BGJ398 (NVP-BGJ398)

 

Biological Activity

Description	BGJ398 (NVP-BGJ398) is a potent and selective FGFR inhibitor for FGFR1/2/3 with IC50 of 0.9 nM/1.4 nM/1 nM, >40-fold selective for FGFR versus FGFR4 and VEGFR2, and little activity to Abl, Fyn, Kit, Lck, Lyn and Yes. Phase 2.
Targets	FGFR1	FGFR2	FGFR3	FGFR4
IC50	0.9 nM	1.4 nM	1 nM	60 nM [1]
In vitro	BGJ398 also prevents VEGFR2 with low potency. The IC50 of BGJ398 for inhibiting VEGFR2 is 0.18 μM. BGJ398 suppresses other kinases including ABL, FYN, KIT, LCK, LYN and YES with IC50 of 2.3 μM, 1.9 μM, 0.75 μM, 2.5 μM, 0.3 μM and 1.1 μM, respectively. At the cellular level, BGJ398 inhibits the proliferation of the FGFR1-, FGFR2-Q, and FGFR3-dependent BaF3 cells with IC50 of 2.9 μM, 2.0 μM and 2 μM, respectively. BGJ398 interferes with autophosphorylation on specific tyrosine residues including FGFR-WT, FGFR2-WT, FGFR3-K650E, FGFR3-S249C and FGFR4-WT with IC50 of 4.6 nM, 4.9 nM, 5 nM, 5 nM and 168 nM, respectively. BGJ398 suppresses proliferation of the cancer cells with wild-type (WT) FGFR3 overexpression such as RT112, RT4, SW780 and JMSU1 with IC50 of 5 nM, 30 nM, 32 nM and 15 nM, respectively. [1]
In vivo	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]
Features

Protocol(Only for Reference)

Kinase Assay: ^[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 μL of a 3-fold concentrated BGJ398 solution or control with 10 μL of the corresponding substrate mixture (peptidic substrate, ATP and [γ33P]ATP). The reactions are initiated by addition of 10 μL 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 μg/mL PEG 20000, 2 μg/mL poly(EY) 4:1, 1% DMSO and 0.5 μM ATP (γ-[33P]-ATP 0.1 μCi). 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 μL including BGJ398. The enzymatic reactions are stopped by the addition of 20 μL of 125 mM EDTA, and the incorporation of 33P into the polypeptidic substrates is quantified as following: 30 μL 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 μL). 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 μL/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.

Cell Assay: ^[1]

Cell lines	Murine BaF3 cell lines
Concentrations	0 μM-0.1 μM
Incubation Time	48 hours
Method	Murine BaF3 cell lines, whose proliferation and survival has been rendered IL-3-independent by stable transduction with tyrosine kinases activated either by mutation or fusion with a dimerizing partner, are cultured in RPMI-1640 media supplemented with 10% FBS, 4.5 g/L glucose, 1.5 g/L sodium bicarbonate, and Pen/Strep. Cells are passaged twice weekly. BGJ398-mediated inhibition of BaF3 cell proliferation and viability is assessed using a Luciferase bioluminescent assay. Exponentially growing BaF3 or BaF3 Tel-TK cells are seeded into 384-well plates (4250 cells/well) at 50 μL/well using a μFill liquid dispenser in fresh medium. BGJ398 is serially diluted in DMSO and arrayed in a polypropylene 384-well plate. Then 50 nL of BGJ398 are transferred into the plates containing the cells by using the pintool transfer device, and the plates incubated at 37 °C (5% CO2) for 48 hours. Then 25 μL of Bright-Glo are added, and luminescence is quantified using an Analyst-GT. Custom curve-fitting software is used to produce a logistic fit of percent cell viability as a function of the logarithm of inhibitor concentration. The IC50 value is determined as the concentration of BGJ398 needed to reduce cell viability to 50% of a DMSO control.

Animal Study: ^[1]

Animal Models	Athymic nude-nu mice bearing parental RT112 cell line
Dosages	10 mg/kg/qd and 30 mg/kg/qd
Administration	Oral administration
Solubility	30% PEG400/0.5% Tween80/5% propylene glycol, 30 mg/mL

1

References

Clinical Trial Information( data from http://clinicaltrials.gov)

NCT Number	Recruitment	Conditions	Sponsor /Collaborators	Start Date	Phases
NCT01004224	Recruiting	Advanced Solid Tumors With FGFR1 Amplication,|Advanced Solid Tumors With FGFR2 Amplication,|Advanced Solid Tumors With FGFR3 Mutation	Novartis|Novartis Pharmaceuticals	2009-12	Phase 1
NCT01697605	Recruiting	Tumor With Alterations of the FGF-R	Novartis|Novartis Pharmaceuticals	2012-10	Phase 1
NCT01820364	Recruiting	Melanoma	Novartis|Novartis Pharmaceuticals	2013-09	Phase 2
NCT01928459	Recruiting	Advanced Solid Tumors,|Metastatic Solid Tumors	Novartis|Novartis Pharmaceuticals	2013-10	Phase 1
NCT01975701	Not yet recruiting	Recurrent Glioblastoma	Novartis|Novartis Pharmaceuticals	2013-11	Phase 2

Chemical Information

Download BGJ398 (NVP-BGJ398) SDF

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

Customer Reviews (2)

Click to enlarge	Rating
	Source	Hepatology, 2013, doi: 10.1002/hep.26890. BGJ398 (NVP-BGJ398) purchased from Selleck
	Method	Western blot
	Cell Lines	NIN3T3 cells
	Concentrations	0.2, 1 uM
	Incubation Time	2 h
	Results	BGJ398 significantly inhibited the phosphorylation of MAPK.

Product Citations (2)

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  FGFR2 tyrosine kinase fusions define a unique molecular subtype of cholangiocarcinoma. [Arai Y, et al. Hepatology 2013;10.1002/hep.26890]

  
  PubMed: 24122810


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  Heregulin induces resistance to lapatinib-mediated growth inhibition of HER6-amplified cancer cells. [Sato Y, et al. Cancer Sci 2013;104(12):1618-25]

  
  PubMed: 24112719