Recent advancements in coordination chemistry and its biological applications have made a significant impact in chemical biology. Motivated by this progress, we have synthesized a new benzimidazole-based imine ligand, 2-((E)-((1H-benzo[d]-2-yl)methylimino)methyl)-4-fluorophenol (HBMF), and investigated its complexes with Co(III) and Cu(II) ions. The metal complexes (C1–C4) were prepared in two distinct stoichiometries: a 2:1 (HBMF: metal ion) ratio and a 1:1:1 ratio (HBMF: metal ion: 1,10-phenanthroline). Comprehensive structural characterization of these compounds was performed using FT-IR, UV-Vis, NMR, mass spectrometry, and elemental analysis. To further understand their properties, we employed both first-principles computational methods and molecular dynamics simulations. Quantum mechanical calculations using density functional theory (DFT) were applied to explore the reactive and spectroscopic features of the HBMF ligand and its metal complexes, while molecular dynamics simulations with the OPLS4 force field were used to investigate the ligand's reactivity in aqueous environments. The biological activity of the compounds was assessed in vitro against several cancer cell lines, including A549, Ehrlich-Lettre ascites carcinoma (EAC), SIHA, and NIH3T3. Among the metal complexes, compound C4 (C27H23Cl2CoFN5O3) demonstrated the strongest anti-proliferative effect against EAC cells, with an impressive IC50 of 10 µM, outperforming both the parent ligand and other metal complexes. Hematological assessments, including the measurement of alkaline phosphatase, creatinine, urea, RBC, and WBC levels, revealed notable results. Additionally, C4’s effect on angiogenesis was evaluated by stimulating neovascularization through rVEGF165, with comparisons to non-tumor models. In vivo studies were conducted on EAC cells, where animals were treated with doses of 50 and 75 mg/kg, and the tumor parameters were carefully analyzed.
