title: Involvement of NADPH: cytochrome P450 reductase in the activation of indoloquinone EO9 to free radical and DNA damaging species.
abstract: Evidence suggests that DT-diaphorase is involved in the activation and mechanism of cytotoxicity of the investigational indoloquinone anticancer drug EO9 under aerobic conditions. Data also implicate a role for other enzymes including NADPH: cytochrome P450 reductase, especially in low DT-diaphorase tumour cells and under hypoxic conditions. Here, we used purified rat NADPH: cytochrome P450 reductase to provide additional evidence in support of a role for this enzyme in activation of EO9 to generate free radical and DNA-damaging species. Electron spin resonance spectrometry studies showed that NADPH: cytochrome P450 reductase reduced EO9 to a free radical species, including a drug radical (most likely the semiquinone) and reactive oxygen species. Plasmid DNA experiments showed that reduction of EO9 catalysed by NADPH: cytochrome P450 reductase results in single-strand breaks in DNA. The information obtained may contribute to the understanding of the molecular mechanism of DNA damage and cytotoxicity exerted by EO9 and may be useful in the design of future bioreductive drugs.
authors.affiliation: CRC Department of Medical Oncology, CRC Beatson Laboratories, Garscube Estate, Switchback Road, Bearsden, Glasgow G61 1BD, Scotland, UK.
authors.forename: S M
authors.initials: SM
authors.lastname: Bailey
authors.forename: A D
authors.initials: AD
authors.lastname: Lewis
authors.forename: L H
authors.initials: LH
authors.lastname: Patterson
authors.forename: G R
authors.initials: GR
authors.lastname: Fisher
authors.forename: R J
authors.initials: RJ
authors.lastname: Knox
authors.forename: P
authors.initials: P
authors.lastname: Workman
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000378:metabolism / Q000494:pharmacology; D001388:Aziridines / Q000378:metabolism* / Q000494:pharmacology; D002384:Catalysis; D004247:DNA / Q000187:drug effects* / Q000378:metabolism; D004249:DNA Damage; D004578:Electron Spin Resonance Spectroscopy; D005609:Free Radicals / Q000494:pharmacology*; D045563:Indolequinones; D007211:Indoles / Q000378:metabolism* / Q000494:pharmacology; D009251:NADPH-Ferrihemoprotein Reductase / Q000378:metabolism*; D051381:Rats
chemical_list: D000970:Antineoplastic Agents; D001388:Aziridines; D005609:Free Radicals; D045563:Indolequinones; D007211:Indoles; D004247:DNA; D009251:NADPH-Ferrihemoprotein Reductase; C060817:apaziquone