Tubulin is a molecular target of the Wnt-activating chemical probe.
Tubulin is a molecular target of the Wnt-activating chemical probe.
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DOI：
10.1186/s12858-016-0066-9
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发表时间：
2016-05-20
期刊：
影响因子：
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通讯作者：
Matsui J
中科院分区：
文献类型：
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作者：
Fukuda Y;Sano O;Kazetani K;Yamamoto K;Iwata H;Matsui J
在药物发现研究中，基于细胞的表型筛选是获得潜在候选药物的重要方法。揭示作用机制是药物发现道路上的关键一步。然而，从表型筛选活动中阐明命中化合物的靶分子仍然是一个困难和麻烦的过程。用于鉴定靶分子的简单而有效的方法是必不可少的。2-氨基-4-（3，4-（亚甲二氧基）苄氨基）-6-（3-甲氧基苯基）嘧啶（AMBMP）从表型筛选活动中被鉴定为衰老诱导剂。该化合物被广泛用作Wnt激动剂，尽管其靶分子仍有待澄清。为了鉴定其靶蛋白，我们将该化合物的一系列细胞测定结果与我们的途径分析数据库进行了比较。该数据库包括来自细胞报告基因和细胞增殖的简单测定的化合物的活性。在该数据库中，化合物根据其活性的统计分析进行分类，这对应于代表性化合物的作用机制。此外，可以使用数据库预测感兴趣的化合物的作用机制。基于我们的数据库分析，该化合物预计是微管蛋白破坏剂，随后通过其抑制微管蛋白聚合的活性证实了这一点。这些结果表明，微管蛋白首次被鉴定为Wnt活化小分子的靶分子，这可能误导了一些先前研究的结论。此外，本研究还强调，我们的途径分析数据库是一个简单而有效的工具，揭示了从表型筛选和脱靶化学探针获得的命中化合物的作用机制。本文的在线版本（doi：10.1186/s12858-016-0066-9）包含补充材料，可供授权用户使用。
In drug discovery research, cell-based phenotypic screening is an essential method for obtaining potential drug candidates. Revealing the mechanism of action is a key step on the path to drug discovery. However, elucidating the target molecules of hit compounds from phenotypic screening campaigns remains a difficult and troublesome process. Simple and efficient methods for identifying the target molecules are essential. 2-Amino-4-(3,4-(methylenedioxy)benzylamino)-6-(3-methoxyphenyl)pyrimidine (AMBMP) was identified as a senescence inducer from a phenotypic screening campaign. The compound is widely used as a Wnt agonist, although its target molecules remain to be clarified. To identify its target proteins, we compared a series of cellular assay results for the compound with our pathway profiling database. The database comprises the activities of compounds from simple assays of cellular reporter genes and cellular proliferations. In this database, compounds were classified on the basis of statistical analysis of their activities, which corresponded to a mechanism of action by the representative compounds. In addition, the mechanisms of action of the compounds of interest could be predicted using the database. Based on our database analysis, the compound was anticipated to be a tubulin disruptor, which was subsequently confirmed by its inhibitory activity of tubulin polymerization. These results demonstrate that tubulin is identified for the first time as a target molecule of the Wnt-activating small molecule and that this might have misled the conclusions of some previous studies. Moreover, the present study also emphasizes that our pathway profiling database is a simple and potent tool for revealing the mechanisms of action of hit compounds obtained from phenotypic screenings and off targets of chemical probes. The online version of this article (doi:10.1186/s12858-016-0066-9) contains supplementary material, which is available to authorized users.