Inhibition of in vitro lipid peroxidation by stable steroidic nitroxyl radicals.
4',4'-dimethylspiro (5 alpha-cholestane-3,2'-oxazolidin)-3'-yloxy (IK-1) and 7 alpha,12 alpha-dihydroxy-4',-4'-dimethylspiro (5 beta-cholan-24-oic-3,2'-oxazolidin)-3'-yloxy acid (IK-2), two stable steroidic nitroxyl radicals, were newly synthesized and tested as possible inhibitors of lipid peroxidation, induced by Fenton's reagent in both rat liver microsomes and egg phosphatidylcholine liposomes. The inhibitory activity, evaluated through the formation of thiobarbituric acid reactive substances (TBARS) and the conjugated diene, was compared with that of alpha-tocopherol and 2,2,6,6-tetramethylpiperidine-1-yloxy (TEMPO). In each model system IK-1 and IK-2 exhibited an IC50 of 8 microM and reduced the formation of TBARS and conjugated diene, showing IK-1 a potency comparable to alpha-tocopherol and higher than TEMPO. Moreover IK-1 and, to a lesser extent IK-2, reduced the lipid peroxidation induced in the microsomes by the water-soluble azo-initiator 2,2'-Azobis (2-methylpropionamidine) dihydrochloride (AMPH), indicating the IK-1 and IK-2 ability as chain-breaking antioxidants. The hydroxylamine 4',4'-dimethylspiro (5 alpha-cholestane-3,2'-oxazolidin)-3'-hydroxide (IK-3), obtained by chemical reduction of IK-1, was completely inactive as an inhibitor of lipid peroxidation in heat pre-treated microsomes and in liposomes. However in microsomes it was active since it was oxidized to the corresponding nitroxyl radical IK-1.
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Department of Medical Chemistry and Biochemistry, Faculty of Medicine, University of Milan, Italy. skinski@imiucca.csi.unimi.it
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D000578:Amidines / Q000494:pharmacology; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D001391:Azo Compounds / Q000378:metabolism; D002793:Cholic Acids / Q000138:chemical synthesis / Q000494:pharmacology*; D003497:Cyclic N-Oxides / Q000138:chemical synthesis / Q000378:metabolism / Q000494:pharmacology*; D005609:Free Radicals / Q000378:metabolism; D006861:Hydrogen Peroxide; D007501:Iron; D015227:Lipid Peroxidation / Q000187:drug effects*; D008081:Liposomes / Q000378:metabolism; D008862:Microsomes, Liver / Q000378:metabolism; D015394:Molecular Structure; D016877:Oxidants / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D017392:Thiobarbituric Acid Reactive Substances / Q000378:metabolism; D014810:Vitamin E / Q000378:metabolism
C109128:7,12-dihydroxy-4',4'-dimethylspiro(5-cholan-24-oic-3,2'-oxazolidin)-3'-yloxy acid; D000578:Amidines; D000975:Antioxidants; D001391:Azo Compounds; D002793:Cholic Acids; D003497:Cyclic N-Oxides; C045076:Fenton's reagent; D005609:Free Radicals; D008081:Liposomes; D016877:Oxidants; D017392:Thiobarbituric Acid Reactive Substances; D014810:Vitamin E; C002296:3-doxylcholestane; C046728:2,2'-azobis(2-amidinopropane); D006861:Hydrogen Peroxide; D007501:Iron; C003959:TEMPO