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E. coli nitroreductase/CB1954 gene-directed enzyme prodrug therapy: role of arylamine N-acetlytransferase 2

Cancer Gene Therapy volume 15, pages 758–764 (2008)Cite this article

Abstract

Gene-directed enzyme prodrug therapy is a promising approach to the local management of cancer and a number of gene prodrug combinations have entered clinical trials. The antitumor activity of Escherichia coli nitroreductase (NTR) in combination with the prodrug CB1954 relies on the reduction of the nitro groups to reactive N-hydroxylamine intermediates that are toxic in proliferating and nonproliferating cells. We examined whether secondary metabolic activation of the N-hydroxylamines by sulfotransferases or acetyltransferases altered cell responsiveness to the drug. We evaluated the coexpression of NTR with the human cytosolic sulfotransferases SULT1A1, 1A2, 1A3, 1E1 and 2A1, or the human arylamine N-acetyltransferases NAT1 and NAT2 on SKOV3 cell survival. Only NAT2 significantly altered the toxicity of CB1954, decreasing the IC50 16-fold from 0.61 to 0.04 μM. These results suggest that one or more of the N-hydroxyl metabolites are a substrate for O-acetylation by NAT2. We also examined the bystander effect of SKOV3 cells expressing NTR or NTR plus NAT2. Addition of the acetyltransferase resulted in a significant decreased bystander effect (P>0.01), possibly due to a lower concentration of reactive metabolites in the culture medium. These results suggest that a combination of bacterial NTR and NAT2 may provide a greater clinical response at therapeutic concentrations of CB1954 provided the reduction in bystander effect is not clinically significant. Moreover, endogenous NAT2, which is localized predominantly in the liver and gut, may be involved in the dose-limiting hepatic toxicity and gastrointestinal side effects seen in patients treated with the higher doses of CB1954.

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Abbreviations

CB1954:

5-(aziridin-1-yl)-2,4-dinitrobenzamide

GDEPT:

gene-directed enzyme prodrug therapy

NAT1:

N-acetyltransferase

NTR:

nitroreductase

SULT:

sulfotransferase

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Acknowledgements

This study was supported by the National Health and Medical Research Council. CB1954 was a kind gift from Professor William Denny, University of Auckland. The parental SKOV3 cells and the SKOV3-NTR cells were kindly provided by Dr Peter Searle, University of Birmingham.

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  1. School of Biomedical Sciences, University of Queensland, St Lucia, Queensland, Australia

    D J Mitchell & R F Minchin

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  1. D J Mitchell
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Correspondence to R F Minchin.

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Mitchell, D., Minchin, R. E. coli nitroreductase/CB1954 gene-directed enzyme prodrug therapy: role of arylamine N-acetlytransferase 2. Cancer Gene Ther 15, 758–764 (2008). https://doi.org/10.1038/cgt.2008.47

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