Four Schiff base ligands (L1–L4) were synthesized by reacting 1, 3 – Bis(4-aminophenoxy) benzene with 2-hydroxy-benzaldehyde, 5-Chloro-2-hydroxybenzaldehyde, 5-Bromo-2-hydroxybenzaldehyde, 2-hydroxy-5-methylbenzaldehyde. By using 1H NMR, 13C NMR, UV–Visible, FT-IR, HRMS technique synthesized ligand were characterized. These ligands exhibited efficient antibacterial activity against four different human pathogenic bacterial strains such as gram-negative Escherichia coli, Pseudomonas aeruginosa, and gram-positive Staphylococcus aureus, Bacillus substilis whereas antifungal activity against Candida albicans and Aspergillus niger fungal strains. The ligand of L3 act as the most active antibacterial agent and show better antibacterial activity with MIC 135 ± 0.39, 122 ± 0.12, and 130± 0.54 respectively against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Bacillus subtilis.The results of the molecular docking analysis for the synthesized compounds indicate that L1, L2, L3 and L4 exhibit the strong antibacterial activity when compared to standard ampilcillin