title: Alprazolam as an in vivo probe for studying induction of CYP3A in cynomolgus monkeys.
abstract: Induction of the cytochrome P450 (P450) enzyme is a major concern in the drug discovery processes. To predict the clinical significance of enzyme induction, it is helpful to investigate pharmacokinetic alterations of a coadministered drug in a suitable animal model. In this study, we focus on the induction of CYP3A, which is involved in the metabolism of approximately 50% of marketed drugs and is inducible in both the liver and intestine. As a marker substrate for CYP3A activity, alprazolam (APZ) was selected and characterized using recombinant CYP3A enzymes expressed in Escherichia coli. Both human CYP3A4 and its cynomolgus P450 ortholog predominantly catalyzed APZ 4-hydroxylation with sigmoidal kinetics. When administered intravenously and orally to cynomolgus monkeys, APZ had moderate clearance; its first-pass extraction ratio after oral dosing was estimated to be 0.09 in the liver and 0.45 in the intestine. Pretreatment with multiple doses of rifampicin (20 mg/kg p.o. for 5 days), a known CYP3A inducer, significantly decreased plasma concentrations of APZ after intravenous and oral administrations (0.5 mg/kg), and first-pass extraction ratios were increased to 0.39 in the liver and 0.63 in the intestine. The results were comparable to those obtained in clinical drug-drug interaction (DDI) reports related to CYP3A induction, although the rate of recovery of CYP3A activity seemed to be slower than rates estimated in clinical studies. In conclusion, pharmacokinetic studies using APZ as a probe in monkeys may provide useful information regarding the prediction of clinical DDIs due to CYP3A induction.
authors.affiliation: DMPK Research Laboratory, 2-2-50 Kawagishi, Toda, Saitama, Japan. ohtsuka.tatsuyuki@ma.mt-pharma.co.jp
authors.forename: Tatsuyuki
authors.initials: T
authors.lastname: Ohtsuka
authors.forename: Takahiro
authors.initials: T
authors.lastname: Yoshikawa
authors.forename: Kazumasa
authors.initials: K
authors.lastname: Kozakai
authors.forename: Yumi
authors.initials: Y
authors.lastname: Tsuneto
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Uno
authors.forename: Masahiro
authors.initials: M
authors.lastname: Utoh
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yamazaki
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Kume
mesh_terms: D000284:Administration, Oral; D000525:Alprazolam / Q000008:administration & dosage / Q000097:blood / Q000378:metabolism / Q000493:pharmacokinetics*; D000818:Animals; D051544:Cytochrome P-450 CYP3A / Q000096:biosynthesis* / Q000235:genetics; D004353:Drug Evaluation, Preclinical; D004347:Drug Interactions; D004790:Enzyme Induction; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D022781:Hepatocytes / Q000201:enzymology / Q000378:metabolism; D006801:Humans; D007275:Injections, Intravenous; D007421:Intestine, Small / Q000201:enzymology* / Q000378:metabolism; D008099:Liver / Q000201:enzymology* / Q000378:metabolism; D008252:Macaca fascicularis; D008297:Male; D008657:Metabolic Clearance Rate; D008861:Microsomes / Q000201:enzymology / Q000378:metabolism; D023421:Models, Animal; D011237:Predictive Value of Tests; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection
chemical_list: D011994:Recombinant Proteins; C104464:CYP3A protein, human; D051544:Cytochrome P-450 CYP3A; D000525:Alprazolam