Infigratinib (BGJ398)

Athymic nude-nu mice bearing parental RT112 cell line

3-(2, 6-dichloro-3, 5-dimethoxyphenyl)-1-(6-(4-(4-ethylpiperazin-1-yl)phenylamino)pyrimidin-4-yl)-1-methylurea

0 uM-0.1 uM

10 mg/kg/qd and 30 mg/kg/qd

0.9 nM, 0.9 nM, 0.9 nM, 0.9 nM, 0.9 nM, 0.9 nM

In this orthotopic xenograft bladder cancer model, BGJ398 induces tumor growth inhibition and stasis after oral administration for 12 consecutive days at the doses of 10 and 30 mg/kg, respectively. Interestingly, the animals that received BGJ398 exhibits either no body weight loss (10 mg/kg) or 10% body weight gain (30 mg/kg), a further indication of efficacy. RT112 tumor-bearing and female Rowett rats receive a single oral administration of the monophosphate salt of BGJ398 at the doses of 4.25 and 8.51 mg/kg. BGJ398 significantly decreases the levels of pFRS2 and pMAPK in a dose-dependent manner. BGJ398 inhibits significantly bFGF-stimulated angiogenesis in a dose-dependent manner. However, BGJ398 does not impair VEGF-induced blood vessel formation. [1]

Radiometric kinase assay, The enzymatic kinase activity is assessed by measuring the phosphorylation of a synthetic substrate by the purified GST-fusion FGFR3-K650E kinase domain, in the presence of radiolabeled ATP. Enzyme activities are measured by mixing 10 uL of a 3-fold concentrated BGJ398 solution or control with 10 uL of the corresponding substrate mixture (peptidic substrate, ATP and [gamma33P]ATP). The reactions are initiated by addition of 10 uL of a 3-fold concentrated solution of the enzyme in assay buffer. The final concentrations of the assay components are as following: 10 ng of GST-FGFR3-K650E, 20 mM Tris-HCl, pH 7.5, 3 mM MnCl2, 3 mM MgCl2, 1 mM DTT, 250 ug/mL PEG 20000, 2 ug/mL poly(EY) 4:1, 1% DMSO and 0.5 uM ATP (gamma-[33P]-ATP 0.1 uCi). The assay is carried out according to the filter binding (FB) method in 96-well plates at room temperature for 10 minutes in a final volume of 30 uL including BGJ398. The enzymatic reactions are stopped by the addition of 20 uL of 125 mM EDTA, and the incorporation of 33P into the polypeptidic substrates is quantified as following: 30 uL of the stopped reaction mixture are transferred onto Immobilon-PVDF membranes previously soaked for 5 minutes with methanol, rinsed with water, soaked for 5 min with 0.5% H3PO4, and mounted on vacuum manifold with disconnected vacuum source. After spotting, vacuum is connected, and each well rinsed with 0.5% H3PO4 (200 uL). Free membranes are removed and ished four times on a shaker with 1% H3PO4 and once with ethanol. Membranes are dried and overlaid with addition of 10 uL/well of a scintillation fluid. The plates are eventually sealed and counted in a microplate scintillation counter. IC50 values are calculated by linear regression analysis of the percentage inhibition of the BGJ398.

560, 48

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