GSK923295 is a selective allosteric inhibitor of CENP-E kinesin motor ATPase(Ki=3.2 nM).

CENP-E:3.2 nM(Ki)

在多种实体瘤模型中,如尤因肉瘤,杆状瘤,横纹肌肉瘤移植瘤,GSK923295均能产生显著地抗肿瘤活性.Colo205移植瘤中,GSK923295（125 mg/kg）能够抑制肿瘤生长,使核分裂和游离的凋亡小体明显增加.

在癌细胞系中,GSK923295具有广谱抗肿瘤活性,对于 SW48（IC50=17.2 nM）, RKO (BRAF mutant)（ IC50=55.6 nM）, SW620 (KRAS mutant)（IC50=42 nM）, and HCT116 (KRAS mutant)（IC50=51.9 nM）。在人类神经母细胞瘤细胞系中,GSK923295平均生长IC50为41 nM。

Enzymology: Kinesin motor domains are expressed in Escherichia coli BL21(DE3) and purified. CENP-E proteins includes residues 2–340 with a carboxyl-terminal 6-his tag. All studies using MT are conducted in PEM25 buffer [25 mM PipesK+ (pH 6.8), 2 mM MgCl2, 1 mM EGTA] supplemented with 10 μM paclitaxel. The IC50 for steady-state inhibition is determined at 500 μM ATP, 5 μM MT, and 1 nM CENP-E in PEM25 buffer.

Cell-growth inhibition assays are performed by MDS in 384-well plates, and DNA content of fixed cells stained with DAPI using an Incell 1000 is analyzed. DNA content is determined 24 h after seeding (T0) and after exposure to varying concentrations of drug for an additional 72 h (T72). All T72 measurements are normalized to T0. Curves are analyzed using the XLfit curve-fitting tool to determine the concentration of GSK923295 yielding 50% growth inhibition relative to T0 and Ymax values (GI50).(Only for Reference)