Preparation of Alginate/Chitosan Microcapsules and Enteric Coated Granules of Mistletoe Lectin

  • Lyu, Su-Yun (School of Pharmaceutical Sciences, Boots Science Building, University of Nottingham) ;
  • Kwon, Young-Ju (College of Natural Sciences, Seoul Womens University, Seoul) ;
  • Joo, Hye-Jin (College of Natural Sciences, Seoul Womens University, Seoul) ;
  • Park, Won-Bong (College of Natural Sciences, Seoul Womens University, Seoul)
  • Published : 2004.01.01


The aqueous extract of European mistletoe (Viscum album, L.) has been used in cancer therapy. The purified mistletoe lectins, main components of mistletoe, have demonstrated cytotoxic and immune-system-stimulating activities. Korean mistletoe (Viscum album L. coloratum), a subspecies of European mistletoe, has also been reported to possess anticancer and immunological activities. A galactose- and N-acetyl-D-galactosamine-specific lectin (Viscum album L. coloratum agglutinin, VCA) with Mr 60 kDa was isolated from Korean mistletoe. Mistletoe preparations have been given subcutaneously due to the low stability of lectin in the gastrointestinal (GI) tract. In the present study, we investigated the possibility of alginate/chitosan microcapsules as a tool for oral delivery of mistletoe lectin. In addition, our strategy has been to develop a system composed of stabilizing cores (granules), which contain mistletoe lectin, extract or powder, coated by a biodegradable polymer wall. Our results indicated that successful incorporation of VCA into alginate/chitosan microcapsules has been achieved and that the alginate/chitosan microcapsule protected the VCA from degradation at acidic pH values. And coating the VCA with polyacrylic polymers, Eudragit, produced outstanding results with ideal release profiles and only minimal losses of cytotoxicity after manufacturing step. The granules prepared with extract or whole plant produced the best results due to the stability in the extract or whole plant during manufacturing process.


  1. Bantel, H., Engels, I. H., Voelter, W., Schulze-Osthoff, K., and Wesselborg, S., Mistletoe lectin activates caspase-8/FLICE independently of death receptor signaling and enhances anticancer drug-induced apoptosis. Cancer Res., 59, 2083-2090 (1999)
  2. Bowersock, T. L., HogenEsch, H., Suckow, M., Guimond, P., Martin, S., Borie, D., Torregrosa, S., Park, H., and Park, K., Oral vaccination of animals with antigens encapsulated in alginate microspheres. Vaccine, 17, 1804-1811 (1999)
  3. B$\ddot{u}$ssing, A., Suzar, K., Bergmann, J., PfUller, U., Schietzel, M., and Schweizer, K., Induction of apoptosis in human lymphocytes treated with Viscum album L. is mediated by the mistletoe lectins. Cancer Lett., 99, 59-72 (1996)
  4. Bussing, A., Biological and pharmacological properties of Viscum a/bum L.; From tissue flask to man. In A Bussing (eds.). Mistletoe, genus Viscum & other genera, Medicinal and aromatic plantslndustrial profiles. Harwood Academic Publishers, Netherland, pp 45-60 (2000)
  5. Clark, M. A., Hirst, B. H., and Jepson, M., A., Lectin-mediated mucosal delivery of drugs and microparticles. Adv. Drug Deliv. Rev., 43, 207-223 (2000)
  6. Delgado, A., Lavelle, E. C., Hartshorne M., and Davis, S. S., PLG microparticles stabilised using enteric coating polymers as oral vaccine delivery systems. Vaccine, 16, 2927-2938 (1999)
  7. Esquisabel, A., Hernandez, R. M., Igartua, M., Gascon, A. R., Calvo, B., and Pedraz, J. L., Production of BCG alginate-PLL microcapsules by emulsification/internal gelation. J. Microencapsul., 14, 627-638 (1997)
  8. Florence,A .T., The oral absorption of micro- and nanoparticulates: neither exceptional nor unusual. Pharm. Res., 14, 259-266 (1997)
  9. Gebert, A and Posselt, W., Glycoconjugate expression defines the origin and differentiation pathway of intestinal M-cells. J. Histochem. Cylochem., 45,1341-1350 (1997)
  10. Giannasca, K. T., Giannasca, P., Falk, J. I., and Neutra, M. R., Regional differences in glycoconjugates of intestinal M cells in mice: potential targets for mucosal vaccines. Am. J. Physiol., 267, G1108-G1121 (1994)
  11. Hajto, T., Hostanska, K., Frey, K., Rordorf, C., and Gabius, H. J., Increased secretion of tumor necrosis fator-$\alpha$, interleukin-6 by human mononuclear cells exposed to $\beta$-galactoside-specific lectin from clinically applied mistletoe extract. Cancer Res., 50, 3322-3326 (1990)
  12. Hari, P. R., Chandy, T., and Sharma, C. P., Chitosan/calcium alginate microcapsules for intestinal delivery of nitrofurantoin. J. Microencapsul., 13, 319-329 (1996)
  13. Jung, M. L., Baudino, S., and Beck, J. P., Charaterization of cytotoxic proteins from mistletoe (Viscum album L.). Cancer Lett., 51, 103-108 (1990)
  14. Jepson, M. A., Clark, M. A., Foster, N., Mason, C. M., Bennett, M. K., Simmons, N. L., and Hirst, B. H., Targeting to intestinal M cells. J. Anat., 189, 507-516 (1996)
  15. Li, S., Wang, X. T., Zhang, X. B., Yang, R. J., Zhang, H. Z., Zhu, L. Z., and Hou, X. P., Studies on alginate/chitosan microcapsules and renal arterial embolization in rabbits. J. Control Release, 5, 87-98 (2002)
  16. Lyu, S. Y., Park, S. M., Choung, B. Y., and Park, W. B., Comparative study of Korean (Viscum album, var. coloratum) and European mistletoes (Viscum album). Arch. Pharm. Res., 23, 592-598 (2000)
  17. Lyu, S. Y., Park, W. B., Choi, K. H., and Kim, W. H., Involvement of caspase-3 in apoptosis induced by Viscum album var. coloratum agglutinin in HL-60 cells. Biosci. Biotechnol. Biochem., 65, 534-541 (2001)
  18. Lyu, S. Y., Choi S. H., and Park, W. B., Korean mistletoe lectin-induced apoptosis in hepatocarcinoma cells is associated with inhibition of telomerase via mitochondrial controlled pathway independent of p53, Arch. Pharm. Res., 25(1) (2002)
  19. Marschutz, M. K. and Bernkop-Schnurch, A., Oral peptide drug delivery: polymer-inhibitor conjugates protecting insulin from enzymatic degradation in vitro. Biomaterials., 21, 1499-1507 (2000)
  20. McClean, S., Prosser, E., Meehan, E., O'Malley, D., Clarke, N., Ramtoola, Z., and Brayden, D., Binding and uptake of biodegradable poly-D,L-Iactide micro- and nanoparticles in intestinal epithelia. Eur. J. Pharm. Sci., 6, 153-163 (1998)
  21. Nakane, S., Kakumoto, M., Yukimatsu, K., and Chien, Y. W., Oramucosal delivery of LHRH: pharmacokinetic studies of controlled and enhanced transmucosal permeation. Pharm. Dev. Technol., 96, 251-259 (1996)
  22. Park, W. B., Ju, Y. J., and Han, S. K., Isolation and characterization of $\beta$
  23. Peumans, W. J. and Van Damme, E. J. M., Recent advances in the purification and characterization of plant lectins. In COST 98: Effects of Antinutrients on the Nutritional value of Legume Diets, Bardocz S., Gelencser E., Pusztai A (Eds.), European Commission, Brussels, Belgium, 1-7 (1996)
  24. Polk, A .E., Amsden, B., Scarratt, D. J., Gonzal, A., Okhamafe, A. O., and Goosen, M. F. A., Oral delivery in aquaculture: controlled release of proteins from chitosanalginate microcapsules. Aquacull. Eng., 13, 311-323 (1994)
  25. Pryme, I. F., Bardocz, S., Pusztai, A., and Ewen, S. w., Dietary mistletoe lectin supplementation and reduced growth of a murine non-Hodgkin lymphoma. Histol. Histopathol., 17, 261-71 (2002)
  26. Pusztai, A., Grant, G., Gelencser, E., Ewen, S. W. B., U. Pf$\ddpt{u}$ller, U., Eifler, R., and Bardocz, S., Effects of an orally administered mistletoe (type-2 RIP) lectin on growth, body composition, small intestinal structure, and insulin levels in young rats, J. Nutr. Biochem., 9, 31-36 (1998)
  27. Quong D. and Neufeld, R. J.,DNA protection from extracapsular nucleases, within chitosan- or poly-L-Iysine-coated alginate beads. Biotechnol. Bioeng., 60, 124-134 (1998)<124::AID-BIT14>3.0.CO;2-Q
  28. Reddy, S. M., Sinha, V. R., and Reddy, D. S., Novel oral colon-specific drug delivery systems for pharmacotherapy of peptide and nonpeptide drugs. Drugs Today (Bare)., 35, 537-580 (1999)
  29. Sharma, E. J. M., Peumans, D. W. J., Sarsfield, P., and Schumacher, U., Lectin binding reveals divergent carbohydrate expression in human and mouse Peyer's patches. Histochem. Cell Biol., 105, 459-465 (1996)
  30. Schellens, J. H. M., Mallnqre, M. M., Kruijtzer, M. C. F., Bardelmeijer, H. A., Tellingen, O., Schinkel, A. H., and Beijnen, J. H., Modulation of oral bioavailability of anticancer drugs: from mouse to man. Eur. J of Pharm. Sci., 12, 103-110 (2000)
  31. Schumacher U., Adam E., and Flavell D. J., Glycosylation patterns of the human colon cancer cell line HT-29 detected by Helix pomatia agglutinin and other lectins in culture, in primary tumors and in metastasis in SCID mice. Clin. Exp. Metastasis, 12, 398-404 (1994)
  32. Schumacher U., Stamouli A., Adam E., Peddie M., and PfUlier U., Biochemical, histochemical and cell biological investigations on the actions of mistletoe lectin I, II and III with human breast cancer cell lines. Glycocojugate J., 12, 250-257 (1995)
  33. Sezer, A. D. and Akbuga, J., Release characteristics of chitosan treated alginate beads: I. Sustained release of a macromolecular drug from chitosan treated alginate beads. J. Microencapsul., 16, 195-203 (1999)
  34. Singh, M. and O'Hagan, D., The preparation and characterization of polymeric antigen delivery systems for oral administration. Adv. Drug Deliv. Rev., 34, 285-304 (1998)
  35. Takka, S., Ocak, O. H., and Acart$\ddot{u}$rk, F., Formulation and investigation of nicardipine HCI-alginate gel beads with factorial design-based studies. Eur. J Pharmaceut. Sci., 6, 241-246 (1998)
  36. Taylor-Papadimitriou J. and Epenetos A. A., Exploiting altered glycosylation pattern in cancer: progress and challenges in diagnosis and therapy. TlBTECH., 12, 227-233 (1994)
  37. Tiourina, O. P. and Sukhorukov, G. B., Multilayer alginate/protamine microsized capsules: encapsulation of $\alpha$-chymotrypsin and controlled release study Int. J. Pharm., 21, 155-161 (2002)
  38. Yoon, T. J., Yoo, Y. C., Kang, T. B., K. Shimazki, Song, S. K., Lee K. H., Kim, S. H., Park, C. H., Azuma, I., and Kim, J. B., Lectins isolated from Korean mistletoe (Viscum album coloratum) induce apoptosis in tumor cells, Cancer Lett., 136, 33-40 (1999)
  39. Zhou, S., Deng, X., and Li, X., Investigation on a novel core-coated microspheres protein delivery system. J. Control Release, 75, 27-36 (2001)