title: Potent inhibition of endopeptidase 24.16 and endopeptidase 24.15 by the phosphonamide peptide N-(phenylethylphosphonyl)-Gly-L-Pro-L-aminohexanoic acid.
abstract: A phosphonamide peptide, N-(phenylethylphosphonyl)-Gly-L-Pro-L-aminohexanoic acid, previously shown to block Clostridium histolyticum collagenases, was examined as a putative inhibitor of endopeptidase 24.16 and endopeptidase 24.15. Hydrolysis of two endopeptidase 24.16 substrates, i.e. 3-carboxy-7-methoxycoumarin (Mcc)-Pro-Leu-Gly-Pro-D-Lys-dinitrophenyl (Dnp) and neurotensin, were completely and dose-dependently inhibited by the phosphonamide inhibitor with KI values of 0.3 and 0.9 nM respectively. In addition, the phosphonamide peptide inhibited the hydrolysis of benzoyl (Bz)-Gly-Ala-Ala-Phe-(pAB) p-aminobenzoate and neurotensin by endopeptidase 24.15 with about a 10-fold lower potency (KI values of 5 and 7.5 nM respectively). The selectivity of this inhibitor towards several exo- and endo-peptidases belonging to the zinc-containing metallopeptidase family established that a 1 microM concentration of this inhibitor was unable to affect leucine aminopeptidase, carboxypeptidase A, angiotensin-converting enzyme and endopeptidase 24.11. The present paper therefore reports on the first hydrophilic highly potent endopeptidase 24.16 inhibitor and describes the most potent inhibitory agent directed towards endopeptidase 24.15 developed to date. These tools should allow one to assess the contribution of endopeptidase 24.16 and endopeptidase 24.15 to the physiological inactivation of neurotensin as well as other neuropeptides.
authors.affiliation: Institut de Pharmacologie Moléculaire et Cellulaire, UPR 411 du CNRS, Université de Nice Sophia Antipolis, Valbonne, France.
authors.forename: H
authors.initials: H
authors.lastname: Barelli
authors.forename: V
authors.initials: V
authors.lastname: Dive
authors.forename: A
authors.initials: A
authors.lastname: Yiotakis
authors.forename: J P
authors.initials: JP
authors.lastname: Vincent
authors.forename: F
authors.initials: F
authors.lastname: Checler
mesh_terms: D000595:Amino Acid Sequence; D000614:Aminocaproates / Q000494:pharmacology*; D002268:Carboxypeptidases / Q000187:drug effects / Q000378:metabolism; D043422:Carboxypeptidases A; D004151:Dipeptides / Q000494:pharmacology*; D006868:Hydrolysis; D007700:Kinetics; D007931:Leucyl Aminopeptidase / Q000187:drug effects / Q000378:metabolism; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D008969:Molecular Sequence Data; D015260:Neprilysin / Q000187:drug effects / Q000378:metabolism; D009842:Oligopeptides; D007703:Peptidyl-Dipeptidase A / Q000187:drug effects / Q000378:metabolism
chemical_list: D000614:Aminocaproates; D004151:Dipeptides; D009842:Oligopeptides; C077650:N-(phenylethylphosphonyl)-glycyl-prolyl-aminohexanoic acid; D002268:Carboxypeptidases; D007931:Leucyl Aminopeptidase; D007703:Peptidyl-Dipeptidase A; D043422:Carboxypeptidases A; D008666:Metalloendopeptidases; D015260:Neprilysin; C022510:thimet oligopeptidase; C062316:neurolysin