1. Design and Synthesis of Quenched Activity-based Probes for Diacylglycerol Lipase and α,β-Hydrolase Domain Containing Protein 6.
- Author
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van Rooden EJ, Kohsiek M, Kreekel R, van Esbroeck ACM, van den Nieuwendijk AMCH, Janssen APA, van den Berg RJBHN, Overkleeft HS, and van der Stelt M
- Subjects
- Aniline Compounds chemistry, Animals, Brain metabolism, Catalysis, Cell Line, Tumor, Copper chemistry, Cycloaddition Reaction, Fluorescent Dyes chemistry, Fluorescent Dyes metabolism, Heterocyclic Compounds, 3-Ring chemistry, Heterocyclic Compounds, 3-Ring metabolism, Humans, Lipoprotein Lipase chemistry, Mice, Monoacylglycerol Lipases chemistry, Triazoles chemistry, Triazoles metabolism, Drug Design, Fluorescent Dyes chemical synthesis, Heterocyclic Compounds, 3-Ring chemical synthesis, Lipoprotein Lipase metabolism, Monoacylglycerol Lipases metabolism, Triazoles chemical synthesis
- Abstract
Diacylglycerol lipases (DAGL) are responsible for the biosynthesis of the endocannabinoid 2-arachidonoylglycerol. The fluorescent activity-based probes DH379 and HT-01 have been previously shown to label DAGLs and to cross-react with the serine hydrolase ABHD6. Here, we report the synthesis and characterization of two new quenched activity-based probes 1 and 2, the design of which was based on the structures of DH379 and HT-01, respectively. Probe 1 contains a BODIPY-FL and a 2,4-dinitroaniline moiety as a fluorophore-quencher pair, whereas probe 2 employs a Cy5-fluorophore and a cAB40-quencher. The fluorescence of both probes was quenched with relative quantum yields of 0.34 and 0.0081, respectively. The probes showed target inhibition as characterized in activity-based protein profiling assays using human cell- and mouse brain lysates, but were unfortunately not active in living cells, presumably due to limited cell permeability., (© 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Published
- 2018
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