A series of 7-chloro-2,3-disubstituted-4(3H)-quinazolinones (4a-j) were synthesized by the reaction of 3-amino-7-chloro-2-phenylquinazolin-4(3H)-one (3) with various amines in presence of formaldehyde. The starting material for the compounds was 2-amino-4-chloro benzoic acid. The structures of the compounds were characterized by IR, 1 H NMR, elemental analyses. Docking analysis of quinazolinone derivatives was done with DNA gyrase protein which was carried out by means of the AutoDockTools (ADT) v 1.5.4 and AutoDock v 4.2 programs. All compounds interacted with DNA gyrase protein. Preclinical evaluation of the compounds was ascertained by in silico toxicity, blood-brain barrier and drug like properties. All compounds were investigated for their antibacterial activity. Overall, compound (4b) showed better properties as a drug like candidate.