Several physicochemical techniques were used to evaluate the synthesis and structural analysis of Fe(III) complex obtained from {3,4-Bis-[(3-ethoxy-2-hydroxy-benzylidene)-amino]-phenyl}-phenyl-methanone (L ligand). The methods encompassed theoretical investigations as well as elemental analysis (CHN), conductivity, magnetic susceptibility, spectroscopic (IR, NMR, mass spectrometry), decomposition point determination, and UV-Vis spectrum analysis. IR spectra suggested which the ligand coordinates through the metal ions in a tetra-dentate fashion, utilizing the (N & O) donor sites from the (C=N & C-O) groups in the ligand moiety. Job's approach and analytical results from solution complexation indicated a molar ratio of 1:1 for the metal to ligand. In accordance with the value of the stability constant (Kf), the complex's stability order was ascertained. According to the pH profile, the compound under study is stable over a broad pH range, usually between (pH= 4:10). The electronic structures of the investigated ligand and its complex is analyzed using quantum chemical calculations via the DFT approach. The anti-pathogenic activity of the generated materials was experimentally proven against a subset of gram (+) and gram (-) bacteria and several fungi using the agar well diffusion method. Additionally, the ability of the L ligand and its metal chelate to cause harm to cells of the liver, breast, and colon was investigated. Furthermore, the examined compounds' ability to suppress the DPPH radical was examined. In addition, landing of molecule simulations was finished to ascertain the mechanism by which the produced compounds bound to the intended protein binding sites.