BI-847325 is a selective dual inhibitor of MEK and aurora kinases (AK) with IC50 values of 4 and 15 nM for human MEK2 and AK-C, respectively.

Aurora kinase:15 nM, MEK:4nM

BI-847325 在BRAF突变和对vemurafenib耐药的黑色素瘤细胞上显示出生长抑制效果，其IC50值在0.3 nM到2 μM范围内，并且在六个BRAF突变的黑色素瘤细胞系中阻止了集落形成。BI-847325 还通过减少Mcl-1表达来诱导凋亡。[1]

在携带1205Lu和1205LuR异种移植瘤的小鼠中，BI-847325（75 mg/kg，口服）明显抑制了肿瘤的生长，而且没有显著改变体重。[1]

Assays are run in the presence of 100 μM ATP using 10 μM of substrate. 30 μL PROTEIN-MIX in 25% DMSO and incubated for 15 min at room temperature. 10 μL PEPTIDE-MIX is added, the mixture is incubated for 60 min at RT and stopped by adding 180 μL 6.4% TCA (final concentration: 5%). Incorporated phosphate is measured in a scintillation counter and IC50 values are calculated using a sigmoidal curve analysis program with variable hill slope[1].

Cells are plated at a density of 2.5 × 103 cells per 100 μL and left to grow overnight before being treated with increasing concentrations of BI-847325 for 72 hours. The metabolic activity is determined using Alamar blue reagent as per the manufacturer's protocol.(Only for Reference)