Phe-AMC (L-Phenylalanine 7-amido-4-methylcoumarin Hydrochloride) is a synthetic fluorogenic substrate and protease inhibitor used in enzymatic assays targeting aminopeptidases and proteases—particularly those that cleave N-terminal L-phenylalanine residues, such as serine and cysteine proteases. It binds to the active site of these enzymes, blocking their function, and is especially suited for aminopeptidase and chymotrypsin-like protease assays, including α-chymotrypsin activity monitoring. Upon enzymatic hydrolysis, Phe-AMC releases 7-amido-4-methylcoumarin (AMC), a strongly fluorescent compound with excitation at 341 nm and emission at 441 nm. This enables sensitive, real-time kinetic measurements across diverse assay formats, from microplate and cuvette setups to automated high-throughput screening (HTS) systems. Beyond its analytical applications, Phe-AMC has been shown to have biological functions, including the prevention of carcinogenesis, making it a valuable tool in medicinal chemistry, drug discovery, and functional protease studies.