| Because of its high selectivity, low cytotoxicity and antimetastatic properties,aryl ruthenium complexes with thiosemicarbazones have attracted much attention.The bioactivity effects of some Ru(II) arene complexes are exerted by the inhibition of the replication and amplification of DNA, which lead to the death of cancer cells.But beyond that, as the assured potential of ruthenium(II) arene complexes of anticancer drugs in therapy, their absorption, distribution, metabolism and excretion properties in organism are still unknown right now.The investigation of the interaction of Ru(II)-arene anticancer drugs and some important biomacromolecules HSA been an active area of the research,which can make us better understand the mechanism of their antibacterial, antitumor, and anticancer activities, as same as the directional design and the efficient target synthesis of novel Ru-based anticancer drugs.In this article,the interaction between ruthenium complexes with thiosemicarbazones anticancer drug and macromolecule was investigated systematically by using spectroscopic methods,cyclic voltammetry and mass spectroscopy.Thesis is divided into following five parts:The first chapter: The antitumor metal complexes and the development of the interaction of biological macromolecules with bioactive small molecules were briefly introduced. the physiological functions and structures of HSA and DNA were also described. Besides, the methods about investigating the binding of drugs to proteins and DNA were reviewed in detail.The second chapter: In this contribution, the interaction between[(?6-pcymene)RuII(acetone-N4-phenylthiosemicarbazone)Cl]Cl(Ru-TSC-1)anticancer drug and human serum albumin(HSA) was investigated by spectroscopic and electrochemical techniques. The fluorescence spectra results indicated that Ru-TSC-1 anticancer drug could quench the intrinsic fluorescence of HSA through dynamic quenching mode. The calculated corresponding activation energy of the interaction between Ru-TSC-1anticancer drug and HSA was 35.62 kJ mol-1. The distance between HSA and Ru-TSC-1 anticancer drug was obtained according to fluorescence resonance energy transfer. The results of synchronous fluorescence spectra, three-dimensional fluorescence spectra, Fourier transform infraredspectroscopy(FTIR)spectra, and circular dichroism(CD)spectra indicated that the microenvironment and the confor-mation of HSA were all changed in the presence of Ru-TSC-1 anticancer drug. The results of cyclic voltammetry further validated the interaction between Ru-TSC-1 and HSA. These results indicated that the biological activity of HSA was affected by Ru-TSC-1 anticancer drug dramaticallyThe third chapter: the interaction between [(?6-p-cymene)RuII(benzaldehyde-N4-phenylthiosemicarbazone)Cl]Cl(Ru-TSC-2)anticancer drug and HSA was investigated systematically under physiological conditions by using some spectroscopic methods, mass spectroscopy and cyclic voltammetry. The experimental results indicated that this anticancer drug could quench the intrinsic fluorescence of HSA through static quenching mechanism. The Stern-Volmer quenching constants together with the modified Stern-Volmer quenching constants at different temperatures were also calculated. The corresponding thermodynamic parameters ?H, ?G and ?S were also calculated. The binding of this anticancer drug and HSA resulted in the formation of drug-HSA complex, and the electrostatic interaction played a major role in the complex stabilization. The distance r between the donor(HSA)and the acceptor(drug) was obtained through fluorescence resonance energy transfer theory. Competitive experiments indicated that the binding site of this anticancer drug to HSA was located at site I. The results of synchronous fluorescence spectra, three-dimensional fluorescence spectra, FT-IR spectra and CD spectra indicated that the microenvironment and the conformation of HSA were changed noticeably due to the presence of this anticancer drug. The results of mass spectra and cyclic voltammetry further confirmed the interaction between HSA and this anticancer drug. These results indicated that the biological activity of HSA was dramatically affected by the Ru-TSC-2 anticancer drug.The four chapter: the interaction between the Ru-TSC-2 anticancer drug and calf thymus DNA(ct DNA) was investigated systematically by using multispectroscopic and electrochemical methods along with viscosity measurements and a ctDNA melting technique. The absorption spectra of the drug with ctDNA showed a slight blue shift and weak hypochromic effect. The relative viscosity and the melting temperature of ctDNA all increased after the addition of the drug. The Stern-Volmer quenching constants together with the modified Stern-Volmer quenching constants at different temperatures were also calculated. The corresponding thermodynamic parameters were also calculated.Using berberine(BH) as a fluorescence probe, this drug statically quenched the fluorescence of the ct DNA-BH complex, and hydrogen bonding and van der Waals interactions played important roles in such an interaction. All the competitive experimental evidence indicated that the binding mode of the drug with ctDNA was intercalative binding.The influences of ionic strength, chemical denaturants, hyperthermia and pH on the binding of the drug with ctDNA were also investigated. The results of FT-IR and CD spectra indicated that the B-form conformation of ctDNA was constant even after the binding interaction of the drug with G-C base pairs of ctDNA. The results of electrochemical experiments further verified the intercalative interaction between ct DNA and the drug. All these results indicated that the biological activity of ctDNA was affected by the Ru-TSC-2 anticancer drug dramatically.The five chapter : the interaction between the ctDNA and three anticancer drugs(Ru-TSC-3,4,5) with different structures were investigated systematically by using multispectroscopic along with viscosity measurements and a ctDNA melting technique. And emphatically discusses the structure-activity relationship for the DNA interactions of three anticancer drugs. The experimental results indicated that the absorption spectra of the drug with ctDNA showed a slight shift and weak hypochromic effect. The relative viscosity and the melting temperature of ctDNA all increased after the addition of the drug. The Stern-Volmer quenching constants together with the modified Stern-Volmer quenching constants at different temperatures were also calculated. The corresponding thermodynamic parameters were also calculated.Using berberine(BH) as a fluorescence probe, hydrogen bonding and van der Waals interactions played important roles in such an interaction.All the competitive experimental evidence indicated that the binding mode of the drug with ctDNA was intercalative binding. The influences of ionic strength,chemical denaturants, hyperthermia and pH on the binding of the drug with ctDNA were also investigated. The results of FT-IR and CD spectra indicated that ctDNA double helixthe was changed after the binding interaction of three drugs. This research could provide valuable information of biological action of this anticancer drug in organism, and might serve as important strategy for pharmacological and toxicological research of novel ruthenium(II) arene complexes with TSC anticancer drugs. |
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