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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 811-818 |
| Number of pages | 8 |
| Journal | Pharmaceutical Research |
| Volume | 21 |
| Issue number | 5 |
| DOIs | |
| State | Published - May 2004 |
| Externally published | Yes |
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Keywords
- Central nervous system
- Drug transport
- Indinavir
- P-glycoprotein
- Saquinavir
ASJC Scopus subject areas
- Chemistry(all)
- Pharmaceutical Science
- Pharmacology
Cite this
Involvement of P-glycoprotein in the transport of saquinavir and indinavir in rat brain microvessel endothelial and microglia cell lines. / Ronaldson, Patrick T; Lee, Gloria; Dallas, Shannon; Bendayan, Reina.
In: Pharmaceutical Research, Vol. 21, No. 5, 05.2004, p. 811-818.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Involvement of P-glycoprotein in the transport of saquinavir and indinavir in rat brain microvessel endothelial and microglia cell lines
AU - Ronaldson, Patrick T
AU - Lee, Gloria
AU - Dallas, Shannon
AU - Bendayan, Reina
PY - 2004/5
Y1 - 2004/5
N2 - 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.
AB - 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.
KW - Central nervous system
KW - Drug transport
KW - Indinavir
KW - P-glycoprotein
KW - Saquinavir
UR - http://www.scopus.com/inward/record.url?scp=2442673340&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=2442673340&partnerID=8YFLogxK
U2 - 10.1023/B:PHAM.0000026433.27773.47
DO - 10.1023/B:PHAM.0000026433.27773.47
M3 - Article
C2 - 15180339
AN - SCOPUS:2442673340
VL - 21
SP - 811
EP - 818
JO - Pharmaceutical Research
JF - Pharmaceutical Research
SN - 0724-8741
IS - 5
ER -