Bioreductive activation of a series of analogues of 5-aziridinyl-3-hydroxymethyl-1-methyl-2-[1H-indole-4, 7-dione] prop-beta-en-alpha-ol (EO9) by human DT-diaphorase.
The enzyme DT-diaphorase (NAD(P)H:quinone acceptor oxidoreductase, EC 1.6.99.2.; DTD) is believed to be a good target for enzyme-directed bioreductive drug development because elevated levels of enzyme activity have been described in several human tumour types and it plays a key role in the bioreductive activation of several quinone-based anticancer drugs. As part of an ongoing program to develop new bioreductive drugs, the ability of a series of indoloquinone compounds to serve as substrates for and to be bioreductively activated by purified recombinant human DTD was investigated. Of the seven compounds evaluated, EO9, EO68 and EO4 were substrates for human DTD, but only EO4 was reduced to a DNA cross-linking species, and this DNA damage was both concentration dependent and inhibited by dicoumarol. A broad spectrum of chemosensitivity was observed in the H460 non-small cell lung cancer cell line, with the most potent compounds being EO4 (IC50 = 23.9 nM), EO9 (IC50 = 34.5 nM) and EO68 (IC50 = 37.8 nM). Relatively minor structural changes resulted in major changes in both substrate specificity and cytotoxic potency. Comparative chemosensitivity studies demonstrated that EO4, EO9 and EO68 are preferentially toxic towards DTD-rich H460 cells compared with DTD-deficient H596 cells (ratio of IC50 values for H596 cells to H460 cells were 113.8, 92.2 and 103.9 respectively). In conclusion, this study has identified two new compounds that are substrates for human DTD, one of which (EO4) is reduced to a DNA cross-linking species. Further studies in a broad panel of cell lines and human tumour xenografts are warranted for EO4 and EO68 based upon the result of this study.
Phillips
R M
RM
Clinical Oncology Unit, University of Bradford, West Yorkshire, U.K.
D000970:Antineoplastic Agents / Q000378:metabolism*; D001388:Aziridines / Q000378:metabolism* / Q000494:pharmacology; D001711:Biotransformation; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000378:metabolism; D006801:Humans; D045563:Indolequinones; D007211:Indoles / Q000378:metabolism* / Q000494:pharmacology; D007700:Kinetics; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000378:metabolism*; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D001388:Aziridines; D045563:Indolequinones; D007211:Indoles; D004247:DNA; D016660:NAD(P)H Dehydrogenase (Quinone); C060817:apaziquone