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Click-generated triazole ureas as ultrapotent in vivo–active serine hydrolase inhibitors

Nature Chemical Biology volume 7, pages 469–478 (2011)Cite this article

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A Corrigendum to this article was published on 15 February 2012

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Abstract

Serine hydrolases are a diverse enzyme class representing ∼1% of all human proteins. The biological functions of most serine hydrolases remain poorly characterized owing to a lack of selective inhibitors to probe their activity in living systems. Here we show that a substantial number of serine hydrolases can be irreversibly inactivated by 1,2,3-triazole ureas, which show negligible cross-reactivity with other protein classes. Rapid lead optimization by click chemistry–enabled synthesis and competitive activity-based profiling identified 1,2,3-triazole ureas that selectively inhibit enzymes from diverse branches of the serine hydrolase class, including peptidases (acyl-peptide hydrolase, or APEH), lipases (platelet-activating factor acetylhydrolase-2, or PAFAH2) and uncharacterized hydrolases (α,β-hydrolase-11, or ABHD11), with exceptional potency in cells (sub-nanomolar) and mice (<1 mg kg−1). We show that APEH inhibition leads to accumulation of N-acetylated proteins and promotes proliferation in T cells. These data indicate 1,2,3-triazole ureas are a pharmacologically privileged chemotype for serine hydrolase inhibition, combining broad activity across the serine hydrolase class with tunable selectivity for individual enzymes.

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Figure 1: Competitive ABPP with clickable NHU activity–based probes AA6 to AA10.
Figure 2: Comparative ABPP of piperidine-based carbamate (AA38-3) and triazole urea (AA26-9) inhibitors.
Figure 3: Rapid optimization of triazole urea inhibitors by click chemistry–enabled synthesis and competitive ABPP.
Figure 4: In vitro and in situ characterization of triazole urea inhibitors AA74-1, AA39-2 and AA44-2 in mouse T cells.
Figure 5: Characterization of the activity and selectivity of APEH inhibitor AA74-1 in vivo.
Figure 6: Proteomic characterization of endogenous APEH substrates using N-terminal labeling and enrichment.

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  • 15 February 2012

    In the version of this article initially published, the authors concluded, on the basis of the substantial (approximately five-fold) N1 regioselectivity observed for reactions that form the unsubstituted triazole ureas shown in Figures 1–3, that the major regioisomeric product for the 4-substituted triazole ureas shown in Figures 1 and 3 was also the N1 regioisomer. They have since determined by X-ray crystallography (provided as Supplementary Data Sets 1 and 2) that the N2 regioisomer is the major product for the 4-substituted triazole ureas. The structures have been corrected in the HTML and PDF versions of the article and in the chemical probe table associated with the article.

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Acknowledgements

We thank J. Savas and D. McClatchy (The Scripps Research Institute) for generously providing 15N-labeled mice and T. Ji for technical assistance. This work was supported by the US National Institutes of Health (DA028845, CA151460 (K99 award to B.R.M.)), the Deutscher Akademischer Austausch Dienst (postdoctoral fellowship to A.A.), the US National Science Foundation (predoctoral fellowship to D.A.B.) and the Skaggs Institute for Chemical Biology.

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Authors and Affiliations

  1. The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, California, USA

    Alexander Adibekian, Brent R Martin, Chu Wang, Ku-Lung Hsu, Daniel A Bachovchin & Benjamin F Cravatt

  2. Department of Chemical Physiology, The Scripps Research Institute, La Jolla, California, USA

    Alexander Adibekian, Brent R Martin, Chu Wang, Ku-Lung Hsu, Daniel A Bachovchin, Sherry Niessen, Heather Hoover & Benjamin F Cravatt

  3. Center for Physiological Proteomics, The Scripps Research Institute, La Jolla, California, USA

    Sherry Niessen & Heather Hoover

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Contributions

A.A. B.R.M. and B.F.C. designed the experiments; A.A., B.R.M. and K.-L.H. performed the experiments; A.A., D.A.B., S.N. and H.H. contributed new reagents and analytic tools; A.A., C.W. and B.F.C. analyzed data; and A.A. and B.F.C. wrote the manuscript.

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Correspondence to Benjamin F Cravatt.

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Supplementary information

Supplementary Text and Figures

Supplementary Methods and Supplementary Results (PDF 1915 kb)

Supplementary Table 1

Supplementary Table 1 (XLS 1249 kb)

Supplementary Data Set 1

Crystal structure data for compound AA80-1 (CIF 18 kb)

Supplementary Data Set 2

Crystal structure data for compound KT117 (CIF 15 kb)

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Adibekian, A., Martin, B., Wang, C. et al. Click-generated triazole ureas as ultrapotent in vivo–active serine hydrolase inhibitors. Nat Chem Biol 7, 469–478 (2011). https://doi.org/10.1038/nchembio.579

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