title: Inhibitors of tripeptidyl peptidase II. 2. Generation of the first novel lead inhibitor of cholecystokinin-8-inactivating peptidase: a strategy for the design of peptidase inhibitors.
abstract: The cholecystokinin-8 (CCK-8)-inactivating peptidase is a serine peptidase which has been shown to be a membrane-bound isoform of tripeptidyl peptidase II (EC 3.4.14.10). It cleaves the neurotransmitter CCK-8 sulfate at the Met-Gly bond to give Asp-Tyr(SO(3)H)-Met-OH + Gly-Trp-Met-Asp-Phe-NH(2). In seeking a reversible inhibitor of this peptidase, the enzymatic binding subsites were characterized using a fluorimetric assay based on the hydrolysis of the artificial substrate Ala-Ala-Phe-amidomethylcoumarin. A series of di- and tripeptides having various alkyl or aryl side chains was studied to determine the accessible volume for binding and to probe the potential for hydrophobic interactions. From this initial study the tripeptides Ile-Pro-Ile-OH (K(i) = 1 microM) and Ala-Pro-Ala-OH (K(i) = 3 microM) and dipeptide amide Val-Nvl-NHBu (K(i) = 3 microM) emerged as leads. Comparison of these structures led to the synthesis of Val-Pro-NHBu (K(i) = 0.57 microM) which served for later optimization in the design of butabindide, a potent reversible competitive and selective inhibitor of the CCK-8-inactivating peptidase. The strategy for this work is explicitly described since it illustrates a possible general approach for peptidase inhibitor design.
authors.affiliation: Department of Chemistry, University College London, 20 Gordon Street, London WC1H 0AJ, England.
authors.forename: C R
authors.initials: CR
authors.lastname: Ganellin
authors.forename: P B
authors.initials: PB
authors.lastname: Bishop
authors.forename: R B
authors.initials: RB
authors.lastname: Bambal
authors.forename: S M
authors.initials: SM
authors.lastname: Chan
authors.forename: J K
authors.initials: JK
authors.lastname: Law
authors.forename: B
authors.initials: B
authors.lastname: Marabout
authors.forename: P M
authors.initials: PM
authors.lastname: Luthra
authors.forename: A N
authors.initials: AN
authors.lastname: Moore
authors.forename: O
authors.initials: O
authors.lastname: Peschard
authors.forename: P
authors.initials: P
authors.lastname: Bourgeat
authors.forename: C
authors.initials: C
authors.lastname: Rose
authors.forename: F
authors.initials: F
authors.lastname: Vargas
authors.forename: J C
authors.initials: JC
authors.lastname: Schwartz
mesh_terms: D000626:Aminopeptidases; D000818:Animals; D002540:Cerebral Cortex / Q000378:metabolism; D004151:Dipeptides / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; D015195:Drug Design; D066298:In Vitro Techniques; D009842:Oligopeptides / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D051381:Rats; D012697:Serine Endopeptidases / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D012844:Sincalide / Q000378:metabolism*; D013329:Structure-Activity Relationship
chemical_list: D004151:Dipeptides; D009842:Oligopeptides; D015842:Serine Proteinase Inhibitors; D000626:Aminopeptidases; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; C048001:tripeptidyl-peptidase 2; D012697:Serine Endopeptidases; D012844:Sincalide