Identification of small molecule inhibitors of beta-amyloid cytotoxicity through a cell-based high-throughput screening platform.
Identification of small molecule inhibitors of beta-amyloid cytotoxicity through a cell-based high-throughput screening platform.
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DOI：
10.1177/1087057108323909
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发表时间：
2008-10
影响因子：
--
通讯作者：
Glicksman MA
中科院分区：
文献类型：
--
作者：
Seyb KI;Schuman ER;Ni J;Huang MM;Michaelis ML;Glicksman MA
钙蛋白酶激活被假设为导致神经退行性变的事件序列中的早期发生，以及连接Aβ肽细胞外蓄积和神经元缠结细胞内形成的信号通路中的早期发生。为了鉴定通过早期干预细胞死亡级联反应预防阿尔茨海默病神经变性的小分子，开发了分化的Sh-SY 5 Y细胞中基于细胞的测定，利用钙蛋白酶活性作为暴露于细胞外Aβ的细胞死亡早期阶段的读数。该测定针对高通量筛选进行了优化，并且测试了约120，000种化合物的文库。预计被鉴定为钙蛋白酶抑制剂的化合物将包括直接作用于酶的化合物和通过阻断途径中的上游步骤来防止钙蛋白酶活化的化合物。事实上，在抑制Aβ激活钙蛋白酶的化合物中，IC 50值< 10 μM，并且在高达30 μM的浓度下显示出很少或没有毒性，没有直接抑制钙蛋白酶。目前正在进行研究，以确定这些化合物在细胞死亡途径中的靶点。
Calpain activation is hypothesized to be an early occurrence in the sequence of events resulting in neurodegeneration, as well as in the signaling pathways linking extracellular accumulation of Aβ peptides and intracellular formation of neurofibrillary tangles. In an effort to identify small molecules that prevent neurodegeneration in Alzheimer’s disease by early intervention in the cell death cascade, a cell-based assay in differentiated Sh-SY5Y cells was developed utilizing calpain activity as a read-out for the early stages of death in cells exposed to extracellular Aβ. This assay was optimized for high-throughput screening, and a library of approximately 120,000 compounds tested. It was expected that the compounds identified as calpain inhibitors would include those that act directly on the enzyme and those that prevented calpain activation by blocking an upstream step in the pathway. In fact, of the compounds that inhibited calpain activation by Aβ with IC50 values < 10 μM and showed little or no toxicity at concentrations up to 30 μM, none inhibit the calpain enzyme directly. Studies to identify the targets of these compounds in the cell death pathway are ongoing.
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影响因子：
64.8
作者：
TSAI, LH;DELALLE, I;HARLOW, E
通讯作者：
HARLOW, E
影响因子：
3.5
作者：
Taniguchi, S;Fujita, Y;Hasegawa, M
通讯作者：
Hasegawa, M
DOI：
10.1006/bbrc.2000.2919
发表时间：
2000-06-24
影响因子：
3.1
作者：
Chen, M;Durr, J;Fernandez, HL
通讯作者：
Fernandez, HL
影响因子：
4.8
作者：
Kusakawa, G;Saito, T;Hisanaga, S
通讯作者：
Hisanaga, S
DOI：
10.1073/pnas.90.7.2628
发表时间：
1993-04-01
影响因子：
11.1
作者：
SAITO, KI;ELCE, JS;NIXON, RA
通讯作者：
NIXON, RA