Synthesis Of Novel Polypyridyl Ruthenium(Ⅱ) Complexes And Their Application As Cancer Diagnosis And Treatment Agents

Objective: The focus of this research to design and synthesize polypyridyl ruthenium(Ⅱ)complexes modified with a series of functionalized groups,and to further study their mechanism of developing as small molecule phosphorescence probes or targeting tumor diagnostic and therapeutic agent.Methods:(1)Microwave-assisted synthesis of polypyridinium(Ⅱ)complexes 1 and 2 containing alkynyl-modified phenylimidazole-[5,6-f][1,10]-phenanthroline(PIP)as the main ligand.The binding ability of the two complexes to c-myc G4 DNA was studied using electron absorption spectroscopy,fluorescence emission spectroscopy,FRET spectroscopic methods,and molecular docking.Analytical ultracentrifugation and atomic force microscopy were used to study the morphology of the two complexes combinding with c-myc G4 DNA in liquid and solid form,respectively.(2)Synthesis of polypyridyl ruthenium(Ⅱ)complexes with Dipyrido[3,2-a:2’,3’-c]phenazine(DPPZ)Derivatives by microwave-assisted synthesis and heat reflux reaction.MTT assay was used to study the cytotoxicity of 3-7 against different tumor cell lines.The effects of 7 on the formation of invadopodia of breast cancer were studied by the wound healing test,gelatin invasion and immunofluorescence experiments.In addition,vascular endothelial cell-labeled zebrafish Tg(flila:EGFP)was used as an animal model to investigate the effect of 7 on neovascularization.Cellular uptake and localization,TUNEL and DAPI double staining,comet assay,and immunofluorescence were used to study the mechanism of the inhibition of the proliferation of breast cancer cells induced by 7.Finally,spectroscopic methods,molecular docking and western blot experiments were used to explore the possibility of c-myc G4 DNA as an antitumor target.Results:(1)Polypyridinium(Ⅱ)complexes 1 and 2 have been successfully synthesized.It was found that both complexes can selectivey bind and stabilize c-myc G4 DNA in a groove-binding manner.Among them,the binding ability of 2 was stronger,and it was found that the phosphorescence was significantly enhanced after binding to c-myc G4 DNA.Furthermore,the both complexes can polymerize to different extents with c-myc G4 DNA in the solution state.In the solid state,it was found that 1 and 2 induced c-myc G4 DNA assembly into nanonet and nanowire structures,respectively.(2)Polypyridinium(Ⅱ)complexes 3-7 have been successfully synthesized.7 was found to be highly sensitive to MDA-MB-231 cells,and also can inhibit the formation of invadopodia and the SIVs of zebrafish to inhibit the growth and metastasis of breast cancer cells.In addition,it was found that 7 can enter the nucleus in a time-dependent manner to promote DNA damage and inhibit the proliferation of breast cancer cells.Finally,it was found that 7 can selectively bind to and stabilize c-myc G4 DNA and down-regulate the expression of c-Myc,which as the potential antitmor target of 7.Conclutions: In summary,different types of polypyridyl ruthenium(Ⅱ)complexes have been designed and synthesized as phosphorescence probe or diagnostic reagent,2 could be developed as c-myc G4 DNA structural phosphorescence probe,and 7 could be used as a targeted therapeutic agent for inhibiting the proliferation and metastasis of breast cancer MDA-MB-231 cells.