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Anti-diabetic activity of insulin-degrading enzyme inhibitors mediated by multiple hormones

Nature volume 511, pages 9498 (03 July 2014) | Download Citation

Subjects

Abstract

Despite decades of speculation that inhibiting endogenous insulin degradation might treat type-2 diabetes1,2, and the identification of IDE (insulin-degrading enzyme) as a diabetes susceptibility gene3,4, the relationship between the activity of the zinc metalloprotein IDE and glucose homeostasis remains unclear. Although Ide–/– mice have elevated insulin levels, they exhibit impaired, rather than improved, glucose tolerance that may arise from compensatory insulin signalling dysfunction5,6. IDE inhibitors that are active in vivo are therefore needed to elucidate IDE’s physiological roles and to determine its potential to serve as a target for the treatment of diabetes. Here we report the discovery of a physiologically active IDE inhibitor identified from a DNA-templated macrocycle library. An X-ray structure of the macrocycle bound to IDE reveals that it engages a binding pocket away from the catalytic site, which explains its remarkable selectivity. Treatment of lean and obese mice with this inhibitor shows that IDE regulates the abundance and signalling of glucagon and amylin, in addition to that of insulin. Under physiological conditions that augment insulin and amylin levels, such as oral glucose administration, acute IDE inhibition leads to substantially improved glucose tolerance and slower gastric emptying. These findings demonstrate the feasibility of modulating IDE activity as a new therapeutic strategy to treat type-2 diabetes and expand our understanding of the roles of IDE in glucose and hormone regulation.

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Accessions

Primary accessions

Protein Data Bank

Data deposits

The coordinates and the structure factors of the IDE·6b complex have been deposited in the Protein Data Bank under the accession code 4LTE.

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Acknowledgements

This research was supported by NIH/NIGMS (R01 GM065865 (D.R.L.), R00 GM080097 (M.A.S.), R01 GM81539 (W.-J.T.), T32 GM008444 (Z.H.F.), F30 CA174152 (Z.H.F.), DP2 OD002374 (A.S.)), Howard Hughes Medical Institute (D.R.L.), Diabetes and Cancer Centers of Albert Einstein College of Medicine (M.J.C.), American Diabetes Association no. 7-11-CD-06 (M.A.L.), Burroughs Wellcome Fund CABS (A.S.), and the Searle Scholars Program (A.S.). The Fonds de Recherche en Santé du Québec (FRSQ) and the Alfred Bader Fund supported J.P.M. We thank C. Russ and H. Spurling (Broad Institute) and C. Daly (FAS Center for Systems Biology) for DNA sequencing assistance. We are grateful to S. Johnston and C. Mosher (Broad Institute) for 6bK stability measurements. W. Nolte provided mouse IDE, L. McCord purified CF-IDE and Y.-G. Kim performed CGRP cleavage assays. We are grateful to A. Badran, E. Homan, A. M. Lone and M. Leidl (Harvard University) for experimental assistance. We thank B. Kahn and N. Gray for discussions.

Author information

Affiliations

  1. Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, Massachusetts 02138, USA

    • Juan Pablo Maianti
    • , Amanda McFedries
    • , Ralph E. Kleiner
    • , Alan Saghatelian
    •  & David R. Liu
  2. Department of Pharmacological Sciences, Stony Brook University, 1 Circle Road, Stony Brook, New York 11794, USA

    • Zachariah H. Foda
    •  & Markus A. Seeliger
  3. Department of Biochemistry, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, New York 10461, USA

    • Xiu Quan Du
    •  & Maureen J. Charron
  4. Institute for Memory Impairments and Neurological Disorders, University of California, Irvine, 3204 Biological Sciences III, Irvine, California 92697, USA

    • Malcolm A. Leissring
  5. Ben-May Department for Cancer Research, University of Chicago, 929 East 57th Street, Chicago, Illinois 60637, USA

    • Wei-Jen Tang
  6. Howard Hughes Medical Institute, Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, Massachusetts 02138, USA

    • David R. Liu

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Contributions

J.P.M., A.M., Z.H.F., R.E.K. and X.Q.D. performed the experiments. M.A.L. provided the Ide–/– mice. W.-J.T. provided IDE protein for structural studies. M.J.C. supervised the Gcgr–/– studies. M.A.S. supervised the IDE·6b structural studies. A.S. supervised the pharmacological validation of 6bK and the in vivo studies. D.R.L. supervised the discovery of IDE inhibitors, the pharmacological studies and the in vivo studies. All authors analysed the data and wrote the manuscript.

Competing interests

J.P.M., A.S. and D.R.L. are co-inventors on a provisional patent application that describes the discovery and properties of 6bK.

Corresponding authors

Correspondence to Alan Saghatelian or David R. Liu.

Extended data

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    This file contains Supplementary results and Discussion, Supplementary Methods, Supplementary Tables 1-5, Supplementary Sequence and Supplementary References.

Videos

  1. 1.

    Animation of the X-ray co-crystal structure of IDE bound to macrocyclic inhibitor 6b.

    The IDE domains 1, 2, 3, and 4 are colored green, blue, yellow, and red, respectively (2.7 Å resolution, pdb: 4LTE). Macrocycle 6b is represented as a ball-and-stick model, and the catalytic zinc atom is represented as an orange sphere. The macrocycle 6b is seen interacting within a 10 Å-deep hydrophobic pocket. The p-benzoyl-phenylalanine is shown in red, the cyclohexylalanine in blue, the fumarate linker in grey, and the D-lysine backbone in purple. The final scene shows a superimposition of 6b (shown as a surface rendering) on the co-crystal structure of insulin (shown in orange) within the IDE cavity (pdb: 2WBY), demonstrating that 6b binding to IDE is predicted to preclude substrate binding.

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DOI

https://doi.org/10.1038/nature13297

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