Capmatinib (INCB28060)

Biological Activity

Capmatinib (INCB28060) is a potent, orally available selective inhibitor of the c-MET receptor tyrosine kinase, a clinically validated cancer target. Capmatinib (INCB28060) exhibits picomolar enzymatic potency and is highly specific for c-MET with more than 10,000-fold selectivity over a large panel of human kinases. INCB28060 potently blocks c-MET phosphorylation and activation of its key downstream effectors in c-MET-dependent tumor cell lines. Capmatinib (INCB28060) potently inhibits c-MET-dependent tumor cell proliferation and migration and effectively induces apoptosis in vitro. Oral dosing of INCB28060 results in time- and dose-dependent inhibition of c-MET phosphorylation and tumor growth in c-MET-driven mouse tumor models, and INCB28060 is well tolerated at doses that achieve complete tumor inhibition.

Product Citations

• 

  Mol Cancer Ther. 2017 Nov;16(11):2399-2409.

  Dual inhibition of Hedgehog and c-Met pathways for pancreatic cancer treatment
  Capmatinib (INCB28060) purchased from AbMole

• 

  Cancer Res. 2017 Jan 1;77(1):41-52.

  Heterogeneous Stromal Signaling within the Tumor Microenvironment Controls the Metastasis of Pancreatic Cancer
  Capmatinib (INCB28060) purchased from AbMole

• 

  Nature. 2015 Jun 18;522(7556):349-53.

  MET is required for the recruitment of anti-tumoural neutrophils.
  Capmatinib (INCB28060) purchased from AbMole

Customer Product Validations & Biological Datas

	Source	Mol Cancer Ther (2017). Figure 4. INCB28060 (Abmole Bioscience. Houston, TX, USA)
	Method	oral gavage
	Cell Lines	KPC and orthotopic transplant model mice
	Concentrations	1 mg/kg
	Incubation Time	7, 14 or 21 days
	Results	Importantly, the group with dual inhibitor treatment in addition to gemcitabine showed no metastases. This result suggests that the combination of c-Met and Hh inhibitors in addition to gemcitabine has significantly suppressed metastasis formation.

	Source	Mol Cancer Ther (2017). Figure 3. INCB28060 (Abmole Bioscience. Houston, TX, USA)
	Method	oral gavage
	Cell Lines	KPC and orthotopic transplant model mice
	Concentrations	1 mg/kg
	Incubation Time	7, 14 or 21 days
	Results	These results suggest that the resistance to the c-Met and Hh inhibitor is developed following a prolonged treatment of each individual inhibitor; however, the resistance may be overcome by combining both c-Met and Hh inhibitors.

	Source	Mol Cancer Ther (2017). Figure 1. INCB28060 (Abmole Bioscience. Houston, TX, USA)
	Method	oral gavage
	Cell Lines	KPC and orthotopic transplant model mice
	Concentrations	1 mg/kg
	Incubation Time	7, 14 or 21 days
	Results	The data suggest that combination of both inhibitors with gemcitabine shows a trend in shrinkage of primary tumor volume in both mouse models of PDA after one week of treatment regimen.

	Source	Can Res (2017). Figure 3.INC280 (Abmole Bioscience).
	Method
	Cell Lines	PDAC cells
	Concentrations
	Incubation Time
	Results	As expected, neither the Hh inhibitor nor the HGF/c-Met inhibitor affected the levels of total AnxA2 or P-Y23-AnxA2 in Panc10.05 cells in the singly cultured serum-reduced media (Fig. 3A and B; Supplementary Fig. S5E and S5F). It is likely the concentration of HGFand IGF-1 in serum-reduced media is lower than the effective thresholds.

	Source	Nature (2015). Figure 3. PF-04217903 and INCB28060 (AbMole Bioscience)
	Method	mice treatment in vivo and histological evaluation
	Cell Lines	B16F10 cells
	Concentrations	40mg/kg PF-04217903 and 50mg/kg INCB28060
	Incubation Time	2 weeks
	Results	Systemic treatment of WT mice carrying B16F10 melanomas (which are dependent on MET14) with three different MET tyrosine-kinase inhibitors (PF-04217903, INCB28060 and JNJ-38877605), strongly reduced TAN recruitment .

	Source	Nature (2015). Figure 2. PF-04217903 and INCB28060 (AbMole Bioscience)
	Method	mice treatment in vivo and histological evaluation
	Cell Lines	B16F10 cells
	Concentrations	40mg/kg PF-04217903
	Incubation Time	2 weeks
	Results	"Systemic administration of PF-04217903 decreased the weight and volume of B16F10 melanomas by 36% and 54%, respectively. Instead, MET knockdown in cancer cells only, led to 58% and 75% inhibition of tumour growth and volume.TAN inhibition by PF-04217903 was comparable in both Met-silenced and scrambled B16F10 melanomas."

Protocol (for reference only)

Cell Experiment
Cell lines	SUN-5 and SUN-1 cells
Preparation method	Optimal cell density used in the viability assay was predetermined for individual cell lines. To determine compound potency, cells were seeded into 96-well microplates at the appropriate density in media containing 1% to 2% FBS and supplemented with serial dilutions of INCB28060 in a final volume of 100 μL per well. After 72-hour incubation, 24 μL of CellTiter 96 AQueous One Solution (Promega) was added to each well, and the plates were incubated for 2 hours in a 37°C incubator. The optical density was measured in the linear range using a microplate reader at 490 nm with wavelength correction at 650 nm. IC50 values were calculated using the GraphPad Prism Software.
Concentrations	0~333 nM
Incubation time	3 days

Animal Experiment
Animal models	B16F10 tumour-bearing mice
Formulation	0.5% methylcellulose in saline
Dosages	50mg/kg INCB28060
Administration	via oral gavage every day once a day starting from day 2 after tumour injection and twice a day from day 11 until the end of the experiment(2 weeks)

Chemical Information

Molecular Weight	412.42
Formula	C23H17FN6O
CAS Number	1029712-80-8
Solubility (25°C)	DMSO 15 mg/mL
Storage	Powder          -20°C   3 years ;  4°C   2 years In solvent       -80°C   6 months ;  -20°C   1 month

Conversion of different model animals based on BSA (PMID: 27057123)

Species	Mouse	Rat	Rabbit	Guinea pig	Hamster	Dog
Weight (kg)	0.02	0.15	1.8	0.4	0.08	10
Body Surface Area (m2)	0.007	0.025	0.15	0.05	0.02	0.5
Km factor	3	6	12	8	5	20

Animal A (mg/kg) = Animal B (mg/kg) multiplied by	Animal B Km
	Animal A Km

For example, to modify the dose of Compound A used for a mouse (20 mg/kg) to a dose based on the BSA for a rat, multiply 20 mg/kg by the K_m factor for a mouse and then divide by the K_m factor for a rat. This calculation results in a rat equivalent dose for Compound A of 10 mg/kg.

References

[1] Sung-Hwa Sohn, et al. BMC Res Notes. INC280 inhibits Wnt/β-catenin and EMT signaling pathways and its induce apoptosis in diffuse gastric cancer positive for c-MET amplification

[2] Sabrina Baltschukat, et al. Clin Cancer Res. Capmatinib (INC280) Is Active Against Models of Non-Small Cell Lung Cancer and Other Cancer Types with Defined Mechanisms of MET Activation

[3] Liu X, et al. Clin Cancer Res. A novel kinase inhibitor, INCB28060, blocks c-MET-dependent signaling, neoplastic activities, and cross-talk with EGFR and HER-3.