Synthesis, Biological Evaluation, Molecular Docking, and DFT Analysis of Potential Bioactive Azo‐Imines

Azo‐Schiff base compounds attracted considerable attention due to their unique properties arising from N═N and C═N functional groups. This study aimed to synthesize four substituted imines containing azo moieties, 4‐((4‐X‐phenyl) diazinyl)‐2‐(( Z )‐((( R )‐2‐hydroxy‐1‐phenylethyl)imino)methyl)phenol, and evaluate their biological potential through acute toxicity (LD 50 ), antibacterial, and antioxidant assays. The substituent X includes H (M1), CH 3 (M2), Cl (M3), and CN (M4). LD 50 results showed compound M4 had the lowest toxicity of 71.86 mg/kg. Antibacterial tests revealed selective activity against Gram‐positive bacteria with strong inhibition by M1 and M2. Antioxidant assays demonstrated significant antioxidant properties in synthesized compounds, in particular for M4 in radical scavenging. The results indicate that azo‐imine showed promise as antimicrobial and antioxidant agents. According to density functional theory calculations, compounds M1 and M2 were found to be most stable, while M4 was more reactive based on its electronic properties. Molecular docking analysis was conducted to investigate the binding mode and energy of the ligand to the Staphylococcus aureus receptor. Compound M1 binds via donor hydrogen bonding with its phenolic group; compounds M2 and M3 interact through both phenolic and alcohol hydroxyl groups; and compound M4 forms an acceptor hydrogen bond via nitrile group.