A systematic interaction map of validated kinase inhibitors with Ser/Thr kinases
A systematic interaction map of validated kinase inhibitors with Ser/Thr kinases
复制标题
DOI：
10.1073/pnas.0708800104
复制
发表时间：
2007-12-18
影响因子：
11.1
通讯作者：
Knapp, Stefan
中科院分区：
文献类型：
--
作者：
Fedorov, Oleg;Marsden, Brian;Knapp, Stefan
蛋白激酶在细胞信号转导中起着关键作用，许多蛋白激酶的失调与疾病的发展有关。因此，大量的激酶抑制剂目前正在进行临床试验研究，到目前为止，已有7种抑制剂被批准为抗癌药物。此外，激酶抑制剂被广泛用作研究细胞信号转导的特异性探针，但描述这些试剂对一系列不同激酶的选择性的系统研究在很大程度上是缺乏的。在这里，我们使用热稳定性变化分析评估了156种有效的激酶抑制剂，包括临床试验中使用的抑制剂，对60个人类丝氨酸/苏氨酸激酶的特异性。我们的分析揭示了许多意想不到的交叉反应，被认为是特定靶点的抑制剂。我们还发现，ATP结合位点上的某些激活位残基的组合与检测到的配体混杂有关，并且一些激酶对使用不同化学类型的抑制高度敏感，这表明它们是首选的干预点。我们的结果还发现了可能导致替代临床应用的抑制剂交叉反应。例如，目前处于第三阶段临床试验的PKCβ抑制剂LY333‘531，在我们的筛选中有效地抑制了PIM1激酶，这是建议用于治疗白血病的靶点。我们用X射线结晶学确定了该抑制物的结合方式，并发现LY333‘531可诱导细胞死亡，并显著抑制急性髓系白血病患者白血病细胞的生长。
Protein kinases play a pivotal role in cell signaling, and dysregulation of many kinases has been linked to disease development. A large number of kinase inhibitors are therefore currently under investigation in clinical trials, and so far seven inhibitors have been approved as anti-cancer drugs. In addition, kinase inhibitors are widely used as specific probes to study cell signaling, but systematic studies describing selectivity of these reagents across a panel of diverse kinases are largely lacking. Here we evaluated the specificity of 156 validated kinase inhibitors, including inhibitors used in clinical trials, against 60 human Ser/Thr kinases using a thermal stability shift assay. Our analysis revealed many unexpected cross-reactivities for inhibitors thought to be specific for certain targets. We also found that certain combinations of activesite residues in the ATP-binding site correlated with the detected ligand promiscuity and that some kinases are highly sensitive to inhibition using diverse chemotypes, suggesting them as preferred intervention points. Our results uncovered also inhibitor cross-reactivities that may lead to alternate clinical applications. For example, LY333'531, a PKC beta inhibitor currently in phase III clinical trials, efficiently inhibited PIM1 kinase in our screen, a suggested target for treatment of leukemia. We determined the binding mode of this inhibitor by x-ray crystallography and in addition showed that LY333'531 induced cell death and significantly suppressed growth of leukemic cells from acute myeloid leukemia patients.