Abstract
In vitro studies were conducted to elucidate the metabolic profiles of and the enzymes responsible for the metabolism of (R)- and (S)-tofisopam (1-(3,4-dimethoxyphenyl)-5-ethyl-7,8-dimethoxy-4-methyl-5H-2,3-benzodiazepine). Large differences were observed between the two enantiomers. The major metabolite in incubations of 500 ng/ml (≈1.3 μM) (R)-tofisopam in human liver microsomes corresponded to demethylation of the methoxy group at the 4-position of the phenyl ring (M3). Incubating (R)-tofisopam with recombinant cytochrome P450 (P450) or with human liver microsomes and isoform-selective P450 chemical inhibitors indicated that M3 was primarily catalyzed by CYP2C9. Similar incubations with S-tofisopam indicated that the primary metabolite was due to demethylation of the methoxy group at the 7-position of the benzodiazepine ring (M1), and this reaction was catalyzed primarily by CYP3A4. The primary metabolites of both enantiomers were further demethylated to form a common didemethylated metabolite (M5) where the methoxy groups at positions 4 and 7 are demethylated. Analysis of plasma and urine samples from human clinical trials confirmed the in vitro observations. Subjects orally treated with 200 mg b.i.d. (R)-tofisopam had a 2-h M1/M3 plasma ratio of 1:29 and a ratio of 1:123 in urine, whereas patients orally administered (S)-tofisopam at 150 mg/kg t.i.d. had opposite M1 to M3 ratios of 8:1 in plasma and 6:1 in urine.
Footnotes
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doi:10.1124/dmd.107.015875.
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ABBREVIATIONS: P450, cytochrome P450; LC-MS/MS, liquid chromatography-tandem mass spectrometry.
- Received March 20, 2007.
- Accepted July 19, 2007.
- The American Society for Pharmacology and Experimental Therapeutics
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