Capmatinib

MET

412, 42

Eight-week-old female Balb/c nu/nu mice (Charles River) are inoculated subcutaneously with 4, 106 tumor cells (S114 model) or with 5, 106 tumor cells (U-87MG glioblastoma model).

INCB28060 is now under Phase I clinical trial for a dose-escalation study in subjects with advanced malignancies.

3, 10, 30 mg/kg

0.13 nM [1], 0.13 nM [1], 0.13 nM [1], 0.13 nM [1], 0.13 nM [1], 0.13 nM [1]

INCB28060 shows strong antitumor activity in c-MET-dependent mouse tumor models, even oral treatment of 0.03 mg/kg INCB28060 causes approximately 50% inhibition of c-MET-phosphorylation. Dose-dependent inhibition of tumor growth is observed in tumor-bearing mice. [1]

c-Met Kinase Assay, The assay buffer contains 50 mM Tris-HCl, 10 mM MgCl2, 100 mM NaCl, 0.1 mg/ml BSA, 5mM DTT, pH 7.8. For HTS 0.8 uL of 5 mM of INCB28060 dissolved in DMSO are dotted on 384-well plates. DMSO titration suggests that the maximum tolerated concentration of the solvent is 4%. To measure IC50s the INCB28060 plate is prepared by 3-fold and 11-point serial dilutions. 0.8 uL of INCB28060 in DMSO is transferred from INCB28060 plate to the assay plate. The final concentration of DMSO is 2%. Solutions of 8 nM unphosphorylated c-Met or 0.5 nM phosphorylated c-Met are prepared in assay buffer. A 1 mM stock solution of peptide substrate Biotin-EQEDEPEGDYFEWLE-amide dissolved in DMSO is diluted to 1 uM in assay buffer containing 400 uM ATP (unphosphorylated c-Met) or 160 uM ATP (phosphorylated c-Met). A 20 uL volume of enzyme solution (or assay buffer for the enzyme blank) is added to the appropriate wells in each plate and then 20 uL/well of substrate solution to initiate the reaction. The plate is protected from light and incubated at 25 C for 90 minutes. The reaction is stopped by adding 20 uL of a solution containing 45 mM EDTA, 50 mM Tris-HCl, 50 mM NaCl, 0.4 mg/ml BSA, 200 nM SA-APC and 3 nM EUPy20. The plate is incubated for 15-30 minutes at room temperature and HTRF (homogenous time resolved fluorescence) is measured on a Perkin Elmer Fusion alpha-FP instrument. The HTRF program settings used are as follows: Primary excitation filter 330/30, Primary window: 200 uSec, Primary delay: 50 uSec, Number of flashes: 15, Well read time: 2000

DMSO 2 mg/mL, Water <1 mg/mL, Ethanol <1 mg/mL

2-fluoro-N-methyl-4-(7-(quinolin-6-ylmethyl)imidazo[1, 2-b][1, 2, 4]triazin-2-yl)benzamide