A triple exon-skipping luciferase reporter assay identifies a new CLK inhibitor pharmacophore.
A triple exon-skipping luciferase reporter assay identifies a new CLK inhibitor pharmacophore.
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三重外显子跳跃荧光素酶报告基因检测鉴定出一种新的 CLK 抑制剂药效团。
DOI：
10.1016/j.bmcl.2016.12.056
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发表时间：
2017
影响因子：
2.7
通讯作者：
Webb,ThomasR
中科院分区：
文献类型：
--
作者：
Shi,Yihui;Park,Jaehyeon;Lagisetti,Chandraiah;Zhou,Wei;Sambucetti,LidiaC;Webb,ThomasR
mRNA 前体的剪接是正常细胞中的一个关键过程，但在癌症中却不受控制。最近，调节这一过程的化合物已被证明可以针对肿瘤中的特定脆弱性。我们开发了一种新型的基于细胞的检测方法，可以特异性激活暴露于 SF3B1 靶向化合物（例如苏德霉素 D6）的细胞中的荧光素酶。该测定用于筛选已批准的药物和生物活性化合物的组合。这种筛选方法确定了几种活性靶标，其中最有效的是 CGP-74514A 和氨基嘌呤醇 A，据报道，两者都是细胞周期蛋白依赖性激酶 (CDK) 抑制剂。我们发现这些化合物及其类似物在 CLK 上表现出显着的 cdc2 样激酶 (CLK) 抑制作用和清晰的构效关系 (SAR)。我们制备了一组类似物，能够“调出”CDK 活性，同时开发出具有低纳摩尔活性的 CLK 抑制剂。因此，我们展示了外显子跳跃测定的实用性，并鉴定了对 CLK 表现出效力和选择性的新分子，以及一些结构相关的双 CLK/CDK 抑制剂。
The splicing of pre-mRNA is a critical process in normal cells and is deregulated in cancer. Compounds that modulate this process have recently been shown to target a specific vulnerability in tumors. We have developed a novel cell-based assay that specifically activates luciferase in cells exposed to SF3B1 targeted compounds, such as sudemycin D6. This assay was used to screen a combined collection of approved drugs and bioactive compounds. This screening approach identified several active hits, the most potent of which were CGP-74514A and aminopurvalanol A, both have been reported to be cyclin-dependent kinases (CDKs) inhibitors. We found that these compounds, and their analogs, show significant cdc2-like kinase (CLK) inhibition and clear structure-activity relationships (SAR) at CLKs. We prepared a set of analogs and were able to ‘dial out’ the CDK activity and simultaneously developed CLK inhibitors with low nanomolar activity. Thus, we have demonstrated the utility of our exon-skipping assay and identified new molecules that exhibit potency and selectivity for CLK, as well as some structurally related dual CLK/CDK inhibitors.