Abstract
The enzyme nitroreductase from E. coli can reduce the relatively non-toxic prodrug 5-(aziridin-1-yl)-2,4-dinitrobenzamide (CB1954) to the 2- or 4-hydroxylamino derivatives; the latter is a potent cytotoxic agent, which following further non-enzymatic reaction with thioesters, leads to the generation of interstrand crosslinks in DNA [1–4] (Figure 1a). These cannot be efficiently repaired by the cell, and lead to cell death. Nitroreductase and CB1954 are therefore of interest as an enzyme-prodrug combination for “virus directed enzyme-prodrug therapy” (VDEPT) of cancer [5, 6]. We describe here the use of retroviral and adenoviral vectors to deliver the gene to human ovarian cancer cells in vitro, and their consequent sensitisation to CB1954.
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Abbreviations
- CB1954:
-
5-(aziridin-1-yl)-2,4-dinitrobenzamide
- cfu:
-
colony forming units
- CMVIE:
-
cytomegalovirus immediate-early (promoter)
- IC50 :
-
concentration of prodrug giving 50% cell survival
- m.o.i.:
-
multiplicity of infection
- pfu:
-
plaque forming units
- VDEPT:
-
virus directed enzyme/prodrug therapy
References
R. J. Knox, et al., 1992: The bioactivation of 5-(aziridin-1-yl)-2,4-dinitrobenzamide (CB 1954). II. A comparison of an Escherichia coli nitroreductase and Walker DT diaphorase. Biochem. Pharmacol. 44, 2297–2301
R. J. Knox, et al., 1988: A new cytotoxic, DNA interstrand crosslinking agent, 5-(aziridin-1-yl)-4-hydroxy-lamino-2-nitrobenzamide, is formed from 5-(aziridin-1-yl)-2,4-dinitrobenzamide (CB 1954) by a nitroreductase enzyme in Walker carcinoma cells. Biochem. Pharmacol. 37, 4661–4669
G. M. Anlezark, et al., 1992: The bioactivation of 5-(aziridin-1-yl)-2,4-dinitrobenzamide (CB 1954)—I. Purification and properties of a nitroreductase enzyme from Escherichia coli- a potential enzyme for antibody-directed enzyme prodrug therapy (ADEPT). Biochem. Pharmacol. 44, 2289–2295
R. J. Knox, et al., 1993: Identification, synthesis and properties of 5-(aziridin-1-yl)-2-nitro-4-nitrosoben-zamide, a novel DNA crosslinking agent derived from CB1954. Biochem. Pharmacol. 46, 797–803
J. A. Bridgewater, et al., 1995: Expression of the bacterial nitroreductase enzyme in mammalian cells renders them selectively sensitive to killing by the prodrug CB1954. Eur. J. Cancer 31A, 2362–2370
N. K. Green, et al., 1997: Sensitization of colorectal and pancreatic cancer cell lines to the prodrug 5-(aziridin-1-yl)-2,4-dinitrobenzamide (CB1954) by retroviral transduction and expression of the E. coli nitroreductase gene. Cancer Gene Therapy 4, 229–238
D. Markowitz, S. Goff and A. Bank, 1988: Construction and use of a safe and efficient amphotropic packaging cell line. Virology 167, 400–406
F.-L. Cosset, et al., 1995: High-titer packaging cells producing recombinant retroviruses resistant to human serum. J. Virol. 69, 7430–7436
N. E. Bowles, et al., 1996: A simple and efficient method for the concentration and purification of recombinant retrovirus for increased hepatocyte transduction in vivo. Human Gene Therapy 7, 1735–1742
G. W. G. Wilkinson and A. Akrigg, 1992: Constitutive and enhanced expression from the CMV major IE promoter in a defective adenovirus vector. Nucleic Acids Res. 20, 2233–2239
E. Frei, et al., 1988: Preclinical studies and correlation of the effects of alkylating dose. Cancer Res. 48, 6417–6423
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© 1998 Springer Science+Business Media New York
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Searle, P.F. et al. (1998). Sensitisation of Human Ovarian Cancer Cells to Killing by the Prodrug CB1954 Following Retroviral or Adenoviral Transfer of the E. coli Nitroreductase Gene. In: Walden, P., Trefzer, U., Sterry, W., Farzaneh, F., Zambon, P. (eds) Gene Therapy of Cancer. Advances in Experimental Medicine and Biology, vol 451. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-5357-1_17
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DOI: https://doi.org/10.1007/978-1-4615-5357-1_17
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