SB 216763 (SB216763) is an effective and specific GSK-3α/β inhibitor (IC50: 34.3 nM).

GSK-3α:34.3 nM, GSK-3β:34.3 nM

SB-216763通过对肺泡上皮细胞损伤的抑制作用显著降低博来霉素诱导的肺泡.在患博来霉素诱导肺部炎症和纤维化的小鼠体内,SB-216763（20 mg/kg）通过对炎症细胞因子MCP-1和TNF-α产量的显著抑制作用,从而明显抑制肺部炎症和纤维化,且明显提高用博来霉素处理的小鼠寿命.

在人肝脏细胞中,SB-216763可刺激糖原合成（EC50：3.6 μM）。在HEK293 细胞在,其可剂量依赖性诱导β-catenin-LEF/TCF相关报告基因的转录,5 μM可诱导达到最大值（增大2.5倍）。在小脑颗粒神经元中,SB-216763可使之免受LY-294002或钾丧失诱导的细胞凋亡,该作用呈浓度依赖性,3 μM时保护神经达到最高值。SB-216763（3 μM）可完全抑制LY-294002诱导的鸡背根神经节感觉神经元死亡。SB-216763（10 μM）除抑制GSK-3α外,对糖原合成酶激酶-3也同样有效（96%）,对其余24种蛋白激酶包括PKBα和PDK1的活性很小（IC50>10 μM）。在前列腺癌细胞系,包括MIA-PaCa2,BXPC-3,PANC1,ASPC1和CFPAC中,SB-216763（25-50 μM）可剂量依赖性降低细胞活力,且显著促进凋亡,处理72 h后凋亡达50%,但不影响HMEC或WI38细胞系。

GSK-3 activity assay: GSK-3 kinase activity is measured, in the presence of various concentrations of SB 216763, in a reaction mixture containing final concentrations of 1 nM human GSK-3α, 50 mM MOPS pH 7.0, 0.2 mM EDTA, 10 mM Mg-acetate, 7.5 mM β-mercaptoethanol, 5% (w/v) glycerol, 0.01% (w/v) Tween-20, 10% (v/v) DMSO, and 28 μM GS-2 peptide substrate. The GS-2 peptide sequence corresponds to a region of glycogen synthase that is phosphorylated by GSK-3. The assay is initiated by the addition of 0.34 μCi [33P]γ-ATP. The total ATP concentration is 10 μM. Following 30 minutes incubation at room temperature the assay is stopped by the addition of one third assay volume of 2.5% (v/v) H3PO4 containing 21 mM ATP. Samples are spotted onto P30 phosphocellulose mats and washed six times in 0.5% (v/v) H3PO4. The filter mats are sealed into sample bags containing Wallac betaplate scintillation fluid. 33P incorporation into the substrate peptide is determined by counting the mats in a Wallac microbeta scintillation counter.

Cells are exposed to various concentrations of SB 216763 for 24, 48 and 72 hours. Relative cell viability is measured using the MTS assay. Apoptotic cells are determined by staining with Hoechst.(Only for Reference)