New series of thiosemicarbazone derivatives incorporating a hippuric acid moiety have been synthesized through the reaction of N-(4-hippuric acid)thiosemicarbazide with various carbonyl compounds such as aldehydes, ketones and cyclic ketones. The synthesized compounds were tested against bacterial and fungal strains. Most of compounds, such as 2-(4-(2-(4-fluorobenzylidene)hydrazinecarbothioamido)benzamido)acetic acid showed significant antibacterial and antifungal activities. These compounds comprise a new class of promising broad-spectrum antibacterial and antifungal agents.