The influence of donor and reservoir additives on Caco-2 permeability and secretory transport of HIV protease inhibitors and other lipophilic compounds

Purpose. To optimize the conditions for determining Caco-2 permeation of HI V protease inhibitors and other lipophilic compounds, and to compare cyclic urea HIV protease inhibitors with marketed compounds. Methods. Absorptive and secretory Caco-2 membrane permeation studies were p erformed with HIV protease inhibitors and various reference compounds, exam ining the effects of adding the solubilizing agents dimethylacetamide (DMAC ) and albumin in donor and reservoir compartments, respectively. Results. DMAC was useful as an additive in the donor vehicles, increasing t he dissolved concentrations of poorly water-soluble HIV protease inhibitors , and enabling more reliable determination of P-app values. Donor vehicles containing up to 5% DMAC could be used without altering Caco-2 barrier func tion, as indicated by the lack of effect on permeabilities of reference com pounds with diverse absorption characteristics. The utilization of a reserv oir containing albumin resulted in marked increases in absorptive P-app val ues for some HIV protease inhibitors as well as other lipophilic, highly pr otein bound compounds, consistent with albumin increasing the release of th ese compounds from the cell monolayer. Conclusions. Poorly soluble, lipophilic, highly bound compounds may require using solubilizing agents in the donor and reservoir compartments of Caco- 2 permeation experiments for estimating in vivo absorption potential. If th e reservoir does not provide adequate sink conditions, cellular retention c ould over-emphasize the contributions of secretory transport. The cyclic ur eas, DMP 450, DMP 850, and DMP 851, have Caco-2 permeabilities suggestive o f moderate-to-high oral absorption potential in humans.