Metallocene dichlorides (Cp2M(IV)Cl2) are the first class of small and hydrophobic organometallic compounds classified as anticancer agents against numerous cancer cell lines and tumors. In this study, the antiproliferative activities of Cp2VCl2,Cp2NbCl2, Cp2HfCl2 and Cp2ZrCl2were assessed on two human cancer cell lines (HT-29 and MCF-7) using MTT assay. Spectroscopic studies were also conducted using these and other known metallocene dichlorides on apo-human transferrin (apo-hTf) at pH 7.4. UV-Vis and CD showed that their interaction with apo-hTf could induce conformational changes of its secondary structure during binding process. In fluorescence, a decrease in intensity of the emission peak was observed when the apo-hTf:Cp2M(IV)Cl2 complex is being formed, probably due to changes in the microenvironment of its tyrosine and tryptophan residues. Among all metallocene dichlorides studied, Cp2VCl2 has the strong ability to quench the intrinsic fluorescence of apo-hTf through a static quenching mechanism. The association constants for each protein-compound complex were also determined at different temperatures (296 K, 303 K, 310 K, and 317 K) based on fluorescence quenching results. Positive enthalpy changes (ΔH) and entropy changes (ΔS) as well as negative free energies (ΔG) suggest that hydrophobic interactions are the main intermolecular forces involved in the binding process, probably via an endothermic and spontaneous reaction mechanism. The distance, r, between donor (apo-hTf) and acceptor (Cp2M(IV)Cl2) obtained according to Forster's theory of non-radiation energy transfer suggest that the energy transfer from apo-hTf to Cp2M(IV)Cl2 occurs with high probability and distances obtained by FRET with high accuracy.
Keywords: anticancer activity; apo human transferrin; metallocene dichlorides; protein-ligand interactions; spectroscopic analysis.