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Metabolic stability of superoxide adducts derived from newly developed cyclic nitrone spin traps

Latest updated: May 29, 2020

Authors: Nicolas Bézière, MicaelHardy, FlorentPoulhès, HakimKaroui, PaulTordo, OlivierOuari, Yves-MichelFrapart, AntalRockenbauer, Jean-LucBoucher, Daniel Mansuy , Fabienne Peyrot

Ref: Free Radical Biology and Medicine, Volume 67, February 2014, Pages 150-158



Reactive oxygen species are by-products of aerobic metabolism involved in the onset and evolution of various pathological conditions. Among them, the superoxide radical is of special interest as the origin of several damaging species such as H2O2, hydroxyl radical, or peroxynitrite (ONOO−). Spin trapping coupled with ESR is a method of choice to characterize these species in chemical and biological systems and the metabolic stability of the spin adducts derived from reaction of superoxide and hydroxyl radicals with nitrones is the main limit to the in vivo application of the method. Recently, new cyclic nitrones bearing a triphenylphosphonium or permethylated β-cyclodextrin moiety have been synthesized and their spin adducts demonstrated increased stability in buffer. In this article, we studied the stability of the superoxide adducts of four new cyclic nitrones in the presence of liver subcellular fractions and biologically relevant reductants using an original setup combining a stopped-flow device and an ESR spectrometer. The kinetics of disappearance of the spin adducts were analyzed using an appropriate simulation program. Our results highlight the interest of the new spin trapping agents CD-DEPMPO and CD-DIPPMPO for specific detection of superoxide with high stability of the superoxide adducts in the presence of liver microsomes.


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EPR stopped-flow EPR flow resonator