National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, USA.
SLAS Discov. 2020 Mar;25(3):253-264. doi: 10.1177/2472555219883625. Epub 2019 Oct 29.
Histone deacetylases (HDACs) are epigenetic modulators linked to diseases including cancer and neurodegeneration. Given their therapeutic potential, highly sensitive biochemical and cell-based profiling technologies have been developed to discover small-molecule HDAC inhibitors. Ultimately, the therapeutic action of these inhibitors is dependent on a physical engagement with their intended targets in cellular and tissue environments. Confirming target engagement in the cellular environment is particularly relevant for HDACs since they function as part of cell type-specific multiprotein complexes. Here we implemented two recently developed high-throughput target engagement technologies, NanoBRET and SplitLuc CETSA, to profile 349 compounds in the Epigenetic-Focused collection for HDAC1 binding. We found that the two HDAC1 target engagement assays correlated well with each other and with orthogonal activity-based assays, in particular those carried out in cellular environments rather than with isolated HDAC proteins. The assays detected a majority of the previously described HDAC1 inhibitors in the collection and, importantly, triaged HDAC inhibitors known to target other HDACs.
组蛋白去乙酰化酶（HDACs）是与癌症和神经退行性疾病等疾病相关的表观遗传调节剂。鉴于其治疗潜力，已经开发出高度敏感的生化和基于细胞的分析技术来发现小分子 HDAC 抑制剂。最终，这些抑制剂的治疗作用取决于它们在细胞和组织环境中与预期靶点的物理结合。在细胞环境中确认靶标结合对于 HDAC 尤为重要，因为它们作为细胞类型特异性多蛋白复合物的一部分发挥作用。在这里，我们实施了两种最近开发的高通量靶标结合技术，NanoBRET 和 SplitLuc CETSA，以分析 Epigenetic-Focused 集合中的 349 种化合物与 HDAC1 的结合。我们发现，两种 HDAC1 靶标结合测定法彼此之间以及与正交基于活性的测定法相关性良好，特别是在细胞环境中而不是在分离的 HDAC 蛋白中进行的那些测定法。这些测定法检测到集合中大多数先前描述的 HDAC1 抑制剂，重要的是，对已知针对其他 HDAC 的 HDAC 抑制剂进行了分类。
