A monoclonal antibody-β-glucuronidase conjugate as activator of the prodrug epirubicin-glucuronide for specific treatment of cancer

H. J. Haisma, E. Boven, M. Van Muijen, J. De Jong, W. J F Van der Vijgh, H. M. Pinedo

Research output: Contribution to journalArticle

Abstract

The anti-pan carcinoma monoclonal antibody (MAb) 323/A3, linked to E. coli-derived β-glucuronidase (GUS) was used to study the tumour-site-selective activation of the prodrug Epirubicin-glucuronide (Epi-glu). Epi-glu was isolated from the urine of patients treated with Epirubicin (Epi) by reversed phase chromatography on a silica-C18 column. Epi-glu was stable in human blood and was not converted into Epi by A2780, MCF-7, or OVCAR-3 cancer cells, despite the presence of intracellular GUS. The stability of the prodrug was confirmed in BALB/c mice. MAb 323/A3 and GUS were linked through a stable thioether bond. The conjugate (1:1) was purified by ion exchange and gel filtration chromatography. Binding to target cells revealed an immunoreactivity of at least 60% and good retention of enzyme activity. A protein dye (sulforhodamine B) assay was used to analyse cytotoxicity. Epi (IC50 of 0.003-0.21 μM) was 100-1000 times more toxic than Epi-glu (IC50 of >20 μM), when cancer cells were exposed for 4 or 24h to the drugs. The low cytotoxicity of Epi-glu was most likely due to the reduced cellular uptake rate of the prodrug (2.7 pmol 10-6 cells min-1) as compared to that of the parent compound (25 pmol 10-6 cells min-1). Pretreatment of antigen-positive cells with the 323/A3-GUS conjugate prior to prodrug exposure completely restored cytotoxicity as a result from hydrolysis of Epi-glu into Epi. Our results demonstrate that the 323/A3-GUS conjugate can specifically activate the stable non-toxic prodrug Epi-glu at the tumour cell level.

Original languageEnglish (US)
Pages (from-to)474-478
Number of pages5
JournalBritish Journal of Cancer
Volume66
Issue number3
StatePublished - 1992
Externally publishedYes

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

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    Haisma, H. J., Boven, E., Van Muijen, M., De Jong, J., Van der Vijgh, W. J. F., & Pinedo, H. M. (1992). A monoclonal antibody-β-glucuronidase conjugate as activator of the prodrug epirubicin-glucuronide for specific treatment of cancer. British Journal of Cancer, 66(3), 474-478.