Regional Peptide Uptake Study in the Rat Intestinal Mucosa: Glatiramer Acetate as a Model Drug
- 51 Downloads
Purpose. To identify regions of the rat intestine that are able to internalize from the lumen oligopeptides, using the model drug glatiramer acetate (GA).
Methods. GA was introduced into rat intestinal sacs and the integrity of GA during uptake was monitored using antibody detection. Sodium docecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting of intestinal homogenates that had been exposed to GA were performed to identify GA presence. An enzyme-linked immunosorbent assay (ELISA) protocol was adapted for GA quantification. Immunohistochemistry was undertaken to examine the rat colonic wall for GA uptake, and confocal microscopy was used to differentiate adsorbed and internalized peptide in cultured colorectal adenocarcinoma cells.
Results. The colon and the ileum, respectively, were identified to be the intestinal regions in which GA was maximally preserved during uptake from the lumen. GA was identified to cross the colonic wall from the epithelium to the serosa. Internalization of GA into cultured colonic epithelial cells was demonstrated.
Conclusions. The rat colonic wall was identified to be less proteolytically active toward GA compared to the wall of the more proximal regions of the small intestine. GA has the capacity to penetrate from the lumen into the colonic wall. The maintenance of GA integrity within the wall of the colon offers the potential for local biological activity of the drug.
Unable to display preview. Download preview PDF.
- 1.J. F. Woodley. Enzymatic barriers for GI peptide and protein delivery. Crit. Rev. Ther. Drug Carrier Syst. 11:61–95 (1994).Google Scholar
- 2.B. G. Arnason. Immunologic therapy of multiple sclerosis. Annu. Rev. Med. 50:291–302 (1999).Google Scholar
- 3.D. Teitelbaum, R. Arnon, and M. Sela. Immunomodulation of experimental autoimmune encephalomyelitis by oral administration of copolymer 1. Proc. Natl. Acad. Sci. U S A 96:3842–3847 (1999).Google Scholar
- 4.D. M. Weir. Labelling Proteins and Polypeptides, 2nd edition, Blackwell Scientific, Abingdon, Oxon, United Kingdom 1973.Google Scholar
- 5.L. Barthe, J. Woodley, and G. Houin. Gastrointestinal absorption of drugs: Methods and studies. Fundam. Clin. Pharmacol. 13:154–168 (1999).Google Scholar
- 6.A. A. Abbas, A. H. Lichtman, and J. S. Pober. Cellular and Molecular Immunology. W.B. Saunders Co., Philadelphia, London, Toronto, 1994.Google Scholar
- 7.R. L. Bacon and N. R. Niles. Medical Histology, Springer-Verlag, New York, Heidelber, Berlin, 1983.Google Scholar
- 8.P. Onori, A. Franchitto, R. Sferra, A. Vetuschi, and E. Gaudio. Peyer's patches epithelium in the rat: A morphological, immunohistochemical, and morphometrical study. Dig. Dis. Sci. 46:1095–1104 (2001).Google Scholar
- 9.A. Rubinstein, B. Tirosh, M. Baluom, T. Nassar, A. David, I. Gliko-Kabir, R. Radai, and M. Friedman. The rationale for peptide drugs delivery to the colon and the potential of polymeric carriers as effective tools. J. Control. Release 46:59–73 (1997).Google Scholar
- 10.W. Wang. Oral protein drug delivery. J. Drug Target. 4:195–232 (1996).Google Scholar
- 11.K. Ikesue, P. Kopeckova, and J. Kopecek. Degradation of proteins by Guinea pig intestinal enzymes. Int. J. Pharm. 95:171–179 (1993).Google Scholar
- 12.R. H. Lu, P. Kopeckova, and J. Kopecek. Degradation and aggregation of human calcitonin in vitro. Pharm. Res. 16:359–367 (1999).Google Scholar
- 13.Y. Sai, M. Kajita, I. Tamai, J. Wakama, T. Wakamiya, and A. Tsuji. Adsorptive-mediated endocytosis of a basic peptide in enterocyte-like Caco-2 cells. Am. J. Physiol. 275:G514–G520 (1998).Google Scholar
- 14.Y. Sai, M. Kajita, I. Tamai, J. Wakama, T. Wakamiya, and A. Tsuji. Adsorptive-mediated transcytosis of a synthetic basic peptide, 001-C8 in Caco-2 cells. Pharm. Res. 15:1305–1309 (1998).Google Scholar
- 15.M. Heyman, A. M. Crain-Denoyelle, S. K. Nath, and J. F. Desjeux. Quantification of protein transcytosis in the human colon carcinoma cell line CaCo-2. J. Cell. Physiol. 143:391–395 (1990).Google Scholar
- 16.J. A. Atchison, W. E. Grizzle, and D. J. Pillion. Colonic absorption of insulin: an in vitro and in vivo evaluation. J. Pharmacol. Exp. Ther. 248:567–572 (1989).Google Scholar
- 17.Y. Zheng, Y. Qiu, M. F. Lu, D. Hoffman, and T. L. Reiland. Permeability and absorption of leuprolide from various intestinal regions in rabbits and rats. Int. J. Pharm. 185:83–92 (1999).Google Scholar
- 18.L. Steidler, W. Hans, L. Schotte, S. Neirynck, F. Obermeier, W. Falk, W. Fiers, and E. Remaut. Treatment of murine colitis by Lactococcus lactis secreting interleukin-10. Science 289:1352–1355 (2000).Google Scholar
- 19.L. Y. Xu, J. S. Yang, Y. M. Huang, M. Levi, H. Link, and B. G. Xiao. Combined nasal administration of encephalitogenic myelin basic protein peptide 68-86 and IL-10 suppressed incipient experimental allergic encephalomyelitis in Lewis rats. Clin. Immunol. 96:205–211 (2000).Google Scholar