Biological Activity

GSK126 is a potent, highly selective, S-adenosyl-methionine-competitive, small-molecule inhibitor of EZH2 methyltransferase activity. GSK126 decreases global H3K27me3 levels and reactivates silenced PRC2 target genes. GSK126 effectively inhibits the proliferation of EZH2 mutant DLBCL cell lines and markedly inhibits the growth of EZH2 mutant DLBCL xenografts in mice.

Customer Product Validations & Biological Datas

	Source	Oncotarget (2017). Figure 2. GSK126
	Method	Clonogenicity
	Cell Lines	multiple myeloma cells
	Concentrations	10 μM
	Incubation Time	24 h
	Results	The results showed that after exposure to GSK126 for 24 h, the colonyformation ability of MM cells were significantly reduced with IC50 < 10 μM

Protocol (for reference only)

Cell Experiment
Cell lines	EZH2 wildtype or mutant B-cell lymphoma cell lines
Preparation method	Cell proliferation assay. The optimal cell seeding was determined empirically for all cell lines by examining the growth of a wide range of seeding densities in a 384-well format to identify conditions that permitted proliferation for 6 days. Cells were then plated at the optimal seeding density 24 h before treatment (in duplicate) with a 20-point twofold dilution series of GSK126 or 0.15% DMSO. Plates were incubated for 6 days at 37℃ in 5% CO2. Cells were then lysed with CellTiter-Glo (CTG) (Promega) and chemiluminescent signal was detected with a TECAN Safire2 microplate reader. In addition, an untreated plate of cells was harvested at the time of compound addition (T0) to quantify the starting number of cells. CTG values obtained after the 6 day treatment were expressed as a percent of the T0 value and plotted against compound concentration. Data were fit with a four-parameter equation to generate a concentration response curve and the concentration of GSK126 required to inhibit 50% of growth (growth IC50) was determined.
Concentrations	0~100µM
Incubation time	6 days

Animal Experiment
Animal models	Pfeiffer or KARPAS-422 cells tumour xenograft in mice
Formulation	20% captisol adjusted to pH 4–4.5 with 1 N acetic acid.
Dosages	0.2 ml per 20 g body weight
Administration	intraperitoneally

Chemical Information

Molecular Weight	526.67
Formula	C31H38N6O2
CAS Number	1346574-57-9
Solubility (25°C)	DMSO 10 mg/mL
Storage	Powder          -20°C   3 years ;  4°C   2 years In solvent       -80°C   6 months ;  -20°C   1 month

References

[1] Sato T, et al. Sci Rep. PRC2 overexpression and PRC2-target gene repression relating to poorer prognosis in small cell lung cancer.

[2] McCabe MT, et al. Nature. EZH2 inhibition as a therapeutic strategy for lymphoma with EZH2-activating mutations.