Characterization of an Etoposide-Glutathione Conjugate Derived from Metabolic Activation by Human Cytochrome P450

Author(s): Naiyu Zheng, Shaokun Pang, Tomoyuki Oe, Carolyn A. Felix, Suzanne Wehrli, Ian A. Blair.

Journal Name: Current Drug Metabolism

Volume 7 , Issue 8 , 2006


Etoposide (VP-16), a DNA topoisomerase II poison widely used as an antineoplastic agent is also known to cause leukemia. One of its major metabolic pathways involves O-demethylation to etoposide catechol (etoposide-OH) by cytochrome P450 3A4 (CYP3A4). The catechol metabolite can undergo sequential one- and two-electron oxidations to form etoposide semi-quinone (etoposide-SQ) and etoposide quinone (etoposide-Q), respectively, which have both been implicated as cytotoxic metabolites. However, etoposide-Q is known to react with glutathione (GSH), which can protect DNA from oxidative damage by this reactive metabolite. In this study, etoposide-Q was reacted with GSH and the two etoposide-GSH conjugates were characterized. The major conjugate was etoposide-OH-6-SG and the minor product was etoposide-OH-2-SG. Etoposide-OH-6-SG, which arose from Michael addition of GSH to etoposide-Q, was characterized by mass spectrometry and 2-D NMR. It was identified as the sole product from in vitro metabolism experiments using recombinant human CYP3A4 or liver microsomes incubated with etoposide in the presence of GSH. Etoposide-OH- 6-SG was also detected from incubations of etoposide-OH and GSH alone. Therefore, the presence of etoposide-OH, which can be formed from etoposide metabolism by CYP3A4, is essential for formation of the GSH conjugate. The oxidation of etoposide-OH to a quinone intermediate is likely the precursor in the formation of etoposide-OH-6-SG.

Keywords: Etoposide metabolism, glutathione conjugate, etoposide catechol, etoposide quinone, human liver microsomes, cytochrome P450, mass spectrometry, NMR spectroscopy

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Article Details

Year: 2006
Page: [897 - 911]
Pages: 15
DOI: 10.2174/138920006779010638
Price: $58

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