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Development of Selective Bisubstrate-Based Inhibitors Against Protein Kinase C (PKC) Isozymes By Using Dynamic Peptide Microarrays

Authors

  • Alex J. Poot,

    1. Department of Medicinal Chemistry and Chemical Biology, Utrecht Institute for Pharmaceutical Sciences, Faculty of Science, Utrecht University, Sorbonnelaan 16, 3584 CA, Utrecht (The Netherlands), Fax: (+31) (0)30 2536655
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  • Jeroen van Ameijde Dr.,

    1. Department of Medicinal Chemistry and Chemical Biology, Utrecht Institute for Pharmaceutical Sciences, Faculty of Science, Utrecht University, Sorbonnelaan 16, 3584 CA, Utrecht (The Netherlands), Fax: (+31) (0)30 2536655
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  • Monique Slijper Dr.,

    1. Biomolecular Mass Spectrometry and Proteomics Group, Utrecht Institute for Pharmaceutical Sciences and Bijvoet Centre for Biomolecular Research, Faculty of Science, Utrecht University, Sorbonnelaan 16, 3584 CA, Utrecht (The Netherlands)
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  • Adriënne van den Berg,

    1. Pamgene International BV, P.O. Box 1345, 5200 BJ, ‘s-Hertogenbosch (The Netherlands)
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  • Riet Hilhorst Dr.,

    1. Pamgene International BV, P.O. Box 1345, 5200 BJ, ‘s-Hertogenbosch (The Netherlands)
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  • Rob Ruijtenbeek Dr.,

    1. Pamgene International BV, P.O. Box 1345, 5200 BJ, ‘s-Hertogenbosch (The Netherlands)
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  • Dirk T. S. Rijkers Dr.,

    1. Department of Medicinal Chemistry and Chemical Biology, Utrecht Institute for Pharmaceutical Sciences, Faculty of Science, Utrecht University, Sorbonnelaan 16, 3584 CA, Utrecht (The Netherlands), Fax: (+31) (0)30 2536655
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  • Rob M. J. Liskamp Prof. Dr.

    1. Department of Medicinal Chemistry and Chemical Biology, Utrecht Institute for Pharmaceutical Sciences, Faculty of Science, Utrecht University, Sorbonnelaan 16, 3584 CA, Utrecht (The Netherlands), Fax: (+31) (0)30 2536655
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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 PKCθ amongst three isozymes.

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