First Bioanalytical Evaluation of Nonpeptidic Cage Dimeric HIV-1 Protease InhibitorN-Benzyl 4-Aryl-1,4-dihydropyridine H17: Biotransformation and Toxicity on Hep G2 Cells

Bioanalysis ofSyn Dimeric HIV-1 Protease InhibitorN-Benzyl 4-Aryl-1,4-dihydropyridine H19: Metabolic and Cytotoxic Properties in Hep G2 Cells

“…This appears plausible on comparing the conformation of H19 in the reported X-ray structure (with ester instead of hydroxymethylene groups) [13] to that of H17 after molecular modeling with similar orientations of the functional groups [14] . These functional group orientations were considered responsible for the poor phase-II conjugation of H17 [6] . Moreover, equal shielding orientations of corresponding aromatic substituents in cyclic ureas may explain why no phase-II metabolites of their hydroxy functions are reported [12] .…”

“…The influence on cell proliferation of H19 at different concentrations is shown in Figure 2a and 2b by the changes of characterizing protein and DNA content, which are both certain indicators of cell proliferation [6] .…”

“…Apoptosis Analysis [6] 2 × 10 6 cells were incubated with 5 × 10 -5 mol/l of H19 for 24 h. Then cells were washed with PBS buffer, removed from the flask with a cell scraper, suspended in 2 ml PBS buffer and centrifuged for 5 min at 1500 g. Supernatant was withdrawn and cells lysed with 400 µl of a buffer containing 10 mM Tris/HCl, 10 mM EDTA, 0.2% Triton X-100, pH 7.5.…”

“…Thus, low metabolic rates and necessary non-toxicity of both drugs and their metabolites accompanied by a satisfactory absorption give realistic prospects for further drug development of novel PI [6] .…”

“…We recently demonstrated the advantages of cell cultures for such early investigations comparing to hepatocyte cultures [6,7] . Among these cultures Hep G2 monolayers have been favoured as they were shown to produce the same metabolic spectrum as hepatocyte cultures [8] .…”