Characterization of novel anthracycline prodrugs activated by human β-glucuronidase for use in antibody-directed enzyme prodrug therapy

Pieter H J Houba, Ruben G G Leenders, Epie Boven, Johannes W. Scheeren, Herbert M. Pinedo, Hidde J. Haisma

Research output: Contribution to journalArticlepeer-review


Antibody-directed enzyme prodrug therapy (ADEPT) aims at the specific activation of a prodrug by an enzyme-immuoconjugate localized in tumor tissue. The use of an enzyme of human origin is preferable in ADEPT because it might not be immunogenic when administered to patients. In the case of human β-glucuronidase, prodrugs should be designed that are rapidly and completely activated at a neutral pH. Four new daunorubicin glucuronides were synthesized by coupling a glucuronide group to daunorubicin via an aliphatic (GA1 and GB1) or an aromatic (GA3, GB6) carbamate spacer, to be released by electron shift (A-type) or by ring closure (B-type). These prodrugs were characterized in vitro for their usefulness in ADEPT and were compared with the previously described prodrugs epirubicin-glucuronide and doxorubicin-nitrophenylglucuronide. The four new prodrugs were stable in serum, hydrophilic when compared to the lipophilic daunorubicin, and at least 20-fold less toxic than the parent compound. The hydrolysis rate at clinically relevant enzyme and prodrug concentrations (1 μg/mL human β-glucuronidase, 100 μM prodrug) at pH 6.8 were similar for GA3 (T( 1/4 ) 160 min) and higher for GB6 (T( 1/4 ) 40 min) when compared to that of doxorubicin-nitrophenylglucuronide (T( 1/4 ) 170 min). Epirubicin-glucuronide, GA1, and GB1 showed a low hydrolysis rate (T( 1/4 ) > 400 min). GA1 and GA3, but not GB1 or GB6, were activated to the parent compound. Complete activation was confirmed in OVCAR-3 cells pretreated with a specific antibody-human β-glucuronidase conjugate, where GA3 had similar antiproliferative effects to those of daunorubicin.

Original languageEnglish (US)
Pages (from-to)455-463
Number of pages9
JournalBiochemical Pharmacology
Issue number3
StatePublished - Aug 9 1996
Externally publishedYes


  • anthracycline-glucuronide prodrugs
  • daunorubicin
  • human β-glucuronidase

ASJC Scopus subject areas

  • Pharmacology


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