Structural Genomics Consortium, UNC Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA.
Promega Corporation, 2800 Woods Hollow Road, Madison, WI, 53711, USA.
Nat Commun. 2020 Jun 2;11(1):2743. doi: 10.1038/s41467-020-16559-0.
Concerted multidisciplinary efforts have led to the development of Cyclin-Dependent Kinase inhibitors (CDKi's) as small molecule drugs and chemical probes of intracellular CDK function. However, conflicting data has been reported on the inhibitory potency of CDKi's and a systematic characterization of affinity and selectivity against intracellular CDKs is lacking. We have developed a panel of cell-permeable energy transfer probes to quantify target occupancy for all 21 human CDKs in live cells, and present a comprehensive evaluation of intracellular isozyme potency and selectivity for a collection of 46 clinically-advanced CDKi's and tool molecules. We observed unexpected intracellular activity profiles for a number of CDKi's, offering avenues for repurposing of highly potent molecules as probes for previously unreported targets. Overall, we provide a broadly applicable method for evaluating the selectivity of CDK inhibitors in living cells, and present a refined set of tool molecules to study CDK function.
协同的多学科努力导致了细胞周期蛋白依赖性激酶抑制剂 (CDKi) 的发展，作为小分子药物和细胞内 CDK 功能的化学探针。然而，关于 CDKi 的抑制效力已经报道了相互矛盾的数据，并且缺乏针对细胞内 CDK 的亲和力和选择性的系统表征。我们开发了一组细胞通透性能量转移探针，以定量测定所有 21 种人类 CDK 在活细胞中的靶标占有率，并对一组 46 种临床先进的 CDKi 和工具分子对细胞内同工酶效力和选择性进行了全面评估。我们观察到一些 CDKi 的细胞内活性谱出乎意料，为高活性分子重新用作以前未报道的靶标的探针提供了途径。总的来说，我们提供了一种广泛适用于评估细胞内 CDK 抑制剂选择性的方法，并提出了一组经过改进的工具分子来研究 CDK 功能。
