N-Carbobenzoxyglycyl-prolyl-4-methylcoumarinyl amide → HCID 131726966, compound

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Reaction Steps

To examine whether FAP could cleave other N-substituted-Gly-Pro-based substrates, N-methyl-, formyl-, succinyl-, benzyloxycarbonyl-(Z-) and biotinyl-Gly-Pro-AMCC substrates 6 were synthesized and reacted with FAP (37 nM) and DPP-4 (6.8 nM). With the exception of succinyl-Gly-Pro-AMCC, FAP cleaved all N-substituted-Gly-Pro-AMCC substrates 6 at rates 35-150% of the normalized rate for Gly-Pro-AMCC hydrolysis (Table 2), indicating that the protease tolerates other N-terminal blocking groups. Kinetic analysis with commercially available Z-Gly-Pro-AMC (Table 5) showed a catalytic efficiency about 3-fold lower than that for Gly-Pro-AFC (Table 4), consistent with these results. In contrast with FAP, DPP-4 cleaved only N-methyl-Gly-Pro-AMCC at a rate comparable to Gly-Pro-AMCC. A low rate of hydrolysis was obtained with formyl-Gly-Pro-AMCC substrates but no cleavage of succinyl-, Z-, or biotinyl-Gly-Pro-AMCC substrates was observed at concentrations up to 1 mM, indicating that DPP-4 does not tolerate N-acyl-Gly-Pro-based substrates. DPP-4 also showed little activity against commercially available Z-Gly-Pro-AMC (Table 5).

Provenance & Evidence