ABSTRACT
Recent studies have highlighted the indispensable role of oxidized lipids in inflammatory responses, cell death, and disease pathogenesis. Consequently, inhibitors targeting oxidized lipids, particularly lipid-derived radicals critical in lipid peroxidation, which are known as radical-trapping antioxidants (RTAs), have been actively pursued. We focused our investigation on nitroxide compounds that have rapid second-order reaction rate constants for reaction with lipid-derived radicals. A novel screening system was developed by employing competitive reactions between library compounds and a newly developed profluorescence nitroxide probe with lipid-derived radicals to identify RTA compounds. A PubMed search of the top hit compounds revealed their wide application as repositioned drugs. Notably, the inhibitory efficacy of methyldopa, selected from these compounds, against retinal damage and bilateral common carotid artery stenosis was confirmed in animal models. These findings underscore the efficacy of our screening system and suggest that it is an effective approach for the discovery of RTA compounds.
Subject(s)
Antioxidants , Lipid Peroxidation , Animals , Humans , Antioxidants/pharmacology , Antioxidants/chemistry , Lipid Peroxidation/drug effects , Retinal Diseases/drug therapy , Retinal Diseases/metabolism , Cerebrovascular Disorders/drug therapy , Cerebrovascular Disorders/metabolism , Free Radicals/metabolism , Disease Models, Animal , Drug Evaluation, Preclinical , Mice , Lipids/chemistryABSTRACT
Age-related macular degeneration (AMD) is the leading cause of blindness worldwide. Although the cause of AMD remains unknown, lipid peroxidation (LPO) end-products are critical molecules for its development. Herein, we report the imaging of lipid radicals, which are key factors in the LPO reaction, and therapeutic information using animal models.
Subject(s)
Lipids/analysis , Macular Degeneration/pathology , Retina/pathology , Animals , Disease Models, Animal , Humans , Lipid Peroxidation , Macular Degeneration/diagnostic imaging , Macular Degeneration/metabolism , Mice , Optical Imaging , Retina/diagnostic imaging , Retina/metabolismABSTRACT
Lipids and their metabolites are easily oxidized in chain reactions initiated by lipid radicals, forming lipid peroxidation products that include the electrophiles 4-hydroxynonenal and malondialdehyde. These markers can bind cellular macromolecules, causing inflammation, apoptosis and other damage. Methods to detect and neutralize the initiating radicals would provide insights into disease mechanisms and new therapeutic approaches. We describe the first high-sensitivity, specific fluorescence probe for lipid radicals, 2,2,6-trimethyl-4-(4-nitrobenzo[1,2,5]oxadiazol-7-ylamino)-6-pentylpiperidine-1-oxyl (NBD-Pen). NBD-Pen directly detected lipid radicals in living cells by turn-on fluorescence. In a rat model of hepatic carcinoma induced by diethylnitrosamine (DEN), NBD-Pen detected lipid radical generation within 1 h of DEN administration. The lipid radical scavenging moiety of NBD-Pen decreased inflammation, apoptosis and oxidative stress markers at 24 h after DEN, and liver tumor development at 12 weeks. Thus, we have developed a novel fluorescence probe that provides imaging information about lipid radical generation and potential therapeutic benefits in vivo.
