DNA Binding And Photodynamic Antitumor Activity Of Hydroxyl Corrole Phosphorus(?) Complex

The application of corrole and its metal complexes in biochemistry field is getting attractive for many research groups.As a member of porphyrin family,corrole has the similar tetrapyrrole macrocyclic structure,it owns the ability to stabilize high-valent transition metal and unique spectral characteristics.A great number of literatures have reported the research of metal corrole complexes in the contents of DNA probes,photonuclease activitiy,and photodynamic therapy.However,the related research on non-metallic corrole complexes is relatively fewer,whose syhthesis methodology and application research is urgently needed.Based on the above background,a kind of mono-hydroxy corrole is employed as the main research object,its synthetic characterization,interaction with DNA,photonuclease activity,phototoxicity towards tumor cells,and photodynamic antitumor activity in vivo are investigated.The research content is as followed:Firstly,10-?3-hydroxylphenyl?-5,15-bis?pentafluorophenyl?corrole?1 or mono-hydroxy corrole?and its phosphorus complex 10-?3-hydroxylphenyl?-5,15-bis?pentafluorophenyl?corrolato?trans-dihydroxo?phosphorus?V??1-P or mono-hydroxy phosphorus corrole?were synthesized and characterized by UV-Vis absorption spectroscopy,high performance liquid chromatography?HPLC?,nuclear magnetic resonance spectroscopy?NMR?,Fourier transform infrared spectroscopy?FT-IR?,X-ray crystals,time resolved fluorescence decay curves,and frontline orbital optimization geometric models?DFT?.Secondly,the interaction between mono-hydroxy phosphorus corrole and calf thymus DNA?ct-DNA?were studied by UV-Vis absorption spectroscopy,fluorescence spectroscopy,circular dichroism spectroscopy,viscosity experiments under fluid mechanics and autodocking calculation,it turned out the corrole bound with ct-DNA in an outside binding mode.The agarose gel electrophoresis experiment was used to investigate the photonuclease activity of mono-hydroxy phosphorus corrole and the reactive oxygen species involved.The results showed mono-hydroxy phosphorus corrole had the good ability of photo-cleavage,and the degree of photo-cleavage was positively correlated with the concentration of corrole.Moreover,singlet oxygen,hydroxyl radical and superoxide anion radical were involved in the process.The existence of the three reactive oxygen species was further confirmed by electron paramagnetic resonance experiments.The singlet oxygen probe was used to measure the singlet oxygen yield in solution.Thirdly,MTT method was used to investigate the cytotoxicity of two kinds of corrole towards four human tumor cell lines?hepatocellular carcinoma cell line Hep G2,nasopharyngeal carcinoma cell line CNE-2,lung cancer cell line A549,and breast cancer cell line MDA-MB-231?,and cisplatin was used as a positive control.It turned out two kinds of corrole showed both low dark toxicity(IC₅₀>100?M)and high phototoxicity(IC₅₀<15?M).The effects of mono-hydroxy phosphorus corrole on the mitochondrial membrane potential of Hep G2 was explored.The results showed it could reduce the mitochondrial membrane potential of tumor cells.Flow cytometry was used to investigate the effects of corrole on the cell cycle of tumor cells,and it was found that mono-hydroxy corrole could block tumor cells in S phase,and mono-hydroxy phosphorus corrole could block tumor cells in G2/M phase.Finally,the acute toxicity of two kinds of corrole to BALB/C mice was investigated through animal experiments.The results showed that 10 mg/kg was the safe dose.