Studies On Synthesis,in Vitro Antitumour Activity And Mechanism Of 3-Estradiol-linked Quinoline Derivatives Of Palladium(Ⅱ)and Platinum(Ⅱ)Complexes

Estrogen,which belong to gonadal hormone in human body,has attracted much attention because of their wide range of biological activities.In 1994,was reported that estrone-3-Osulfamate irreversibly inhibits a new potential hormone-dependent cancer.This dissertation mainly focused on two typical 3-Estradiol-linked quinoline derivatives of ligands and their palladium(Ⅱ)complexes,platinum(Ⅱ)complexes.Four mononuclear palladium(Ⅱ)and platinum(Ⅱ)complexes have been newly-synthesized,and their structures have been characterized by elemental analysis,IR,ESI-MS and NMR analysis,the purity of the tested compounds was determined by HPLC,stabilization of the tested compounds was determined by UV-Vis spectra and HPLC.On the cellular level,antitumor activity of two ligands along with their metal complexes against MDA-MB-231,SK-OV-3 and HL-7702 cell apoptosis induced by them has been examined by MTT assay and flow cytometry(FCM),respectively.On the molecular level,the interaction of two ligands and their metal complexes with DNA were investigated by CD spectroscopy,agraose gel electrophoresis.This research is beneficial to the research and development of metal-based antitumor drug.The main contents of the thesis are as follows:Ⅰ.The progress and achievements of antitumor metal complexes,introducing hormones and quinoline derivatives research progress of anti-tumor drugs.On these bases,the significance and design of the thesis was proposed.Ⅱ.Four mononuclear metal complexes with 3-Estradiol-linked 8-Hydroxyquinoline(HL2)and 3-Estradiol-linked 8-Aminoquinoline(NQE)have been synthesized,by solution methods,respectively.The four compounds are[Pd(HL2)Cl](1),[Pt(HL2)Cl](2),[Pd(NQE)Cl2](3),[Pt(NQE)Cl2](4).All of the 4 metal-based compounds have been synthesized,structurally determined by NMR,elemental analysis,EMI-MS and IR analysis.The results showed that complexes 1-4 have a mononuclear structure and the central metal is four-coordinated.Complexes 1 and 2 have similar structures.The palladium(Ⅱ)and platinum(Ⅱ)are fourcoordinated by two N atoms and one O atom from a HL2 ligand and one Cl-atom.Complexes 3 and 4 are similar structures.The central metal are four-coordinated by two N atoms from a NQE ligand and two Cl-atoms,respectively.III.1.The in vitro antitumor activities of these complexes against a series of human tumor cell lines(NCI-H460,MGC80-3,BEL-7404,T-24,Hep-G2,HeLa,SK-OV-3,MCF-7 and MDA-MB-231 cells)and human normal liver cell line HL-7702 were screeened by MTT assay,and the IC50 values were also evaluated,The results of experiments indicate that all of these compounds exhibited antitumor activity against all of these cell lines in different degrees,the inhibition rate to NCI-H460,T-24,Hep-G2 and MGC80-3 can reach more than 50%.The complexes 1 and 2 are the more cytotoxic compounds that exhibits significant activity against all of the above tumor cell lines,especially to the MDA-MB-231 and SK-OV-3 tumor cell lines,with IC50 values close to 8 μM.At same conditions,The complexes 1 and 2 showed the higher cytotoxicity than cisplatin(IC50>10μM)both the complexes 1 and 2 were examined for their cell cycle arrest and cell apoptosis induction in the MDA-MB-231,SK-OV-3 and HL-7702 cell lines using flow cytometry.The cell cycle results showed that both the complexes 1 and 2 disturb SK-OV-3 cell cycle and induce an obvious arrest in G1 phase.Complexes 1 disturb MDA-MB231 and HL-7702 cell cycle and induce an obvious arrest in G2 phase,And complexe 2 disturb MDA-MB-231 and HL-7702 cell cycle and induce an obvious arrest in S phase.2.The apoptotic mechanism in MDA-MB-231 cell line induced by complexes 1 and 2 was further studied by flow cytometry assay.The ROS level was detected to be enhanced in tumor cells in the presence of complexes 1 and 2,along with the increment on the[Ca2+].Ⅳ.In molecular level,interactions between DNA and these compounds were investigated schemtically using CD spectroscopy and agraose gel electrophoresis assay.The CD spectroscopy result showed that two ligands have no intercalation binding mode with ct-DNA.After two ligands combined with the metal ion of palladium(Ⅱ)and platinum(Ⅱ).Complexes 1-3 interacted with ct-DNA in an intercalative binding mode.Which could be ascribed to the central metal ion who acting the key role to promote the DNA binding.From the results of the agarose gel electrophoresis experiment,the complexes 1-3 could more effectively reduce the shift mobility of supercoiled DNA.Which lay the important clue for the antitumor activity mechanism.