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A series of 1,3-thiazolidin-4-one derivatives were prepared by the reaction of respective aromatic amine, aromatic aldehyde and thioglycolic acid in dry benzene/toluene. The newly synthesized compounds were characterized on the basis of elemental analysis, IR, 1HNMR and mass spectra. The newly synthesized final compounds were evaluated for their in-vitro antibacterial, antifungal and anti-viral activities. Preliminary results indicated that most of the compounds demonstrated moderate to good antimicrobial activity, comparable to standard drugs. Structure-activity relationship studies revealed that the nature of the substituents at the 2nd and 3rd positions of the thiazolidinone nucleus had a significant impact on the in-vitro antimicrobial activity of this class of potent antimicrobial agents.