BLU9931 is the first selective small molecule inhibitor of FGFR4.

FGFR4:3 nM, FGFR3:150 nM

在MDA-MB-453细胞中，BLU9931强效抑制FGFR4信号通路的磷酸化。BLU9931抑制表达完整FGFR4信号复合体的HCC细胞系的增殖，如Hep 3B、HUH-7和JHH-7细胞系，其EC50 <1 μM。BLU9931也抑制具有完整FGFR4信号通路的PDX来源细胞系的增殖。[1]

在携带FGF19扩增的Hep 3B肝癌的小鼠中，BLU9931（300 mg/kg，口服）导致肿瘤缩小并阻止了由肿瘤引起的体重减轻。在携带表达过量FGF19的PDX源LIXC012异种移植物的小鼠中，使用BLU9931（300 mg/kg，口服）同样导致肿瘤缩小。[1]

FGFR1-4 Biochemical Assays: FGFR kinase inhibition assays are performed at KM for ATP. Picomolar to low nanomolar concentrations of FGFR proteins are incubated in 1× Kinase Reaction Buffer (KRB) with 1 μM of CSKtide and 50 to 250 of μM ATP at 25°C for 90 minutes in the presence or absence of a dosed concentration series of inhibitor. All reactions are terminated by the addition of Stop buffer, and plates are read on a Caliper EZReader2. IC50 values are fit with a four-parameter log[Inhibitor] versus response model with floating Hill Slope.

Established and PDX-derived HCC cell lines are seeded in 96-well plates in respective growth media, allowed to attach overnight, and treated with a dilution series of test compounds for two cell-doubling times. Cell viability is determined by CellTiter-Glo, and results represented as background-subtracted relative light units normalized to a DMSO-treated control. Relative EC50 values are determined at 50% inhibition between the top and bottom plateau of the dose–response curve.(Only for Reference)