization.In addition,other eight tumor cell lines were used to test the anti-proliferative activities of compounds 4d,8a,and8e,in which the activity of 8a was still the best one,IC₅₀0 was all less than 7.6?M.Then the experiments of apoptosis,cell cycle arrest and migration of A549 tumor cells were carried out.The results showed that the activity of 8a was stronger than the positive control,SAHA at the same concentration;In addition,the molecular docking of 8a to HDAC6 and microtubule protein were carried out,respectively by GOLD.exe software.Conclusion:a total of 20 compounds were designed and synthesized in this study,all of which showed the ability to inhibit the activity of HDACs,and some of them displayed the abilities to inhibit the proliferation of tumor cells.Among them,8a was the most active compound.It shows strong anti-proliferative ability to 11 tumor cells and is worthy of further activity evaluation.