Used primarily in biochemical research, this compound serves as a substrate for proteolytic enzymes, particularly for studying the activity of proteases like cathepsin B. Its application is crucial in enzyme kinetics assays, where the release of 7-amido-4-methylcoumarin (AMC) upon enzymatic cleavage is monitored through fluorescence. This allows researchers to quantify enzyme activity and screen for potential inhibitors, aiding in drug discovery and the study of disease mechanisms. Additionally, it is employed in cell biology to investigate intracellular protease functions and their roles in processes such as apoptosis and protein degradation.