This study focuses on the synthesis of Imidazole-4-one derivatives (F11–F15) through the reaction of three equivalents of the amino acid alanine with one equivalent of Schiff bases. The structural confirmation of the synthesized compounds was carried out using various physical and spectroscopic techniques, including FT-IR, 1H, 13C-NMR, and elemental analysis. Additionally, melting point measurements and purity assessments were performed, while the reaction progress was monitored using Thin-Layer Chromatography (TLC). The laser activity of the synthesized compounds was evaluated using a helium-neon laser in the visible spectrum, with exposure durations of 15, 30, 45, and 60 seconds, and the resulting physical changes were analyzed. Furthermore, the antibacterial activity of the prepared compounds was tested against two bacterial isolates: Escherichia coli (Gram-negative) and Staphylococcus aureus (Gram-positive). Standard antibiotics—amoxicillin, ampicillin, and ciprofloxacin—served as reference controls. Moreover, molecular docking studies were conducted on selected compounds (F12 and F13) to assess their interaction with E. coli using the Molecular Operating Environment (MOE) software (2009).