Abstract
The prodrug CB1954 (5-[aziridin-1-yl]-2,4-dinitrobenzamide) is a weak monofunctional alkylating agent originally synthesized at the Chester Beatty Laboratories in the late 1960s (1). The antitumor activity of CB1954 was determined by screening a panel of aziridines for cytotoxicity against the rat Walker 256 carcinoma. It is highly selective and effective against Walker tumor cells in vivo and in vitro with a therapeutic index of 70 (2). Based on the efficacy against the Walker carcinoma, a small clinical trial using CB1954 was initiated at the Royal Marsden Hospital in 1970. The results of this study are unpublished; however, a dose-limiting toxicity was reached (the major side effect being diarrhea) without any evidence of tumor regression (Wiltshaw, unpublished data). Recently, a clinical trial to determine the maximum tolerated dose (MTD) of CB1954 by intravenous and intraperitoneal routes has been performed in Birmingham. This study concluded that the MTD for CB1954 is 24 mg/m2 by iv administration and 37.5 mg/m2 by the ip route. Toxicity included diarrhea and elevation of liver transaminases (3).
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References
Khan, A. H. and Ross, W. C. J. (1969/1970) Tumor growth inhibitory nitropheny-laziridines and related compounds: structure activity relationships. Chem. Biol. Interact. 1, 27–47.
Cobb, L. M., Connors, T. A., Elson, L. A., et al. (1969) 2,4-dinitro-5-ethyleneiminobenzamide (CB1954): a potent and selective inhibitor of the growth of the Walker carcinoma 256. Biochem. Pharmacol., 18, 1519–1527.
Chung-Faye, G. A., Palmer, D., Anderson, D., et al. (2001) Virus directed, enzyme prodrug therapy with Nitroimidazole Reductase: A phase I and pharmacokinetic study of its prodrug, CB1954. Clin. Cancer Res., in press.
Knox, R. J., Boland, M., Friedlos, F., Coles, B., Southan, C., and Roberts, J. J. (1988) The nitroreductase enzyme in Walker cells that activates 5-(aziridin-1-yl)-2,4-dinitrobenzamide (CB 1954) to 5-(aziridin-1-yl)-4-hydroxylamino-2-nitrobenzamide is a form of NAD(P)H dehydrogenase (quinone) (EC 1.6.99.2). Biochem. Pharmacol. 37, 4661–4669.
Knox, R. J., Friedlos, F., Marchbank, T. and Roberts, J. J. (1991) Bioactivation of CB1954: reaction of the active 4-hydroxylamino derivative with thioesters to form the ultimate DNA-DNA interstrand crosslinking species. Biochem. Pharmacol. 42, 1691–1697.
Anlezark, G. M., Melton, R. G., Sherwood, R. F., Coles, B., Friedlos, F. and Knox R. J. (1992) The bioactivation of 5-(aziridin-1-yl)-2,4-dinitrobenzamide (CB1954) 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.
Boland, M. P., Knox, R. J., and Roberts, J. J. (1991) The differences in kinetics of rat and human DT-diaphorase result in a differential sensitivity of derived cell lines to CB1954 (5-(aziridin-1-yl)-2,4-dinitrobenzamide). Biochem. Pharmacol. 41, 867–875.
Chen, S., Know, R., Lewis, A. D., et al. (1995) Catalytic properties of NAD(P)H: quinone acceptor oxidoreductase: study involving mouse, rat human and mouse-rat chimeric enzymes. Mol. Pharmacol. 47, 934–939.
Chen, S., Knox, R., Wu, K., et al. (1997) Molecular basis of the catalytic differences among DT-diaphorase of human, rat and mouse. J. Biol. Chem., 272, 1437–1439.
Bridgwater, J. A., Springer, C. J., Knox, R. J., 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.
Miller, A. D. and Rosman, G. J. (1989) Improved retroviral vectors for gene transfer and expression. BioTechniques 7, 980–988.
Green, N. K., Youngs, D. J., Neoptolemos, J. P., 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 Ther. 4, 229–238.
McNeish, I. A., Green, N. K., Gilligan, M. G., et al. Virus directed enzyme prodrug therapy for ovarian and pancreatic cancer using retrovirally delivered E.coli nitroreductase and CB1954. Gene Ther. 5, 1061–1069.
Weedon, S. W., Green, N. K., McNeish, I. A., et al. (2000) Sensitisation of human carcinoma cells to the prodrug CB1954 by adenovirus vector-mediated expression of E. coli nitroreductase. Int. J. Cancer 86, 848–854.
Graham, F. L., Smiley, J., Russell, W. C., and Nairn, R. (1977) Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J. Gen. Virol., 36(1), 59–74.
Cosset, F. L., Takeuchi, Y., Battini, J. L., Weiss, R. A., and Collins, M. K. (1995) High-titer packaging cells producing recombinant retroviruses resistant to human serum. J. Virol, 69(12), 7430–7436.
Kozak, M. (1986) Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell 44, 283–292.
Gerin, P. A., Gilligan, M. G., Searle, P. F., and Al-Rubeai, M. (1999) Improved titers of retroviral vectors from the human FLY RD18 packaging cell line in serum-and proteinfree medium. Hum. Gene Ther. 10(12), 1965–1974.
Bowles, N. E., Eisensmith, R. C., Mohuiddin, R., Pyron, M., and Woo, S. L. (1996) A simple and efficient method for the concentration and purification of recombinant retrovirus for increased hepatocyte transduction in vivo. Hum. Gene Ther. 7(14), 1735–1742.
Gilligan, M. G., Knox, P., Weedon, S., et al. (1998) Adenoviral delivery of B7-1 (CD80) increases the immunogenicity of human ovarian and cervical carcinoma cells. Gene Ther., 5(7), 965–974.
Grove, J. I., Searle, P. F., Weedon, S. J., Green, N. K., McNeish, I. A., and Kerr, D. J. (1999) Virus-directed enzyme prodrug therapy using CB1954. Anticancer Drug Design, 14(6), 461–472.
Miller, A. D. and Buttimore, C. (1986) Redesign of retrovirus packaging lines to avoid recombination leading to helper virus production. Mol. Cell. Biol., 6, 2895–2909.
Danos, O. and Mulligan, R. C. (1998) Safe and efficient generation of recombinant retroviruses with amphotrophic and ecotropic host ranges. Proc. Natl. Acad. Sci. USA, 86, 6460–6464.
Markowitz, D., Goff, S., and Bank, A. (1998) Construction and use of a safe and efficient amphotrophic packaging cell line. Virology 167, 400–406.
Tong-Chuan, H., Zhou, S., Da Costa, L. T., Yu, J., Kinzler, K. W., and Vogelstein, B. (1998) A simplified system for generating recombinant adenoviruses. Proc. Natl. Acad. Sci. USA, 95, 2509–2514.
Fallaux, F. J., Kranenburg, O., Cramer, S. J., et al. (1996) Characterization of 911: a new helper cell line for the titration and propagation of early region 1-deleted adenoviral vectors. Hum. Gene Ther. 7(2), 215–222.
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Green, N.K., Kerr, D.J., Mautner, V., Harris, P.A., Searle, P.F. (2004). The Nitroreductase/CB1954 Enzyme-Prodrug System. In: Springer, C.J. (eds) Suicide Gene Therapy. Methods in Molecular Medicine™, vol 90. Humana Press. https://doi.org/10.1385/1-59259-429-8:459
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DOI: https://doi.org/10.1385/1-59259-429-8:459
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