Region-Dependent Role of Cell-Penetrating Peptides in Insulin Absorption Across the Rat Small Intestinal Membrane

We have reported that the cell-penetrating peptide (CPP) penetratin acts as a potential absorption enhancer in oral insulin delivery systems and that this action occurs through noncovalent intermolecular interactions. However, the region-dependent role of CPPs in intestinal insulin absorption has not been clarified. To identify the intestinal region where CPPs have the most effect in increasing insulin absorption, the region-dependent action of penetratin was investigated using in situ closed intestinal loops in rats. The order of the insulin area under the insulin concentration–time curve (AUC) increase effect by l-penetratin was ileum > jejunum > duodenum > colon. By contrast, the AUC order after coadministration of insulin with d-penetratin was colon > duodenum ≥ jejunum and ileum. We also compared the effects of the l- and d-forms of penetratin, R8, and PenetraMax on ileal insulin absorption. Along with the CPPs used in this study, l- and d-PenetraMax produced the largest insulin AUCs. An absorption study using ilea pretreated with CPPs showed that PenetraMax had no irreversible effect on the intestinal epithelial membrane. The degradation of insulin in the presence of CPPs was assessed in rat intestinal enzymatic fluid. The half-life (t_1/2) of insulin increased from 14.5 to 23.7 and 184.7 min in the presence of l- and d-PenetraMax, respectively. These enzymatic degradation-resistant effects might contribute partly to the increased ileal absorption of insulin induced by d-PenetraMax. In conclusion, this study demonstrated that the ability of the l- and d-forms of penetratin to increase intestinal insulin absorption was maximal in the ileum and the colon, respectively, and that d-PenetraMax is a powerful but transient enhancer of oral insulin absorption.