A high-throughput screen to identify inhibitors of SOD1 transcription.
A high-throughput screen to identify inhibitors of SOD1 transcription.
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DOI：
10.2741/e584
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发表时间：
2012-06-01
期刊：
影响因子：
--
通讯作者：
Brown RH Jr
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文献类型：
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作者：
Wright PD;Wightman N;Huang M;Weiss A;Sapp PC;Cuny GD;Ivinson AJ;Glicksman MA;Ferrante RJ;Matson W;Matson S;Brown RH Jr
肌萎缩侧索硬化症（amyotrophiclateralsclerosis，ALS）是一种致死性的退行性运动神经元疾病。大约20%的家族性ALS病例是由Cu/Zn超氧化物歧化酶（SOD 1）基因突变引起的。表达突变体SOD 1转基因的啮齿动物发展为进行性、致命的运动神经元疾病，并且疾病的发作和进展取决于SOD 1的水平。我们研究了SOD 1蛋白减少可能对ALS有治疗益处的可能性，并筛选了30，000种抑制SOD 1转录的化合物。所鉴定的最有效的抑制剂是N-{4-[4-（4-甲基苯甲酰基）-1-哌嗪基]苯基}-2-噻吩甲酰胺（化合物ID 7687685），其在PC 12细胞中显示抑制SOD 1表达的EC 50为10.6 μ M，LD 50>30 μ M。该化合物随后显示降低HeLa细胞中的内源性SOD 1水平，并表现出小鼠脊髓组织中SOD 1蛋白水平的适度降低。这些数据表明，化合物7687685作为SOD 1基因表达抑制剂的功效不太可能在临床上有用，尽管报道的策略可广泛应用于筛选基因表达的小分子抑制剂。
Amyotrophic lateral sclerosis (ALS) is a fatal degenerative motor neuron disease. Approximately 20% of familial ALS cases are caused by mutations in the Cu/Zn superoxide dismutase (SOD1) gene. Rodents expressing mutant SOD1 transgenes develop progressive, fatal motor neuron disease and disease onset and progression is dependent on the level of SOD1. We investigated the possibility that a reduction in SOD1 protein may be of therapeutic benefit in ALS and screened 30,000 compounds for inhibition of SOD1 transcription. The most effective inhibitor identified was N-{4-[4-(4-methylbenzoyl)-1-piperazinyl]phenyl}-2-thiophenecarboxamide (Compound ID 7687685), which in PC12 cells showed an EC50 of 10.6 microM for inhibition of SOD1 expression and an LD50 >30 microM. This compound was subsequently shown to reduce endogenous SOD1 levels in HeLa cells and to exhibit a modest reduction of SOD1 protein levels in mouse spinal cord tissue. These data suggest that the efficacy of compound 7687685 as an inhibitor of SOD1 gene expression is not likely to be clinically useful, although the strategy reported could be applied broadly to screening for small molecule inhibitors of gene expression.