Abstract
A Cu(II) thiolato complex [CuL(imz)] (1) (H2L = o-HOC6H4C(H)=NC6H4SH-o) and the corresponding water-soluble stable sulfinato-O complex [CuL'(imz)] (2) (H2L' = o-HOC6H4C(H)=NC6H4S(=O)OH) were synthesized and characterized using physicochemical techniques. Compound 2 is found to be a dimer in the solid state as characterized using single-crystal X-ray crystallography. XPS studies clearly showed the differences in the sulfur oxidation states in 1 and 2. Both compounds are found to be monomers in solution as revealed from their four-line X-band electron paramagnetic resonance spectra in CH3CN at room temperature (RT). 1-2 were tested to assess their ability to exhibit DNA binding and cleavage activity. Spectroscopic studies and viscosity experiments suggest that 1-2 bind to CT-DNA through the intercalation mode having moderate binding affinity (Kb ∼ 104 M-1). This is further supported by molecular docking studies of complex 2 with CT-DNA. Both complexes display significant oxidative cleavage of pUC19 DNA. Complex 2 also showed hydrolytic DNA cleavage. The interaction of 1-2 with HSA revealed that they have strong ability to quench the intrinsic fluorescence of HSA by a static quenching mechanism (kq ∼ 1013 M-1 s-1). This is further complemented by Förster resonance energy transfer studies that revealed binding distances of r = 2.85 and 2.75 nm for 1 and 2, respectively, indicating high potential for energy transfer from HSA to complex. 1-2 were capable of inducing conformational changes of HSA at secondary and tertiary levels as observed from synchronous and three-dimensional fluorescence spectroscopy. Molecular docking studies with 2 indicate that it forms strong hydrogen bonds with Gln221 and Arg222 located near the entrance of site-I of HSA. 1-2 showed potential toxicity in human cervical cancer HeLa cells, lung cancer A549 cells, and cisplatin-resistant breast cancer MDA-MB-231 cells and appeared to be most potent against HeLa cells (IC50 = 2.04 μM for 1 and 1.86 μM for 2). In HeLa cells, 1-2 mediated cell cycle arrest in S and G2/M phases, which progressed into apoptosis. Apoptotic features seen from Hoechst and AO/PI staining, damaged cytoskeleton actin viewed from phalloidin staining, and increased caspase-3 activity upon treatment with 1-2 collectively suggested that they induced apoptosis in HeLa cells via caspase activation. This is further supported by western blot analysis of the protein sample extracted from HeLa cells treated with 2.