ML346 is a novel activator of Hsp70.

HSP70:4.6 μM (EC50, HeLa cells)

ML346具有良好的化学稳定性，水溶性显著高，不与过量的谷胱甘肽反应，并能穿透细胞膜。ML346作为探针，将有助于阐明Hsp70和HSF-1在防止和促进癌症发展、细胞衰老、代谢及神经退行性疾病中的作用。ML346是针对蛋白质构象疾病的新型蛋白质稳态调节剂。

ML346在C. elegans模型中激活Hsp70启动子的转录，并抑制多聚谷氨酸的聚集，表明该探针在调节蛋白质聚集及其相关毒性方面具有效力。ML346诱导HSF-1依赖的分子伴侣表达，并在构象疾病模型中恢复蛋白质折叠。

Assays (25.5 μL volume) are carried out robotically at room temperature (21°C) and are linear with respect to time and enzyme concentration under the conditions used. Assays are performed for 30 min using Multidrop Micro reagent dispensers in a 96-well format. The concentration of magnesium acetate in the assays is 10 mM and [γ-33P]ATP (800 c.p.m./pmol) is used at 5, 20 or 50 μM as indicated, in order to be at or below the Kmfor ATP for each enzyme.The assays are initiated with MgATP, stopped by the addition of 5 μL of 0.5 M orthophosphoric acid and spotted on to P81 filter plates using a unifilter harvester. The IC50 values of inhibitors are determined after carrying out assays at ten different concentrations of each compound[2].