title: Human NAD(P)H:quinone oxidoreductase 1 (NQO1)-mediated inactivation of reactive quinoneimine metabolites of diclofenac and mefenamic acid.
abstract: quinone oxidoreductase 1 (NQO1) is an enzyme capable of reducing a broad range of chemically reactive quinones and quinoneimines (QIs) and can be strongly upregulated by Nrf2/Keap1-mediated stress responses. Several commonly used drugs implicated in adverse drug reactions (ADRs) are known to form reactive QI metabolites upon bioactivation by P450, such as acetaminophen (APAP), diclofenac (DF), and mefenamic acid (MFA). In the present study, the reductive activity of human NQO1 toward the QI metabolites derived from APAP and hydroxy-metabolites of DF and MFA was studied, using purified bacterial P450 BM3 (CYP102A1) mutant M11 as a bioactivation system. The NQO1-catalyzed reduction of the QI metabolites was quantified relative to spontaneous glutathione (GSH) conjugation. Addition of NQO1 to the incubations strongly reduced the formation of all corresponding GSH conjugates, and this activity could be prevented by dicoumarol, a selective NQO1 inhibitor. The GSH conjugation was strongly increased by adding human GSTP1-1 in a wide range of GSH concentrations. Still, NQO1 could effectively compete with the GST catalyzed GSH conjugation by reducing the QIs. In conclusion, we identified the QI metabolites of the 4'- and 5-hydroxy-metabolites of DF and MFA as novel substrates for human NQO1. NQO1-mediated reduction proves to be an effective pathway to detoxify these QI metabolites in addition to GSH conjugation. Genetically determined deficiency of NQO1 therefore might be a risk factor for ADRs induced by reactive QI drug metabolites.
authors.affiliation: Division of Molecular Toxicology, Amsterdam Institute for Molecules Medicines and Systems (AIMMS), Faculty of Sciences, VU University Amsterdam , De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands.
authors.forename: Galvin
authors.initials: G
authors.lastname: Vredenburg
authors.forename: Naura S
authors.initials: NS
authors.lastname: Elias
authors.forename: Harini
authors.initials: H
authors.lastname: Venkataraman
authors.forename: Delilah F G
authors.initials: DF
authors.lastname: Hendriks
authors.forename: Nico P E
authors.initials: NP
authors.lastname: Vermeulen
authors.forename: Jan N M
authors.initials: JN
authors.lastname: Commandeur
authors.forename: J Chris
authors.initials: JC
authors.lastname: Vos
mesh_terms: D065767:Activation, Metabolic; D002384:Catalysis; D002460:Cell Line; D004008:Diclofenac / Q000493:pharmacokinetics*; D005978:Glutathione / Q000378:metabolism; D051549:Glutathione S-Transferase pi / Q000378:metabolism; D006801:Humans; D007097:Imines / Q000737:chemistry; D008528:Mefenamic Acid / Q000493:pharmacokinetics*; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000378:metabolism*; D011809:Quinones / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism
chemical_list: D007097:Imines; D011809:Quinones; D004008:Diclofenac; D008528:Mefenamic Acid; D016660:NAD(P)H Dehydrogenase (Quinone); C447479:NQO1 protein, human; C496556:GSTP1 protein, human; D051549:Glutathione S-Transferase pi; D005978:Glutathione