A selective chemical probe for exploring the role of CDK8 and CDK19 in human disease.
A selective chemical probe for exploring the role of CDK8 and CDK19 in human disease.
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DOI：
10.1038/nchembio.1952
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发表时间：
2015-12
影响因子：
14.8
通讯作者：
Blagg J
中科院分区：
文献类型：
--
作者：
Dale T;Clarke PA;Esdar C;Waalboer D;Adeniji-Popoola O;Ortiz-Ruiz MJ;Mallinger A;Samant RS;Czodrowski P;Musil D;Schwarz D;Schneider K;Stubbs M;Ewan K;Fraser E;TePoele R;Court W;Box G;Valenti M;de Haven Brandon A;Gowan S;Rohdich F;Raynaud F;Schneider R;Poeschke O;Blaukat A;Workman P;Schiemann K;Eccles SA;Wienke D;Blagg J
对化学工具的需求尚未得到满足，以探索介体复合物在从癌症到心血管疾病的人类病理学中的作用。在这里，我们确定了通过基于细胞的筛选发现的小分子WNT途径抑制剂CCT 251545是人类介体复合物相关蛋白激酶CDK 8和CDK 19的有效和选择性化学探针，其选择性超过291种其他激酶的100倍。X射线晶体学证明了1型结合模式，涉及CDK 8 C-末端插入配体结合位点。与CDK 8/19的II型抑制剂相比，CCT 251545显示出有效的细胞活性。我们发现，CCT 251545及其类似物改变了WNT通路调节的基因表达和调节CDK 8/19的其他靶向效应，包括STAT 1调节的基因。与此一致，我们发现STAT 1 SER 727的磷酸化是体外和体内CDK 8激酶活性的生物标志物。最后，我们证明了CCT 251545在WNT依赖性肿瘤中的体内活性。
There is unmet need for chemical tools to explore the role of the Mediator complex in human pathologies ranging from cancer to cardiovascular disease. Here we determine that CCT251545, a small molecule WNT-pathway inhibitor discovered through cell-based screening, is a potent and selective chemical probe for the human Mediator complex-associated protein kinases CDK8 and CDK19 with >100-fold selectivity over 291 other kinases. X-ray crystallography demonstrates a Type 1 binding mode involving insertion of the CDK8 C-terminus into the ligand binding site. In contrast to Type II inhibitors of CDK8/19, CCT251545 displays potent cell-based activity. We show that CCT251545 and close analogues alter WNT-pathway regulated gene expression and other on-target effects of modulating CDK8/19 including genes regulated by STAT1. Consistent with this we find that phosphorylation of STAT1SER727 is a biomarker of CDK8 kinase activity in vitro and in vivo. Finally, we demonstrate in vivo activity of CCT251545 in WNT-dependent tumors.