A methylester of the glucuronide prodrug DOX-GA3 for improvement of tumor-selective chemotherapy

Michelle De Graaf, Tapio J. Nevalainen, Hans W. Scheeren, Herbert M. Pinedo, Hidde J. Haisma, Epie Boven

Research output: Contribution to journalArticlepeer-review

32 Scopus citations


The glucuronide prodrug of doxorubicin, DOX-GA3, can be selectively activated in tumors by extracellular human β-glucuronidase, resulting in a better therapeutic index than doxorubicin. DOX-GA3, however, is rapidly excreted by the kidney. We hypothesized that slow release of DOX-GA3 from its methylester, DOX-mGA3, by esterase activity in blood would result in improved circulation half-life (t 1/2) of DOX-GA3. DOX-mGA3 was synthesized more efficiently with an overall yield of 60% as compared to 37% in the case of DOX-GA3. We showed that DOX-mGA3 was enzymatically converted to DOX-GA3 with a t 1/2 of approximately 0.5 min in mouse plasma to 2.5 h in human plasma, which was in agreement with differences in esterase activity between species. DOX-mGA3, similar to DOX-GA3, was at least 37-fold less potent than the parent drug doxorubicin in growth inhibition of four different human malignant cell lines in vitro. Incubation of OVCAR-3 cells with DOX-mGA3 in combination with an excess of human β-glucuronidase (0.05 U mL -1) resulted in a similar growth inhibition to that of doxorubicin. Intravenous administration of DOX-mGA3 in FMa-bearing mice resulted in an area under the concentration versus time curve (AUC) of DOX-GA3 in tumor and most normal tissues that was 2.5- to 3-fold higher than after the same dose of DOX-GA3 itself. In tumor tissue, this was accompanied by a 2.7-fold increase in the AUC of doxorubicin from DOX-mGA3 than from DOX-GA3. In conclusion, an advantage of DOX-mGA3 over DOX-GA3 is that this prodrug can be produced with a higher yield. Another important advantage is the improved pharmacokinetics of the lipophilic DOX-mGA3 as compared to that of the hydrophilic DOX-GA3. This effect may even be more pronounced in man, because of the lower plasma esterase activity than measured in mice.

Original languageEnglish (US)
Pages (from-to)2273-2281
Number of pages9
JournalBiochemical Pharmacology
Issue number11
StatePublished - Dec 1 2004
Externally publishedYes


  • β-Glucuronidase
  • Cancer chemotherapy
  • Doxorubicin
  • Esterases
  • Glucuronide
  • Prodrug

ASJC Scopus subject areas

  • Pharmacology


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