Ru(II)-<i>p</i>-Cymene Complexes of Furoylthiourea Ligands for Anticancer Applications against Breast Cancer Cells

Abstract

Half-sandwich Ru(II) complexes containing nitro-substituted furoylthiourea ligands, bearing the general formula [(<i>η</i>⁶-<i>p</i>-cymene)RuCl₂(L)] (<b>1</b>-<b>6</b>) and [(<i>η</i>⁶-<i>p</i>-cymene)RuCl(L)(PPh₃)]⁺ (<b>7-</b>-<b>12</b>), have been synthesized and characterized. In contrast to the spectroscopic data which revealed monodentate coordination of the ligands to the Ru(II) ion <i>via</i> a "S" atom, single crystal X-ray structures revealed an unusual bidentate N, S coordination with the metal center forming a four-membered ring. Interaction studies by absorption, emission, and viscosity measurements revealed intercalation of the Ru(II) complexes with calf thymus (CT) DNA. The complexes showed good interactions with bovine serum albumin (BSA) as well. Further, their cytotoxicity was explored exclusively against breast cancer cells, namely, MCF-7, T47-D, and MDA-MB-231, wherein all of the complexes were found to display more pronounced activity than their ligand counterparts. Complexes <b>7</b>-<b>12</b> bearing triphenylphosphine displayed significant cytotoxicity, among which complex <b>12</b> showed IC₅₀ values of 0.6 ± 0.9, 0.1 ± 0.8, and 0.1 ± 0.2 μM against MCF-7, T47-D, and MDA-MB-231 cell lines, respectively. The most active complexes were tested for their mode of cell death through staining assays, which confirmed apoptosis. The upregulation of apoptotic inducing and downregulation of apoptotic suppressing proteins as inferred from the western blot analysis also corroborated the apoptotic mode of cell death. The active complexes effectively generated reactive oxygen species (ROS) in MDA-MB-231 cells as analyzed from the 2',7'-dichlorofluorescein diacetate (DCFH-DA) staining. Finally, <i>in vivo</i> studies of the highly active complexes (<b>6</b> and <b>12</b>) were performed on the mice model. Histological analyses revealed that treatment with these complexes at high doses of up to 8 mg/kg did not induce any visible damage to the tested organs.