Two new ruthenium(II) polypyridyl complexes, [Ru(dmp)2(maip)](ClO4)21 (maip = 2-(3-aminophenyl)imizado[4,5-f][1,10]phenanthroline and [Ru(dmp)2(paip)](ClO4)22 (paip = 2-(4-aminophenyl)imidazo[4,5-f][1,10]phenanthroline, dmp = 2, 9-dimethyl-1,10-phenanthroline) have been synthesized and characterized. The DNA-binding behaviors of complexes 1 and 2 were studied by viscosity measurements, thermal denaturation, and absorption titration. The results show that the two complexes intercalate between the base pairs of DNA. The DNA-binding constants Kb for complexes 1 and 2 were determined to be 3.23 ± 0.16 × 104 M−1 (s = 0.97) and 4.34 ± 0.65 × 104 M−1 (s = 1.13). The cytotoxicity of these complexes has been evaluated by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The IC50 values are 35.70, 41.04, 55.25 and 37.50 for complex 1 and 37.02, 103.08, 130.07 and 37.80 for complex 2 against BEL-7402, C-6, HepG-2 and MCF-7 cell lines, respectively. The antioxidant activity against hydroxyl radical (OH•) was also investigated.