Abstract
The indolequinone EO9 demonstrated good preclinical activity but failed to show clinical efficacy against a range of tumours following intravenous drug administration. A significant factor in EO9's failure in the clinic has been attributed to its rapid pharmacokinetic elimination resulting in poor drug delivery to tumours. Intravesical administration of EO9 would circumvent the problem of drug delivery to tumours and the principal objective of this study is to determine whether or not bladder tumours have elevated levels of the enzyme NQO1 (NAD(P)H:quinone oxidoreductase) which plays a key role in activating EO9 under aerobic conditions. Elevated NQO1 levels in human bladder tumour tissue exist in a subset of patients as measured by both immunohistochemical and enzymatic assays. In a panel of human tumour cell lines, EO9 is selectively toxic towards NQO1 rich cell lines under aerobic conditions and potency can be enhanced by reducing extracellular pH. These studies suggest that a subset of bladder cancer patients exist whose tumours possess the appropriate biochemical machinery required to activate EO9. Administration of EO9 in an acidic vehicle could be employed to reduce possible systemic toxicity as any drug absorbed into the blood stream would become relatively inactive due to an increase in pH. © 2001 Cancer Research Campaign http://www.bjcancer.com
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References
Bradford MM (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilising the principle of protein-dye binding. Anal Biochem 72: 248–254
Butler J, Spanswick VJ and Cummings J (1996) The autooxidation of reduced forms of EO9. Free Rad Res 25: 141–148
Connors TA (1996) Bioreductive agents, hypoxic cells and therapy. Eur J Cancer 32A: 1833–1834
Cummings J, Spanswick VJ, Tomaz M and Smyth JF (1998a) Enzymology of MMC metabolic activation in tumour tissue. Implications for enzyme directed bioreductive drug development. Biochem Pharmacol 56: 405–414
Cummings J, Spanswick VJ, Gardiner J, Ritchie A and Smyth JF (1998b) Pharmacological and biochemical determinants of the antitumour activity of the indoloquinone EO9. Biochem Pharmacol 55: 253–260
Dirix LY, Tonnesen F, Cassidy J, Epelbaum R, Huinink WWT, Pavlidis N, Sorio R, Gamucci T, Wolff I, Tevelde A, Lan J and Verweij J (1996) EO9 phase II study in advanced breast, gastric, pancreatic and colorectal carcinoma by the early clinical studies group. Eur J Cancer 32A: 2019–2022
Fitzsimmons SA, Workman P, Grever M, Paull K, Camalier R and Lewis AD (1996) Reductase enzyme expression across the National Cancer Institute tumour cell line panel: Correlation with sensitivity to MMC and EO9. J Natl Cancer Inst 88: 259–269
Gutierrez PL (1989) Mechanism of bioreductive alkylation. The example of diazaquinone (AZQ). Free Radical Bio Med 6: 405–445
Hendriks HR, Piazo PE, Berger DP, Kooistra KL, Bibby MC, Boven E, Dreef-Van Der Meulen HC, Henrar REC, Fiebig HH, Double JA, Hornstra HW, Pinedo HM, Workman P and Swartsmann G (1993) EO9: A novel bioreductive alkylating indolequinone with preferential solid tumour activity and lack of bone marrow toxicity in preclinical models. Eur J Cancer 29A: 897–906
Herr HW (1987) Intravesical therapy – a critical review. Urol Clin N Am 14: 399–404
Hodnick WF and Sartorelli AC (1997) Measurement of dicumarol sensitive NADPH: (menadione cytochrome c) oxidoreductase activity results in an artifactual assay of DT-diaphorase in cell sonicates. Anal Biochem 252: 165–168
Kennedy AS, Raleigh JA, Perez GM, Calkins DP, Thrall DE, Novotny DB and Varia MA (1997) Proliferation and hypoxia in human squamous cell carcinoma of the cervix: first report of combined immunohistochemical assays. Int J Radial Oncol Biol Phys 37: 897–905
Kuin A, Alders M, Lamfers M, Van Zuidam DJ, Essers M, Beijnen JH and Smets LA (1999) Potentiation of anti-cancer activity at low intratumoural pH induced by the mitochondrial inhibitor m -iodobenzylguanidine (MIBG) and its analogue benzylguanidine (BG). Br J Cancer 79: 793–801
Maffezzini M, Simonata A, Zanon M, Raber M and Carmignani G (1996) Up-front chemotherapy for low stage low grade recurrent bladder cancer. J Urol 155: 91–93
Malkinson AM, Siegel D, Forrest GL, Gazdar AF, Oie HK, Chan DC, Bunn PA, Mabry M, Dykes DJ, Harrison SD and Ross D (1992) Elevated DT-diaphorase activity and messenger RNA content in human non small cell lung carcinoma – Relationship to the response of lung tumour xenografts to MMC. Cancer Res 52: 4752–4757
Maliepaard M, Wolf A, Groot SE, De Mol NJ and Janssen LHM (1995) Indoloquinone EO9: DNA interstrand cross linking upon reduction by DT-diaphorase or xanthine oxidase. Br J Cancer 71: 836–839
Mossoba MM, Alizadeh M and Gutierrez PL (1985) Mechanism for the reductive activation of diazaquinone. J Pharm Sci 74: 1249–1254
Newling D (1990) Intravesical therapy in the management of superficial transitional cell carcinoma of the bladder: the experience of the EORTC GU group. Br J Cancer 61: 497–499
Oosterlink W, Kurth KH, Schröder F, Bultinck J, Hammond B and Sylvester R members of the European Organisation for Research and Treatment of Cancer Genitourinary Group (1993) A prospective European Organisation for Research and Treatment of Cancer Genitourinary Group randomised trial comparing transurethral resection followed by a single instillation of epirubicin or water in single stage Ta, T1 papillary carcinoma of the bladder. J Urol 149: 749–752
Pan SS, Yu F and Hipsher C (1993) Enzymatic and pH modulation of MMC induced DNA damage in MMC resistant HCT 116 human colon cancer cells. Mol Pharmacol 43: 870–877
Phillips RM (1996) Bioreductive activation of a series of analogues of 5-aziridinyl-3-hydroxymethyl-1-methyl-2-[1H-indole-4,7-dione] prop-β-en-α-ol (EO9) by human DT-diaphorase. Biochem Pharmacol 52: 1711–1718
Phillips RM (1999) Inhibition of DT-diaphorase (NAD(P)H:quinone oxidoreductase, EC 1.6.99.2) by 5,6-dimethylxanthenone-4-acetic acid (DMXAA) and flavone-8-acetic acid (FAA): Implications for bioreductive drug development. Biochem Pharmacol 58: 303–310
Phillips RM, Hulbert PB, Bibby MC, Sleigh NR and Double JA (1992) In vitro activity of the novel indolequinone EO-9 and the influence of pH on cytotoxicity. Br J Cancer 65: 359–364
Phillips RM, Loadman PM and Cronin BP (1998) Evaluation of a novel in vitro assay for assessing drug penetration into avascular regions of tumours. Br J Cancer 77: 2112–2119
Plumb JA and Workman P (1994) Unusually marked hypoxic sensitisation to indolequinone EO9 and MMC in a human colon tumour cell line that lacks NQO1 activity. Int J Cancer 56: 134–139
Robertson N, Haigh A, Adams GE and Stratford IJ (1994) Factors affecting sensitivity to EO9 in rodent and human tumour cells in vitro: DT-diaphorase activity and hypoxia. Eur J Cancer 30A: 1013–1019
Saunders MP, Jaffar M, Patterson AV, Nolan J, Naylor MA, Phillips RM, Harris AL and Stratford IJ (2000) The relative importance of NADPH:cytochrome c (P450) reductase for determining the sensitivity of human tumour cells to the indoloquinone EO9 and related analogues lacking functionality at the C-2 and C-3 positions. Biochem Pharmacol 59: 993–996
Schellens JHM, Planting AST, Van Acker BAC, Loos WJ, De Boer-Dennert M, Van Der Burg MEL, Koier I, Krediet RT, Stoter G and Verweij J (1994) Phase I and pharmacologic study of the novel indolequinone bioreductive alkylating cytotoxic drug EO9. J Natl Cancer Inst 86: 906–912
Schlager JJ and Powis G (1988) MMC is not metabolised by but is an inhibitor of human kidney NAD(P)H:(quinone acceptor) oxidoreductase. Cancer Chemother Pharmacol 22: 126–130
Siegel D, Gibson NW, Preusch PC and Ross D (1990) Metabolism of MMC by DT-diaphorase. Role in MMC induced DNA damage and cytotoxicity in human colon carcinoma cells. Cancer Res 50: 7483–7489
Siegel D, Beall HD, Kasai M, Gibson NW and Ross D (1993) PH dependent inactivation of DT-diaphorase by MMC and porfiromycin. Mol Pharmacol 44: 1128–1134
Siegel D, Franklin WA and Ross D (1998) Immunohistochemical detection of NAD(P)H:Quinone oxidoreductase in human lung and lung tumours. Clin Cancer Res 4: 2065–2070
Smitkamp-Wilms E, Peters GJ, Pinedo HM, Van Arkotte J and Giaccone G (1994) Chemosensitivity to the indoloquinone EO9 is correlated with DT-diaphorase activity and gene expression. Biochem Pharmacol 47: 1325–1332
Smitkamp-Wilms E, Giaccone G, Pinedo HM, Van Der Laan BFAM and Peters GJ (1995) DT-diaphorase activity in normal and neoplastic human tissues: An indicator of sensitivity to bioreductive agents?. Br J Cancer 72: 917–921
Tolley DA, Parmar MKB, Grigor KM and Lallemand G the Medical Research Council superficial bladder cancer working party (1996) The effect of intravesical MMC on recurrence of newly diagnosed superficial bladder cancer: A further report with 7 years of followup. J Urol 155: 1233–1238
Traver RD, Horikoshi T, Dannenberg KD, Stadlbauer THW, Dannenberg PV, Ross D and Gibson NW (1992) NAD(P)H:quinone oxidoreductase gene expression in human colon carcinoma cells: Characterisation of a mutation which modulates DT-diaphorase activity and mitomycin sensitivity. Cancer Res 52: 797–802
Walton MI, Smith PJ and Workman P (1991) The role of NAD(P)H:quinone reductase (EC 1.6.99.2, DT-diaphorase) in the reductive bioactivation of the novel indoloquinone antitumour agent EO9. Cancer Commun 3: 199–206
Workman P (1994) Enzyme directed bioreductive drug development revisited: A commentary on recent progress and future prospects with emphasis on quinone anticancer drugs and quinone metabolising enzymes, particularly NQO1. Oncol Res 6: 461–475
Yen WC, Schmittgen T and Au JL (1996) Different pH dependency of mitomycin C activity in monolayer and three dimensional cultures. Pharmaceut Res 13: 1887–1891
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Choudry, G., Stewart, P., Double, J. et al. A novel strategy for NQO1 (NAD(P)H:quinone oxidoreductase, EC 1.6.99.2) mediated therapy of bladder cancer based on the pharmacological properties of EO9. Br J Cancer 85, 1137–1146 (2001). https://doi.org/10.1054/bjoc.2001.2056
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DOI: https://doi.org/10.1054/bjoc.2001.2056
Keywords
- bioreductive drugs: EO9
- mitomycin C: bladder cancer
- NQO1
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