Intracellular concentration of the HIV protease inhibitors indinavir and saquinavir in human endothelial cells.

Human vascular endothelial cells may serve as targets and a reservoir for human immunodeficiency virus type 1 (HIV-1). The antiviral activity of HIV protease inhibitors is reported to be related directly to the intracellular amount of the drug. To assess intracellular concentrations of two HIV protease inhibitors, human umbilical venous endothelial cells (HUVECs) were exposed for 3 h and 24 h to 100, 10 and 1 mg/L indinavir and saquinavir. Intracellular drug concentrations and the total drug amount in the supernatant were measured by means of high-performance liquid chromatography (HPLC). Exposure of HUVECs to 10 and 1 mg/L indinavir and saquinavir resulted in undetectable intracellular drug levels in 6 x 10(5) cells/well. Incubation of cells with solutions of 100 mg/L indinavir and saquinavir led to mean intracellular concentrations of indinavir (132 +/- 56 mg/L after 3 h and 150 +/- 29 mg/L after 24 h, respectively) and of saquinavir (96 +/- 10 mg/L after 3 h and 100 +/- 5 mg/L after 24 h) that were comparable to the levels determined for the substances in the supernatant over time (P > 0.001). These data indicate that intracellular concentrations of indinavir and saquinavir correlate well with the extracellular levels. Consequently, measurements of drug concentrations in patient's plasma by HPLC are assumed to be a good means of monitoring the intracellular drug concentration.