The present study was conducted to synthesis of some new imine Cr(III), VO(II), Mn(II) and Ni(II) complexes derived from the condensation of 2-amino phenol with 2-hydroxynapthaldehyde were synthesized. The prepared HNPN imine ligand was analyzed by its melting point, IR, 1H NMR and 13C NMR spectroscopies. The investigated HNPN imine complexes were characterized by elemental analysis, FT IR, UV–vis and thermal analysis (TGA) under nitrogen atmosphere from ambient temperature to 750 °C. The experimental results revealed that the investigated complexes contain hydrated water molecules. The molar conductance values of complexes are relatively low, indicating the non-electrolytic nature of these complexes. Magnetic susceptibility measurements show that the investigated complexes are paramagnetic. Moreover, the stability constants of the preparing complexes were determined spectrophotometrically. All the complexes were found to be monomeric 1:1 (M:L) stoichiometry in nature with octahedral geometry for Cr(III), tetrahedral for Mn(II), square planner for Ni(II) and square pyramidal for VO(II). Moreover, the prepared HNPN imine ligand and its complexes were evaluated for antimicrobial effect against some types of bacteria such as Bacillus subtilis (+ ve)Escherichia coli(− ve) and Staphylococcus aureus (+ ve) and some types of fungi such as AspergillusnigerCandida glabrata and Trichophyton rubrum. The results of these studies indicate that the metal complexes exhibit a stronger antibacterial and antifungal efficiency compared to their corresponding imine ligand. Moreover, the interaction of the investigated complexes with CT-DNA was checked using spectral studies, viscosity measurements and gel electrophoreses. The absorption titration studies revealed that each of these complexes is an avid binder to calf thymus-DNA. Also, there was appreciable changes in the relative viscosity of DNA, which is consistent with enhanced hydrophobic interaction of the aromatic rings and intercalation mode of binding. In addition to, the cytotoxic activity of the prepared imine complexes on human colon carcinoma cells, (HCT-116 cell line), hepatic cellular carcinoma cells, (HepG-2 cell line) and breast carcinoma cells (MCF-7 cell line) has cytotoxicity effect against growth of carcinoma cells compared to the clinically used Vinblastine standard. Furthermore, the molecular docking into TRK (PDB: 1t46) was done for the optimization of the investigated compounds as potential TRK inhibitors