In medicinal chemistry, Schiff bases have garnered considerable interest because of their diverse array of pharmacological properties. Many derivatives of Schiff bases demonstrate antimicrobial, antiviral, antimalarial, and anticancer activities, making them promising candidates for drug development. Their therapeutic potential is largely owing to their capacity to engage with biological molecules, including proteins and nucleic acids. In our research, we synthesized Schiff base intermediate molecules from aniline and utilized 1HNMR, 13CNMR, and LCMS analysis to elucidate the structures of the synthesized compounds. Additionally, we conducted molecular docking studies to analyse the antibacterial efficacy of the most promising compound. The compound (3-(4-chloro-3-nitrophenyl)-2-(3-chlorophenyl)-1, 3 thiazolidin-4-one) demonstrated a strong binding affinity with the protein. Furthermore, in vitro antibacterial activity tests revealed that the synthesized compounds exhibited significant antibacterial activity.