This study was conducted to prepare novel azomethine chelates of Cu(II), Pd(II), Zn(II) and Cr(III) with tri-dentate dianionic azomethine OVAP ligand 2-[(2-Hydroxy-phenylimino)-methyl]-6-methoxy-phenol. The prepared compounds were characterized with the help of elemental analyses, spectral (NMR, FT-IR, UV-vis, mass), conductivity, magnetic and thermal measurements. The spectroscopic data suggest that the parent azomethine ligand bind to the investigated metal ions through both deprotonated phenol oxygen and azomethine nitrogen atoms and adopted distorted octahedral geometry in case for Cr(III) and Cu(II) ions while tetrahedral and square planner geometries for Zn(II)and Pd(II) ions, respectively. In order to confirm the molecular geometry of the investigated azomethine chelator and its complexes, the theoretical DFT calculations were employed. Correlation between experimental observations and theoretical calculations the geometry optimization results are in a good agreement. Absorption titration has been used to explore the interaction of the investigated azomethine metal chelates with Calf thymus DNA (CT-DNA) and the constant of binding (Kb) as well as Gibb's free energy have been evaluated. Viscosity measurement and gel electrophoreses studies suggest that an intercalative and replacement binding modes of the titled azomethine metal chelates with CT-DNA. Additionally, antimicrobial study of the complexes has been screened against some of pathogenic bacteria and fungi. This biological study shows that the subject complexes have shown a remarkable inhibitory effect compared to its corresponding ligand and standard drugs. Furthermore, The effect of the new synthesized compounds as antioxidants was determined by reduction of 1,1-diphenyl-2-picryl hydrazyl (DPPH) and compared with that of Vitamin C. Finally, in vitro cell proliferation via MTT assay was investigated against Colon carcinoma cells, (HCT-116 cell line), hepatic cellular carcinoma cells, (HepG-2( and breast carcinoma cells (MCF-7) to calculate the cytotoxicity of the prepared compounds. The cell proliferation was inhibited for all compounds and in a dose-dependent manner in the sequence of OVAPPd> OVAPCu> OVAPZn> OVAPCr> OVAP azomethine ligand.