Selective Targeting of Extracellular Insulin-Degrading Enzyme by Quasi-Irreversible Thiol-Modifying Inhibitors
| Autor: | A Masson, SO Abdul-Hay, Juliette Bertrand, Umberto Crisafulli, Michael P. McGuire, Franck Madoux, Terrone R. Rosenberry, Michael D. Cameron, Caitlyn L. Topper, Caroline R. Thompson, Thomas D. Bannister, Thomas R. Caulfield, Peter Hodder, Malcolm A. Leissring, H Wang, Erin A. Howard, Stephan C. Schürer |
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| Rok vydání: | 2015 |
| Předmět: |
medicine.medical_treatment
Drug Evaluation Preclinical Biology Insulysin Models Biological Biochemistry Article Inhibitory Concentration 50 Insulin Antagonists Structure-Activity Relationship Cytosol Drug Delivery Systems medicine Insulin-degrading enzyme Extracellular Structure–activity relationship Computer Simulation Sulfhydryl Compounds Enzyme Inhibitors Cellular compartment chemistry.chemical_classification Protease Molecular Structure General Medicine Enzyme chemistry Molecular Medicine Extracellular Space |
| Zdroj: | ACS Chem Biol |
| ISSN: | 1554-8937 1554-8929 |
| DOI: | 10.1021/acschembio.5b00334 |
| Popis: | Many therapeutically important enzymes are present in multiple cellular compartments, where they can carry out markedly different functions, thus there is a need for pharmacological strategies to selectively manipulate distinct pools of target enzymes. Insulin-degrading enzyme (IDE) is a thiol-sensitive zinc-metallopeptidase that hydrolyzes diverse peptide substrates in both the cytosol and the extracellular space, but current genetic and pharmacological approaches are incapable of selectively inhibiting the protease in specific subcellular compartments. Here we describe the discovery, characterization and kinetics-based optimization of potent benzoisothiazolone-based inhibitors that, by virtue of a unique “quasi-irreversible” mode of inhibition, exclusively inhibit extracellular IDE. The mechanism of inhibition involves nucleophilic attack by a specific active-site thiol of the enzyme on the inhibitors, which bear an isothiazolone ring that undergoes irreversible ring opening with the formation of a disulfide bond. Notably, binding of the inhibitors is reversible under reducing conditions, thus restricting inhibition to IDE present in the extracellular space. The identified inhibitors are highly potent (IC(50)(app) = 63 nM), non-toxic at concentrations up to 100 μM and, were found to selectively target a specific cysteine residue within IDE. These novel inhibitors represent powerful new tools for clarifying the physiological and pathophysiological roles of this poorly understood protease, and their unusual mechanism of action should be applicable to other therapeutic targets. |
| Databáze: | OpenAIRE |
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