title: Identification of nitroxide radioprotectors.
abstract: The nitroxide Tempol, a stable free radical, has recently been shown to protect mammalian cells against several forms of oxidative stress including radiation-induced cytotoxicity. To extend this observation, six additional water-soluble nitroxides with different structural features were evaluated for potential radioprotective properties using Chinese hamster V79 cells and clonogenic assays. Nitroxides (10 mM) were added 10 min prior to radiation exposure and full radiation dose-response curves were determined. In addition to Tempol, five of the six nitroxides afforded in vitro radioprotection. The best protectors were found to be the positively charged nitroxides, Tempamine and 3-aminomethyl-PROXYL, with protection factors of 2.3 and 2.4, respectively, compared with Tempol, which had a protection factor of 1.3. 3-Carboxy-PROXYL, a negatively charged nitroxide, provided minimal protection. DNA binding characteristics as studied by nonequilibrium dialysis of DNA with each of the nitroxides demonstrated that Tempamine and 3-amino-methyl-PROXYL bound more strongly to DNA than did Tempol. Since DNA is assumed to be the target of radiation-induced cytotoxicity, differences in protection may be explained by variabilities in affinity of the protector for the target. This study establishes nitroxides as a general class of new nonthiol radioprotectors and suggests other parameters that may be exploited to find even better nitroxide-induced radioprotection.
authors.affiliation: Radiobiology Section, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: S M
authors.initials: SM
authors.lastname: Hahn
authors.forename: L
authors.initials: L
authors.lastname: Wilson
authors.forename: C M
authors.initials: CM
authors.lastname: Krishna
authors.forename: J
authors.initials: J
authors.lastname: Liebmann
authors.forename: W
authors.initials: W
authors.lastname: DeGraff
authors.forename: J
authors.initials: J
authors.lastname: Gamson
authors.forename: A
authors.initials: A
authors.lastname: Samuni
authors.forename: D
authors.initials: D
authors.lastname: Venzon
authors.forename: J B
authors.initials: JB
authors.lastname: Mitchell
mesh_terms: D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects / Q000528:radiation effects; D006224:Cricetinae; D003497:Cyclic N-Oxides / Q000494:pharmacology*; D004307:Dose-Response Relationship, Radiation; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011759:Pyrrolidines / Q000494:pharmacology; D011837:Radiation-Protective Agents / Q000494:pharmacology*; D013113:Spin Labels; D014216:Triacetoneamine-N-Oxyl / Q000494:pharmacology
chemical_list: D003497:Cyclic N-Oxides; D011500:Protein Synthesis Inhibitors; D011759:Pyrrolidines; D011837:Radiation-Protective Agents; D013113:Spin Labels; C015527:4-amino-TEMPO; C032289:2,2,5,5-tetramethyl-1-pyrrolidinyloxy-3-carboxylic acid; C077428:3-cyano-2,2,5,5-tetramethyl-1-pyrrolidinyl-N-oxyl; D014216:Triacetoneamine-N-Oxyl; C077426:3-carbamoyl-2,2,5,5-tetramethyl-1-pyrrolidinyl-N-oxyl; C077429:3-aminomethyl-2,2,5,5-tetramethyl-1-pyrrolidinyl-N-oxyl; C001803:tempol