Ligand Rigidity and π-Surface Modulate Biomolecular Interactions and Cytotoxicity in Ru(II) Polypyridyl Complexes

The complexes cis-[Ru(dmbpy)2Cl(bpy)](PF6) (Rubpy) and cis-[Ru(dmbpy)2Cl(bpe)](PF6) (Rubpe) (dmbpy = 4,4′-Dimethyl-2,2′-dipyridyl, bpy= 4,4′-dipyridyl and bpe = 1,2-bis(4-pyridyl)ethane) were synthesized and spectroelectrochemically characterized. Both Ru(II) complexes exhibited absorption bands assigned to intraligand and metal-to-ligand charge transfer (MLCT) transitions, and their spectral stability in PBS buffer (pH 7.4) supports their suitability for biological studies involving biomolecules or living cells. Fluorescence quenching assays revealed strong interactions with bovine serum albumin (BSA), with binding constants (Kb) values were 2.89 × 105 M−1 for Rubpy and 1.97 × 105 M−1 for Rubpe, and a stoichiometry of one binding site per albumin molecule. DNA-binding studies demonstrated non-covalent interactions with ss-DNA, evidenced by a hyperchromic effect in the MLCT bands, suggesting a partial intercalation or groove-binding mechanism. Cellular uptake assays indicated moderate incorporation of both complexes in tumor cells, with uptake levels of 52% (Rubpy) and 47% (Rubpe) in HeLa cells, and 42% (Rubpy) and 32% (Rubpe) in MDA-MB-231 cells. Despite the similar uptake profiles, cytotoxicity assays showed that Rubpe is approximately 2.4 times more potent than Rubpy, with IC50 values of 9 μM (HeLa) and 12 μM (MDA-MB-231), compared to 22 μM and 29 μM for Rubpy, respectively. These results highlight the relevance of these Ru(II) complexes as molecular platforms for exploring structure–activity relationships in anticancer agents.