Nitroxide stable radical prevents primaquine-induced lysis of red blood cell.

Primaquine (PQ), an antimalarial drug, is known to produce multiple oxidative effects in red blood cells (RBC). Because H2O2, OH and intracellular superoxide are implicated in this oxidation, the effect of cell-permeable nitroxide 2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPO) capable of scavenging O2.- has been studied. PQ caused RBC lysis and facilitated the oxidation of oxyhemoglobin (oxyHb) to methemoglobin (MetHb). The lysis was partially inhibited by catalase and by the metal chelating agent 2,2-dipyridyl. TEMPO blocked PQ-induced RBC lysis in dose-dependent manner (2 mM IC50) but enhanced the oxidation of oxyHb to MetHb. PQ facilitated the lysis also in the presence of CO but without effecting Hb oxidation. This hemolysis, however, was inhibited by TEMPO. The results indicated that: (a) no causative relationship exists between PQ-induced Hb oxidation and RBC lysis; (b) TEMPO can directly oxidize heme-iron without causing membrane injury; (c) the aerobic toxicity of PQ in this system is mediated by O2.- and H2O2 and possibly by redox-active labile metals (d) TEMPO can protect by detoxifying O2.- and oxidizing reduced labile metal ions and thus blocking their participation in Fenton reaction.

Activated oxygen generation by a primaquine metabolite: inhibition by antioxidants derived from Chinese herbal remedies.

Primaquine is an important antimalarial drug which causes hemolytic anemia in patients with glucose-6-phosphate dehydrogenase (G6PDH) deficiency, probably due to oxidant generation by its metabolites. One of primaquine's metabolites, 5,6-dihydroxy-8-aminoquinoline (AQD), was found to cause chemiluminescence (CL) in vitro when incubated in the presence of luminol. This CL is inhibited by catalase and deferoxamine, unaffected by mannitol, and stimulated by superoxide dismutase (SOD), suggesting that it is mediated by H2O2. Three antioxidants (daphnetin, ferulate, and maltol), derived from Chinese herbal remedies, inhibited AQD- and H2O2-mediated CL, whereas a fourth, anisodamine, had no effect. Daphnetin also potently inhibited H2O2-mediated lipid peroxidation as measured by the production of thibarbituric acid reacting substances (TBARS). Thus, the possibility is raised that an antioxidant might be able to mitigate the oxidant hemolytic effects of primaquine.