Cutaneous tolerance to nitroxide free radicals and nitrone spin traps in the guinea pig

Abstract

The attempts to use nitroxide free radicals and nitrone spin traps topically in skin requires analysis of their potential cutaneous adverse effects. The objective of this study was to investigate the skin irritation and sensitizing potential of nitroxides and nitrones in the guinea pig. The following unsubstituted nitroxides were investigated: 2,2,6,6-tetramethyl-1-piperidinoxyl (Tempo), 2,2,5,5-tetramethyl-3-oxazolidinoxyl (Doxo), 2,2,5,5-tetramethyl-1-dihydro-pyrrolinoxyl (Proxo), 2,2,3,4,5,5-hexamethyl-imidazoline-1-yloxyl (Imidazo) and the nitrones: 5,5-dimethyl-1-pyrroline- N-oxide (DMPO) and N- tert.-butyl-phenylnitrone (PBN). Cutaneous irritation was determined following the modified Draize protocol. The response was evaluated clinically as well as by a biophysical method analyzing transepidermal water loss (TEWL). The nitroxides and nitrones were classified clinically from non-irritant (Proxo, Imidazo, DMPO) to slightly irritant (Tempo, Doxo, PBN) according to the Draize protocol. In agreement with the clinical scoring, the TEWL values were significantly increased by Tempo, Doxo and PBN. TOLH, the hydroxylamine of Tempo and its major skin metabolite, did not cause skin irritation. The sensitizing effect was evaluated according to the Magnusson and Kligman test. The results showed no cutaneous hypersensitivity to all nitroxides and nitrones, indicating a weak sensitizing potential. That concludes that the nitroxides and nitrones tested in this study have a low potential of acute skin intolerance.