This chemical is primarily used in biochemical research as a substrate for measuring the activity of proteolytic enzymes, particularly those involved in blood coagulation pathways. It is specifically designed to assess the activity of thrombin, a key enzyme in the clotting process. By cleaving the peptide bond, it releases a chromogenic group (p-nitroaniline), which can be quantified spectrophotometrically to determine enzyme activity. This makes it a valuable tool in studying coagulation disorders, evaluating anticoagulant therapies, and conducting in vitro diagnostic tests. Its application extends to pharmaceutical development, where it aids in screening and characterizing potential inhibitors of thrombin or related enzymes.