Transport of HIV-protease inhibitors across 1 alpha,25di-hydroxy vitamin D3-treated Calu-3 cell monolayers: modulation of P-glycoprotein activity.

PURPOSE: The presence of P-glycoprotein (P-gp) within the lipid bi-layers of the absorptive cells greatly influences drug entry into the HIV-infected sanctuary sites. The objective of this study was to acces the potential role of pulmonary cells expressing high levels of P-gp in the efflux of potent anti-HIV drugs such as protease inhibitors.
METHODS: Human airway epithelium-derived Calu-3 cells grown in the presence of 0.025 mM 1alpha,25di-hydroxy Vitamin D3 (di-OH vit D3) were used as a model to evaluate the effects of p-glycoprotein efflux of HIV protease inhibitors. Cells used as controls were not treated with di-OH vit D3. The anti-HIV agents 3H Ritonavir and Saquinavir (50 microM) were used as model compounds for influx and efflux studies
RESULTS: Di-OH vit D3 treatment enhanced the differentiation c Calu-3 cells indicated by more cilia and mucus secretion. It also caused elevated P-gp expression as demonstrated by Western Bla analysis and enhanced basal to apical transport of cyclosporine as compared with untreated cells. The amount of Saquinavir transported, after 3 h, across untreated Calu-3 cells (A-B) was 3-fold higher (1.62 microg; Papp = 2A (+/- 0.79) x 10(-6) cm/s) than di-OH vi D3-treated cells (0.57 microg with the Papp = 5.02 (+/- 0.62) x 10(-7) cm/s) Similar transport profiles were obtained for 3H ritonavir and a significant increase (p < 0.05) in the A-B transport (2.5-fold) of 3H ritonavir was observed when the cell monolayers were preincubated with testosterone prior to transport studies. However, transport of AZT remained unaltered in di-OH vit D3 treated monolayers.
CONCLUSION: Modulation of P-gp activity may be necessary to increase the therapeutic efficacy of protease inhibitors against HIV-1 reservoirs across alveolar lining cells and fluids.