Metabolic activation of adriamycin by NADPH-cytochrome P450 reductase; overview of its biological and biochemical effects.

• Authors: Agnieszka Bartoszek
• Languages of publication: EN

Abstracts

EN

NADPH-cytochrome P450 reductase (P450 reductase) is one of the enzymes implicated in the metabolism of adriamycin, a very important clinically used antitumour drug. However, apart from the enzyme involvement, so far little was known about the chemical route and biochemical effects of this process. We demonstrated that the application of P450 reductase simultaneously with adriamycin to tumour cells in culture significantly increased cytotoxicity of the drug. Under tissue culture conditions, we noticed also that, in the presence of P450 reductase, adriamycin metabolite(s), displaying an altered spectrum within the visible light range were formed. This observation was taken adavantage of to study the metabolism of adriamycin in cell-free systems, using initially the enzyme isolated from rat liver and the recently obtained recombinant human P450 reductase. The reductive conversion of the drug turned out to be a multi-stage process, which occurred only under aerobic conditions and was accompanied by excessive NADPH consumption. Further research carried out with the aid of radical scavengers and radiolabelled adriamycin revealed that the enhancement of biological activity of adriamycin by P450 reductase stemmed from the formation of alkylating metabolite(s) rather than from the promotion of redox cycling known to be induced in the presence of anthracyclines.

Keywords

EN

covalent binding to macromolecules; adriamycin; oxygen radicals; NADPH-cytochrome P450 reductase

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2002
• Volume: 49
• Issue: 2
• Pages: 323-331

Dates

published

2002

received

2001-09-10

revised

2002-04-15

accepted

2002-05-24

Contributors

author

Agnieszka Bartoszek

• Department of Pharmaceutical Technology and Biochemistry, Gdańsk University of Technology, Gdańsk, Poland

References

• Alegria AA, Samuni A, Mitchell JB, Riesz P, Russo A. (1989) Free radicals induced by adriamycin-sensitive and adriamycin-resistant cells: a spin-trapping study. Biochemistry.; 28: 8653-8.
• Bachur NS, Gordon SL, Gee MV, Kon H. (1979) NADPH cytochrome P450 reductase activation of anticancer agents to free radicals. Proc Natl Acad Sci U S A.; 76: 954-7.
• Bartoszek A, Wolf CR. (1992) Enhancement of doxorubicin toxicity following activation by NADPH cytochrome P450 reductase. Biochem Pharmacol.; 43: 1449-57.
• Bligh HFJ, Bartoszek A, Robson CN, Hickson ID, Kasper CB, Beggs JD, Wolf CR. (1990) Activation of mitomycin C by NADPH: cytochrome P450 reductase. Cancer Res.; 50: 7789-92.
• Cummings J, Allan L, Willmott N, Riley R, Workman P, Smyth JF. (1994) The enzymology of doxorubicin quinone reduction in tumour tissue. Biochem Pharmacol.; 44: 2175-83.
• Cummings J, Bartoszek A, Smyth JF. (1991) Determination of covalent binding to intact DNA, RNA, and oligonucleotides by intercalating anticancer drugs using high-performance liquid chromatography. Studies with doxorubicin and cytochrome P450 reductase. Anal Biochem.; 194: 146-55.
• Doroshow JH. (1986) Prevention of doxorubicin-induced killing of MCF-7 human breast cells by oxygen radical scavengers and iron chelating agents. Biochem Biophys Res Commun.; 135: 330-5.
• Gaudiano G, Koch TH, Lo Bello M, Nuccetelli M, Ravagnan G, Serafino A, Sinibaldi-Vallebona P. (2000) Lack of glutathione conjugation to adriamycin in human breast cancer MCF-7/DOX cells. Biochem Pharmacol.; 60: 1915-23.
• Gerwitz DA. (1999) A critical evaluation of the mechanism of action proposed for the antitumor effects of the anthracycline antibiotics adriamycin and daunomycin. Biochem Pharmacol.; 57: 724-41.
• Konopa J. (1990) Interstrand DNA crosslinking by 1-nitroacridines, anthracyclines and aminoanthraquinones. Pharmacol Ther.; 7(suppl.): 83-94.
• Paur E, Youngman RJ, Lengfelder E, Elstner EF. (1984) Mechanism of adriamycin-dependent oxygen activation catalyzed by NADPH-cytochrome c-(ferrodoxin)-oxidoreductase. Z Naturforsch.; 39c: 261-7.
• Podell ER, Harrington DJ, Taatjes DJ, Koch TH. (1999) Crystal structure of epidoxorubicin-formaldehyde virtual crosslink of DNA and evidence for its formation in human breast-cancer cells. Acta Cryst.; D55: 1516-23.
• Sharples RA, Cullinane C, Phillips DR. (2000) Adriamycin-induced inhibition of mitochondrial-encoded polypeptides as a model system for the identification of hotspots for DNA-damaging agents. Anti-Cancer Drug Design.; 15: 183-90.
• Sinha BK. (1989) Free radicals in anticancer drug pharmacology Chem-Biol Interact.; 69: 293-317.
• Składanowski A, Konopa J. (1994) Relevance of interstrand DNA crosslinking induced by anthracyclines for their biological activity. Biochem Pharmacol.; 47: 2279-87.
• Taatjes DJ, Fenick DJ, Gaudiano G, Koch TH. (1998) A redox pathway leading to the alkylation of nucleic acids by doxorubicin and related anthracyclines: application to the design of antitumor drugs for resistant cancer. Curr Pharmaceut Design.; 4: 203-18.
• Taatjes DJ, Koch TH. (2001) Nuclear targeting and retention of anthracycline antitumor drugs in sensitive and resistant tumor cells. Curr Med Chem.; 8: 15-29.
• Tempczyk A, Tarasiuk J, Ossowski T, Borowski E. (1988) An alternative concept for the molecular nature of the peroxidating ability of anthracycline antitumor antibiotics and anthracenediones. Anti-Cancer Drug Design.; 2: 371-85.
• Tolba KA, Deliargyris EN. (1999) Cardiotoxicity of cancer therapy. Cancer Invest.; 17: 408-22.
• Wallace KB, Johnson AJ. (1987) Oxygen-dependent effect of microsomes on the binding of doxorubicin to rat hepatic nuclear DNA. Mol Pharmacol.; 31: 307-11.
• Zeman SM, Phillips DR, Crothers DM. (1998) Characterization of covalent adriamycin-DNA adducts. Proc Natl Acad Sci U S A.; 95: 11561-5.