A series of new thiazolidinones with barbituric acid scaffolds 5(a-j) were synthesized and evaluated for their antibacterial activity against Gram-positive bacteria (B. subtilis, B. sphaericus, S. aureus), Gram-negative bacteria (P. aeruginosa, K. aerogenes, C. violaceum) and also antifungal activity against C. albicans, A. fumigatus, T. rubrum and T. metagrophytes. The antibacterial activity of compounds containing 4-hydroxyphenyl (5c), 2,6-difluorophenyl (5g) and 4-hydroxy-3-methoxy- phenyl (5i) groups on thizolidionone ring showed considerable activity against tested bacterial strains. Compound, contain 4-chlorophenyl group (5c) and 4-N,N-dimethylaminophenyl (5h) showed better activity against Gram-positive (except B. Subtilis) and Gram-negative bacteria (except C. Violaceum). Similarly, compounds containing 4-methylphenyl (5b), 3-nitrophenyl (5f) and pipernyl (5j) were active against B. sphaericus, and S. Aureus, respectively. The antifungal evaluation of 5(a-j) indicates that the compounds 5e and 5i which contain 4-hydroxyphenyl and 4-hydroxy-3-methoxyphenyl respectively showed considerable activity against all the fungal strains.