Dual cyclooxygenase–fatty acid amide hydrolase inhibitor exploits novel binding interactions in the cyclooxygenase active site

Goodman, Michael C.; Xu, Shu; Rouzer, Carol A.; Banerjee, Surajit; Ghebreselasie, Kebreab; Migliore, Marco; Piomelli, Daniele; Marnett, Lawrence J.

doi:10.1074/jbc.M117.802058

Title: Dual cyclooxygenase–fatty acid amide hydrolase inhibitor exploits novel binding interactions in the cyclooxygenase active site

Journal Article · Thu Mar 01 00:00:00 EST 2018 · Journal of Biological Chemistry

DOI:https://doi.org/10.1074/jbc.M117.802058· OSTI ID:1769434

Goodman, Michael C.; Xu, Shu; Rouzer, Carol A.; Banerjee, Surajit; Ghebreselasie, Kebreab; Migliore, Marco; Piomelli, Daniele; Marnett, Lawrence J.

The cyclooxygenases COX-1 and COX-2 oxygenate arachidonic acid (AA) to prostaglandin H₂ (PGH₂). COX-2 also oxygenates the endocannabinoids 2-arachidonoylglycerol (2-AG) and arachidonoylethanolamide (AEA) to the corresponding PGH₂ analogs. Both enzymes are targets of nonsteroidal anti-inflammatory drugs (NSAIDs), but NSAID-mediated COX inhibition is associated with gastrointestinal toxicity. One potential strategy to counter this toxicity is to also inhibit fatty acid amide hydrolase (FAAH), which hydrolyzes bioactive fatty acid ethanolamides (FAEs) into fatty acids and ethanolamine. Here, we investigated the mechanism of COX inhibition by ARN2508, an NSAID that inhibits both COXs and FAAH with high potency, target selectivity, and decreased gastrointestinal toxicity in mouse models, presumably due to its ability to increase levels of FAEs. A 2.27-Å–resolution X-ray crystal structure of the COX-2·(S)-ARN2508 complex reveals that ARN2508 adopts a binding pose similar to that of its parent NSAID flurbiprofen. However, ARN2508's alkyl tail is inserted deep into the top channel, an active site region not exploited by any previously reported NSAID. As for flurbiprofen, ARN2508's potency is highly dependent on the configuration of the α-methyl group. Furthermore, (S)-ARN2508 is more potent than (R)-ARN2508 for inhibition of AA oxygenation by both COXs and 2-AG oxygenation by COX-2. Also, similarly to (R)-flurbiprofen, (R)-ARN2508 exhibits substrate selectivity for inhibition of 2-AG oxygenation. Site-directed mutagenesis confirms the importance of insertion of the alkyl tail into the top channel for (S)-ARN2508's potency and suggests a role for Ser-530 as a determinant of the inhibitor's slow rate of inhibition compared with that of (S)-flurbiprofen.

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Research Organization:: Argonne National Lab. (ANL), Argonne, IL (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities Division; National Inst. of Health; National Inst. of General Medical Sciences

Grant/Contract Number:: AC02-06CH11357; R01 CA089450; R01 GM030910; P41 GM103403; S10OD021527

OSTI ID:: 1769434

Alternate ID(s):: OSTI ID: 1432870

Journal Information:: Journal of Biological Chemistry, Journal Name: Journal of Biological Chemistry Vol. 293 Journal Issue: 9; ISSN 0021-9258

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 11 works

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59 BASIC BIOLOGICAL SCIENCES
cyclooxygenase (COX)
enzyme catalysis
enzyme inhibitor
enzyme kinetics
enzyme structure
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inflammation
polyunsaturated fatty acid (PUFA)

Title: Dual cyclooxygenase–fatty acid amide hydrolase inhibitor exploits novel binding interactions in the cyclooxygenase active site

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