Dual cyclooxygenase–fatty acid amide hydrolase inhibitor exploits novel binding interactions in the cyclooxygenase active site
Abstract
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)-ARN2508more »
- Authors:
- Publication Date:
- Research Org.:
- Argonne National Lab. (ANL), Argonne, IL (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities Division; National Inst. of Health; National Inst. of General Medical Sciences
- OSTI Identifier:
- 1769434
- Alternate Identifier(s):
- OSTI ID: 1432870
- Grant/Contract Number:
- AC02-06CH11357; R01 CA089450; R01 GM030910; P41 GM103403; S10OD021527
- Resource Type:
- Published Article
- Journal Name:
- Journal of Biological Chemistry
- Additional Journal Information:
- Journal Name: Journal of Biological Chemistry Journal Volume: 293 Journal Issue: 9; Journal ID: ISSN 0021-9258
- Publisher:
- Elsevier
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; cyclooxygenase (COX); enzyme catalysis; enzyme inhibitor; enzyme kinetics; enzyme structure; fatty acid metabolism; inflammation; polyunsaturated fatty acid (PUFA)
Citation Formats
Goodman, Michael C., Xu, Shu, Rouzer, Carol A., Banerjee, Surajit, Ghebreselasie, Kebreab, Migliore, Marco, Piomelli, Daniele, and Marnett, Lawrence J. Dual cyclooxygenase–fatty acid amide hydrolase inhibitor exploits novel binding interactions in the cyclooxygenase active site. United States: N. p., 2018.
Web. doi:10.1074/jbc.M117.802058.
Goodman, Michael C., Xu, Shu, Rouzer, Carol A., Banerjee, Surajit, Ghebreselasie, Kebreab, Migliore, Marco, Piomelli, Daniele, & Marnett, Lawrence J. Dual cyclooxygenase–fatty acid amide hydrolase inhibitor exploits novel binding interactions in the cyclooxygenase active site. United States. https://doi.org/10.1074/jbc.M117.802058
Goodman, Michael C., Xu, Shu, Rouzer, Carol A., Banerjee, Surajit, Ghebreselasie, Kebreab, Migliore, Marco, Piomelli, Daniele, and Marnett, Lawrence J. Thu .
"Dual cyclooxygenase–fatty acid amide hydrolase inhibitor exploits novel binding interactions in the cyclooxygenase active site". United States. https://doi.org/10.1074/jbc.M117.802058.
@article{osti_1769434,
title = {Dual cyclooxygenase–fatty acid amide hydrolase inhibitor exploits novel binding interactions in the cyclooxygenase active site},
author = {Goodman, Michael C. and Xu, Shu and Rouzer, Carol A. and Banerjee, Surajit and Ghebreselasie, Kebreab and Migliore, Marco and Piomelli, Daniele and Marnett, Lawrence J.},
abstractNote = {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.},
doi = {10.1074/jbc.M117.802058},
journal = {Journal of Biological Chemistry},
number = 9,
volume = 293,
place = {United States},
year = {Thu Mar 01 00:00:00 EST 2018},
month = {Thu Mar 01 00:00:00 EST 2018}
}
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https://doi.org/10.1074/jbc.M117.802058
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Works referenced in this record:
The COXIB Experience: A Look in the Rearview Mirror
journal, February 2009
- Marnett, Lawrence J.
- Annual Review of Pharmacology and Toxicology, Vol. 49, Issue 1
(R)-Profens are substrate-selective inhibitors of endocannabinoid oxygenation by COX-2
journal, September 2011
- Duggan, Kelsey C.; Hermanson, Daniel J.; Musee, Joel
- Nature Chemical Biology, Vol. 7, Issue 11
Structural requirements for time-dependent inhibition of prostaglandin biosynthesis by anti-inflammatory drugs.
journal, December 1975
- Rome, L. H.; Lands, W. E.
- Proceedings of the National Academy of Sciences, Vol. 72, Issue 12
Solving structures of protein complexes by molecular replacement with Phaser
journal, December 2006
- McCoy, Airlie J.
- Acta Crystallographica Section D Biological Crystallography, Vol. 63, Issue 1
Iterative model building, structure refinement and density modification with the PHENIX AutoBuild wizard
journal, December 2007
- Terwilliger, Thomas C.; Grosse-Kunstleve, Ralf W.; Afonine, Pavel V.
- Acta Crystallographica Section D Biological Crystallography, Vol. 64, Issue 1
Involvement of Arginine 120, Glutamate 524, and Tyrosine 355 in the Binding of Arachidonate and 2-Phenylpropionic Acid Inhibitors to the Cyclooxygenase Active Site of Ovine Prostaglandin Endoperoxide H Synthase-1
journal, January 1996
- Bhattacharyya, Dipak K.; Lecomte, Marc; Rieke, Caroline Jill
- Journal of Biological Chemistry, Vol. 271, Issue 4
Nonsteroidal Antiinflammatory Drug-Related Injury to the Gastrointestinal Tract: Clinical Picture, Pathogenesis, and Prevention
journal, September 2010
- Scarpignato, Carmelo; Hunt, Richard H.
- Gastroenterology Clinics of North America, Vol. 39, Issue 3
Fatty Acid Substrate Specificities of Human Prostaglandin-endoperoxide H Synthase-1 and −2: FORMATION OF 12-HYDROXY-(9
journal, August 1995
- Laneuville, Odette; Breuer, Debra K.; Xu, Naxing
- Journal of Biological Chemistry, Vol. 270, Issue 33
Endocannabinoids and the gastrointestinal tract: what are the key questions?: Endocannabinoids and the GI tract
journal, November 2007
- Sanger, G. J.
- British Journal of Pharmacology, Vol. 152, Issue 5
Multitarget fatty acid amide hydrolase/cyclooxygenase blockade suppresses intestinal inflammation and protects against nonsteroidal anti-inflammatory drug-dependent gastrointestinal damage
journal, June 2015
- Sasso, Oscar; Migliore, Marco; Habrant, Damien
- The FASEB Journal, Vol. 29, Issue 6
Crystal Structure of Fatty Acid Amide Hydrolase Bound to the Carbamate Inhibitor URB597: Discovery of a Deacylating Water Molecule and Insight into Enzyme Inactivation
journal, July 2010
- Mileni, Mauro; Kamtekar, Satwik; Wood, David C.
- Journal of Molecular Biology, Vol. 400, Issue 4
The X-ray crystal structure of the membrane protein prostaglandin H2 synthase-1
journal, January 1994
- Picot, Daniel; Loll, Patrick J.; Garavito, R. Michael
- Nature, Vol. 367, Issue 6460
Identification and Characterization of Carprofen as a Multitarget Fatty Acid Amide Hydrolase/Cyclooxygenase Inhibitor
journal, September 2012
- Favia, Angelo D.; Habrant, Damien; Scarpelli, Rita
- Journal of Medicinal Chemistry, Vol. 55, Issue 20
Structural basis for selective inhibition of cyclooxygenase-2 by anti-inflammatory agents
journal, December 1996
- Kurumbail, Ravi G.; Stevens, Anna M.; Gierse, James K.
- Nature, Vol. 384, Issue 6610
Nonsteroidal anti-inflammatory drugs and upper and lower gastrointestinal mucosal damage
journal, January 2013
- Sostres, Carlos; Gargallo, Carla J.; Lanas, Angel
- Arthritis Research & Therapy, Vol. 15, Issue Suppl 3
Acetylation of prostaglandin synthase by aspirin.
journal, August 1975
- Roth, G. J.; Stanford, N.; Majerus, P. W.
- Proceedings of the National Academy of Sciences, Vol. 72, Issue 8
Effect of inhibitor time-dependency on selectivity towards cyclooxygenase isoforms
journal, February 1995
- Ouellet, M.; Percival, M. D.
- Biochemical Journal, Vol. 306, Issue 1
Oxygenation of the Endocannabinoid, 2-Arachidonylglycerol, to Glyceryl Prostaglandins by Cyclooxygenase-2
journal, August 2000
- Kozak, Kevin R.; Rowlinson, Scott W.; Marnett, Lawrence J.
- Journal of Biological Chemistry, Vol. 275, Issue 43
Cyclooxygenases: Structural, Cellular, and Molecular Biology
journal, June 2000
- Smith, William L.; DeWitt, David L.; Garavito, R. Michael
- Annual Review of Biochemistry, Vol. 69, Issue 1
XDS
journal, January 2010
- Kabsch, Wolfgang
- Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2
The Productive Conformation of Arachidonic Acid Bound to Prostaglandin Synthase
journal, September 2000
- Malkowski, M. G.
- Science, Vol. 289, Issue 5486
Tyrosine-385 Is Critical for Acetylation of Cyclooxygenase-2 by Aspirin
journal, July 2000
- Hochgesang, G. Phillip; Rowlinson, Scott W.; Marnett, Lawrence J.
- Journal of the American Chemical Society, Vol. 122, Issue 27
Comparison of Cyclooxygenase-1 Crystal Structures: Cross-Talk between Monomers Comprising Cyclooxygenase-1 Homodimers,
journal, August 2010
- Sidhu, Ranjinder S.; Lee, Jullia Y.; Yuan, Chong
- Biochemistry, Vol. 49, Issue 33
NSAIDs and cardiovascular disease: transducing human pharmacology results into clinical read-outs in the general population
journal, May 2010
- Capone, Marta L.; Tacconelli, Stefania; Garcia Rodriguez, Luis
- Pharmacological Reports, Vol. 62, Issue 3
The Molecular Basis for Dual Fatty Acid Amide Hydrolase (FAAH)/Cyclooxygenase (COX) Inhibition
journal, November 2015
- Palermo, Giulia; Favia, Angelo D.; Convertino, Marino
- ChemMedChem, Vol. 11, Issue 12
2.0 Å structure of prostaglandin H 2 synthase-1 reconstituted with a manganese porphyrin cofactor
journal, January 2006
- Gupta, Kushol; Selinsky, Barry S.; Loll, Patrick J.
- Acta Crystallographica Section D Biological Crystallography, Vol. 62, Issue 2
Structural and Functional Basis of Cyclooxygenase Inhibition
journal, April 2007
- Blobaum, Anna L.; Marnett, Lawrence J.
- Journal of Medicinal Chemistry, Vol. 50, Issue 7
13-Methylarachidonic Acid Is a Positive Allosteric Modulator of Endocannabinoid Oxygenation by Cyclooxygenase
journal, February 2015
- Kudalkar, Shalley N.; Nikas, Spyros P.; Kingsley, Philip J.
- Journal of Biological Chemistry, Vol. 290, Issue 12
Oxicams Bind in a Novel Mode to the Cyclooxygenase Active Site via a Two-water-mediated H-bonding Network
journal, January 2014
- Xu, Shu; Hermanson, Daniel J.; Banerjee, Surajit
- Journal of Biological Chemistry, Vol. 289, Issue 10
Mechanism of Free Radical Oxygenation of Polyunsaturated Fatty Acids by Cyclooxygenases
journal, June 2003
- Rouzer, Carol A.; Marnett, Lawrence J.
- Chemical Reviews, Vol. 103, Issue 6
Structural Basis of Fatty Acid Substrate Binding to Cyclooxygenase-2
journal, May 2010
- Vecchio, Alex J.; Simmons, Danielle M.; Malkowski, Michael G.
- Journal of Biological Chemistry, Vol. 285, Issue 29
Features and development of Coot
journal, March 2010
- Emsley, P.; Lohkamp, B.; Scott, W. G.
- Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 4
Action at a Distance: MUTATIONS OF PERIPHERAL RESIDUES TRANSFORM RAPID REVERSIBLE INHIBITORS TO SLOW, TIGHT BINDERS OF CYCLOOXYGENASE-2
journal, March 2015
- Blobaum, Anna L.; Xu, Shu; Rowlinson, Scott W.
- Journal of Biological Chemistry, Vol. 290, Issue 20
The Kinetic Factors That Determine the Affinity and Selectivity for Slow Binding Inhibition of Human Prostaglandin H Synthase 1 and 2 by Indomethacin and Flurbiprofen
journal, February 1996
- Callan, Ondine Harris; So, On-Yee; Swinney, David C.
- Journal of Biological Chemistry, Vol. 271, Issue 7
PHENIX: a comprehensive Python-based system for macromolecular structure solution
journal, January 2010
- Adams, Paul D.; Afonine, Pavel V.; Bunkóczi, Gábor
- Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2, p. 213-221
The structure of ibuprofen bound to cyclooxygenase-2
journal, January 2015
- Orlando, Benjamin J.; Lucido, Michael J.; Malkowski, Michael G.
- Journal of Structural Biology, Vol. 189, Issue 1
Metabolism of the Endocannabinoids, 2-Arachidonylglycerol and Anandamide, into Prostaglandin, Thromboxane, and Prostacyclin Glycerol Esters and Ethanolamides
journal, September 2002
- Kozak, Kevin R.; Crews, Brenda C.; Morrow, Jason D.
- Journal of Biological Chemistry, Vol. 277, Issue 47
Structural Analysis of NSAID Binding by Prostaglandin H 2 Synthase: Time-Dependent and Time-Independent Inhibitors Elicit Identical Enzyme Conformations †
journal, May 2001
- Selinsky, Barry S.; Gupta, Kushol; Sharkey, Caroline T.
- Biochemistry, Vol. 40, Issue 17
The Dynamics of Prostaglandin H Synthases: STUDIES WITH PROSTAGLANDIN H SYNTHASE 2 Y355F UNMASK MECHANISMS OF TIME-DEPENDENT INHIBITION AND ALLOSTERIC ACTIVATION
journal, March 1998
- So, On-Yee; Scarafia, Liliana E.; Mak, Amy Y.
- Journal of Biological Chemistry, Vol. 273, Issue 10
A Novel Mechanism of Cyclooxygenase-2 Inhibition Involving Interactions with Ser-530 and Tyr-385
journal, August 2003
- Rowlinson, Scott W.; Kiefer, James R.; Prusakiewicz, Jeffery J.
- Journal of Biological Chemistry, Vol. 278, Issue 46
Biochemically based design of cyclooxygenase-2 (COX-2) inhibitors: Facile conversion of nonsteroidal antiinflammatory drugs to potent and highly selective COX-2 inhibitors
journal, January 2000
- Kalgutkar, A. S.; Crews, B. C.; Rowlinson, S. W.
- Proceedings of the National Academy of Sciences, Vol. 97, Issue 2
Potent multitarget FAAH-COX inhibitors: Design and structure-activity relationship studies
journal, February 2016
- Migliore, Marco; Habrant, Damien; Sasso, Oscar
- European Journal of Medicinal Chemistry, Vol. 109
Cyclooxygenase in biology and disease
journal, September 1998
- Dubois, Raymond N.; Abramson, Steven B.; Crofford, Leslie
- The FASEB Journal, Vol. 12, Issue 12
Crystal Structure of Aspirin-Acetylated Human Cyclooxygenase-2: Insight into the Formation of Products with Reversed Stereochemistry
journal, February 2016
- Lucido, Michael J.; Orlando, Benjamin J.; Vecchio, Alex J.
- Biochemistry, Vol. 55, Issue 8
Synthesis of Prostaglandin E 2 Ethanolamide from Anandamide by Cyclooxygenase-2
journal, August 1997
- Yu, Ming; Ives, Danial; Ramesha, Chakkodabylu S.
- Journal of Biological Chemistry, Vol. 272, Issue 34
The Binding of Arachidonic Acid in the Cyclooxygenase Active Site of Mouse Prostaglandin Endoperoxide Synthase-2 (COX-2): A PUTATIVE L-SHAPED BINDING CONFORMATION UTILIZING THE TOP CHANNEL REGION
journal, August 1999
- Rowlinson, Scott W.; Crews, Brenda C.; Lanzo, Cheryl A.
- Journal of Biological Chemistry, Vol. 274, Issue 33
Investigating Substrate Promiscuity in Cyclooxygenase-2: THE ROLE OF ARG-120 AND RESIDUES LINING THE HYDROPHOBIC GROOVE
journal, May 2012
- Vecchio, Alex J.; Orlando, Benjamin J.; Nandagiri, Ritwik
- Journal of Biological Chemistry, Vol. 287, Issue 29
Molecular Basis for Cyclooxygenase Inhibition by the Non-steroidal Anti-inflammatory Drug Naproxen
journal, September 2010
- Duggan, Kelsey C.; Walters, Matthew J.; Musee, Joel
- Journal of Biological Chemistry, Vol. 285, Issue 45
Competition and allostery govern substrate selectivity of cyclooxygenase-2
journal, September 2015
- Mitchener, Michelle M.; Hermanson, Daniel J.; Shockley, Erin M.
- Proceedings of the National Academy of Sciences, Vol. 112, Issue 40
B-cell subpopulations in humans and their differential susceptibility to depletion with anti-CD20 monoclonal antibodies
journal, January 2013
- Leandro, Maria J.
- Arthritis Research & Therapy, Vol. 15, Issue Suppl 1
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- Khasabova, Iryna A.; Uhelski, Megan; Khasabov, Sergey G.
- Blood, Vol. 133, Issue 18