Skip to main content
SpringerLink
Log in

Pharmacokinetic and immunogenic behavior of three recombinant human GM-CSF-EPO hybrid proteins in cynomolgus monkeys

  • Research
  • Published:
Molecular Biotechnology Aims and scope Submit manuscript

Abstract

MEN 11300, MEN 11301, and MEN 11303 are three recombinant human hybrid proteins that, as has recently been described, induce in vitro erythroid differentiation. This article provides data on their pharmacokinetic and immunogenic behavior after repeated iv administration to cynomolgus monkeys at 0.8 or 1.6 μg/kg doses. Pharmacokinetic data, obtained after the first administration, showed that the half-life (t 1/2) and clearance (CL) values are dose dependent, with no significant differences among the three hybrid proteins. After the tenth administration, MEN 11300 and MEN 11301, both at high and low dose, and MEN 11303 at high dose were undetectable in plasma, whereas MEN 11303 at the lower dose showed no alteration in its pharmacokinetic profile. Immunologic analyses of plasma provided an explanation for this different pharmacokinetic behavior. In fact, plasma samples from animals treated repeatedly with MEN 11300 and MEN 11301 showed specific antibody formation in response to both the high- and the low-dose regimens. These antibodies exerted in vitro a strong neutralizing activity of the hybrid proteins, with a predominant specificity for the erythropoietin (EPO) portion. By contrast, MEN 11303 at the lower dose did not induce a detectable antibody response whereas the antibodies observed on the high-dose regimen did not exert neutralizing activity against the hybrid proteins nor against granulocyte-macrophage colony-stimulating factor (GM-CSF) or EPO. Hematologic parameters were not affected by the treatments, thus indicating that the anti-EPO neutralizing antibody response does not cross react with the endogenous monkey cytokine. The overall immunogenicity data suggest that among the three fusion proteins, MEN 11303 could have a lower immunogenic potential.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Coscarella, A., Carloni, C., Liddi, R., Mauro, S., Novelli, S., Mele, A., Massella, B., Valtieri, M., and De Santis, R. (1997) Production of recombinant human GM-CSF-EPO hybrid protein:in vitro biological characterization.Eur. J. Haematol. 59, 238–246.

    Article  PubMed  CAS  Google Scholar 

  2. Benedetti Panici, P., Pierelli, L., Scambia, G., Foddai, M. L., Salerno, M. G., Menichella, G., Vittori, M., Maneschi, F., Caracussi, U., Serafini, R., Leone, G., and Mancuso, S. (1997) High-dose carboplatin, etoposide, and melphan (CEM) with peripheral blood progenitor cell support as late intensification for highrisk cancer: nonhaematologic, haematologic toxicities and role of growth factor administration.Br. J. Cancer 75, 1205–1212.

    PubMed  CAS  Google Scholar 

  3. Khan, K. N. M., Kats, A. A., Foutan, M., Monique, M., Snook, S. S., McKearn, J. P., Alden, C. L., and Smith, P. F. (1996) Recombinant human interleukin-3 induces extramedullary hematopoiesis at subcutaneous injection sites in cynomolgus monkeys.Toxicol. Pathol. 24(4), 391–397.

    Article  PubMed  CAS  Google Scholar 

  4. Krumwieh, D., Weinmann, E., Siebold, B., and Seiler, F. R. (1990) Preclinical studies on synergistic effects of IL-1, IL-3, G-CSF and GM-CSF in cynomolgus monkeys.Int. J. Cell. Cloning 8, 229–248.

    Article  PubMed  CAS  Google Scholar 

  5. Donahue, R., Wang, E., Stone, D., Kamen, R., Wong, G. G., Sehgal, P. K., Nathan, D. G., and Clark S. C. (1986) Stimulation of haematopoiesis in primates by continuous infusion of recombinant human GM-CSF.Nature 321, 872–875.

    Article  PubMed  CAS  Google Scholar 

  6. Donahue, R., Seehra, J., Metzger, M., Lefebvre, D., Rock, B., Carbone, S., Nathan, D. G., Garnick, M., Sehgal, P. K., Laston, D., Lavallie, E., McCoy, J., Schendel, P. F., Norton, C., Turner, K., Yang, Y., and Clark, S. C. (1988) Human IL-3 and GM-CSF act synergistically in stimulating hematopoiesis in primates.Science 241, 1820–1823.

    Article  PubMed  CAS  Google Scholar 

  7. Krumwied, D., Siebold, B., Weinmann, E., and Seiler, F. R. (1991) Combined effects of granulocytemacrophage colony stimulating factor with erythropoietin in subhuman primates.Am. J. Clin. Oncol. 14, 1–4.

    Article  Google Scholar 

  8. Gibaldi, M. and Perrier, D. (1982)Pharmacokinetics, 2nd ed. Marcel Dekker, New York.

    Google Scholar 

  9. D'Andrea, A. D., Youssoufian, H., Zon, L., Koo, J. W., and Lodish, H. (1991) The cytoplasmatic region of the erythropoietin receptor contains non overlapping positive and negative growth-regulatory domains.Mol. Cell. Biol. 11, 1980–1987.

    PubMed  Google Scholar 

  10. Lange, B., Valtieri, M., Santoli, D., Caracciolo, D., Mavilio, F., Gemperlin, I., Griffin, C., Emanuel, B., Finan, J., Nowell, P., et al. (1987) Growth factor requirements of childhood acute leukemia: establishment of GM-CSF-dependent cell lines.Blood 70, 192–199.

    PubMed  CAS  Google Scholar 

  11. George, J., Bracco, C., Shannon, K., Davis, J., Smith, I. L., Phibbs, R. H., and Hendrickx, A. G. (1990) Agerelated differences in erythropoietic response to recombinant human erythropoietin: comparison in adult and infant rhesus monkeys.Pediatric Res. 28, 567–571.

    Article  CAS  Google Scholar 

  12. Coscarella, A., Liddi, R., Di Loreto, M., Bach, S., Faiella, A., van der Meide, P. H., Mele, A., and De Santis, R. (1998) The rhGM-CSF-EPO hybrid protein MEN 11300 induces anti-EPO antibodies and severe anemia in rhesus monkeys.Cytokine, in press.

  13. Goochee, C. H., Gramer, M., Andersen, D., Bahr, J., and Rasmussen, J. (1991) The oligosaccharides of glycoproteins: bioprocess factors affecting oligosaccharide structure and their effect on glycoprotein properties.Biotechnology 9, 1347–1355.

    Article  PubMed  CAS  Google Scholar 

  14. Quakyi, I. A., Miller, L. H., Good, M. F., Ahlers, J. D., Isaacs, S. N., Nunberg, J. H., Houghte, R. A., Keister, D. B., Coligan, J. E., Moss, B., et al. (1995) Synthetic peptides from P. falciparum sexual stage 25-kDa protein induce antibodies that react with the native protein: the role of IL-2 and conformational structure on immunogenicity of Pfs25.Pept. Res. 8, 335–344.

    PubMed  CAS  Google Scholar 

  15. Jones, T., Stern, A., and Lin, R. (1994) Potential role of granulocyte-macrophage colony-stimulating factor as vaccine adjuvant.Eur. J. Clin. Microbiol. Infect. Dis. 13, 47–53.

    Article  Google Scholar 

  16. Tao, M. H. and Levy, R. (1993) Idiotype/granulocyte-macrophage colony-stimulating factor fusion protein as a vaccine for B-cell lymphoma.Nature 362, 755–758.

    Article  PubMed  CAS  Google Scholar 

  17. Hopp, T. H. and Woods, K. (1981) Prediction of protein antigenic determinants from amino acid sequences.Proc. Natl. Acad. Sci. 78, 3824–3828.

    Article  PubMed  CAS  Google Scholar 

  18. Amoresano, A., Andolfo, A., Siciliano, R. A., Mele, A., Coscarella, A., De Santis, R., Mauro, S., Pucci, P., and Marino, G. (1998) Structural characterization and independent folding of a chimeric glycoprotein comprising Granulocyte-Macrophage Colony Stimulating Factor and Erythropoietin sequences.Glycobiology 8, 779–790.

    Article  PubMed  CAS  Google Scholar 

  19. Vadhan-Raj, S., Papadopoulos, N. E., Burgess, M., Linke, K. A., Patel, S. R., Hays, C., Arcenas, A., Plager, C., Kudela, A. P., Hittelman, W. N., Broxmeyer, H. E., Williams, D. P., Garrison, L., and Benjamin, R. S. (1994) Effects of PIXY321, a granulocyte-macrophage colony-stimulating factor/interleukin-3 fusion protein, on chemotherapy-induced multilineage myelosuppression in patients with sarcoma.J. Clin. Oncol. 12, 715–724.

    PubMed  CAS  Google Scholar 

  20. Vadhan-Raj, S., Broxmeyer, H., Andreeff, M., Bandres, J. C., Buescher, E. S., Benjamin, R. S., Papadopoulos, N. E., Burgess, A., Patel, S., Plager, C., Hittelman, W. N., McAlister, I., Garrison, L., and Williams, D. E. (1995) In vivo biological effects of PIXY321, a synthetic hybrid protein of recombinant human granulocyte-macrophage colony-stimulating factor and interleukin-3 in cancer patients with normal hematopoiesis: a phase I study.Blood 86, 2098–2105.

    PubMed  CAS  Google Scholar 

  21. O'Shaughnessy, J. A., Tolcher, A., Riseberg, D., Venzon, D., Zujewski, J. A., Noone, M.,Gossard, M., Danforth, D., Jacobson, J., Chang, V., Goldspiel, B., Keegan, P., Giusti, R., Cowan K. H. (1996) Prospective, randomized trial of 5-fluorouracil, Leuconvorin, Doxorubicin, and cyclophosphamide chemotherapy in combination with the interleukin-3/GM-CSF fusion protein (PIXY321) versus GM-CSF in patients with advanced breast cancer.Blood 87, 2205–2211.

    PubMed  Google Scholar 

  22. Wen, D., Boissel, J. P. R., Tracy, T. E., Gruninger, R. H., Mulcahy, L. S., Czelusniak, J., Goodman, M., and Bunn, H. F. (1993) Erythropoietin structure-function relationships: high degree of sequence homology among mammals.Blood 82, 1507–1576.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Biotechnology Research Department, Menarini Ricerche S.p.A., Via Tito Speri 10-00040 Pomezia, Rome, Italy

    Annamaria Coscarella, Roberto Liddi, Simona Bach, Sabrina Zappitelli, Antonio Mele & Rita De Santis

  2. Pharmacokinetics and Metabolism Department, Menarini Ricerche S.p.A., Via Tito Speri 10-00040 Pomezia, Rome, Italy

    Renato Urso

Authors
  1. Annamaria Coscarella
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Roberto Liddi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Simona Bach
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sabrina Zappitelli
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Renato Urso
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Antonio Mele
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Rita De Santis
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Annamaria Coscarella.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Coscarella, A., Liddi, R., Bach, S. et al. Pharmacokinetic and immunogenic behavior of three recombinant human GM-CSF-EPO hybrid proteins in cynomolgus monkeys. Mol Biotechnol 10, 115–122 (1998). https://doi.org/10.1007/BF02760860

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02760860

Index Entries

Search

Navigation