Synthesis And Anti-tumor Activity Of Phenothiazine Compounds Targeting Mitochondria

Objective: To synthesize two phenothiazine derivatives with mitochondrial targeting,in order to investigate the effects of the two compounds on tumor proliferation,migration,invasion,autophagy and apoptosis,and further explore the molecular mechanism of their effects on tumors.Methods: Two positively charged phenothiazine derivatives were synthesized by using some delocalized cationic group and a phenothiazine structure.Two compounds were separated and purified for cell experiments and tumor cells AGS,H520,Lovo,Hela,Hu-7,U87,etc.were selected as research objects.After screening the antitumor activity of the two compounds using the MTT experiment which the optimal administration time of the compounds and the concentration of subsequent experiments were further investigated.Next,used MTT experiments,plate cloning experiments,transwell migration experiments,transwell invasion experiments,autophagy,and apoptosis experiments to investigate the effects of drugs on the monoclonal formation,migration,invasion,autophagy,and apoptosis of tumor cells,and verified whether the compounds can target mitochondria of tumor cells.Then we used molecular docking experiments and Western blotting experiments to explore the molecular mechanism of the compound against tumors.Finally,siRNA and dsRNA transfection experiments were used to further verify the molecular mechanisms discovered by the experiments.Results: Through MTT experiments,it was found that compared with the positive control group,all tumor cells in compound one had better activity,and the appropriate administration time was 24 h as well as 0,2 and 4 ?M were selected as the concentration for subsequent experiments.Compound 2 was far less effective than the positive control group.Once the compound one reached 2 ?M,it could inhibit the formation,migration and invasion of tumor cells AGS,Lovo,Hu-7.And it can kill tumor cells by inducing tumor cell apoptosis instead of autophagy.The mitochondrial probe can prove that compound one can target tumor cell mitochondria.Molecular docking experiments revealed that the possible target of this compound is the oncogene PELP1.Western blot experiments also proved that it can induce tumor cell apoptosis by regulating PELP1 and its related targets.Finally,siRNA and dsRNA transfection experiments further verified that PELP1 is a target of this compound.Conclusion: Mitochondrial targeting compound one can inhibit tumor cells by targeting PELP1 to induce tumor cell apoptosis.