Selective Targeting of Extracellular Insulin-Degrading Enzyme by Quasi-Irreversible Thiol-Modifying Inhibitors

Samer O. Abdul-Hay, Thomas D. Bannister, Hui Wang, Michael D. Cameron, Thomas Caulfield, Amandine Masson, Juliette Bertrand, Erin A. Howard, Michael P. McGuire, Umberto Crisafulli, Terrone R. Rosenberry, Caitlyn L. Topper, Caroline R. Thompson, Stephan C. Schürer, Franck Madoux, Peter Hodder, Malcolm A. Leissring

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

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 (IC50 app = 63 nM), nontoxic at concentrations up to 100 μM, and appear to preferentially 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.

Original languageEnglish (US)
Pages (from-to)2716-2724
Number of pages9
JournalACS Chemical Biology
Volume10
Issue number12
DOIs
StatePublished - Dec 18 2015

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Insulysin
Sulfhydryl Compounds
Extracellular Space
Peptide Hydrolases
Enzymes
Pharmacology
Metalloproteases
Enzyme Inhibitors
Application programs
Disulfides
Cytosol
Inhibitory Concentration 50
Cysteine
Zinc
Catalytic Domain
Peptides
Kinetics
Substrates

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine

Cite this

Abdul-Hay, S. O., Bannister, T. D., Wang, H., Cameron, M. D., Caulfield, T., Masson, A., ... Leissring, M. A. (2015). Selective Targeting of Extracellular Insulin-Degrading Enzyme by Quasi-Irreversible Thiol-Modifying Inhibitors. ACS Chemical Biology, 10(12), 2716-2724. https://doi.org/10.1021/acschembio.5b00334

Selective Targeting of Extracellular Insulin-Degrading Enzyme by Quasi-Irreversible Thiol-Modifying Inhibitors. / Abdul-Hay, Samer O.; Bannister, Thomas D.; Wang, Hui; Cameron, Michael D.; Caulfield, Thomas; Masson, Amandine; Bertrand, Juliette; Howard, Erin A.; McGuire, Michael P.; Crisafulli, Umberto; Rosenberry, Terrone R.; Topper, Caitlyn L.; Thompson, Caroline R.; Schürer, Stephan C.; Madoux, Franck; Hodder, Peter; Leissring, Malcolm A.

In: ACS Chemical Biology, Vol. 10, No. 12, 18.12.2015, p. 2716-2724.

Research output: Contribution to journalArticle

Abdul-Hay, SO, Bannister, TD, Wang, H, Cameron, MD, Caulfield, T, Masson, A, Bertrand, J, Howard, EA, McGuire, MP, Crisafulli, U, Rosenberry, TR, Topper, CL, Thompson, CR, Schürer, SC, Madoux, F, Hodder, P & Leissring, MA 2015, 'Selective Targeting of Extracellular Insulin-Degrading Enzyme by Quasi-Irreversible Thiol-Modifying Inhibitors', ACS Chemical Biology, vol. 10, no. 12, pp. 2716-2724. https://doi.org/10.1021/acschembio.5b00334
Abdul-Hay, Samer O. ; Bannister, Thomas D. ; Wang, Hui ; Cameron, Michael D. ; Caulfield, Thomas ; Masson, Amandine ; Bertrand, Juliette ; Howard, Erin A. ; McGuire, Michael P. ; Crisafulli, Umberto ; Rosenberry, Terrone R. ; Topper, Caitlyn L. ; Thompson, Caroline R. ; Schürer, Stephan C. ; Madoux, Franck ; Hodder, Peter ; Leissring, Malcolm A. / Selective Targeting of Extracellular Insulin-Degrading Enzyme by Quasi-Irreversible Thiol-Modifying Inhibitors. In: ACS Chemical Biology. 2015 ; Vol. 10, No. 12. pp. 2716-2724.
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