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Quantitative Assessment of HIV-1 Protease Inhibitor Interactions with Drug Efflux Transporters in the Blood–Brain Barrier

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Purpose

To quantitatively characterize the drug efflux interactions of various HIV-1 protease inhibitors in an in vitro model of the blood–brain barrier (BBB) and to compare that with HIV-1 protease inhibitor stimulated P-glycoprotein (P-gp)-ATPase activity.

Methods

Cellular accumulation of the P-gp sensitive probe, rhodamine 123 (R123), and the mixed P-gp/multidrug resistance–associated protein (MRP) probe, 2′,7′-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF), were evaluated in primary cultured bovine brain microvessel endothelial cells (BBMEC) in the presence of various concentrations of HIV-1 protease inhibitors. The potency (IC50) and efficacy (Imax) of the drugs in the cell accumulation assays for P-gp and/or MRP was determined and compared to activity in a P-gp ATPase assay.

Results

For R123 (P-gp probe), the rank order potency for inhibiting R123 accumulation in the BBMEC was saquinavir = nelfinavir > ritonavir = amprenavir > indinavir. This correlated well with the rank order affinity in the P-gp ATPase assay. The rank order potency for MRP-related drug efflux transporters, was nelfinavir > ritonavir > saquinavir > amprenavir > indinavir.

Conclusions

HIV-1 protease inhibitors potently interact with both P-gp and MRP-related transporters in BBMEC. Characterization of the interactions between the HIV-1 protease inhibitors and drug efflux transporters in brain microvessel endothelial cells will provide insight into potential drug–drug interactions and permeability issues in the BBB.

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Abbreviations

BBB:

blood–brain barrier

BBMEC:

bovine brain microvessel endothelial cells

BCA:

bicinchoninic acid

BCECF:

2′,7′-bis(2-carboxyethyl)-5(6)-carboxyfluorescein

BCRP:

brain cancer resistance protein

CNS:

central nervous system

HAART:

highly active retroviral therapy

HIV-1:

human immunodeficiency virus

MRP:

multidrug resistance–associated protein

OAT:

organic anion transporter

OATP:

organic anion transporting peptide

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Acknowledgments

These studies were supported by funds from NIH Grant R01-NS42549 (WFE) and R01-CA93558 (DWM). C. J. Bachmeier was supported through the McDonald/Bukey and Blanche Widaman graduate studies fellowships.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, University of Nebraska Medical Center, 986025 Nebraska Medical Center, Omaha, Nebraska, USA

    Corbin J. Bachmeier & Timothy J. Spitzenberger

  2. Department of Pharmaceutics, University of Minnesota, Minneapolis, Minnesota, USA

    William F. Elmquist

  3. Department of Pharmacology and Therapeutics, University of Manitoba, Winnipeg,, MB, R3E 0T6, Canada

    Donald W. Miller

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  1. Corbin J. Bachmeier
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  3. William F. Elmquist
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Correspondence to Donald W. Miller.

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Bachmeier, C.J., Spitzenberger, T.J., Elmquist, W.F. et al. Quantitative Assessment of HIV-1 Protease Inhibitor Interactions with Drug Efflux Transporters in the Blood–Brain Barrier. Pharm Res 22, 1259–1268 (2005). https://doi.org/10.1007/s11095-005-5271-y

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