SB 216763

Biological Activity

SB 216763 is a potent and selective ATP-competitive inhibitor of the serine/threonine protein kinase glycogen synthase kinase-3 (GSK-3) α and β isozymes. Glycogen synthase kinase 3 (GSK3) is a serine/threonine protein kinase that is inhibited by an assortment of extracellular stimuli including insulin, growth factors, cell specification factors, and cell adhesion. SB 216763 displays similar potency for GSK-3β with 96% inhibition at 10 μM while exhibiting minimal activity against 24 other protein kinases (IC50 >10 μM). SB 216763 stimulates glycogen synthesis in Chang human liver cells with an EC50 value of 3.6 µM and induces expression of a β-catenin-LEF/TCF regulated reporter gene in HEK293 cells. 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. 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.

Protocol

Cell Experiment
Cell lines	BXPC-3 cells and MIA-PaCa2 cells
Preparation method	BXPC-3 cells were treated with DMSO or the indicated concentration of GSK-3β inhibitors (AR-A014418 and SB216763) over the indicated time course. Relative cell viability was measured using the colorimetric 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium assay. B, MIA-PaCa2 cells and the normal human mammary epithelial cell line HMEC, and embryonic lung fibroblast cell line WI38 were treated with DMSO or the indicated GSK-3β inhibitor (25 μmol/L).
Concentrations	1, 10, 25, 50µM
Incubation time	24, 48 or 72h

Animal Experiment
Animal models	saline induced pulmonary fibrosis model of C57BL/6N mice (12 weeks old)
Formulation	25% dimethyl sulfoxide, 25% polyethylene glycol, and 50% saline
Dosages	20 mg/kg
Administration	intravenously twice a week until day 28

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

Species	Mouse	Rat	Rabbit	Guinea pig	Hamster	Dog
Weight (kg)	0.02	0.15	1.8	0.4	0.08	10
Body Surface Area (m2)	0.007	0.025	0.15	0.05	0.02	0.5
Km factor	3	6	12	8	5	20

Animal A (mg/kg) = Animal B (mg/kg) multiplied by	Animal B Km
	Animal A Km

For example, to modify the dose of resveratrol used for a mouse (22.4 mg/kg) to a dose based on the BSA for a rat, multiply 22.4 mg/kg by the K_m factor for a mouse and then divide by the K_m factor for a rat. This calculation results in a rat equivalent dose for resveratrol of 11.2 mg/kg.

Chemical Information

Customer Product Validations & Biological Datas

	Source	PLoS One (2012). Figure 1. SB 216763
	Method	dual luciferase reporter activity
	Cell Lines	HEK293 cells
	Concentrations	10 μM
	Incubation Time	24 h
	Results	"As expected, HEK293 cells treated with 1–20 μM SB-216763 exhibited a dose-dependent increase in Super TOPFlash activity"
	Rating

References

Glycogen synthase kinase 3 (GSK3) inhibitor, SB-216763, promotes pluripotency in mouse embryonic stem cells.
Kirby LA, et al. PLoS One. 2012;7(6):e39329. PMID: 22745733.

Selective small molecule inhibitors of glycogen synthase kinase-3 modulate glycogen metabolism and gene transcription.
Coghlan MP, et al. Chem Biol. 2000 Oct;7(10):793-803. PMID: 11033082.