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

The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, California, USA.
Nature Chemical Biology (Impact Factor: 13). 05/2011; 7(7):469-78. DOI: 10.1038/nchembio.579
Source: PubMed


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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Available from: Ku-Lung Hsu, Oct 27, 2014
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    • "We selected one of the more promiscuous compounds, JJH221 (4), for analysis by mass spectrometry (MS)-based ABPP methods to assess the range of SHs targeted by NHH carbamates . JJH221-sensitive SHs were identified using the quantitative MS method ABPP-SILAC (stable isotope labeling by amino acids in cell culture; Mann, 2006) (Adibekian et al., 2011). In brief, proteomes from isotopically heavy-and lightamino acid-labeled human PC3 cells were treated with JJH221 (20 mM) or DMSO, respectively, for 4 hr followed by the biotinylated FP probe FP-biotin (Liu et al., 1999) (2.5 mM, 1 hr). "
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    • "The analysis below uses mice, wholly labeled with 15N, generated by raising postweaned mice for 10 weeks on a diet in which the nitrogen component is exclusively derived from 15N-labeled Spirulina. Greater than 95% of brain proteins within these reference mice are 15N labeled (Adibekian et al., 2011). "
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