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Research Area
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Description
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Cancer |
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Biological Activity
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Description
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SB 216763 is a potent and selective GSK-3α inhibitor with IC50 of 34.3 nM. |
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Targets
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GSK-3α |
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IC50 |
34.3 nM [1] |
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In Vitro
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SB 216763 displays similar potency for GSK-3β with 96% inhibition at 10 μM while exhibiting minimal activity against 24 other protein kinases including PKBα and PDK1 with IC50 of >10 μM. SB 216763 stimulates glycogen synthesis in human liver cells with EC50 of 3.6 μM, and induces dose-dependent transcription of a β-catenin-LEF/TCF regulated reporter gene in HEK293 cells with a maximum 2.5-fold induction at 5 μM. [1] SB 216763 protects the cerebellar granule neurones from apoptotic cell death induced by LY-294002 or potassium-deprivation in a concentration-dependent manner, with a maximal neuroprotection at 3 μM in contrast with the effect of lithium chloride at which 10 mM is required. SB 216763 at 3 μM also completely prevents death of chicken dorsal root ganglion sensory neurones induced by LY-294002 regardless of NGF. SB 216763 treatment at 5 μM markedly inhibits the GSK-3-dependent phosphorylation of neuronal-specific microtubule-associated protein tau in cerebellar granule neurones or recombinant tau in HEK293 cells, and induces increased levels of cytoplasmic β-catenin in both cells mimicking the effect of Wnt-mediated inhibition of GSK-3. [2] In pancreatic cancer cell lines including BXPC-3, MIA-PaCa2, PANC1, ASPC1, and CFPAC, SB 216763 treatment at 25-50 μM reduces cell viability in a dose-dependent manner, and leads to significant increase in apoptosis by 50% at 72 hours due to the specific down regulation of GSK-3β, while has no effect in HMEC or WI38 cell lines. [3] |
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In Vivo
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Administration of SB 216763 at 20 mg/kg significantly prevents lung inflammation and the subsequent fibrosis in bleomycin (BLM)-induced pulmonary inflammation and fibrosis model in mice by significantly blocking the production of inflammatory cytokines MCP-1 and TNF-α by macrophages, and significantly improves the survival of BLM-treated mice. SB 216763 treatment causes a significant reduction in BLM-induced alveolitis by inhibiting alveolar epithelial cell damage. [4] |
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Clinical Trials
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Features
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Protocol
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Kinase Assay
[1]
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| 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. |
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Cell Assay
[3]
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| Cell Lines |
BXPC-3, MIA-PaCa2, PANC1, ASPC1, and CFPAC cells |
| Concentrations |
Dissolved in DMSO, final concentrations ~50 μM |
| Incubation Time |
24, 48, and 72 hours |
| Methods |
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. |
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Animal Study
[4]
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| Animal Models |
C57BL/6N mice with lung inflammation and fibrosis induced by bleomycin (BLM) |
| Formulation |
Dissolved in DMSO, and diluted in saline |
| Doses |
20 mg/kg |
| Administration |
Intravenously |
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References |
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[1] Coghlan MP, et al. Chem Biol, 2000, 7(10), 793-803.
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[2] Cross DA, et al. J Neurochem, 2001, 77(1), 94-102.
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[3] Ougolkov AV, et al. Cancer Res, 2005, 65(6), 2076-2081.
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[4] Gurrieri C, et al. J Pharmacol Exp Ther, 2010, 332(3), 785-794.
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