AZD2858 是具有口服活性的GSK-3抑制剂，可以抑制 GSK-3α (IC50:0.9 nM) 和 GSK-3β (IC50:5 nM) 的活性，可用于骨折愈合的研究。

AZD2858 is a selective GSK-3 inhibitor, inhibiting tau phosphorylation at the S396 site and activating Wnt signaling pathway.

30 μM/kg AZD2858处理大鼠三周后胼胝中矿物质密度(2周时28%,三周时38%)和矿物质含量(两周时81%,三周时93%)增加.大鼠用AZD2858处理28天后,血清中骨更新标志物产生时间依赖性变化,且骨密度增加.大鼠用AZD2858处理7天后,骨形成标志P1NP增加,溶蚀标志物TRAcP-5b降低,表明骨合成代谢增加,大鼠吸收减少.大鼠口服AZD2858治疗两周后,与对照组相比,引起骨密度剂量依赖增加,治疗两周后,在每天20 mg/kg(总BMC: 对照组的172%)剂量下具有最大疗效.AZD2858处理使骨折恢复更快,有骨性骨痂但没有明显的软骨成分.

AZD2858在人类和大鼠间充质干细胞中引起β-连环蛋白稳定,激活体外成骨细胞和成骨矿化。初级分离的人成骨细胞样细胞上AZD2858处理（1 μM,12小时）导致β-连环蛋白水平增加3倍。

Tau phosphorylation assay: NIH-3T3 cells expressing 4-repeat Tau are used to assess functional activity of AZD2858 in vitro. The cells are grown in DMEM media and 2 mM L-glut, and 10% HiFCS, and plated at a concentration of 6×105 cells/well in 6-well plates. In each experiment, AZD2858 is dosed in triplicates at a concentration of 1, 10, 100, 500, 1000, 2000 and 10,000 nM. Cells are treated for 4 h prior to cell lysis using 100 μL ice cold lysis buffer (0.5% NP-40, 10 mM Tris, pH 7.2, 150 mM NaCl, 2 mM EDTA). A suspension is made with addition of protease and phosphatase inhibitors: 50 mM NaF, 0.2 mM NaVO4 and Cocktail Protease inhibitors. The solution is then snap frozen at ? 80 °C for at least 1 h, before thawing on ice and lysate clarification by centrifugation, followed by Western blot according to standard protocols. After blocking, the blots are exposed to the primary antibody, Phospho-Ser396-tau (1:1000) over night, washed and incubated with the secondary antibody (donkey anti-rabbit, 1:5000), followed by a final wash. For re-probing, the primary antibody Tau5 (1:200) and the secondary horseradish peroxidase linked antibody (sheep anti-mouse, 1:10000) are used. All blots are developed using ECL Western blot detection reagents, Kodak X-ray films, quantified using densitometric analysis, and the ratio of S396 tau to total tau (tau5) is calculated.

Human adipose derived stem cells and rat MSCs (isolated from bone marrow of Sprague Dawley rats at less than 8 weeks after gestation) are cultured in a basal media of DMEM containing 5% FBS and 2 mM GlutaMax. Cells are seeded in basal media into 96-well plates (3–5000 cells/well) for 18 h before treatment with AZD2858 (0.3 nM to 20 mM). After 24 h, β-catenin stabilisation is measured.(Only for Reference)

486424-20-8

C21H23N7O3S

453.52

Ethanol:<1 mgml
DMSO:7 mg/mL (15.43 mM)
Powder: -20°C for 3 years
In solvent: -80°C for 2 years