title: Rational optimization of a short human P-selectin-binding peptide leads to nanomolar affinity antagonists.
abstract: P-selectin plays an important role in the development of various diseases, including atherosclerosis and thrombosis. In our laboratory we recently identified a number of specific human P-selectin-binding peptides containing a Glu-Trp-Val-Asp-Val consensus motif, displaying a low micromolar affinity for P-selectin (IC(50) = 2 microm). In search of more potent antagonists for P-selectin, we have optimized the EWVDV pentapeptide core motif via a two-step combinatorial chemistry approach. A dedicated library of peptide derivatives was generated by introducing seven substituents at the N and C termini of the motif. In particular, pentapeptides with gallic acid or 1,3,5-benzenetricarboxylic acid substituents at the N terminus proved to be considerably more potent inhibitors of P-selectin binding than the parental peptide. After removal of the N-terminal glutamic acid from the core sequence, which appeared to be replaceable by a carboxamide function without loss of affinity, a second library was synthesized to map the chemical moieties within the gallic acid or 1,3,5-benzenetricarboxyl acid groups responsible for the enhanced P-selectin binding. Moreover, by varying the length and rigidity of the connective spacer, we have further optimized the spatial orientation of the N-terminal substituent. The combined use of phage display and subsequent combinatorial chemistry led to the design of a number of gallic acid- containing peptides with low nanomolar affinity for P-selectin both under static and dynamic conditions (IC(50) = 15.4 nm). These small synthetic antagonists, which are equally as potent as the natural ligand P-selectin glycoprotein ligand-1, are promising leads in anti-atherothrombotic therapy.
authors.affiliation: Division of Biopharmaceutics, Leiden/Amsterdam Center for Drug Research, Leiden University, Gorlaeus Laboratories, The Netherlands.
authors.forename: Chantal C M
authors.initials: CC
authors.lastname: Appeldoorn
authors.forename: Tom J M
authors.initials: TJ
authors.lastname: Molenaar
authors.forename: Arnaud
authors.initials: A
authors.lastname: Bonnefoy
authors.forename: Steven H
authors.initials: SH
authors.lastname: van Leeuwen
authors.forename: Petra A H
authors.initials: PA
authors.lastname: Vandervoort
authors.forename: Marc F
authors.initials: MF
authors.lastname: Hoylaerts
authors.forename: Theo J C
authors.initials: TJ
authors.lastname: van Berkel
authors.forename: Erik A L
authors.initials: EA
authors.lastname: Biessen
mesh_terms: D000595:Amino Acid Sequence; D002352:Carrier Proteins / Q000378:metabolism / Q000494:pharmacology; D020650:Combinatorial Chemistry Techniques; D015195:Drug Design; D006801:Humans; D008969:Molecular Sequence Data; D019007:P-Selectin / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism / Q000494:pharmacology*; D013329:Structure-Activity Relationship
chemical_list: D002352:Carrier Proteins; D019007:P-Selectin; D010455:Peptides