Abstract

A growing awareness of the risks associated with extensive intestinal metabolism has triggered an interest in developing robust methods for its quantitative assessment. This study explored the utility of intestinal S9 fractions, human liver microsomes, and recombinant cytochromes P450 to quantify CYP3A-mediated intestinal extraction in humans for a selection of marketed drugs that are predominantly metabolized by CYP3A4. A simple competing rates model is used to estimate the fraction of drug escaping gut wall metabolism (f_g) from in vitro intrinsic clearance in humans. The f_g values extrapolated from the three in vitro systems used in this study, together with literature-derived f_g from human intestinal microsomes, were validated against f_g extracted from human in vivo pharmacokinetic (PK) profiles using a generic whole-body physiologically-based pharmacokinetic (PBPK) model. The utility of the rat as a model for human CYP3A-mediated intestinal metabolism was also evaluated. Human f_g from PBPK compares well with that from the grapefruit juice method, justifying its use for the evaluation of human in vitro systems. Predictive performance of all human in vitro systems was comparable [root mean square error (RMSE) = 0.22–0.27; n = 10]. Rat f_g derived from in vivo PK profiles using PBPK has the lowest RMSE (0.19; n = 11) for the prediction of human f_g for the selected compounds, most of which have a fraction absorbed close to 1. On the basis of these evaluations, the combined use of f_g from human in vitro systems and rats is recommended for the estimation of CYP3A4-mediated intestinal metabolism in lead optimization and preclinical development phases.