This study explores the potential of compounds as antibacterial agent by examining its binding interactions with biotin protein ligase and DNA gyrase, essential enzymes in bacterial metabolic pathways. Molecular docking simulations reveal strong affinities between compound 6a and both target proteins. The study also a class of compounds 6(a-e) of substituted azetidinone derivatives of drug. Among of them, Synthesis of novel compound 6(a-e) were showed valuable characteristic and efficacy as antibacterial activity. 6c, 3-chloro-4-(4-chlorophenyl)-1-(3-phenyl-[1,2,4]triazolo[3,4-b][1,3,4] thiadiazol-6-yl)azetidin-2-one, Chloro and nitro substituted of azetidinone 6c and 6d derivatives were showed moderate to better biological activity as compare to standard drugs moxifloxacin, for regarding this observation, heterocyclic compounds of these derivative are very useful in the field of medicinal chemistry. These derivatives of azetidinone nucleus were possesses remarkable, better clinically efficacy with low toxicity. Purity of these derivatives were checked with the help of TLC.The structure of new synthesised compounds was identified and confirmed by elemental analysis, IR, NMR, and activity of new synthesised these drugs were screened for their antibacterial activity.