Used as a fluorogenic substrate in enzymatic assays, particularly for studying protease activity. The compound is designed so that upon cleavage by specific proteases, especially trypsin-like enzymes, the 7-amido-4-methylcoumarin group is released, producing a measurable fluorescent signal. This allows real-time monitoring of enzyme kinetics and inhibition in biochemical research. Commonly applied in high-throughput screening for drug discovery, especially in assays targeting serine proteases involved in coagulation, inflammation, and viral entry pathways. Its high sensitivity and signal-to-noise ratio make it suitable for both in vitro and cell-based assays.