Isatin and its derivatives are important heterocycles found in nature and present in numerous bioactive compounds which possess various biological activities. Moreover, it is an essential building block in organic synthesis. The discovery of novel compounds active against human pathogenic bacteria and fungi is an urgent need, and the isatin may represent the suitable scaffold in the design of biologically relevant antimicrobials. A small library of 18 isatin hybrids was synthetized and evaluated for their antimicrobial potential on three reference strains: , , both important human pathogens infamous for causing community- and hospital-acquired severe systemic infections; and , responsible for devastating invasive infections, mainly in immunocompromised individuals. The study highlighted two lead compounds, and endowed with inhibitory activity against at very low concentrations (39.12 and 24.83 µg/mL, respectively).