Fig. 6.

Formation of dapivirine metabolites by CYP isozymes. A. Ability of individual cDNA-expressed CYPs to produce dapivirine metabolites. CYP isozymes (10 pmol/mL) were incubated individually with dapivirine (10 μM) for 30 min at 37 °C in the presence of an NADPH-regenerating system. B. Inhibition of individual CYPs in order to examine dapivirine metabolite production when multiple CYP isozymes are present. Human liver microsomes (1 mg/mL) were preincubated with small molecule CYP inhibitors in the presence of a NADPH-regenerating system for 5 min at 37 °C prior to the addition of dapivirine (10 μM), then incubated for 30 min at 37°C. Dapivirine metabolites were detected via uHPLC-MS/MS in selected reaction monitoring mode. The experiments were performed in triplicate. * = p ≤ 0.05; ** = p ≤ 0.01; *** = p ≤ 0.001.