[Pd(L 1 )(C,N)]CF 3 SO 3 and [Pd(L 2 )(C,N)]CF 3 SO 3 (L 1 = 2,2′-bipyridine, L 2 = 1,10-phenanthroline and C,N = benzylamine) novel orthopalladated complexes have been synthesized and characterized using various techniques. The binding of the complexes with native calf thymus DNA (CT-DNA) was monitored using UVvisible absorption spectrophotometry, fluorescence spectroscopy and thermal denaturation studies. Our results indicate that these complexes can strongly bind to CT-DNA via partial intercalative mode. In addition, fluorescence spectrometry of bovine serum albumin (BSA) with the complexes shows that the fluorescence quenching mechanism of BSA is a static process. The results of site-competitive replacement experiments with specific site markers clearly indicate that the complexes bind to site I of BSA. Notably, the complexes exhibit significant in vitro cytotoxicity against two human cancer cell lines (Jurkat and MCF-7) with IC 50 values varying from 37 to 53 μM. Finally, a molecular docking experiment effectively proves the binding of the Pd(II) complexes to DNA and BSA.