Masitinib (AB1010)

 

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Biological Activity

Description	Masitinib is a novel inhibitor for Kit and PDGFRα/β with IC50 of 200 nM and 540 nM/800 nM, weak inhibition to ABL and c-Fms. Phase 2/3.
Targets	Kit	PDGFRα/β
IC50	200 nM	540 nM/800 nM [1]
In vitro	Masitinib is a competitive inhibitor against ATP at concentrations ≤500 nM. Masitinib also potently inhibits recombinant PDGFR and the intracellular kinase Lyn, and to a lesser extent, fibroblast growth factor receptor 3. In contrast, Masitinib demonstrates weak inhibition of Abl and c-Fms. Masitinib more strongly inhibits degranulation, cytokine production, and bone marrow mast cell migration than imatinib. In Ba/F3 cells expressing human wild-type Kit, Masitinib inhibits SCF (stem cell factor)-induced cell proliferation with an IC50 of 150 nM, while the IC50 for inhibition of IL-3-stimulated proliferation is at approximately >10 µM. In Ba/F3 cells expressing PDGFRα, Masitinib inhibits PDGF-BB-stimulated proliferation and PDGFRα tyrosine phosphorylation with IC50 of 300 nM. Masitinib also causes inhibition of SCF-stimulated tyrosine phosphorylation of human Kit in mastocytoma cell-lines and BMMC. Masitinib inhibits Kit gain-of-function mutants, including V559D mutant and Δ27 mouse mutant with IC50 of 3 and 5 nM in Ba/F3 cells. Masitinib inhibits the cell proliferation of mastocytoma cell lines including HMC-1α155 and FMA3 with IC50 of 10 and 30 nM, respectively. [1] Masitinib inhibits cell growth and PDGFR phosphorylation in two novel ISS cell lines, which suggest that Masitinib displays activity against both primary and metastatic ISS cell line and may aid in the clinical management of ISS. [2]
In vivo	Masitinib inhibits tumour growth and increases the median survival time in Δ27-expressing Ba/F3 tumor models at 30 mg/kg, without cardiotoxicity or genotoxicity. [1] Masitinib (12.5 mg/kg/d PO) increases overall TTP (time-to-tumor progression) compared with placebo in dogs. [3] The combination of masitinib/gemcitabine shows synergy in vitro on proliferation of gemcitabine-refractory cell lines Mia Paca2 and Panc1, and to a lesser extent on Mia Paca-2 pancreatic tumours in Nogç’–CID mice. [4]
Features	Potential low side-effect profile.

Protocol(Only for Reference)

Kinase Assay: ^[1]

In vitro enzyme-linked immunoassay with recombinant protein kinases

A 96-well microtitre plateis coated overnight with 0.25 mg/ml poly(Glu,Tyr 4:1), rinsed twice with 250 µL of washing buffer (10 mM phosphate-buffered saline [pH 7.4] and 0.05% Tween 20) and dried for 2 hours at room temperature. Assays are performed at room temperature with a final volume of 50 µL in kinase buffer (10 mM MgCl2, 1 mM MnCl2, 1 mM sodium orthovanadate, 20 mM HEPES, pH 7.8) containing ATP at a concentration of at least twice the Km for each enzyme and an appropriate amount of recombinant enzyme to ensure a linear reaction rate. Reactions are initiated upon introduction of the enzyme and terminated with the addition of one reaction volume (50 μL) of 100 mM EDTA per 5 M urea mix. Plates are washed three times and incubated with 1:30,000 horseradish peroxidase-conjugated anti-phosphotyrosine monoclonal antibody, then washed three times and incubated with tetramethylbenzidine. The final reaction product is quantified by spectrophotometry at 450 nm.

Cell Assay: ^[1]

Cell lines	Ba/F3 cells expressing wild-type or mutant human Kit, HMC1, HMC-1α155
Concentrations	0.1 nM - 10 μM
Incubation Time	48 hours
Method	For the assay of Ba/F3 cell proliferation, microtitre plates are seeded with a total of 104 cells/well in 100 μL of RPMI 1640 medium with 10% foetal bovine serum at 37 °C. These are supplemented, or not, with either 0.1% conditioned medium from X63-IL-3 cells or 250 ng/mL murine SCF. The murine SCF, which activates Kit, is purified from the conditioned medium of SCF-producing CHO cells. Cells are grown for 48 hours at 37 °C with Masitinib and then incubated with 10 μL/well of WST-1 reagent for 3 hours at 37 °C. The amount of formazan dye formed is quantified by its absorbance at 450 nm using a scanning multiwell spectrophotometer. A blank well without cells is used as a background control for the spectrophotometer.

Animal Study: ^[1]

Animal Models	Ba/F3 Δ27 tumour model in female MBRI Nu/Nu mice
Dosages	30 mg/kg (intraperitoneal) or 10, 30, or 45 mg/kg (orally).
Administration	Intraperitoneal or orally administered.
Solubility	1% DMSO/30% polyethylene glycol/1% Tween 80, 30 mg/mL

1

References

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

NCT Number	Recruitment	Conditions	Sponsor /Collaborators	Start Date	Phases
NCT01449162	Recruiting	Asthma	AB Science	2011-06	Phase 3
NCT01410695	Recruiting	Rheumatoid Arthritis (RA)	AB Science	2011-07	Phase 2|Phase 3
NCT01433497	Recruiting	Multiple Sclerosis, Secondary Progressive|Multiple Sclerosis, Primary Progressive|Multiple Sclerosis, Relapse Free	AB Science	2011-08	Phase 2|Phase 3
NCT01872598	Recruiting	Alzheimer Disease	AB Science	2012-01	Phase 3
NCT01694277	Recruiting	Gastrointestinal Stromal Tumors	AB Science	2012-09	Phase 3

Chemical Information

Download Masitinib (AB1010) SDF

Chemical Name	N-(4-methyl-3-(4-(pyridin-3-yl)thiazol-2-ylamino)phenyl)-4-((4-methylpiperazin-1-yl)methyl)benzamide

Preparing Stock Solutions

Stock Solution (1ml DMSO)	1mM	10mM	20mM	30mM
Mass(mg)	0.49864	4.9864	9.9728	14.9592

Customer Reviews (1)

Click to enlarge	Rating
	Source	Biomaterials, 2013, 34(38):9737-46. Masitinib (AB1010) purchased from Selleck
	Method	H&E staining
	Cell Lines	wild-type C57BL/6 mice
	Concentrations
	Incubation Time	14, 21, 28 days
	Results	H&E-stained tissue preparations exhibit variable distributions of different inflammatory and wound-recruited cell densities for both control and masitinib-releasing implant sites at each time point as shown in Figure. Densities of PMNs, macrophages, FBGCs, and fibroblasts were quantified as a function of distance from the implant surfaces. Cell densities up to distances of 300 μm from implant interfaces were considered in this analysis.

Product Citations (2)

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  Modulation of the foreign body response to implanted sensor models through device-based delivery of the tyrosine kinase inhibitor, masitinib. [Avula MN, et al. Biomaterials 2013;34(38):9737-46]

  
  PubMed: 24060424


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  Foreign Body Response To Subcutaneous Biomaterial Implants in a Mast Cell-deficient Kitw-Sh Murine Model. [Avula M ,et al. Acta Biomater 2013;10.1016/j.actbio.2013.12.056]

  
  PubMed: 24406200