4-Hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl (Tempol) Inhibits Peroxynitrite-Mediated Phenol Nitration

• Published in: Chemical research in toxicology 2000-04, Vol.13 (4), p.294-300
• Main Authors: Carroll, Richard T, Galatsis, Paul, Borosky, Susan, Kopec, Karla K, Kumar, Vikram, Althaus, John S, Hall, Edward D
• Format: Article
• Language: English
• Subjects: 4-Hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl; Animals; Antioxidants - pharmacology; Cyclic N-Oxides - pharmacology; Hydrogen-Ion Concentration; Hydroxylation; L-Lactate Dehydrogenase - secretion; Molsidomine - analogs & derivatives; Molsidomine - antagonists & inhibitors; Nitrates - antagonists & inhibitors; Nitrates - metabolism; Nitro Compounds - metabolism; Nitroso Compounds - metabolism; PC12 Cells; peroxynitrite; Phenol - metabolism; Rats; Spin Labels
• ISSN: 0893-228X; 1520-5010
• DOI: 10.1021/tx990159t

Peroxynitrite (PN), a very reactive oxidant formed by the combination of superoxide and nitric oxide, appears to play a role in producing tissue damage in a number of inflammatory conditions. Pharmacological scavenging and decomposition of PN within these areas has therapeutic value in several tissue injury models. Recently, we have been interested in nitroxide free radical-containing compounds as possible scavengers of PN decomposition products. Nitroxides can undergo redox reactions to the corresponding hydroxylamine anion or oxoammonium cation in biological systems as shown by its ability to react with superoxide, leading to the formation of hydrogen peroxide and molecular oxygen. We found that 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl (Tempol) inhibits PN-mediated nitration of phenolic compounds in the presence of a large molar excess of PN, suggesting a catalytic-like mechanism. In these experiments, Tempol inhibited PN-mediated nitration over the pH range of 6.5−8.5. This inhibition was specific for nitration and had no effect on hydroxylation. After the inhibition of PN-mediated nitration, Tempol was recovered from the reaction mixtures unmodified. In addition, Tempol was effective in protecting PC-12 cells from death induced by SIN-1, a PN-generating compound. The exact mechanism of Tempol's interaction with PN is not clear; however, we propose that an intermediate in this reaction may be a nitrogen dioxide radical−Tempol complex. This complex could react with water to form either nitrite or nitrate, or with a phenol radical to produce nitrophenol or nitrosophenol products and regenerate the nitroxide.