S-Ruxolitinib

Biological Activity

S-Ruxolitinib is the chirality of INCB018424, which is the first potent, selective, JAK1/2 inhibitor to enter the clinic with IC50 of 3.3 nM/2.8 nM, >130-fold selectivity for JAK1/2 versus JAK3.

Product Citations

• 

  Blood Cancer J. 2016 Oct 7;6(10):e481.

  Expression of CALR mutants causes mpl-dependent thrombocytosis in zebrafish.
  S-Ruxolitinib purchased from AbMole

• 

  Nature. 2014 Aug 7;512(7512):78-81.

  Neuropathy of haematopoietic stem cell niche is essential for myeloproliferative neoplasms.
  S-Ruxolitinib purchased from AbMole

Customer Product Validations & Biological Datas

	Source	Blood cancer journal (2016) . Figure 4. ruxolitinib (Abmole Bioscience, Houston, TX, USA) and fedratinib (Abmole Bioscience)
	Method	Western blotting
	Cell Lines	Embroys of wild-type zebrafish
	Concentrations	1 μM ~ 8 μM
	Incubation Time	24 h
	Results	The expression of CALRdel52 mRNA significantly increased stat5 phosphorylation (Figure 4a, lane 2). Furthermore, treatment with ruxolitinib and fedratinib significantly ameliorated the enhanced stat5 phosphorylation induced by CALR-del52 mRNA (Figure 4a, lane 3 and 4). In addition, treatment with ruxolitinib significantly decreased the numbers of CD41+ thrombocytes in uninjected control as well as CALR-del52-injected embryos in a dose-dependent manner (Figure 4b). Whereas treatment with fedratinib only had minimal inhibitory effect on the number of CD41+ thrombocytes in uninjected control embryos, and had a modest and significant dose-independent inhibitory effect on mutant CALR-induced thrombocytosis (Figure 4c). Our results demonstrated that mutant CALR-mediated pathogenic thrombopoiesis involves jak-stat activation that can be blocked by JAK inhibitors.

	Source	Nature (2014). Figure 1. The JAK inhibitor INCB018424 (S-ruxolitinib, Abmole Bioscience)
	Method	automated blood counter, flowcytometry
	Cell Lines	haematopoietic cell and BM MSCs
	Concentrations	oral gavage (30mg/kg, twice per day separated 10–12h)
	Incubation Time	4 weeks
	Results	Nestin1 MSC reduction was instead explained by a threefold increased apoptotic rate in mutant mice, and was not prevented by the JAK inhibitor ruxolitinib. Treatment with the JAK1/2 inhibitor ruxolitinib reduces haematopoietic cell expansion in MPN but it does not rescue BM MSCs.

Protocol (for reference only)

Cell Experiment
Cell lines	Ba/F3-EpoR-JAK2V617F or HEL cells
Preparation method	Cells were seeded at 2000/well of white bottom 96-well plates, treated with compounds from DMSO stocks (0.2% final DMSO concentration), and incubated for 48 hours at 37°C with 5% CO2. Viability was measured by cellular ATP determination using the Cell-Titer Glo (Promega) luciferase reagent or viable cell counting. Values were transformed to percent inhibition relative to vehicle control, and IC50 curves were fitted according to nonlinear regression analysis of the data using PRISM GraphPad.
Concentrations	0~10µM
Incubation time	48h

Animal Experiment
Animal models	6- to 8-week-old female BALB/c mice Ba/F3-JAK2V617F cells model
Formulation	5% dimethyl acetamide, 0.5% methocellulose
Dosages	180mg/kg, twice a day
Administration	orally

Chemical Information

Molecular Weight	306.37
Formula	C17H18N6
CAS Number	941685-37-6
Solubility (25°C)	DMSO 51 mg/mL
Storage	Powder          -20°C   3 years ;  4°C   2 years In solvent       -80°C   6 months ;  -20°C   1 month