Zwick Vincent, Simões-Pires Claudia, Cuendet Muriel
a School of Pharmaceutical Sciences, University of Geneva, University of Lausanne , Geneva , Switzerland.
J Enzyme Inhib Med Chem. 2016;31(sup1):209-214. doi: 10.1080/14756366.2016.1180595. Epub 2016 May 5.
Histone deacetylases (HDAC) are involved in several diseases including cancer, cardiovascular and neurodegenerative disorders, and the search for inhibitors is a current topic in drug discovery. Four HDAC inhibitors have already been approved by the FDA for cancer therapy and others are under clinical studies. However, the clinical utility of some of them is limited because of unfavorable toxicities associated with their broad range of HDAC inhibitory effects. Toxicity could be decreased by using HDAC inhibitors with improved specificity. To date, the most popular screening assays are based on fluorescence-labeled substrates incubated with an enzymatic source (cells extracts or recombinant isoforms). Here, we describe a high-throughput cell-based UHPLC-ESI-MS/MS assay able to rapidly predict activity against HDAC1 and HDAC6 in a cell environment. This method is predicted to be a useful tool to accelerate the search for class-selective HDAC inhibitors in drug discovery.
组蛋白去乙酰化酶（HDAC）与多种疾病有关，包括癌症、心血管疾病和神经退行性疾病，寻找抑制剂是药物研发领域当前的一个热门话题。四种HDAC抑制剂已获美国食品药品监督管理局（FDA）批准用于癌症治疗，其他抑制剂正处于临床研究阶段。然而，其中一些抑制剂的临床应用受到限制，因为其广泛的HDAC抑制作用会带来不良毒性。使用特异性更高的HDAC抑制剂可以降低毒性。迄今为止，最常用的筛选方法是基于与酶源（细胞提取物或重组亚型）一起孵育的荧光标记底物。在此，我们描述了一种基于细胞的高通量超高效液相色谱-电喷雾串联质谱（UHPLC-ESI-MS/MS）分析方法，该方法能够在细胞环境中快速预测对HDAC1和HDAC6的活性。预计该方法将成为加速药物研发中寻找选择性HDAC抑制剂的有用工具。
