Cytochrome P4502B6 and 2C9 do not metabolize midazolam: kinetic analysis and inhibition study with monoclonal antibodies N Hamaoka, Y Oda, I Hase, A.

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Cytochrome P4502B6 and 2C9 do not metabolize midazolam: kinetic analysis and inhibition study with monoclonal antibodies N Hamaoka, Y Oda, I Hase, A Asada British Journal of Anaesthesia Volume 86, Issue 4, Pages 540-544 (April 2001) DOI: 10.1093/bja/86.4.540 Copyright © 2001 British Journal of Anaesthesia Terms and Conditions

Fig 1 Lineweaver–Burk plots of midazolam 1′-hydroxylation by recombinant CYP2B6, 3A4 and 3A5. Midazolam (0.5–20 µmol litre−1) was incubated at 37°C for 10 mins with recombinant P450 (30 pmol) and 0.4 mmol litre−1 reduced NADP in 0.1 mol litre−1 potassium phosphate buffer (pH 7.4) in a final volume of 500 µl. Each plot depicts the mean of four experiments. British Journal of Anaesthesia 2001 86, 540-544DOI: (10.1093/bja/86.4.540) Copyright © 2001 British Journal of Anaesthesia Terms and Conditions

Fig 2 Correlations between midazolam 1′-hydroxylation and S-mephenytoin N-demethylation, a marker of CYP2B6 activity (a) and testosterone 6β-hydroxylation, a marker of CYP3A4 activity (b) with human liver microsomes obtained from 11 individuals. Concentrations of midazolam, S-mephenytoin and testosterone were 10 µmol litre−1, 1 mmol litre−1 and 500 µmol litre−1, respectively. Each plot is the mean of three experiments. British Journal of Anaesthesia 2001 86, 540-544DOI: (10.1093/bja/86.4.540) Copyright © 2001 British Journal of Anaesthesia Terms and Conditions

Fig 3 Effects of monoclonal antibodies against CYP2B6 and 3A4 on midazolam 1′-hydroxylation. Human liver microsomes (100 µg of protein) obtained from five individuals were preincubated with antibodies at room temperature for 20 min followed by incubation with midazolam (10 µmol litre−1) and reduced NADP (0.4 mmol litre−1) in 0.1 mol litre−1 potassium phosphate buffer (pH 7.4) at 37°C for 2 min in a final volume of 500 µl. Midazolam 1′-hydroxylation activity without antibodies was 6.8 (2.0) nmol min−1 mg protein−1. Each plot depicts the mean±sd of five experiments. British Journal of Anaesthesia 2001 86, 540-544DOI: (10.1093/bja/86.4.540) Copyright © 2001 British Journal of Anaesthesia Terms and Conditions