Tideglusib (NP031112), a non-ATP competitive inhibitor of glycogen synthase kinase 3 (GSK-3), is with anti-inflammatory and neuroprotective activities.

GSK-3β:60 nM

在成年雄性Wistar大鼠海马体中,注射 Tideglusib（50 mg/kg）能够改善炎症反应,并对损伤区域起到保护作用.在APP/tau双转基因小鼠中,口服 Tideglusib（200 mg/kg）能够下调 tau蛋白磷酸化水平,抑制淀粉样蛋白沉积和斑块相关的星形胶质细胞增殖.

在大鼠原代星形胶质细胞或小胶质细胞中,Tideglusib（2.5 μM ）能够抑制谷氨酸盐诱导的细胞激活。在人的成神经细胞瘤细胞和鼠的原代神经元中,Tideglusib不可逆的抑制GSK-3,因此tau蛋白磷酸化水平降低并防止细胞凋亡。

[35S]Tideglusib (207 Bq/nmol) at 55 μM is incubated with 5 μM GSK-3β for 1 h at 25°C in 315 μL of 50 mM Tris-HCl, pH 7.5, containing 150 mM NaCl and 0.1 mM EGTA. The incubation is extended for another 30 min after having added 35 μL of the same buffer with or without 100 mM DTE. Samples are then processed in three different ways. First, an aliquot of 125 μL of each sample is mixed with 375 μL of 8 M GdnHCl in Water and heated at 80°C for 5 min. A second aliquot of 125 μL is diluted up to 500 μL with Water and left at room temperature for 5 min. In both cases, the free drug is removed afterwards by gel filtration through Sephadex G-25, and the amount of bound drug is determined by liquid scintillation counting on a 1450-MicroBeta TriLux counter. Finally, a third 40 μL aliquot of each original sample is mixed with 10 μL of denaturing electrophoresis sample buffer without reducing agents, and 35 μL of this mixture is loaded onto a 10% polyacrylamide gel and subjected to SDS-PAGE (again in the absence of reducing agents except for the DTE already included in the corresponding sample), followed by fluorography of the dried gel[1].