Histone deacetylases (HDAC) are enzymes that regulate the concentration of acetylated histones which, in turns, interact with the bromodomain (BRD) of BET (Bromodomain and Extracellular domain) proteins to affect transcriptional activity. Simultaneous blockade of both epigenetic players has shown synergistic effects in a variety of cancer cell lines. In this paper we report the design, synthesis and activity of new dual inhibitors, obtained by adding a methyltriazole moiety to some HDAC inhibitors carrying a benzodiazepine core, which were previously developed by us. An Alphascreen FRET assay showed that the compounds were able to interact with BRD4-1 and BRD4-2 proteins, with some selectivity for the latter, while the HDAC inhibiting properties were measured by means of an immunoprecipitation assay. The antiproliferative activity was tested on C26 adenocarcinoma, SSMC2 melanoma and SHSY5Y neuroblastoma cells. Interestingly, both compounds were endowed with antihyperalgesic activity in the mouse Spared Nerve Injury (SNI) model.

组蛋白去乙酰化酶 (HDAC) 是调节乙酰化组蛋白浓度的酶，乙酰化组蛋白反过来又与 BET（溴结构域和胞外结构域）蛋白的溴结构域 (BRD) 相互作用以影响转录活性。同时阻断两种表观遗传因素已在多种癌细胞系中显示出协同作用。在本文中，我们报告了新的双重抑制剂的设计、合成和活性，这些抑制剂是通过在我们之前开发的一些带有苯二氮卓核心的 HDAC 抑制剂中添加甲基三唑部分而获得的。Alphascreen FRET 试验表明，这些化合物能够与 BRD4-1 和 BRD4-2 蛋白相互作用，对后者具有一定的选择性，而 HDAC 抑制特性是通过免疫沉淀试验测量的。在 C26 腺癌细胞、SSMC2 黑色素瘤和 SHSY5Y 神经母细胞瘤细胞上测试了抗增殖活性。有趣的是，这两种化合物在小鼠备用神经损伤 (SNI) 模型中都具有抗痛觉过敏活性。