title: Development of selective bisubstrate-based inhibitors against protein kinase C (PKC) isozymes by using dynamic peptide microarrays.
abstract: Kinase inhibitors are increasingly important in drug development. Because the majority of current inhibitors target the conserved ATP-binding site, selectivity might become an important issue. This could be particularly problematic for the potential drug target protein kinase C (PKC), of which twelve isoforms with high homology exist in humans. A strategy to increase selectivity is to prepare bisubstrate-based inhibitors that target the more selective peptide-binding site in addition to the ATP-binding site. In this paper a generally applicable, rapid methodology is presented to discover such bisubstrate-based leads. Dynamic peptide microarrays were used to find peptide-binding site inhibitors. These were linked with chemoselective click chemistry to an ATP-binding site inhibitor, and this led to novel bisubstrate structures. The peptide microarrays were used to evaluate the resulting inhibitors. Thus, novel bisubstrate-based inhibitors were obtained that were both more potent and selective compared to their constituent parts. The most promising inhibitor has nanomolar affinity and selectivity towards PKCtheta amongst three isozymes.
authors.affiliation: Department of Medicinal Chemistry and Chemical Biology, Utrecht Institute for Pharmaceutical Sciences, Faculty of Science, Utrecht University, 3584 CA, Utrecht (The Netherlands).
authors.forename: Alex J
authors.initials: AJ
authors.lastname: Poot
authors.forename: Jeroen
authors.initials: J
authors.lastname: van Ameijde
authors.forename: Monique
authors.initials: M
authors.lastname: Slijper
authors.forename: Adriënne
authors.initials: A
authors.lastname: van den Berg
authors.forename: Riet
authors.initials: R
authors.lastname: Hilhorst
authors.forename: Rob
authors.initials: R
authors.lastname: Ruijtenbeek
authors.forename: Dirk T S
authors.initials: DT
authors.lastname: Rijkers
authors.forename: Rob M J
authors.initials: RM
authors.lastname: Liskamp
mesh_terms: D001665:Binding Sites; D006801:Humans; D007527:Isoenzymes / Q000037:antagonists & inhibitors*; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D040081:Protein Array Analysis / Q000379:methods*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors*; D047428:Protein Kinase Inhibitors / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D013329:Structure-Activity Relationship
chemical_list: D007527:Isoenzymes; D010455:Peptides; D047428:Protein Kinase Inhibitors; D011493:Protein Kinase C