Investigation On DNA Interaction And Antitumor Activity In Vitro Of Novel Water Soluble Copper Porphyrin Compounds

In this work,nine A₃B type water-soluble copper porphyrin compounds with novel structures have been designed and synthesized by modifying functional groups.Through the study of interaction with CT-DNA,screened the porphyrin compounds with best results,and then studied the antitumor activity in vitro in cell level,which will provides important research basis for further study of water-soluble copper porphyrin compunds in biological activities of medical research.1.The research progress of the synthesis methods of A₃B type,water-soluble porphyrin compounds,and the interaction of porphyrin compounds with DNA,as well as porphyrin compounds'antitumor effects,respectively2.We designed and synthesized a series of water-soluble bromine-containing copper porphyrin and their bromine substituted derivatives.The interaction of water-soluble bromine-containing copper porphyrin and their bromine substituted derivatives with CT-DNA have been studied by UV spectrum,EtBr-DNA fluorescence quenching,viscosity method and circular dichroism spectrum.The results showed that all of them bind to CT-DNA via outside binding mode,and the affinity of the water-soluble bromine-containing copper porphyrin with CT-DNA is better than its'derivatives.3.We designed and synthesized a series of water-soluble nitro-containing copper porphyrin and its'Schiff base complexes.The interaction of the water-soluble nitro-containing copper porphyrin and its'Schiff base complexes with CT-DNA have been investigated by UV spectrum,EtBr-DNA fluorescence quenching,viscosity method and circular dichroism spectrum.The results showed that all of them bind to CT-DNA via insertion binding mode,and the water-soluble nitro-containing copper porphyrin has the highest affinity.The antitumor activity in vitro of the water-soluble nitro-containing copper porphyrin has been studied,based on the affinity of the water-soluble nitro-containing copper porphyrin with CT-DNA is the best one.The in vitro inhibitory activities of the water-soluble nitro-containing copper porphyrin against carcinoma of Hela cell,MDA cell and TCA8113 cell were evaluated by MTT under different concentration and different time.The cell apoptosis of tumor cells which interacted with the water-soluble nitro-containing copper porphyrin have been studied by using of Annexin V-FITC/PI double marking flow cytometry after intervened 48 h.The cell cycles of tumor cells have been studied by PI single staining flow cytometry after intervened 48 h.The morphological changes of tumor cells have been observed by using Hoechst33342 staining method after intervened 48 h.The results showed that the water-soluble nitro-containing copper porphyrin inhibit the growth of Hela,MDA and TCA8113 cell,and showing concentration and time-dependent.The order of the inhibit the growth of tumor cells is as follows:TCA8113 cell>MDA cell>Hela cell.The mechanism of inhibiting the growth of tumor cell in vitro may realized by inducing tumor cells apoptosis and inhibiting cell cycle of G₁/S stage.Hoechst33342 staining results showed that the experimental group cells became shriveled,smaller in volume,chromosome concentrated with different levels,and which exhibited the typical apoptosis morphology of tumor cells.4.We designed and synthesized water-soluble 2-nitrophenol-containing copper porphyrin and its'Schiff base complexes.The interaction of the water-soluble2-nitrophenol-containing copper porphyrin and its'Schiff base with CT-DNA have been researched by UV spectrum,Et Br-DNA fluorescence quenching,viscosity method and circular dichroism spectrum.The results showed that the three water-soluble copper porphyrin compounds all bind to CT-DNA via insertion binding mode,and all of them have the better affinity.The antitumor activities in vitro of the three water-soluble copper porphyrin compounds have been studied,because of the better affinity of them with CT-DNA.The in vitro inhibitory activities of three water-soluble copper porphyrin compounds against carcinoma of Hela cell,MDA cell and TCA8113 cell were evaluated by MTT under different concentration and different time.The cell apoptosis of tumor cells which interacted with three water-soluble copper porphyrin compounds have been studied by using of Annexin V-FITC/PI double marking flow cytometry after intervened 48 h.The cell cycles of tumor cells have been studied by PI single staining flow cytometry after intervened 48 h.The morphological changes of tumor cells have been observed by using Hoechst33342staining method after intervened 48 h.The results showed that three water-soluble copper porphyrin compounds inhibit the growth of Hela,MDA and TCA8113 cell,and showing concentration and time-dependent.The order of the inhibit the growth of tumor cells is as follows:TCA8113 cell>MDA cell>Hela cell.The mechanism of inhibiting the growth of tumor cell in vitro may realized by inducing tumor cells apoptosis and inhibiting cell cycle of G₁/S stage.Hoechst33342 dyeing results showed that the experimental group cells became shriveled,smaller in volume,chromosome concentrated with different levels,and which exhibited the typical apoptosis morphology of tumor cells.In addition,antitumor activity in vitro of the2-nitrophenol-containing copper porphyrin is much better than its'Schiff base complexes.