Then synthesis of Ni (L) (1) and VO (L) (2) [H₂L = N-2-hydroxyacetophenon-Ń-2, 4-dihydroxbenzaldehyde-1, 2-phenylenediimine)] novel unsymmetrical heterocyclic Schiff base complexes is reported. Elemental analysis, UV-Vis, IR spectroscopy, multi-nuclear spectroscopy (¹H and ¹³C NMR) and single crystal X-ray diffraction analysis have been used to characterize the products. UV-Vis spectroscopy, emission titration, viscosity measurement, helix melting methods and circular dichroism (CD) spectroscopy have been applied to investigate the interactions of the complexes with CT-DNA. Intercalative binding mode between the complexes and DNA is suggested by the binding constant (K_b) values of 2.9 × 10⁴ and 5.4 × 10⁴ for complexes 1 and 2, respectively. In particular, the in vitro cytotoxicity of the complexes on two cancer cells lines (MDA-MB 231, SKOV-3 cell line) showed that the compounds had …