title: Molecular basis for cyclooxygenase inhibition by the non-steroidal anti-inflammatory drug naproxen.
abstract: Naproxen ((S)-6-methoxy-α-methyl-2-naphthaleneacetic acid) is a powerful non-selective non-steroidal anti-inflammatory drug that is extensively used as a prescription and over-the-counter medication. Naproxen exhibits gastrointestinal toxicity, but its cardiovascular toxicity may be reduced compared with other drugs in its class. Despite the fact that naproxen has been marketed for many years, the molecular basis of its interaction with cyclooxygenase (COX) enzymes is unknown. We performed a detailed study of naproxen-COX-2 interactions using site-directed mutagenesis, structure-activity analysis, and x-ray crystallography. The results indicate that each of the pendant groups of the naphthyl scaffold are essential for COX inhibition, and only minimal substitutions are tolerated. Mutation of Trp-387 to Phe significantly reduced inhibition by naproxen, a result that appears unique to this inhibitor. Substitution of S or CH(2) for the O atom of the p-methoxy group yielded analogs that were not affected by the W387F substitution and that exhibited increased COX-2 selectivity relative to naproxen. Crystallization and x-ray analysis yielded structures of COX-2 complexed to naproxen and its methylthio analog at 1.7 and 2.3 Å resolution, respectively. The combination of mutagenesis, structure analysis, and x-ray crystallography provided comprehensive information on the unique interactions responsible for naproxen binding to COX-2.
authors.affiliation: AB Hancock Jr Memorial Laboratory for Cancer Research, Department of Biochemistry, Vanderbilt Institute for Chemical Biology, the Center in Molecular Toxicology, Nashville, Tennessee 37232-0146.
authors.forename: Kelsey C
authors.initials: KC
authors.lastname: Duggan
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Walters
authors.forename: Joel
authors.initials: J
authors.lastname: Musee
authors.forename: Joel M
authors.initials: JM
authors.lastname: Harp
authors.forename: James R
authors.initials: JR
authors.lastname: Kiefer
authors.forename: John A
authors.initials: JA
authors.lastname: Oates
authors.forename: Lawrence J
authors.initials: LJ
authors.lastname: Marnett
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000737:chemistry*; D018360:Crystallography, X-Ray; D051546:Cyclooxygenase 2 / Q000737:chemistry*; D006801:Humans; D051379:Mice; D016297:Mutagenesis, Site-Directed; D020125:Mutation, Missense; D009288:Naproxen / Q000737:chemistry*; D011485:Protein Binding
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D009288:Naproxen; C077623:Ptgs2 protein, mouse; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human
references.citation: Nature. 1996 Dec 19-26;384(6610):644-8
references.pmid: 8967954
references.citation: J Biol Chem. 1999 Jun 11;274(24):17109-14
references.pmid: 10358065
references.citation: J Biol Chem. 2000 Mar 24;275(12):8501-7
references.pmid: 10722687
references.citation: Acta Crystallogr D Biol Crystallogr. 2006 Feb;62(Pt 2):151-6
references.pmid: 16421446
references.citation: J Pharmacol Exp Ther. 2007 Aug;322(2):453-60
references.pmid: 17473175
references.citation: Proc Natl Acad Sci U S A. 2010 Jan 12;107(2):657-62
references.pmid: 20080731
references.citation: Nature. 1994 Jan 20;367(6460):243-9
references.pmid: 8121489
references.citation: Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
references.pmid: 15572765
references.citation: Biochemistry. 2004 Dec 14;43(49):15439-45
references.pmid: 15581355
references.citation: J Pharmacol Exp Ther. 1994 Nov;271(2):927-34
references.pmid: 7965814
references.citation: Science. 1998 May 22;280(5367):1268-70
references.pmid: 9596581
references.citation: Biochemistry. 1996 Jun 11;35(23):7330-40
references.pmid: 8652509
references.citation: J Biol Chem. 1999 Aug 13;274(33):23305-10
references.pmid: 10438506
references.citation: Acta Crystallogr D Biol Crystallogr. 1997 May 1;53(Pt 3):240-55
references.pmid: 15299926
references.citation: Proc Natl Acad Sci U S A. 2002 May 14;99(10):7130-5
references.pmid: 12011469
references.citation: Bioorg Med Chem Lett. 2006 Apr 15;16(8):2219-23
references.pmid: 16455248
references.citation: Carbohydr Res. 2007 May 21;342(7):970-4
references.pmid: 17324385
references.citation: BMJ. 2006 Jun 3;332(7553):1302-8
references.pmid: 16740558
references.citation: J Biol Chem. 1996 Jan 26;271(4):2179-84
references.pmid: 8567676
references.citation: Biochemistry. 2001 May 1;40(17):5172-80
references.pmid: 11318639
references.citation: Biophys J. 2008 Sep;95(5):2415-22
references.pmid: 18375516
references.citation: J Med Chem. 2007 Apr 5;50(7):1425-41
references.pmid: 17341061
references.citation: Biochem J. 1999 May 1;339 ( Pt 3):607-14
references.pmid: 10215599
references.citation: J Med Chem. 1970 Mar;13(2):203-5
references.pmid: 5418489
references.citation: J Urol. 2010 Apr;183(4):1598-603
references.pmid: 20172542
references.citation: Biochem Pharmacol. 1996 Apr 26;51(8):1003-8
references.pmid: 8866821
references.citation: J Med Chem. 2005 Sep 8;48(18):5705-20
references.pmid: 16134939
references.citation: Circ Cardiovasc Qual Outcomes. 2009 May;2(3):155-63
references.pmid: 20031832
references.citation: Proc Natl Acad Sci U S A. 1975 Dec;72(12):4863-5
references.pmid: 1061075
references.citation: Lancet. 2005 Feb 5-11;365(9458):475-81
references.pmid: 15705456
references.citation: JAMA. 2006 Oct 4;296(13):1633-44
references.pmid: 16968831
references.citation: Inorg Chem. 2003 May 5;42(9):2988-91
references.pmid: 12716192
references.citation: J Biol Chem. 2007 Jun 1;282(22):16379-90
references.pmid: 17434872
references.citation: J Biol Chem. 2003 Nov 14;278(46):45763-9
references.pmid: 12925531
references.citation: J Biol Chem. 1996 Jun 28;271(26):15810-4
references.pmid: 8663121