The Study On The Synthesis And Antitumor Activity Of Arene Ruthenium(?) Complexes,Polypyridyl Ruthenium(?) Complexes

To date,cancer remains one of humanity's greatest challenges,and according to the World Health Organization(WHO),cancer is the second leading cause of death worldwide.Most of the FDA-approved anticancer drugs are organic molecules,and metal drugs are very scarce.As the first metallic antitumor drug,cis-platin,due to its shortcomings such as non-cancer cell toxicity,drug resistance and serious adverse reactions,its clinical application are limited.Ruthenium complexes are of great interest in the design of new metallic anticancer drugs.Recently,many metal ruthenium(?)complexes have been designed and developed into anticancer drugs,and arene ruthenium(?)complexes and polypyridyl ruthenium(?)complexes have received extensive attention due to their excellent antitumor activity.In this research,there are an arene ruthenium(?)complex and four polypyridyl ruthenium(?)complexes were synthesized successfully.Ruthenium(?)complexes with good antitumor activity were screened out and their antitumor mechanisms was explored.The main research of this paper is as follows:(1)Arene ruthenium(?)complex RAP07 was successfully synthesized and MTT assay was used to study the inhibitory effects of RAP07 on different tumor cells and normal cell.It was found that RAP07 was sensitive to MDA-MB-231 cells.Through the tumor zebrafish model experiment,the results showed that RAP07 can inhibit the cell proliferation,invasion and metastasis.Flow cytometry assay was used to detect the effects of RAP07 on cell cycle and apoptosis.The results showed that at low concentration,S phase arrest occurred,and at high concentration,apoptosis occurred.Both comet assay and immunofluorescence assay demonstrated that RAP07 can cause DNA damage.The abeling vascular endothelial cells in zebrafish(flila:EGFP)experiment and tube formation assay have found that RAP07 can block angiogenesis.The results of VEGF factor secreted by the cells were detected by ELISA and qPCR experiments showed that RAP07 can regulate the expression of VEGF.The wound healing assay,FITC-gelatin invasion and immunofluorescence experiments showed that RAP07 can inhibit the metastasis of breast cancer cells by inhibiting the formation of invadopodia.The binding ability of RAP07 to VEGF G4 DNA was investigated by spectroscopy.The toxicity assessment of RAP07 on zebrafish and the safe concentration to zebrafish was less than 8 ?M.Western Blot analysis was used to detect the expression levels of proteins related to different mechanisms.(2)Four polypyridyl ruthenium(?)complexes were successfully synthesized.MTT assay was used to detect the anti-tumor activity of complexes 1-4 on different tumor cells and normal cell lines,the results showed that 1 had better antitumor activity against HepG2 cells and less toxicity to HaCaT cells.Through the tumor zebrafish model experiment,the study showed that 1 can inhibit the proliferation of HepG2 cells.Flow cytometry assay demonstrated that 1 inhibited the proliferation of HepG2 cells mainly by causing cells apoptosis.The localization assay was used to detect the localization of 1 in the cells,and it was found that 1 was mainly enriched in mitochondria.The decrease of mitochondrial membrane potential also demonstrates the damage of mitochondrial function.Detection of reactive oxygen species(ROS)revealed that the levels of ROS were rised.DNA damage is caused by impaired mitochondrial function and increased ROS levels.Therefore,comet assay and immunofluorescence assay have shown that 1 can cause DNA damage.Finally,the zebrafish was used to evaluate the toxicity of 1,and the safe concentration to zebrafish was found to be less than 25 ?M.In summary,arene ruthenium(?)complexes was synthesized successfully can be used as an inhibitor against the invasion and migration of MDA-MB-231 cells through blocking the formation of angiogenesis via regulating the expression of VEGF.In this paper,four polypyridyl ruthenium(?)complexes 1-4 were successfully synthesized,and 1 could be developed into an inhibitor of liver cancer through mitochondria mediated apoptosis.