title: Development and optimization of piperidyl-1,2,3-triazole ureas as selective chemical probes of endocannabinoid biosynthesis.
abstract: We have previously shown that 1,2,3-triazole ureas (1,2,3-TUs) act as versatile class of irreversible serine hydrolase inhibitors that can be tuned to create selective probes for diverse members of this large enzyme class, including diacylglycerol lipase-β (DAGLβ), a principal biosynthetic enzyme for the endocannabinoid 2-arachidonoylglycerol (2-AG). Here, we provide a detailed account of the discovery, synthesis, and structure-activity relationship (SAR) of (2-substituted)-piperidyl-1,2,3-TUs that selectively inactivate DAGLβ in living systems. Key to success was the use of activity-based protein profiling (ABPP) with broad-spectrum and tailored activity-based probes to guide our medicinal chemistry efforts. We also describe an expanded repertoire of DAGL-tailored activity-based probes that includes biotinylated and alkyne agents for enzyme enrichment coupled with mass spectrometry-based proteomics and assessment of proteome-wide selectivity. Our findings highlight the broad utility of 1,2,3-TUs for serine hydrolase inhibitor development and their application to create selective probes of endocannabinoid biosynthetic pathways.
authors.affiliation: The Skaggs Institute for Chemical Biology and ‡Department of Chemical Physiology, The Scripps Research Institute , SR107 10550 North Torrey Pines Road, La Jolla, California 92037, United States .
authors.forename: Ku-Lung
authors.initials: KL
authors.lastname: Hsu
authors.forename: Katsunori
authors.initials: K
authors.lastname: Tsuboi
authors.forename: Landon R
authors.initials: LR
authors.lastname: Whitby
authors.forename: Anna E
authors.initials: AE
authors.lastname: Speers
authors.forename: Holly
authors.initials: H
authors.lastname: Pugh
authors.forename: Jordon
authors.initials: J
authors.lastname: Inloes
authors.forename: Benjamin F
authors.initials: BF
authors.lastname: Cravatt
mesh_terms: D004305:Dose-Response Relationship, Drug; D055808:Drug Discovery; D063388:Endocannabinoids / Q000096:biosynthesis* / Q000737:chemistry; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D008071:Lipoprotein Lipase / Q000037:antagonists & inhibitors / Q000378:metabolism; D015394:Molecular Structure; D010880:Piperidines / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D014230:Triazoles / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D014508:Urea / Q000031:analogs & derivatives / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D063388:Endocannabinoids; D004791:Enzyme Inhibitors; D010880:Piperidines; D014230:Triazoles; D014508:Urea; C480281:DAGLB protein, human; D008071:Lipoprotein Lipase