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Midazolam Exhibits Characteristics of a Highly Permeable P-Glycoprotein Substrate

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Abstract

Purpose. The purpose of this study was to investigate whether midazolam exhibits characteristics of a highly permeable P-glycoprotein (P-gp) substrate and to evaluate the potential influence of P-gp inhibition on 1-OH midazolam formation during midazolam transport.

Methods. P-gp interaction was investigated by P-gp ATPase assay, efflux inhibition studies, and transport studies of midazolam across MDR1-MDCK and 1-α,25-dihydroxy vitamin D3-induced Caco-2 monolayers with and without the P-gp inhibitor GF120918.

Results. Midazolam was highly permeable and transport appeared essentially unpolarized. In MDR1-MDCK, the basolateral-to-apical (B-to-A) permeability was slightly higher (16%) than apical-to-basolateral (A-to-B) permeability (p = 0.04); GF120918 increased A-to-B permeability by 27% (p = 0.01), and increased cellular midazolam accumulation during A-to-B transport by 45% (p = 0.01). Midazolam (200 μM) decreased rhodamine123 and vinblastine B/A ratios 3-fold (p < 0.006), while increasing their cellular accumulation (p < 0.003). P-gp ATPase activation by midazolam was dose-dependent and saturable [K m=11.5(±4.0) μM; V max = 41.1(±7.4) nmol/mg/min]. P-gp inhibition increased 1-OH midazolam formation in A-to-B studies 1.3-fold when midazolam donor ≥ μM (p < 0.03). In B-to-A studies, P-gp inhibition did not significantly increase metabolite formation (p = 0.06). Midazolam's extraction ratio was not influenced by P-gp (p = 0.2).

Conclusions. The results indicate that midazolam exhibited characteristics of a highly permeable P-gp substrate. 1-OH midazolam formation during A-to-B midazolam transport increased slightly when P-gp was inhibited.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, University of Maryland, 20 N Pine Street, Baltimore, Maryland

    Sanna Tolle-Sander & James E. Polli

  2. Department of Pharmaceutical Chemistry, University of Kuopio, Kuopio, Finland

    Jarkko Rautio

  3. Preclinical Drug Metabolism and Pharmacokinetics, Trimeris, Durham, North Carolina

    Steve Wring

  4. Preclinical Drug Metabolism and Pharmacokinetics, GlaxoSmithKline, Research Triangle Park, North Carolina

    Joseph W. Polli

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  1. Sanna Tolle-Sander
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  2. Jarkko Rautio
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Correspondence to James E. Polli.

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Tolle-Sander, S., Rautio, J., Wring, S. et al. Midazolam Exhibits Characteristics of a Highly Permeable P-Glycoprotein Substrate. Pharm Res 20, 757–764 (2003). https://doi.org/10.1023/A:1023433502647

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