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Involvement of P-Glycoprotein in the Transport of Saquinavir and Indinavir in Rat Brain Microvessel Endothelial and Microglia Cell Lines

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Abstract

Purpose. Membrane-bound efflux transporters, such as P-glycoprotein (P-gp), may limit the brain entry and distribution of HIV-1 protease inhibitors and be in part responsible for HIV-1-associated dementia treatment failure. The purpose of this study was to characterize the transport properties of saquinavir and indinavir in a brain microvessel endothelial cell line and in microglia, the immune cells of the brain and primary HIV-1 cellular target.

Methods. Biochemical and transport studies were performed in an immortalized rat brain endothelial cell line (RBE4), a rat microglia cell line (MLS-9), and a P-gp overexpressing Chinese hamster ovary cell line (CHRC5).

Results. Western blot analysis using the P-gp monoclonal antibody C219 detected a single band at approximately 170 to 180 kDa (a size previously reported for P-gp) in all cell lines. Cellular accumulation of [14C]saquinavir and [3H]indinavir by RBE4, MLS-9, and CHRC5 monolayers was significantly enhanced in the presence of P-gp inhibitors, HIV-1 protease inhibitors, the ATPase inhibitor sodium azide, and the ATP depleting agent 2',4'-dinitrophenol respectively. [14C]Saquinavir and [3H]indinavir efflux from both cell systems was rapid and significantly reduced in the presence of PSC833.

Conclusions. These results provide evidence for P-gp mediated transport of saquinavir and indinavir in RBE4 and MLS-9 and suggest that this transporter can restrict, at least in part, the permeation of HIV-1 protease inhibitors at both the brain barrier site and in brain parenchyma.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, Ontario, Canada

    Patrick T. Ronaldson, Gloria Lee, Shannon Dallas & Reina Bendayan

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  1. Patrick T. Ronaldson
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Correspondence to Reina Bendayan.

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Ronaldson, P.T., Lee, G., Dallas, S. et al. Involvement of P-Glycoprotein in the Transport of Saquinavir and Indinavir in Rat Brain Microvessel Endothelial and Microglia Cell Lines. Pharm Res 21, 811–818 (2004). https://doi.org/10.1023/B:PHAM.0000026433.27773.47

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  • DOI: https://doi.org/10.1023/B:PHAM.0000026433.27773.47

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