In vitro and in vivo evaluations of intestinal barriers for the zwitterion L-767,679 and its carboxyl ester prodrug L-775,318. Roles of efflux and metabolism.

The barriers to oral delivery of the hydrophilic zwitterion L-767, 679 (I) and its carboxyl ester prodrug L-775,318 (II) were examined. In the Caco-2 cell model, transport of II, but not I, was strongly oriented in the secretory direction. The basal-to-apical transport of II displayed saturable kinetics and was markedly inhibited by verapamil and quinidine, known P-glycoprotein inhibitors. In Caco-2 cells, metabolism of I was not observed, whereas hydrolysis of II was modest (</=20%). In the in situ rat intestinal loop model, verapamil did not affect the absorption of I but significantly increased the absorption of II. I was resistant to intestinal metabolism, whereas II underwent hydrolysis partially in rat lumen but more extensively in rat intestinal tissue and blood. In vitro metabolism studies indicated that verapamil also inhibited the hydrolysis of II in rats. The inhibition was relatively specific for the intestinal and not the luminal esterases. These results suggested that the intestinal absorption of I was limited not by intestinal efflux or metabolism but more likely by the low lipophilicity of I. However, an efflux system, likely mediated by P-glycoprotein, played an important role in limiting the absorption of II. In rats, metabolism served as an additional barrier to the absorption of II. Verapamil increased the intestinal absorption of the prodrug by inhibiting the efflux system in the two models studied, as well as possibly inhibiting metabolism in rats. For the first time, secretory transport was identified as a cause of the failure to increase the absorption of a lipophilic and cationic prodrug developed to overcome the absorption problem.