Identification of benzothiazoles as potential polyglutamine aggregation inhibitors of Huntington's disease by using an automated filter retardation assay
Identification of benzothiazoles as potential polyglutamine aggregation inhibitors of Huntington's disease by using an automated filter retardation assay
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DOI：
10.1073/pnas.182426599
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发表时间：
2002-12-10
影响因子：
11.1
通讯作者：
Wanker, EE
中科院分区：
文献类型：
--
作者：
Heiser, V;Engernann, S;Wanker, EE
防止神经元中不溶性的含多聚谷氨酰胺的蛋白质聚集体的形成可能代表改善亨廷顿病（HD）的有吸引力的治疗策略。因此，筛选抑制亨廷顿蛋白自组装的小分子的能力将具有潜在的临床和重要的研究应用。我们已经开发了一种自动化的过滤阻滞试验，用于快速鉴定在体外阻止HD外显子1蛋白聚集的化合物。利用这种方法，从大约184，000个小分子的文库中发现了25个以剂量依赖性方式抑制亨廷顿蛋白原纤维形成的苯并噻唑衍生物。通过免疫印迹、电子显微镜和质谱法证实了通过过滤测定获得的结果。此外，细胞培养研究显示，2-氨基-4，7-二甲基-苯并噻唑-6-醇（一种与利鲁唑相似的化合物）可显著抑制HD外显子1的体内聚集。这些发现可能为一种新的治疗方法提供基础，以防止亨廷顿病和相关谷氨酰胺重复障碍中不溶性蛋白质聚集体的积累。
Preventing the formation of insoluble polyglutamine containing protein aggregates in neurons may represent an attractive therapeutic strategy to ameliorate Huntington's disease (HD). Therefore, the ability to screen for small molecules that suppress the self-assembly of huntingtin would have potential clinical and significant research applications. We have developed an automated filter retardation assay for the rapid identification of chemical compounds that prevent HD exon 1 protein aggregation in vitro. Using this method, a total of 25 benzothiazole derivatives that inhibit huntingtin fibrillogenesis in a dose-dependent manner were discovered from a library of approximate to184,000 small molecules. The results obtained by the filter assay were confirmed by immuno-blotting, electron microscopy, and mass spectrometry. Furthermore, cell culture studies revealed that 2-amino-4,7-dimethyl-benzothiazol-6-ol, a chemical compound similar to riluzole, significantly inhibits HD exon 1 aggregation in vivo. These findings may provide the basis for a new therapeutic approach to prevent the accumulation of insoluble protein aggregates in Huntington's disease and related glutamine repeat disorders.