In this work, to contribute to research in the area of new chemotherapeutic agents, we attempted at obtaining new metal-based compounds as alternatives to the existing metal-based anticancer drugs. In this context, Cu(II), Zn(II) and Pt(II) metal-based complexes of pharmaceutically active compound, effective antineoplastic methotrexate (MTX), were synthesized, and their structures were elucidated by analytical (melting point, elemental analysis, conductivity, and solubility), spectroscopic (UV-Vis, IR, 1H NMR, LC-MS and ICP-OES) and thermal (TGA, DTA) methods. UV-Vis spectroscopy examined interactions of MTX pharmaceutically active substances and their metal-based compounds with the fish sperm double-stranded DNA (FS-dsDNA). Regarding the data obtained from spectroscopic measurements, it is understood that all compounds interact with FS-dsDNA. The antiproliferative activity of the active agent MTX and the newly synthesized metal-based compounds was investigated on C6 and He-La cells in comparison to the active anticancer agents present in the market using real-time cell analyzer with four different concentrations. Also, interactions of all compounds obtained with the double-stranded FS-dsDNA were studied using pencil graphite electrode (PGE) by monitoring changes in the signal of the guanine base. The associated surface morphology was examined by differential pulse voltammetry (DPV) and scanning electron microscopy (SEM) techniques.