Abstract：
The PCTAIRE subfamily belongs to the CDK (cyclin-dependent kinase) family and represents an understudied class of kinases of the dark kinome. They exhibit a highly conserved binding pocket and are activated by cyclin Y binding. CDK16 is targeted to the plasma membrane after binding to N-myristoylated cyclin Y and is highly expressed in post-mitotic tissues, such as the brain and testis. Dysregulation is associated with several diseases, including breast, prostate, and cervical cancer. Here, we used the N-(1H-pyrazol-3-yl)pyrimidin-4-amine moiety from the promiscuous inhibitor 1 to target CDK16, by varying different residues. Further optimization steps led to 43d, which exhibited high cellular potency for CDK16 (EC50 = 33 nM) and the other members of the PCTAIRE and PFTAIRE family with 20-120 nM and 50-180 nM, respectively. A DSF screen against a representative panel of approximately 100 kinases exhibited a selective inhibition over the other kinases. In a viability assessment, 43d decreased the cell count in a dose-dependent manner. A FUCCI cell cycle assay revealed a G2/M phase cell cycle arrest at all tested concentrations for 43d, caused by inhibition of CDK16.
摘要：
PCTAIRE亚家族属于CDK（细胞周期蛋白依赖性激酶）家族，代表了一类未被研究的暗激酶。它们表现出高度保守的结合口袋，并被细胞周期蛋白Y结合激活。CDK16与N-肉豆蔻酰化细胞周期蛋白Y结合后靶向质膜，并在有丝分裂后组织中高度表达，比如大脑和睾丸。失调与几种疾病相关，包括乳房,前列腺,还有宫颈癌.这里,我们使用来自混杂抑制剂1的N-(1H-吡唑-3-基)嘧啶-4-胺部分,通过改变不同的残基靶向CDK16.进一步的优化步骤导致43d，它对CDK16（EC50=33nM）和PCTAIRE和PFTAIRE家族的其他成员具有20-120nM和50-180nM的高细胞效力，分别。针对大约100种激酶的代表性小组的DSF筛选显示出优于其他激酶的选择性抑制。在可行性评估中，43d以剂量依赖性方式减少细胞计数。FUCCI细胞周期分析显示，在所有测试浓度下，G2/M期细胞周期停滞43d，由CDK16的抑制引起。
