This compound is primarily used in biochemical research as a substrate for studying enzyme activity, particularly for proteases and peptidases. It helps in understanding the mechanisms of enzyme-catalyzed reactions and their specificity. The 4-nitroaniline group, released upon enzymatic cleavage, can be easily detected spectrophotometrically, making it a valuable tool for kinetic assays and enzyme inhibition studies. It is also employed in drug discovery to screen and evaluate potential inhibitors of proteolytic enzymes, aiding in the development of therapeutic agents targeting diseases where protease activity is dysregulated.