A new hydrazone ligand; 4,6-bis(2-hydroxynaphthalen-1-yl)methyl-ene)hydrazono)ethyl)benzene-1,3-diol (H4L; DHNAPH) was synthesized by the reaction of 4,6-bis(1-hydrazonoethyl)benzene-1,3-diol (NH2DAR) with 2-hydroxy 1-naphthaldehyde. Co(II), Ni(II) and Cu(II)- complexes ([M2L(H2O)6].nH2O.mEtOH; M = Co, Ni or Cu, n = 0.5 or 1 and m = 0.5 or 0) of DHNAPH have been successfully synthesized. Characterization of DHNAPH and its complexes was performed by analytical, spectral (IR, mass, UV-Vis, 1H NMR and ESR), magnetic susceptibility, molar conductivity and thermal gravimetric analysis (TGA) techniques. The scanning electron microscopy (SEM) was used for detection of the morphology of DHNAPH and its Co- and Ni-complexes, which refers that the DHNAPH complexes are in the nano scale. The analytical data, magnetic moments and spectral studies recognized octahedral geometries for DHNAPH complexes. DHNAPH acts as a bis(dibasic triadentate) via (C=Nazomethine and 2 OH) with metal in complexes. The optimized structure of DHNAPH and its complexes has been done theoretically by Hyperchem program and structural parameters were linked with the IR experimental data. The activity of DHNAPH and its complexes against Hepatocellular carcinoma, fungi and bacteria has been tested. The new complexes are more active than DHNAPH and the highest antitumor activity was given by copper(II) complex. The DNA-binding of DHNAPH and Cu- DHNAPH has been investigated. Molecular docking studies showed that all the tested compounds show good binding score revealing good fitting in between the DNA strands. There is agreement between docking data and DNA binding results.