Abstract
The enhancement of paracellular transport of heparin disaccharide using several absorption enhancers across Caco-2 cell monolayers was tested. The cytotoxicity of these enhancers was also examined. The enhancing effects byQuillaja saponin, dipotassium glycyrrhizinate, 18p-glycyrrhetinic acid, sodium caprate and taurine were determined by changes in transepithelial electrical resistance (TEER) and the amount of heparin disaccharide transported across Caco-2 cell monolayers. Among the absorption enhancers, 18β-glycyrrhetinic acid and taurine decreased TEER and increased the permeability of heparin disaccharide in a dose-dependent and time-dependent manner with little or negligible cytotoxicity. Our results indicate that these absorption enhancers can widen the tight junction, which is a dominant paracellular absorption route of hydrophilic compounds. It is highly possible that these absorption enhancers can be applied as pharmaceutical excipients to improve the transport of macromolecules and hydrophilic drugs having difficulty in permeability across the intestinal epithelium.
Similar content being viewed by others
References
Anderson, J. M. and van Itallie, C. M., Tight junctions and the molecular basis for regulation of paracellular permeability.Am. J. Physiol., 269, G467-G475 (1995).
Baughman, R. A., Kapoor, S. C., Agarwal, R. K., Kisicki, J., Catella-Lawson, F., and FitzGerald, G. A., Oral delivery of anticoagulant doses of heparin. A randomized, double- blind, controlled study in humans.Circulation, 98, 1610–1615 (1998).
Björk I., Olson, S. T., and Shore J. D., Molecular mechanisms of the accelerating efffect of heparin on the reactions between antithrombin and clotting proteinases. In: Lane D., Lindahl U. (Eds.).Heparin: Chemical and biological properties, clinical applications. CRC Press, Boca Raton, pp. 229–255 (1989).
Brayden, D., Creed, E., O’Connell, A., Leipold, H., Agarwal, R., and Leone-Bay, A., Heparin absorption across the intestine: effects of sodiumN-[8-(2-hydroxybenzoyl)amino]caprylate in rat in situ intestinal instillations and in Caco-2 monolayers.Pharm. Res., 14, 1772–1779 (1997).
Cahalon, L., Lider, O., Schor, H., Avron, A., Gilat, D., Hershkoviz, R., Margalit, R., Eshel, A., Shoseyev O., and Cohen, I.R., Heparin disaccharides inhibit tumor necrosis factor-alpha production by macrophages and arrest immune inflammation in rodents.Int. Immunol., 9, 1517–1522 (1997).
Choksakulnimitr S., Masuda, S., Tokuda H., Takakura Y., and Hashida M.,In vitro cytotoxicity of macromolecules in different cell culture system.J. Control. Release, 34, 233–241 (1995).
Chowers, Y., Lider, O., Schor, H., Barshack, I., Tal, R., Ariel, A., Bar-Meir, S., Cohen I.R., and Cahalon, L., Disaccharides derived from heparin or heparan sulfate regulate IL-8 and IL-1 beta secretion by intestinal epithelial cells.Gastroenterology, 120, 449–459 (2001).
Clarke, H., Marano, C. W., Peralta Soler, A., and Mullin, J. M., Modification of tight junction function by protein kinase C isoforms.Adv. Drug Deliv. Rev., 41, 283–301 (2000).
Duffey, M. E., Hainau, B., Ho, S., and Bentzel, C. J., Regulation of epithelial tight junction permeability by cyclic AMP.Nature, 294, 451–453 (1981).
Fasano, A., Novel approaches for oral delivery of macromolecules.J. Pharm. Sci., 87, 1351–1356 (1998).
Fasano, A. and Uzzau, S., Modulation of intestinal tight junctions byZonula occludens toxin permits enteral administration of insulin and other macromolecules in an animal model.J. Clin. Invest., 99, 1158–1164 (1997).
Harenberg, J. and Casu, B., Nonanticoagulant actions of glycosaminoglycans, Plenum Press, New York (1996).
Hershkoviz, R., Schor, H., Ariel, A., Hecht, I., Cohen, I.R., Lider O., and Cahalon, L., Disaccharides generated from heparan sulphate or heparin modulate chemokine-induced T-cell adhesion to extracellular matrix.Immunology, 99, 87–93 (2000).
Hiebert, L. M., Wice, S. M., Ping, T., Hileman, R. E., Capila, I., and Linhardt, R. J., Tissue distribution and antithrombotic activity of unlabeled or14C- labeled porcine intestinal mucosal heparin following administration to rats by the oral route.Can. J. Physiol. Pharmacol., 78, 307–320 (2000).
Hopkins, A. M., Li, D., Mrsny, R. J., Walsh, S. V., and Nusrat, A., Modulation of tight junction function by G protein-coupled events.Adv. Drug Deliv. Rev., 41, 329–340 (2000).
Kim, B. T., Kim, W. S., Kim, Y. S., Linhardt, R. J., and Kim, D. H., Purification and characterization of a novel heparinase fromBacteroides stercoris HJ-15.J. Biochem. (Tokyo), 128, 323–328 (2000).
Leone-Bay, A., Paton, D. R., Freeman, J., Lercara, C., O’Toole, D., Gschneidner, D., Wang, E., Harris, E., Rosado, C., Rivera, T., DeVincent, A., Tai, M., Mercogliano, F., Agarwal, R., Leipold, H., and Baughman, R.A., Synthesis and evaluation of compounds that facilitate the gastrointestinal absorption of heparin.J. Med. Chem., 41, 1163–1171 (1998).
Lindmark, T., Kimura, Y., Artursson, P., Absorption enhancement through intracellular regulation of tight junction permeability by medium chain fatty acids in Caco-2 cells.J. Pharmacol. Exp. Ther., 284, 362–369 (1998)
Liu, D. Z., LeCluyse, E. L., and Thakker, D. R., Dodecylphosphocholine-mediated enhancement of paracellular permeability and cytotoxicity in Caco-2 cell monolayers.J. Pharm. Sci., 88, 1161–1168 (1999).
Ma, T. Y., Nguyen, D., Bui, V., Nguyen, H., and Hoa, N., Ethanol modulation of intestinal epithelial tight junction barrier. Am. J. Physiol. 276, G965-G974 (1999).
Madara, J. L.., Increases in guinea pig small intestinal transepithelial resistance induced by osmotic loads are accompanied by rapid alterations in absorptive-cell tight-junction structure.J. Cell. Biol., 97, 125–136 (1983).
Madara, J. L., Barenberg, D., and Carlson, S., Effects of cytochalasin D on occluding junctions of intestinal absorptive cells: further evidence that the cytoskeleton may influence paracellular permeability and junctional charge selectivity.J. Cell. Biol., 102, 2125–2136 (1986).
Merchant, Z. M., Kim, Y. S., Rice, K. G., and Linhardt, R. J., Structure of heparin-derived tetrasaccharides.Biochem. J., 229, 369–377 (1985).
Nelson, R. M., Cecconi, O., Roberts, W. G., Aruffo, A., Linhardt, R. J., and Bevilacqua, M. P., Heparin oligosaccharides bind L-selectin and P-selectin and inhibit acute inflammation.Blood, 82, 3253–3258 (1993).
Palant, C. E., Duffey, M. E., Mookerjee, B. K., HO, S., and Bentzel, C.J., Ca2+ regulation of tight-junction permeability and structure in Necturus gall bladder.Am. J. Physiol., 245, C203-C212 (1983).
Palmi, M., Youmbi, G. T., Fusi, F., Sgaragli, G. P., Dixon, H. B. F., Frosini, M., and Tipton, K. F., Potentiation of mitochondrial Ca2+ sequestration by taurine.Biochem. Pharmacol., 58, 1123–1131 (1999).
Pillion, D. J., Amsden, J. A., Kensil, C. R., and Recchia, J., Structure-function relationship amongQuillaja saponins serving as excipients for nasal and ocular delivery of insulin.J. Pharm. Sci., 85, 518–524 (1996).
Rivera, T. M., Leone-Bay, A., Paton, D. R., Leipold, H. R., and Baughman, R. A., Oral delivery of heparin in combination with sodium N-[8-(2-hydroxybenzoyl) amino]caprylate: pharmacological considerations.Pharm. Res., 14, 1830–1834 (1997).
Sakai, M., Imai, T., Ohtake, H., Azuma, H., and Otagiri, M., Effects of absorption enhancers on cytoskeletal actin filaments in Caco- 2 cell monolayers.Life Sci., 63, 45–54 (1998).
Sakai, M., Imai, T., Ohtake, H., Azuma, H., and Otagiri, M., Simultaneous use of sodium deoxycholate and dipotassium glycyrrhizinate enhances the cellular transport of poorly absorbed compounds across Caco-2 cell monolayers.J. Pharm. Pharmacol., 51, 27–33 (1999).
Stenson, W. F., Easom, R. A., Riehl, T. E., and Turk, J., Regulation of paracellular permeability in Caco-2 cell monolayers by protein kinase C.Am. J. Physiol., 265, G955-G962 (1993).
Walsh, S. V., Hopkins, A. M., and Nusrat, A., Modulation of tight junction structure and function by cytokines.Adv. Drug Deliv. Rev., 41, 303–313 (2000).
Ward, P. D., Tippin, T. K., and Thakker, D. R., Enhancing paracellular permeability by modulating epithelial tight junctions.Pharm. Sci. Tech. Today, 3, 346–358 (2000).
Xie, X., Rivier, A. S., Zakrzewicz, A., Bernimoulin, M., Zeng, X. L., Wessel, H. P., Schapira, M., and Spertini, O., Inhibition of selectin-mediated cell adhesion and prevention of acute inflammation by nonanticoagulant sulfated saccharides.J. Biol. Chem., 275, 34818–34825 (2000).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Cho, S.Y., Kim, J.S., Li, H. et al. Enhancement of paracellular transport of heparin disaccharide across Caco-2 cell monolayers. Arch Pharm Res 25, 86–92 (2002). https://doi.org/10.1007/BF02975268
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF02975268