title: Evidence for the bioactivation of zomepirac and tolmetin by an oxidative pathway: identification of glutathione adducts in vitro in human liver microsomes and in vivo in rats.
abstract: Although zomepirac (ZP) and tolmetin (TM) induce anaphylactic reactions and form reactive acyl glucuronides, a direct link between the two events remains obscure. We report herein that, in addition to acyl glucuronidation, both drugs are subject to oxidative bioactivation. Following incubations of ZP with human liver microsomes fortified with NADPH and glutathione (GSH), a metabolite with an MH+ ion at m/z 597 was detected by LC/MS/MS. On the basis of collision-induced dissociation and NMR evidence, the structure of this metabolite was determined to be 5-[4'-chlorobenzoyl]-1,4-dimethyl-3-glutathionylpyrrole-2-acetic acid (ZP-SG), suggesting that the pyrrole moiety of ZP had undergone oxidation to an epoxide intermediate, followed by addition of GSH and loss of the elements of H2O to yield the observed conjugate. The oxidative bioactivation of ZP most likely is catalyzed by cytochrome P450 (P450) 3A4, since the formation of ZP-SG was reduced to approximately 10% of control values following pretreatment of human liver microsomes with ketoconazole or with an inhibitory anti-P450 3A4 IgG. A similar GSH adduct, namely 5-[4'-methylbenzoyl]-1-methyl-3-glutathionylpyrrole-2-acetic acid (TM-SG), was identified when TM was incubated with human liver microsomal preparations. The relevance of these in vitro findings to the in vivo situation was established through the detection of the same thiol adducts in rats treated with ZP and TM, respectively. Taken together, these data suggest that, in addition to the formation of acyl glucuronides, oxidative metabolism of ZP and TM affords reactive species that may haptenize proteins and thereby contribute to the drug-mediated anaphylactic reactions.
authors.affiliation: Department of Drug Metabolism, Merck Research Laboratories, Rahway, NJ, USA. qing_chen@merck.com
authors.forename: Qing
authors.initials: Q
authors.lastname: Chen
authors.forename: George A
authors.initials: GA
authors.lastname: Doss
authors.forename: Elaine C
authors.initials: EC
authors.lastname: Tung
authors.forename: Wensheng
authors.initials: W
authors.lastname: Liu
authors.forename: Yui S
authors.initials: YS
authors.lastname: Tang
authors.forename: Matthew P
authors.initials: MP
authors.lastname: Braun
authors.forename: Varsha
authors.initials: V
authors.lastname: Didolkar
authors.forename: John R
authors.initials: JR
authors.lastname: Strauss
authors.forename: Regina W
authors.initials: RW
authors.lastname: Wang
authors.forename: Ralph A
authors.initials: RA
authors.lastname: Stearns
authors.forename: David C
authors.initials: DC
authors.lastname: Evans
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Baillie
authors.forename: Wei
authors.initials: W
authors.lastname: Tang
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods; D005260:Female; D005978:Glutathione / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D022781:Hepatocytes / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008297:Male; D008862:Microsomes, Liver / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D009249:NADP / Q000378:metabolism / Q000494:pharmacology; D010084:Oxidation-Reduction / Q000187:drug effects; D051381:Rats; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D014046:Tolmetin / Q000031:analogs & derivatives* / Q000378:metabolism*; D014316:Tritium; D014217:Troleandomycin / Q000378:metabolism / Q000494:pharmacology
chemical_list: D014316:Tritium; D009249:NADP; C020549:zomepirac; D014217:Troleandomycin; D014046:Tolmetin; D005978:Glutathione