An in vitro screening cascade to identify neuroprotective antioxidants in ALS.
Amyotrophic lateral sclerosis (ALS) is an adult-onset neurodegenerative disease, characterized by progressive dysfunction and death of motor neurons. Although evidence for oxidative stress in ALS pathogenesis is well described, antioxidants have generally shown poor efficacy in animal models and human clinical trials. We have developed an in vitro screening cascade to identify antioxidant molecules capable of rescuing NSC34 motor neuron cells expressing an ALS-associated mutation of superoxide dismutase 1. We have tested known antioxidants and screened a library of 2000 small molecules. The library screen identified 164 antioxidant molecules, which were refined to the 9 most promising molecules in subsequent experiments. Analysis of the in silico properties of hit compounds and a review of published literature on their in vivo effectiveness have enabled us to systematically identify molecules with antioxidant activity combined with chemical properties necessary to penetrate the central nervous system. The top-performing molecules identified include caffeic acid phenethyl ester, esculetin, and resveratrol. These compounds were tested for their ability to rescue primary motor neuron cultures after trophic factor withdrawal, and the mechanisms of action of their antioxidant effects were investigated. Subsequent in vivo studies can be targeted using molecules with the greatest probability of success.
Academic Neurology Unit and Sheffield Care and Research Centre for Motor Neuron Disorders, University of Sheffield, Sheffield, UK.
Siân C
SC
Barber
Adrian
A
Higginbottom
Richard J
RJ
Mead
Stuart
S
Barber
Pamela J
PJ
Shaw
D000690:Amyotrophic Lateral Sclerosis / Q000188:drug therapy* / Q000473:pathology / Q000503:physiopathology; D000818:Animals; D000975:Antioxidants / Q000737:chemistry / Q000627:therapeutic use*; D017209:Apoptosis / Q000187:drug effects; D001812:Blood-Brain Barrier / Q000187:drug effects; D002109:Caffeic Acids / Q000737:chemistry / Q000627:therapeutic use; D018929:Cell Culture Techniques; D002460:Cell Line; D002490:Central Nervous System / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D009046:Motor Neurons / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D009154:Mutation; D018384:Oxidative Stress / Q000187:drug effects; D000077185:Resveratrol; D054852:Small Molecule Libraries / Q000737:chemistry / Q000627:therapeutic use; D013267:Stilbenes / Q000737:chemistry / Q000627:therapeutic use; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism*; D000072105:Superoxide Dismutase-1; D019076:Transgenes; D014468:Umbelliferones / Q000737:chemistry / Q000378:metabolism
D000975:Antioxidants; D002109:Caffeic Acids; C000606290:SOD1 protein, human; D054852:Small Molecule Libraries; D013267:Stilbenes; D014468:Umbelliferones; C000606291:Sod1 protein, mouse; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1; D000077185:Resveratrol; C007628:esculetin; C040048:caffeic acid