Abstract
Purpose. To investigate the usefulness of wheat germ agglutinin as a targeting carrier protein for an acid-labile chemotherapeutic prodrug directed against colon carcinoma cells in vitro.
Methods. Cis-aconityl-linked doxorubicin-wheat germ agglutinin was prepared by a two step procedure and the conjugate-binding capacity of target- and non-target cells was assayed by flow cytometry. The antiproliferative activity of the prodrug on Caco-2 and MOLT-4 cells was determined by the XTT- and BrdU-test and compared with that of the parent drug and the lectin alone.
Results. At pH 4.0, about 50% of the conjugated doxorubicin were released within 24 h from the water soluble prodrug exhibiting a conjugation number of 24 (mol doxorubicin/mol WGA). The prodrug-binding capacity of colon carcinoma cells exceeded that of human colonocytes and lymphoblastic MOLT-4 cells 4.5-fold. Additionally, the antiproliferative effect of the conjugate on Caco-2 cells was 39% as opposed to 5% in case of MOLT-4 cells. As the unmodified carrier protein inhibited or stimulated Caco-2 cell growth in a concentration-dependent manner, the cytostatic activity of the conjugate was determined at WGA concentrations without an effect on cell-proliferation. Considering 50% release of conjugated drug at the most, the prodrug yielded 160% of the cytostatic activity of free doxorubicin.
Conclusions. WGA-prodrug targeting offers new perspectives for site-specific, cytoinvading drug delivery in colon cancer chemotherapy.
Similar content being viewed by others
REFERENCES
R. Duncan, T. A. Connors, and H. Meada. Drug targeting in cancer therapy: the magic bullet, what next? J. Drug Targeting 3:317–319 (1996).
G. R. Thrush, L. R. Lark, B. C. Clinchy, and E. S. Vitetta. Immunotoxins: an update. Ann. Rev. Immunol. 14:49–71 (1996).
F. Gabor, I. Haberl, M. Wirth, K. Richter, G. Theyer, G. Baumgartner, E. Wenzl, and G. Hamilton. In vitro antitumor activity of MIC2 protein-doxorubicin conjugates. Int. J. Onc. 9:527–531 (1996).
S. S. Davis and L. Illum. Colloidal carriers and drug targeting. Acta Pharm. Technol. 32:4–9 (1986).
J. Kreuter. Nanoparticulate systems in drug delivery and targeting. J. Drug Targeting. 3:171–173 (1995).
N. Sharon and H. Lis. Lectins: cell agglutination and sugar specific proteins. Science 177:949–959 (1972).
C. M. Lehr in Lectins—Biomedical Perspectives. A. Pusztai and S. Bardocz (eds.) Taylor & Francis Ltd., London 1995.
A. Shanghal and S. Hakamori. Molecular changes in carbohydrate antigens associated with cancer. Bioessays 12:223–230 (1990).
I. E. Liener, N. Sharon, and I. E. Goldstein, The lectins: properties, functions and applications in biology and medicine, Academic Press, Orlando, Florida, 1986.
A. Pusztai, S. W. B. Ewen, G. Grant, D. S. Brown, J. C. Stewart, W. J. Peumans, E. J. M. van Damme, and S. Bardocz. Antinutritive effects of wheat-germ agglutinin and other N-acetylglucosamine-specific lectins. Br. J. Nutr. 70:313–321 (1993).
F. Gabor, M. Wirth, G. Walcher, and G. Hamilton. Lectin-mediated bioadhesion: Gastrointestinal stability and binding-characteristics of wheat germ agglutinin and Solanum tuberosum lectin on Caco-2, HT-29 and human colonocytes. J. Contr. Rel. 49:27–37 (1997).
H. S. Yang and R. A. Reisfeld. Doxorubicin conjugated with a monoclonal antibody directed to a human melanoma-associated proteoglycan suppresses the growth of established tumor xenografts in nude mice. Proc. Natl. Acad. Sci. USA 85:1189–1193 (1988).
M. Wilchek, T. Miron, and J. Kohn. Affinity chromatography. Methods Enzymol. 104:3–55 (1984).
R. O. Dillman, D. E. Johnson, D. L. Shawler, and J. A. Koziol. Superiority of an acid-labile daunorubicin-monoclonal antibody immunoconjugate compared to free drug. Cancer Res. 48:6079–6102 (1988).
W. C. Shen and H. J.-P. Ryser. Cis-aconityl spacer between daunomycin and macromolecular carriers: a model of pH-sensitive linkage releasing drug from a lysosomotropic conjugate. Biochem. Biophys. Res. Comm. 102:1048–1054 (1981).
M. Ramanathan. Flow cytometry applications in pharmacodynamics and drug delivery. Pharm. Res. 14:1106–1114 (1997).
K. A. Kronis and J. P. Carver. Specificity of isolectins of wheat germ agglutinin for sialyloligosaccharides: a 360-Mhz proton nuclear magnetic resonance binding study. Biochemistry 21:3050–3057 (1982).
Z. Brich, S. Ravel, T. Kissel, J. Fritsch, and A. Schoffmann. Preparation and characterization of a water soluble dextran immunoconjugate of doxorubicin and the monoclonal antibody (ABL364). J. Contr. Rel. 19:245–258 (1992).
S. D. Ryder, J. A. Smith, E. G. Rhodes, N. Parker, and J. M. Rhodes. Proliferative responses of HT-29 and Caco-2 human colorectal cancer cells to a panel of lectins. Gastroenterology 106:85–93 (1994).
T. Kawamoto, J. D. Sato, A. Le, J. Polikoff, G. H. Sato, and J. Mendelsohn. Growth stimulation of A431 cells by epidermal growth factor: Identification of high-affinity receptors for epidermal growth factor by an anti-receptor monoclonal antibody. Proc. Natl. Acad. Sci. USA 80:1337–1341 (1983).
F. Y. Zeng, A. Benguría, S. Kafert, S. André, H. J. Gabius, and A. Villalobo. Differential response of the epidermal growth factor receptor tyrosine kinase activity to several plant and mammalian lectins. Mol. Cell Biochem. 142:117–124 (1995).
W. Yin and P. W. Cheng. Lectin conjugate-directed gene transfer to airway epithelial cells. Biochem. Biophys. Res. Comm. 205:826–833 (1994).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Wirth, M., Fuchs, A., Wolf, M. et al. Lectin-Mediated Drug Targeting: Preparation, Binding Characteristics, and Antiproliferative Activity of Wheat Germ Agglutinin Conjugated Doxorubicin on Caco-2 Cells. Pharm Res 15, 1031–1037 (1998). https://doi.org/10.1023/A:1011926026653
Issue Date:
DOI: https://doi.org/10.1023/A:1011926026653