title: Inhibition of myeloperoxidase-mediated hypochlorous acid production by nitroxides.
abstract: Tissue damage resulting from the extracellular production of HOCl (hypochlorous acid) by the MPO (myeloperoxidase)-hydrogen peroxide-chloride system of activated phagocytes is implicated as a key event in the progression of a number of human inflammatory diseases. Consequently, there is considerable interest in the development of therapeutically useful MPO inhibitors. Nitroxides are well established antioxidant compounds of low toxicity that can attenuate oxidative damage in animal models of inflammatory disease. They are believed to exert protective effects principally by acting as superoxide dismutase mimetics or radical scavengers. However, we show here that nitroxides can also potently inhibit MPO-mediated HOCl production, with the nitroxide 4-aminoTEMPO inhibiting HOCl production by MPO and by neutrophils with IC50 values of approx. 1 and 6 microM respectively. Structure-activity relationships were determined for a range of aliphatic and aromatic nitroxides, and inhibition of oxidative damage to two biologically-important protein targets (albumin and perlecan) are demonstrated. Inhibition was shown to involve one-electron oxidation of the nitroxides by the compound I form of MPO and accumulation of compound II. Haem destruction was also observed with some nitroxides. Inhibition of neutrophil HOCl production by nitroxides was antagonized by neutrophil-derived superoxide, with this attributed to superoxide-mediated reduction of compound II. This effect was marginal with 4-aminoTEMPO, probably due to the efficient superoxide dismutase-mimetic activity of this nitroxide. Overall, these data indicate that nitroxides have considerable promise as therapeutic agents for the inhibition of MPO-mediated damage in inflammatory diseases.
authors.affiliation: The Heart Research Institute, 114 Pyrmont Bridge Rd, Camperdown, Sydney, NSW 2050, Australia. reesm@hri.org.au
authors.forename: Martin D
authors.initials: MD
authors.lastname: Rees
authors.forename: Steven E
authors.initials: SE
authors.lastname: Bottle
authors.forename: Kathryn E
authors.initials: KE
authors.lastname: Fairfull-Smith
authors.forename: Ernst
authors.initials: E
authors.lastname: Malle
authors.forename: John M
authors.initials: JM
authors.lastname: Whitelock
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Davies
mesh_terms: D006418:Heme / Q000378:metabolism; D006801:Humans; D006898:Hydroxylamines / Q000737:chemistry / Q000494:pharmacology; D006997:Hypochlorous Acid / Q000378:metabolism*; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism; D009589:Nitrogen Oxides / Q000737:chemistry / Q000494:pharmacology*; D010084:Oxidation-Reduction; D009195:Peroxidase / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: D006898:Hydroxylamines; D009589:Nitrogen Oxides; D006418:Heme; D006997:Hypochlorous Acid; D009195:Peroxidase