title: Detection of a new N-oxidized metabolite of flutamide, N-[4-nitro-3-(trifluoromethyl)phenyl]hydroxylamine, in human liver microsomes and urine of prostate cancer patients.
abstract: Flutamide (2-methyl-N-[4-nitro-3-(trifluoromethyl)phenyl]-propanamide), a nonsteroidal antiandrogen, is used in the treatment of prostate cancer but is occasionally associated with hepatic dysfunction. In the present study, the metabolism of flutamide including the formation of the possible reactive toxic metabolites was investigated using human liver microsomes and 10 isoforms of recombinant human cytochrome P450 (P450). 2-Hydroxyflutamide (OH-flutamide) and 4-nitro-3-(trifluoromethyl)phenylamine (FLU-1) were the main products of flutamide metabolism in human liver microsomes. The formation of OH-flutamide was markedly inhibited by ellipticine, an inhibitor of CYP1A1/1A2, and was mainly catalyzed by the recombinant CYP1A2. FLU-1 was also produced from OH-flutamide, but its metabolic rate was much less than that from flutamide. An inhibitor of carboxylesterase, bis-(p-nitrophenyl)phosphoric acid, completely inhibited the formation of FLU-1 from flutamide in human liver microsomes. A new metabolite, N-[4-nitro-3-(trifluoromethyl)phenyl]hydroxylamine (FLU-1-N-OH), was detected as a product of the reaction of FLU-1 with human liver microsomes and identified by comparison with the synthetic standard. The formation of FLU-1-N-OH was markedly inhibited by the addition of miconazole, an inhibitor of CYP3A4, and was mediated by recombinant CYP3A4. Furthermore, FLU-1-N-OH was detected mostly as the conjugates (glucuronide/sulfate) in the urine of prostate cancer patients collected for 3 h after treatment with flutamide. The formation of FLU-1-N-OH, however, did not differ between patients with and without abnormalities of hepatic functions among a total of 29 patients. The lack of an apparent association of the urinary excretion of FLU-1-N-OH and hepatic disorder may suggest the involvement of an additional unknown factor in the mechanisms of flutamide hepatotoxicity.
authors.affiliation: R&D Division, Pharmaceuticals Group, Nippon Kayaku Co., Ltd., 3-31-12 Shimo, Tokyo 115-8588, Japan. rika.goda@nipponkayaku.co.jp
authors.forename: Rika
authors.initials: R
authors.lastname: Goda
authors.forename: Daichi
authors.initials: D
authors.lastname: Nagai
authors.forename: Yuji
authors.initials: Y
authors.lastname: Akiyama
authors.forename: Kiyohiro
authors.initials: K
authors.lastname: Nishikawa
authors.forename: Isao
authors.initials: I
authors.lastname: Ikemoto
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Aizawa
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Nagata
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Yamazoe
mesh_terms: D000726:Androgen Antagonists / Q000493:pharmacokinetics* / Q000652:urine; D018931:Antineoplastic Agents, Hormonal / Q000493:pharmacokinetics* / Q000652:urine; D002851:Chromatography, High Pressure Liquid; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005485:Flutamide / Q000493:pharmacokinetics* / Q000652:urine; D006801:Humans; D007202:Indicators and Reagents; D008297:Male; D008862:Microsomes, Liver / Q000378:metabolism*; D009249:NADP / Q000494:pharmacology; D011471:Prostatic Neoplasms / Q000652:urine*; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D000726:Androgen Antagonists; D018931:Antineoplastic Agents, Hormonal; D004791:Enzyme Inhibitors; D007202:Indicators and Reagents; D009249:NADP; D005485:Flutamide; D003577:Cytochrome P-450 Enzyme System