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Inhibition of CYP3A4 and CYP3A5 catalyzed metabolism of alprazolam and quinine by ketoconazole as racemate and four different enantiomers

  • Pharmacokinetics and Disposition
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European Journal of Clinical Pharmacology Aims and scope Submit manuscript

Abstract

Objective

The antifungal drug ketoconazole (KTZ) is known as an inhibitor of, especially, the CYP3A subfamily, which catalyzes the metabolism of a large variety of drugs. Interactions between KTZ and CYP3A substrates have been reported both in vivo and in vitro. Most of them, however, involved the KTZ racemate. KTZ racemate and the separate enantiomers, 2R,4R; 2R,4S; 2S,4S, and 2S,4R, were evaluated for their selectivity in inhibiting alprazolam and quinine metabolism.

Methods

The inhibition of alprazolam and quinine metabolism was studied in an in vitro system of human liver microsomes (HLM), recombinant of CYP3A4 and CYP3A5. The concentrations of formed 3-hydroxyquinine and 4- and α-hydroxyalprazolam were measured by HPLC and LC-MS, respectively.

Results

Quinine 3-hydroxylation was catalyzed to a similar extent by CYP3A4 and CYP3A5. The formation rate of 4-hydroxyalprazolam was higher than that of α-hydroxyalprazolam for each HLM, CYP3A4 and CYP3A5. KTZ racemate and enantiomers showed differential inhibitory effects of quinine and alprazolam metabolism. Quinine metabolism catalyzed by HLM, CYP3A4 and CYP3A5 was potently inhibited by the trans-enantiomer KTZ 2S,4S, with IC50 value of 0.16 μM for HLM, 0.04 μM for CYP3A4 and 0.11 μM for CYP3A5. The same enantiomer showed the lowest IC50 values of 0.11 μM for HLM and 0.04 μM for CYP3A5 with respect to alprazoalm 4-hydroxylation and also the same pattern for alprazolamα-hydroxylation, 0.13 μM for HLM and 0.05 μM for CYP3A5. Alprazolam metabolism (both α- and 4- hydroxylations) catalyzed by CYP3A4 was inhibited potently by the cis-enantiomer KTZ 2S,4R, with IC50 values of 0.03 μM .

Conclusions

Alprazolam and quinine metabolism is catalyzed by both CYP3A4 and CYP3A5. The present study showed that different KTZ enantiomers inhibit CYP3A4 and CYP3A5 to different degrees, indicating that structural differences among the enantiomers would be related to their inhibitory potency on these two enzymes.

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Acknowledgements

The study was financially supported by the Swedish Research Council, Medicine (3902), the National Institutes of Health, USA (R01 GM60548), Karolinska Institutet and Pfizer Ltd. We thank Dr. Jack Spira of InSpira Medical, Tyresö, Sweden, for supplying ketoconazole enantiomers and Anna Nordmark and Johanna Kenas for their collaboration with the quinine incubations.

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Author notes

  1. Rajaa A. Mirghani

    Present address: Department of Clinical Toxicology, Central Laboratories & Blood Bank, King Fahad Medical City, Riyadh, 11525, Saudi Arabia

Authors and Affiliations

  1. Karolinska Institutet, Department of Laboratory Medicine, Division of Clinical Pharmacology at Karolinska University Hospital, Huddinge, Stockholm, Sweden

    Annika Allqvist, Jun Miura, Leif Bertilsson & Rajaa A. Mirghani

  2. Division of Clinical Pharmacology, C1-68, Karolinska University Hospital, Huddinge, 141 86, Stockholm, Sweden

    Annika Allqvist

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  1. Annika Allqvist
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Correspondence to Annika Allqvist.

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Allqvist, A., Miura, J., Bertilsson, L. et al. Inhibition of CYP3A4 and CYP3A5 catalyzed metabolism of alprazolam and quinine by ketoconazole as racemate and four different enantiomers. Eur J Clin Pharmacol 63, 173–179 (2007). https://doi.org/10.1007/s00228-006-0230-z

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