Nitroarenes pp 231-244 | Cite as

Activation of Carcinogenic N-Arylhydroxamic Acids by Peroxidase/H2O2/Halide Systems: Route to C-Nitroso Aromatics

  • Danuta Malejka-Giganti
  • Clare L. Ritter
  • Lauri J. Sammartano
Part of the Environmental Science Research book series (ESRH, volume 40)


Evidence that locally carcinogenic N-arylhydroxamic acids may be activated by oxidants generated by leukocytes has been obtained from model systems including tissue peroxidases, myeloperoxidase (MPO)/H2O2/ halide (X-) or hypohalous acids (HOX). Peroxidative oxidations of N-hydroxy-N-2-fluorenylacetamide (N-OH-2-FAA) via one electron (le-) to equimolar 2-nitrosofluorene (2-NOF) and N-acetoxy-2-FAA, and via oxidative cleavage to 2-NOF were determined. The latter oxidation predominated with peroxidase-rich extracts of rat uterus and mammary gland and with eosinophils from intraperitoneal fluid in the presence of H2O2, Br- and cationic detergent. Contribution to 2-NOF by le- oxidation was up to 50% at 0.1 mM Br- and pH 7.4, but negligible at 10 mM Br- and pH 5.5. Oxidation of N-OH-2-FAA by MPO of human neutrophils in the presence of physiologic concentrations of Cl- (0.1 M) or Br- (0.1 mM) or their mixture was examined in the pH range of 4 to 6.5. At the respective pH optima (4 for Br- and 5 for Cl- or Cl- + Br-), oxidation of N-OH-2-FAA to 2-NOF by MPO/H2O2 was much more rapid with Br- and Br- + Cl- than with C1-. Since HOBr oxidized N-OH-2-FAA to 2-NOF much more rapidly than did HOC1, MPO/H2O2-catalyzed oxidation in the presence of C1-+Br- was possibly due in part to HOBr. le- oxidation of N-OH-2-FAA occurred to a lesser extent in chemical (HOX) than enzymatic systems (MPO/H2O2/X-), in which it appeared to be stimulated by Br- at pH > 5.5. In the presence of taurine, a scavenger of hypohalous acids in vivo, oxidation of N-OH-2-FAA to 2-NOF bv MPO/H2O2 was unaffected with Br-, inhibited with Cl-, and partially inhibited with Cl- + Br-. These results were linked to N-halotaurine formation since it was found that N-bromotaurine, but not N-chloro-taurine, oxidized N-OH-2-FAA chiefly to 2-NOF. The possibility that 2-NOF through its interactions with unsaturated lipids may initiate lipid peroxidation was investigated by measurements of malondialdehyde (MDA), a lipid degradation product, following aerobic incubations of 2-NOF with arachidonic, linolenic or linoleic acid at a molar ratio of 1:1. The amounts of MDA were optimal after 4 hr and were increased by 50% when 2-NOF was preincubated with the fatty acids for 24 hr under anaerobic conditions. Our studies indicate potential significance of: 1) Br--derived oxidants in activation of carcinogenic N-arylhydroxamic acids to C-nitroso aromatics, and 2) interactions of C-nitroso aromatics with unsaturated lipids leading to lipid peroxidation and thus, genotoxicity.


Electron Spin Resonance Mammary Gland Hydroxamic Acid Unsaturated Lipid Peroxidative Oxidation 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1..
    E. C. Miller, J. A. Miller, and H. A. Hartmann, N-Hydroxy-2-acetylaminofluorene: a metabolite of 2-acetylaminofluorene with increased carcinogenic activity in the rat, Cancer Res. 21: 815–824 (1961).PubMedGoogle Scholar
  2. 2..
    J. A. Miller, C. S. Wyatt, E. C. Miller, and H. A. Hartmann, The N-hydroxylation of 14-acetylaminobiphenyl by the rat and dog and the strong carcinogenicity of N-hydroxy-Ц-acetylaminobiphenyl in the rat, Cancer Res. 21:1465–1473 (1961).Google Scholar
  3. 3..
    H. R. Gutmann, S. B. Galitski, and W. A. Foley, The conversion of noncarcinogenic aromatic amides to carcinogenic arylhydroxamic acids by synthetic N-hydroxylation, Cancer Res. 27:1443–1455 (1967).PubMedGoogle Scholar
  4. 4.
    H. R. Gutmann, D. S. Leaf, Y. Yost, R. E. Rydell, and C. C. Chen, Structure-activity relationships of N-acylhydroxylamines in the rat, Cancer Res. 30:1485–1498 (1970).PubMedGoogle Scholar
  5. 5.
    D. Malejka-Giganti, and C. L. Ritter, Peroxidase-mediated metabolism of N-arylhydroxamic acids in relation to tumorigenesis, in: “Carcinogenic and Mutagenic Responses to Aromatic Amines and Nitroarenes”, C. M. King, L. J. Romano, and D. Schuetzle, eds., Elsevier Science Publishing Co., Inc., New York, pp. 109–209 (1988).Google Scholar
  6. 6.
    Y. Tsuruta, V. V. Subrahmanyam, W. Marshall, and P. J. O’Brien, Peroxidase-mediated irreversible binding of arylamine carcinogens to DNA in intact polymorphonuclear leukocytes activated by a tumor promoter, Chem. -Biol. Interact. 53:25-35 (1985).CrossRefPubMedGoogle Scholar
  7. 7.
    M. D. Corbett, B. R. Corbett, M. -H. Hannothiaux, and S. J. Quintana, Metabolic activation and nucleic acid binding of acetaminophen and related arylamine substrates by the respiratory burst of human granulocytes, Chem. Res. Toxicol. 2:260–266 (1989).CrossRefPubMedGoogle Scholar
  8. 8.
    M. A. Trush, J. L. Seed, and T. W. Kensler, Oxidant-dependent metabolic activation of polycyclic aromatic hydrocarbons by phorbol ester-stimulated human polymorphonuclear leukocytes: Possible link between inflammation and cancer, Proc. Natl. Acad. Sci. USA 82:5194–5198 (1985).CrossRefPubMedGoogle Scholar
  9. 9.
    S. J. Klebanoff, Oxygen metabolism and the toxic properties of phagocytes, Ann. Int. Med. 93:480–489 (1980).PubMedGoogle Scholar
  10. 10.
    S. J. Weiss, and A. F. LoBuglio, Biology of disease. Phagocyte-generated oxygen metabolites and cellular injury, Lab. Invest. 47:5-18 (1982).Google Scholar
  11. 11.
    E. C. Jong, W. R. Henderson, and S. J. Klebanoff, Bactericidal activity of eosinophil peroxidase. J. Immunol. 124:1378–1382 (1980).PubMedGoogle Scholar
  12. 12.
    A. R. J. Bakkenist, J. E. G. De Boer, H. Plat, and R. Weyer, The halide complexes of myeloperoxidase and the mechanism of the halogenation reactions, Biochim. Biophys. Acta 613:337–348 (1980).CrossRefPubMedGoogle Scholar
  13. 13.
    R. Cramer, M. R. Soranzo, and P. Patriarca, Evidence that eosinophils catalyze the bromide-dependent decarboxylation of amino acids, Blood 58:1112–1118 (1981).PubMedGoogle Scholar
  14. 14.
    S. J. Weiss, S. T. Test, C. M. Eckmann, D. Roos, and S. Regiani, Brominating oxidants generated by human eosinophils, Science 234: 200–203 (1986).CrossRefPubMedGoogle Scholar
  15. 15.
    A. N. Mayeno, A. J. Curran, R. L. Roberts, and C. S. Foote, Eosinophils preferentially use bromide to generate halogenating agents, J. Biol. Chem. 264:5660–5668 (1989).PubMedGoogle Scholar
  16. 16.
    H. Bartsch, J. A. Miller, and E. C. Miller, N-Acetoxy-N-acetylaminoarenes and nitrosoarenes. One-electron non-enzymatic and enzymatic oxidation products of various carcinogenic aromatic acethydroxamic acids, Biochim. Biophys. Acta 273:40–51 (1972).CrossRefPubMedGoogle Scholar
  17. 17.
    H. Bartsch, M. Traut, and E. Hecker, On the metabolic activation of N-hydroxy-N-2-acetylamiпofluorene. II. Simultaneous formation of 2-nitrosofluorene and N-acetoxу-N-2-acetylamiпofluorene from N-hydroxy-N-2-acetylaminofluorene via a free radical intermediate, Biochim. Bophys. Acta 237:556–566 (1971).CrossRefGoogle Scholar
  18. 18.
    H. Bartsch, and E. Hecker, On the metabolic activation of the carcinogen N-hydroxy-N-2-acetуlaminofluorene. III. Oxidation with horseradish peroxidase to yield 2-nitrosofluorene and N-acetoxу-N-2-acetylaminofluorene. Biochim. Biophys. Acta 237: 567–578 (1971).CrossRefPubMedGoogle Scholar
  19. 19.
    R. A. Floyd, L. M. Soong, and P. L. Culver, Horseradish peroxidase/ hydrogen peroxide-catalyzed oxidation of the carcinogen N-hydroxy-N-acetyl-2-aminofluorene as effected by cyanide and ascorbate, Cancer Res. 36:1510–1519 (1976).PubMedGoogle Scholar
  20. 20.
    R. A. Floyd, L. M. Soong, R. N. Walker, and M. Stuart, Lipid hydro-peroxide activation of N-hydroxy-N-acetylaminofluorene via a free radical route, Cancer Res. 36:2761–2767 (1976).PubMedGoogle Scholar
  21. 21.
    R. A. Floyd, and L. M. Soong, Obligatory free radical intermediate in the oxidative activation of the carcinogen N-hydroxy-2-acetylaminofluorene, Biochim. Biophys. Acta 498:244–249 (1977).CrossRefPubMedGoogle Scholar
  22. 22.
    C. L. Ritter, D. Malejka-Giganti, and C. F. Polnaszek, Cytochrome c/H2O2-mediated one electron oxidation of carcinogenic Nfluorenylacetohydroxamic acids to nitroxyl free radicals, Chem. -Biol. Interact. 46:317–334 (1983).CrossRefPubMedGoogle Scholar
  23. 23.
    M. D. Corbett, and B. R. Corbett, HRP-catalyzed bioactivation of carcinogenic hydroxamic acids. The greater reactivity of glycolyl-versus acetyl-derived hydroxamic acids, Chem. -Biol. Interact. 63:249–264 (1987).CrossRefPubMedGoogle Scholar
  24. 24.
    C. L. Ritter, and D. Malejka-Giganti, A novel oxidation of the carcinogen N-hydroxy-N-2-fluoreпylacetamide catalyzed by peroxidase/H;2O2/Br-, Biochem. Biophys. Res. Commun. 131:174–181 (1985).CrossRefPubMedGoogle Scholar
  25. 25.
    C. L. Ritter, and D. Malejka-Giganti, Oxidations of the carcinogen N-hydroxy-N-2-fluoreпylacetamide by enzymatically or chemically generated oxidants of chloride and bromide, Chem. Res. Toxicol. 2:325–333 (1989).CrossRefPubMedGoogle Scholar
  26. 26.
    E. M. Faustman-Watts, J. C. Greenaway, M. J. Namkung, A. G. Fantel, and M. R. Juchau, Teratogenicity in vitro of two deacetylated metabolites of N-hydroxy-2-acetylaminofluorene, Toxicol. Appl. Pharmacol. 76:161–171 (1984).CrossRefPubMedGoogle Scholar
  27. 27.
    D. Malejka-Giganti, C. L. Ritter, R. W. Decker, and J. M. Suilman, Peroxidative metabolism of a carcinogen, N-hydroxy-N-2-fluorenylacetamide, by rat uterus and mammary gland in vitro, Cancer Res. 46:6200–6206 (1986).PubMedGoogle Scholar
  28. 28.
    J. M. Strum, Hormonal activation of mammary gland peroxidase, Tissue & Cell 10:505–514 (1978).Google Scholar
  29. 29.
    D. Malejka-Giganti, R. W. Decker, C. L. Ritter, and M. R. Polovina, Microsomal metabolism of the carcinogen, N-2-fluoreпylacetamide, by the mammary gland and liver of female rats. I. Ring-and N- hydroxylations of N-2-fluorenylacetamide, Carcinogenesis 6:95–103 (1985).CrossRefPubMedGoogle Scholar
  30. 30.
    R. Arriagada, C. Unda, E. Sentis, H. Kong, and A. N. Tchernitchin, Estrogen-induced tissue eosinophilia in the rat mammary gland, Med. Sci. Res. 15:1505–1506 (1987).Google Scholar
  31. 31.
    M. Kh. Dabbous, R. Walker, L. Haney, L. M. Carter, G. L. Nicolson, and D. E. Woolley, Mast cells and matrix degradation at sites of tumor invasion in rat mammary adenocarcinoma, Br. J. Cancer 54:459–465 (1986).CrossRefPubMedGoogle Scholar
  32. 32.
    K. Diem, and C. Lentner, eds., “Scientific Tables”, Documenta Geigy, 7th Ed., Geigy Pharmaceuticals, Ardsley, NY, pp. 563, 644, 663 (1970)Google Scholar
  33. 33.
    M. S. Jensen, and D. F. Bainton, Temporal changes in pH within the phagocytic vacuole of the polymorphonuclear neutrophilic leukocyte, J. Cell. Biol. 56:379–388 (1973).CrossRefPubMedGoogle Scholar
  34. 34.
    M. Pelecanou, and M. Novak, Oxidation-reduction reactions of N-sulfonoxyacetanilides: mechanisms of the halide-induced reduction of models for carcinogenic metabolites of aromatic amides. J. Am. Chem. Soc. 107:4499–4503 (1985).CrossRefGoogle Scholar
  35. 35.
    J. A. Sturman, and K. C. Hayes, The biology of taurine in nutrition and development, in: “Advances in Nutritional Research”, Vol. 3, Plenum Press, New York, pp. 231–299 (1980).Google Scholar
  36. 36.
    M. B. Grisham, M. M. Jefferson, D. F. Melton, and E. L. Thomas, Chlorination of endogenous amines by isolated neutrophils. Ammonia-dependent bactericidal, cytotoxic, and cytolytic activities of the chloramines, J. Biol. Chem. 259:10404–10413 (1984).PubMedGoogle Scholar
  37. 37.
    C. E. Wright, T. T. Lin, Y. Y. Lin, J. A. Sturman, and G. E. Gauli, Taurine scavenges oxidized chlorine in biological systems, in: “Taurine: Biological Actions and Clinical Perspectives”, Alan R. Liss, Inc., New York, pp. 137–146 (1985).Google Scholar
  38. 38.
    F. X. R. Van Leeuwen, and B. Sangster, The toxicology of bromide ion, CRC Critical Reviews in Toxicology 18:189–213 (1987).CrossRefPubMedGoogle Scholar
  39. 39.
    P. D. Lotlikar, E. C. Miller, J. A. Miller, and A. Margreth, The enzymatic reduction of the N-hydroxy-derivatives of 2-acetylaminofluorene and related carcinogens by tissue preparations, Cancer Res. 25:1743–1752 (1965).PubMedGoogle Scholar
  40. 40.
    L. A. Sternson, Detection of arylhydroxylamines as intermediates in the metabolic reduction of nitro compounds, Experientia 31: 268–269 (1975).CrossRefPubMedGoogle Scholar
  41. 41.
    G. J. Mulder, L. E. Unruh, F. E. Evans, B. Ketterer, and F. F. Kadlubar, Formation and identification of glutathione conjugates from 2-nitrosofluorene and N-hydroxy-2-aminofluorene, Chem. -Biol. Interact. 39:111–127 (1982).CrossRefPubMedGoogle Scholar
  42. 42.
    E. Kriek, On the interaction of N-2-fluorenylhydroxylamine with nucleic acids in vitro, Biochem. Biophys. Res. Commun. 20:793–799 (1965).CrossRefPubMedGoogle Scholar
  43. 43.
    D. T. Beranek, G. L. White, R. H. Heflich, and F. A. Beland, Aminofluorene-DNA adduct formation in Salmonella typhimurium exposed to the carcinogen N-hydroxy-2-acetylaminofluorene, Proc. Natl. Acad. Sci. USA 79:5175–5178 (1982).CrossRefPubMedGoogle Scholar
  44. 44.
    R. A. Floyd, L. M. Soong, M. A. Stuart, and D. L. Reigh, Free radicals and carcinogenesis. Some properties of the nitroxyl free radicals produced by covalent binding of 2-nitrosofluorene to unsaturated lipids of membranes, Arch. Biochem. Biophys. 185: 450–457 (1978).CrossRefPubMedGoogle Scholar
  45. 45.
    R. Sridhar, M. J. Hampton, J. E. Steward, and R. A. Floyd, Studies on the mutagenicity and electron spin resonance spectra of nitrosofluorene-lipid adducts, Applied Spectroscopy 34:289–293 (1980).CrossRefGoogle Scholar
  46. 46.
    R. A. Floyd, Free radicals in arylamine carcinogenesis, in: “Free Radicals in Biology”, Vol. IV, W. A. Pryor, ed., Academic Press, Inc., New York, pp. 187–208 (1980).Google Scholar
  47. 47.
    R. Sridhar, and R. A. Floyd, An electron paramagnetic resonance study of the reaction of nitrosobenzene with cholesterol, Can.J. Chem. 60:1574–1576 (1982).CrossRefGoogle Scholar
  48. 48.
    A. B. Sullivan, Electron spin resonance studies of a stable aryl-nitroso-olefin adduct free radical, J.Org.Chem. 31:2811–2817 (1966).CrossRefGoogle Scholar
  49. 49.
    D. J. Kornburst, and R. D. Mavis, Relative susceptibility of microsomes from lung, heart, liver, kidney, brain and testes to lipid peroxidation. Correlation with vitamin E content, Lipids 15:315–322 (1979).CrossRefGoogle Scholar
  50. 50.
    A. Stier, R. Clauss, A. Lücke, and I. Reitz, Redox cycle of stable mixed nitroxides formed from carcinogenic aromatic amines, Xenobiotica 10:661–673 (1980).CrossRefPubMedGoogle Scholar
  51. 51.
    B. R. Brooks, and O. L. Klamerth, Interaction of DNA with bifunctional aldehydes, Eur. J. Biochem. 5:178–182 (1968).CrossRefPubMedGoogle Scholar
  52. 52.
    C. E. Vaca, and M. Harms-Ringdahl, Nuclear membrane lipid peroxidation products bind to nuclear macromolecules, Arch. Biochem. Biophys. 269:548–554 (1989).CrossRefPubMedGoogle Scholar
  53. 53.
    V. Nair, G. A. Turner, and R. J. Offerman, Novel adducts from the modification of nucleic acid bases by malondialdehyde, J. Am. Chem. Soc. 106:3370–3371 (1984).CrossRefGoogle Scholar
  54. 54.
    L. J. Marnett, A. K. Basu, S. M. O’Hara, P. E. Weller, A. F. M. M. Rahman, and J. P. Oliver, Reaction of malondialdehyde with guanine nucleosides: formation of adducts containing oxadiazbicyclononene residues in the base-pairing region, J. Am. Chem. Soc. 108:1348–1350 (1986).CrossRefGoogle Scholar
  55. 55.
    C.K. Winter, H. J. Segall, and W. F. Haddon, Formation of cyclic adducts of deoxyguanosine with the aldehydes trans-4-hydroxy-2hexenal and trans-4-hydroxy-2-nonenal in vitro, Cancer Res. 46: 5682–5686 (1986).PubMedGoogle Scholar
  56. 56.
    A. K. Basu, S. M. O’Hara, P. Valladier, K. Stone, O. Mols, and L. J. Marnett, Identification of adducts formed by reaction of guanine nucleosides with malondialdehyde and structurally related aldehydes, Chem. Res. Toxicol. 1:53–59 (1988).CrossRefPubMedGoogle Scholar
  57. 57.
    H. Krokan, R. C. Grafstrom, K. Sundqvist, H. Esterbauer, and C. C. Harris, Cytotoxicity, thiol depletion and inhibition of O6-methylguanine-DNA methyltransferase by various aldehydes in cultured human bronchial fibroblasts, Carcinogenesis 6:1755–1759 (1985).CrossRefPubMedGoogle Scholar
  58. 58.
    E. J. Barry, D. Malejka-Giganti, and H. R. Gutmann, Interaction of aromatic amines with rat-liver proteins in vivo. III. On the mechanism of binding of the carcinogens, N-2-fluorenylacetamide and N-hydroxy-2-fluorenylacetamide, to the soluble proteins, Chem. -Biol. Interact. 1:139–155 (1969/70).CrossRefPubMedGoogle Scholar
  59. 59.
    B. Dölle, W. Töpner, and H.-G. Neumann, Reaction of arylnitroso compounds with mercaptans, Xenobiotica 10:527–536 (1980).CrossRefPubMedGoogle Scholar
  60. 60.
    K. Saito, and R. Kato, Glutathione conjugation of arylnitroso compound: detection and monitoring labile intermediates in situ inside a fast atom bombardment mass spectrometer, Biochem. Biophys. Res. Commun. 124:1–5 (1984).CrossRefPubMedGoogle Scholar
  61. 61.
    N. Takahashi, V. Fischer, J. Schreiber, and R. P. Mason, An ESR study of nonenzymatic reactions of nitroso compounds with biological reducing agents, Free Rad. Res. Commun. 4:351–358 (1988).CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1990

Authors and Affiliations

  • Danuta Malejka-Giganti
    • 1
  • Clare L. Ritter
    • 1
  • Lauri J. Sammartano
    • 1
  1. 1.Veterans Administration Medical CenterUniversity of MinnesotaMinneapolisUSA

Personalised recommendations