Preparation of alginate/chitosan microcapsules and enteric coated granules of mistletoe lectin

  • Su-Yun Lyu
  • Young-Ju Kwon
  • Hye-Jin Joo
  • Won-Bong ParkEmail author
Research Articles Articles


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 cyto-toxic 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- andN-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 (Gl) 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.

Key words

Mistletoe Lectin Microcapsules Enteric coating Oral administration 


  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).PubMedGoogle Scholar
  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).PubMedCrossRefGoogle Scholar
  3. Bussing, A., Suzar, K., Bergmann, J., Pfüller, U., Schietzel, M., and Schweizer, K., Induction of apoptosis in human lymphocytes treated withViscum album L. is mediated by the mistletoe lectins.Cancer Lett., 99, 59–72 (1996).PubMedCrossRefGoogle Scholar
  4. Bussing, A., Biological and pharmacological properties ofViscum album L; From tissue flask to man. In A. Büssing (eds.), Mistletoe, genus Viscum & other genera, Medicinal and aromatic plantslndustrial profiles. Harwood Academic Publishers, Netherland, pp 45–60 (2000).Google Scholar
  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).PubMedCrossRefGoogle Scholar
  6. Delgado, A., Lavelle, E. C., Hartshome M., and Davis, S. S., PLG microparticles stabilised using enteric coating polymers as oral vaccine delivery systems.Vaccine, 16, 2927–2938 (1999).CrossRefGoogle Scholar
  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).PubMedCrossRefGoogle Scholar
  8. Florence, A. T., The oral absorption of micro- and nanoparticulates: neither exceptional nor unusual.Pharm. Res., 14, 259–266 (1997).PubMedCrossRefGoogle Scholar
  9. Gebert, A. and Posselt, W., Glycoconjugate expression defines the origin and differentiation pathway of intestinal M-cells.J. Histochem. Cytochem., 45, 1341–1350 (1997).PubMedGoogle Scholar
  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).PubMedGoogle Scholar
  11. Hajto, T., Hostanska, K., Frey, K., Rordorf, C., and Gabius, H. J., Increased secretion of tumor necrosis fator-α, interleukin-6 by human mononuclear cells exposed to β-galactoside-specific lectin from clinically applied mistletoe extract.Cancer Res., 50, 3322–3326 (1990).PubMedGoogle Scholar
  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).PubMedCrossRefGoogle Scholar
  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).PubMedCrossRefGoogle Scholar
  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. Anal., 189, 507–516 (1996).Google Scholar
  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).CrossRefGoogle Scholar
  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).PubMedGoogle Scholar
  17. Lyu, S. Y., Park, W. B., Choi, K. H., and Kim, W. H., Involvement of caspase-3 in apoptosis induced byViscum album var.coloratum agglutinin in HL-60 cells.Biosci. Biotechnol. Biochem., 65, 534–541 (2001).PubMedCrossRefGoogle Scholar
  18. Lyu, S. Y., Choi S. H., and Park, W. B., Korean mistletoe lectininduced apoptosis in hepatocarcinoma cells is associated with inhibition of telomerasevia mitochondrial controlled pathway independent of p53,Arch. Pharm. Res., 25(1) (2002).Google Scholar
  19. Marschutz, M. K. and Bernkop-Schnurch, A., Oral peptide drug delivery: polymer-inhibitor conjugates protecting insulin from enzymatic degradationin vitro.Biomaterials., 21, 1499–1507 (2000).PubMedCrossRefGoogle Scholar
  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-lactide micro- and nanoparticles in intestinal epithelia.Eur. J. Pharm. Sci., 6, 153–163 (1998).PubMedCrossRefGoogle Scholar
  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).CrossRefGoogle Scholar
  22. Park, W. B., Ju, Y. J., and Han, S. K., Isolation and characterization of β-galactoside specific lectin from Korean mistletoe (Viscum album L, var.coloratum) with lactose-BSA-Sepharose 4B and changes of lectin conformation.Arch. Pharm. Res., 21, 429–435 (1998).PubMedGoogle Scholar
  23. Peumans, W. J. and Van Damme, E. J. M., Recent advances in the purification and characterization of plant lectins. InCOST 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).Google Scholar
  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.Aquacult. Eng., 13, 311–323 (1994).CrossRefGoogle Scholar
  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).PubMedGoogle Scholar
  26. Pusztai, A., Grant, G., Gelencsér, E., Ewen, S. W. B. U., Pfüller, 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).CrossRefGoogle Scholar
  27. Quong D. and Neufeld, R. J., DNA protection from extracapsular nucleases, within chitosan- or poly-L-lysine-coated alginate beads.Biotechnol. Bioeng., 60, 124–134 (1998).PubMedCrossRefGoogle Scholar
  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 (Barc)., 35, 537–580 (1999).Google Scholar
  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).PubMedCrossRefGoogle Scholar
  30. Schellens, J. H. M., Malingre, 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).CrossRefGoogle Scholar
  31. Schumacher U., Adam E., and Flavell D. J., Glycosylation patterns of the human colon cancer cell line HT-29 detected byHelix pomatia agglutinin and other lectins in culture, in primary tumors and in metastasis in SCID mice.Clin. Exp. Metastasis, 12, 398–404 (1994).PubMedCrossRefGoogle Scholar
  32. Schumacher U., Stamouli A., Adam E., Peddie M., and Pfüller 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).CrossRefGoogle Scholar
  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).PubMedCrossRefGoogle Scholar
  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).PubMedCrossRefGoogle Scholar
  35. Takka, S., Ocak, O. H., and Acartiirk, F., Formulation and investigation of nicardipine HCI-alginate gel beads with factorial design-based studies.Eur. J. Pharmaceut. Sci., 6, 241–246 (1998).CrossRefGoogle Scholar
  36. Taylor-Papadimitriou J. and Epenetos A.A., Exploiting altered glycosylation pattern in cancer: progress and challenges in diagnosis and therapy.TIBTECH., 12, 227–233 (1994).Google Scholar
  37. Tiourina, O. P. and Sukhorukov, G. B., Multilayer alginate/protamine microsized capsules: encapsulation of α-chymotrypsin and controlled release study.Int. J. Pharm., 21, 155–161 (2002).CrossRefGoogle Scholar
  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).PubMedCrossRefGoogle Scholar
  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).PubMedCrossRefGoogle Scholar

Copyright information

© The Pharmaceutical Society of Korea 2004

Authors and Affiliations

  • Su-Yun Lyu
    • 1
  • Young-Ju Kwon
    • 2
  • Hye-Jin Joo
    • 2
  • Won-Bong Park
    • 2
    Email author
  1. 1.College of Natural SciencesSeoul Womens UniversitySeoulKorea
  2. 2.School of Pharmaceutical Sciences, Boots Science BuildingUniversity of NottinghamNottinghamUK

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