Abstract
In this study, induction and inhibition of rhesus monkey CYP3A64 versus human CYP3A4 were characterized in vitro, and the corresponding pharmacokinetic consequences were evaluated in rhesus monkeys. In monkey hepatocytes, rifampin markedly induced CYP3A64 mRNA (EC50 = 0.5 μM; Emax = 6-fold) and midazolam (MDZ) 1′-hydroxylase activity (EC50 = 0.2 μM; Emax = 2-fold). Compound A (N-[2(R)-hydroxy-1(S)-indanyl-5-[2(S)-(1,1-dimethylethylaminocarbonyl)-4-[(furo[2,3-b]pyridin-5-yl)-methyl]piperazin-1-yl]-4(S)-hydroxy-2(R)-phenylmethylpentanamide), a known potent and mechanism-based inhibitor of CYP3A4, strongly inhibited the formation of 1′-hydroxy MDZ by recombinant CYP3A64 in a concentration- and time-dependent manner (KI = 0.25 μM; kinact = 0.4 min–1). Similar corresponding results also were obtained with human CYP3A4 in the presence of rifampin or compound A. In rhesus monkeys, MDZ exhibited a relatively high metabolic clearance (primarily via 1′-hydroxylation followed by glucuronidation) and a low hepatic availability (Fh = 16%). Consistent with the induction of hepatic metabolism of a high-clearance compound, pretreatment with rifampin (18 mg/kg p.o. for 5 days) did not significantly affect the i.v. kinetics of MDZ, but caused a pronounced reduction (∼10-fold) in the systemic exposure to MDZ and, consequently, its Fh following intrahepatic portal vein administration (i.pv.) of MDZ. A comparable extent of the pharmacokinetic interaction also was obtained after a 1.8 mg/kg rifampin dose. Also consistent with the in vitro CYP3A64 inhibition finding, compound A (6 mg/kg i.v.) markedly increased (10-fold) the i.pv. administered MDZ exposure. At the doses studied, plasma concentrations of rifampin or compound A reached or exceeded their respective in vitro EC50 or KI values. These findings suggest the potential applicability of the in vitro-in vivo relationship approach in rhesus monkeys for studying CYP3A-mediated interactions in humans.
Footnotes
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Article, publication date, and citation information can be found at http://dmd.aspetjournals.org.
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This work was conducted at Merck Research Laboratories, West Point, PA.
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doi:10.1124/dmd.106.009878.
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ABBREVIATIONS: MDZ, midazolam; 1′-OH MDZ, 1′-hydroxy midazolam; 4-OH MDZ, 4-hydroxy midazolam; OR, oxidoreductase; DMSO, dimethyl sulfoxide; PCR, polymerase chain reaction; AUC, area under the plasma concentration-time curve; Cmax, peak plasma concentration; CL, plasma clearance; Fh, hepatic availability; Vdss, volume of distribution at steady state; t1/2, half-life; i.pv., intrahepatic portal vein; LC-MS/MS, liquid chromatography coupled with tandem mass spectrometry.
- Received February 22, 2006.
- Accepted June 14, 2006.
- The American Society for Pharmacology and Experimental Therapeutics
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