Bystander killing of tumour cells by antibody-targeted enzymatic activation of a glucuronide prodrug

Summary

RHI-βG-PEG, formed by linking poly(ethylene glycol)-modified β-glucuronidase to Mab RH1, was employed to examine bystander killing of antigen-negative N1S1 rat hepatoma cells by activation of a glucuronide prodrug (BHAMG) of p-hydroxyaniline mustard (pHAM) at antigen-positive AS-30D rat hepatoma cells. Sequential treatment of cells with 10 μg ml–1 RH1-βG-PEG and 20 μM BHAMG was not toxic to N1S1 cells but killed 99% of AS-30D cells. Over 98% of N1S1 cells, however, were killed in mixed populations containing as few as 2% AS-30D cells after identical treatment, demonstrating an in vitro bystander effect. Subcutaneous injection of AS-30D and N1S1 cells in BALB/c nu/nu mice produced solid tumours containing both cells. Uptake of radiolabelled RH1-βG-PEG in solid AS-30D and mixed AS-30D/N1S1 tumours was 11.6 and 9.3 times greater than a control antibody conjugate 120 h after i.v. injection. Intravenous treatment with RH1-βG-PEG and BHAMG cured seven of seven nude mice bearing solid s.c. AS-30D tumours and significantly delayed, compared with control conjugate and prodrug treatment, the growth of mixed N1S1/AS-30D tumours with one cure, showing that targeted activation of BHAMG kills bystander tumour cells in vivo.

Change history

  • 16 November 2011

    This paper was modified 12 months after initial publication to switch to Creative Commons licence terms, as noted at publication

References

  1. Bagshawe, K. D., Springer, C. J., Searle, F., Antoniw, P., Sharma, S. K., Melton, R. G. & Sherwood, R. F. (1988). A cytotoxic agent can be generated selectively at cancer sites. Br J Cancer 58: 700–703.

    CAS  Article  Google Scholar 

  2. Baxter, L. T., Yuan, F. & Jain, R. K. (1992). Pharmacokinetic analysis of the perivascular distribution of bifunctional antibodies and haptens: comparison with experimental data. Cancer Res 52: 5838–5844.

    CAS  PubMed  Google Scholar 

  3. Chari, R. V. J., Jackel, K. A., Bourret, L. A., Derr, S. M., Tadayoni, B. M., Mattocks, K. M., Shah, S. A., Liu, C., Blättler, W. A. & Goldmacher, V. S. (1995). Enhancement of the selectivity and antitumor efficacy of a CC-1065 analogue through immunoconjugate formation. Cancer Res 55: 4079–4084.

    CAS  PubMed  Google Scholar 

  4. Chen, B. M., Chan, L. Y., Wang, S. M., Wu, M. F., Chern, J. W. & Roffler, S. R. (1997). Cure of malignant ascites and generation of protective immunity by monoclonal antibody targeted activation of a glucuronide prodrug in rats. Int J Cancer 73: 392–402.

    CAS  Article  Google Scholar 

  5. Cheng, T. L., Chen, B. M., Chan, L. Y., Wu, P. Y., Chern, J. W. & Roffler, S. R. (1997). Poly(ethylene glycol) modification of β-glucuronidase-antibody conjugates for solid tumor therapy by targeted activation of glucuronide prodrugs. Cancer Immun Immunother 44: 305–315.

    CAS  Article  Google Scholar 

  6. Douillard, J. Y., Lehur, P. A., Aillet, G., Kremer, M., Bianco-Arco, A., Peltier, P. & Chatal, J. F. (1986). Immunohistochemical antigenic expression and in vivo tumor uptake of monoclonal antibodies with specificity for tumors of the gastrointestinal tract. Cancer Res 46: 4221–4224.

    CAS  PubMed  Google Scholar 

  7. Ejigu Retta, B. M., Burke, P. J., Photiou, A., Melton, R. & Eno-Amooquaye, E. (1996). Antibody-directed enzyme prodrug therapy (ADEPT): evidence for a bystander effect in vitro. Int J Oncol 9: 567–570.

    Google Scholar 

  8. Fujimori, K., Covell, D. G., Fletcher, J. E. & Weinstein, J. N. (1990). A modeling analysis of monoclonal antibody percolation through tumors: a binding-site barrier. J Nucl Med 31: 1191–1198.

    CAS  PubMed  Google Scholar 

  9. Gerlowski, L. E. & Jain, R. K. (1986). Microvascular permeability of normal and neoplastic tissues. Microvascular Res 31: 288–305.

    CAS  Article  Google Scholar 

  10. Jain, R. K. (1990). Physiological barriers to delivery of monoclonal antibodies and other macromolecules in tumors. Cancer Res 50: 814–819.

    Google Scholar 

  11. Less, J. R., Posner, M. C., Boucher, Y., Borochovitz, D., Wolmark, N. & Jain, R. K. (1992). Interstitial hypertension in human breast and colorectal tumors. Cancer Res 52: 6371–6374.

    CAS  PubMed  Google Scholar 

  12. Roffler, S. R., Wang, S. M., Chern, J. W., Yeh, M. Y. & Tung, E. (1991). Anti-neoplastic glucuronide prodrug treatment of human tumor cells targeted with a monoclonal antibody–enzyme conjugate. Biochem Pharmacol 42: 2062–2065.

    CAS  Article  Google Scholar 

  13. Roffler, S. R., Chan, J. & Yeh, M. Y. (1994). Potentiation of radioimmunotherapy by inhibition of topoisomerase I. Cancer Res 54: 1276–1285.

    CAS  PubMed  Google Scholar 

  14. Rosenblum, M. G., Kavanagh, J. J., Burke, T. W., Wharton, J. T., Cunningham, J. E., Shanken, L. J., Silva, E. G., Thompson, L., Cheung, L., Lamki, L. & Murray, J. L. (1991). Clinical pharmacology, metabolism, and tissue distribution of 90Y-labeled monoclonal antibody B72.3 after intraperitoneal administration. J Natl Cancer Inst 83: 1629–1636.

    CAS  Article  Google Scholar 

  15. Sahin, U., Hartmann, F., Senter, P., Pohl, C., Engert, A., Diehl, V. & Pfreundschuh, M. (1990). Specific activation of the prodrug mitomycin phosphate by a bispecific anti-CD30/anti-alkaline phosphatase monoclonal antibody. Cancer Res 50: 6944–6948.

    CAS  PubMed  Google Scholar 

  16. Senter, P. D., Saulnier, M. G., Schreiber, G. J., Hirschberg, D. L., Brown, J. P., Hellström, I. & Hellström, K. E. (1988). Anti-tumor effects of antibody-alkaline phosphatase conjugates in combination with etoposide phosphate. Proc Natl Acad Sci USA 85: 4842–4846.

    CAS  Article  Google Scholar 

  17. Shockley, T. R., Lin, K., Sung, C., Nagy, J. A., Tompkins, R. G., Dedrick, R. L., Dvorak, H. F. & Yarmush, M. L. (1992). A quantitative analysis of tumor specific monoclonal antibody uptake by human melanoma xenografts: effects of antibody immunological properties and tumor antigen expression levels. Cancer Res 52: 357–366.

    CAS  PubMed  Google Scholar 

  18. Smith, D. F., Walborg, E. F. & Chang, J. P. (1970). Establishment of a transplantable ascites variant of a rat hepatoma induced by 3′-methyl-4-dimethylaminoazobenzene. Cancer Res 30: 2306–2309.

    CAS  PubMed  Google Scholar 

  19. Sung, C., Dedrick, R. L., Hall, W. A., Johnson, P. A. & Youle, R. J. (1993). The spatial distribution of immunotoxins in solid tumors: assessment by quantitative autoradiography. Cancer Res 53: 2092–2099.

    CAS  PubMed  Google Scholar 

  20. Tomlinson, E. (1990). Biological dispersion and the design of site-specific protein therapeutic systems. In Targeting of Drugs 2, Optimization Strategies. Gregoriadis G, Allison AC and Poste G (eds), pp. 1–19. Plenum Press: New York

    Google Scholar 

  21. Wallace, P. M., Macmaster, J. F., Smith, V. F., Kerr, D. E., Senter, P. D. & Cosand, W. L. (1994). Intratumoral generation of 5-fluorouracil mediated by an antibody-cytosine deaminase conjugate in combination with 5-fluorocytosine. Cancer Res 54: 2719–2723.

    CAS  PubMed  Google Scholar 

  22. Wang, S. M., Chern, J. W., Yeh, M. Y., Ng, J. C., Tung, E. & Roffler, S. R. (1992). Specific activation of glucuronide prodrugs by antibody-targeted enzyme conjugates for cancer therapy. Cancer Res 52: 4484–4491.

    CAS  PubMed  Google Scholar 

Download references

Author information

Affiliations

Authors

Rights and permissions

From twelve months after its original publication, this work is licensed under the Creative Commons Attribution-NonCommercial-Share Alike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/

Reprints and Permissions

About this article

Cite this article

Cheng, T., Wei, S., Chen, B. et al. Bystander killing of tumour cells by antibody-targeted enzymatic activation of a glucuronide prodrug. Br J Cancer 79, 1378–1385 (1999). https://doi.org/10.1038/sj.bjc.6690221

Download citation

Keywords

  • bystander effect
  • antibody-directed enzyme prodrug therapy
  • monoclonal antibody
  • prodrugs
  • β-glucuronidase
  • immunoconjugates

Further reading