Ross-Macdonald Petra, de Silva Heshani, Guo Qi, Xiao Hong, Hung Chen-Yi, Penhallow Becky, Markwalder Jay, He Liqi, Attar Ricardo M, Lin Tai-an, Seitz Steven, Tilford Charles, Wardwell-Swanson Judith, Jackson Donald
Bristol-Myers Squibb Research and Development, Princeton, NJ 08543-5400, USA.
Mol Cancer Ther. 2008 Nov;7(11):3490-8. doi: 10.1158/1535-7163.MCT-08-0826.
In developing inhibitors of the LIM kinases, the initial lead molecules combined potent target inhibition with potent cytotoxic activity. However, as subsequent compounds were evaluated, the cytotoxic activity separated from inhibition of LIM kinases. A rapid determination of the cytotoxic mechanism and its molecular target was enabled by integrating data from two robust core technologies. High-content assays and gene expression profiling both indicated an effect on microtubule stability. Although the cytotoxic compounds are still kinase inhibitors, and their structures did not predict tubulin as an obvious target, these results provided the impetus to test their effects on microtubule polymerization directly. Unexpectedly, we confirmed tubulin itself as a molecular target of the cytotoxic kinase inhibitor compounds. This general approach to mechanism of action questions could be extended to larger data sets of quantified phenotypic and gene expression data.
在开发LIM激酶抑制剂时，最初的先导分子将强效的靶点抑制与强效的细胞毒性活性结合在一起。然而，在评估后续化合物时，细胞毒性活性与LIM激酶的抑制作用分离了。通过整合来自两项强大核心技术的数据，能够快速确定细胞毒性机制及其分子靶点。高内涵分析和基因表达谱分析均表明对微管稳定性有影响。尽管细胞毒性化合物仍是激酶抑制剂，且其结构并未预测微管蛋白是一个明显的靶点，但这些结果促使人们直接测试它们对微管聚合的影响。出乎意料的是，我们证实微管蛋白本身就是细胞毒性激酶抑制剂化合物的分子靶点。这种针对作用机制问题的通用方法可以扩展到更大的定量表型和基因表达数据集。
