Bifunctional coumarin derivatives in solution and solid phase synthesis of fluorogenic enzyme substrates

Abstract

AbstractThe use of the fluorescent bifunctional compounds 7‐amino‐4‐coumarinyl‐acetic acid 1, 7‐hydroxy‐4‐coumarinyl‐acetic acid 2 and ethyl 7‐amino‐4‐coumarinyl‐acetate 3 in solution and solid phase synthesis of fluorogenic enzyme substrates was examined. The intramolecularly quenched fluorogenic substrate N‐(7‐amino‐4‐coumarinyl‐acetyl)‐L‐phenylalanyl‐p‐nitroanilide 5, and the fluorogenic one ethyl 7‐(glutaryl‐L‐phenylalanilamido)‐4‐coumarinyl‐acetate 8, both suitable for chymotrypsin and/or chymotrypsin like enzymes determination, were prepared in solution. The substrates 7‐oleyloxy‐4‐coumarinyl‐acetic acid 13 and 7‐palmitoyloxy‐4‐coumarinyl‐acetic acid 14, suitable for the enzymatic study of lipases, were prepared by solid phase technique using 2‐chloro‐chlorotrityl‐resin. The study of the fluorescence properties of the fluorophores 1, 2, 3, and substrates 5, 8,13,14 showed that the examined bifunctional coumarin derivatives are suitable markers for solution and solid phase synthesis of fluorogenic enzyme substrates.