Ruxolitinib (INCB018424)

 

Distributor Tel: +886-2-2827-1197jimmy@new.abscience.com.tw

Biological Activity

Description	Ruxolitinib (INCB018424) 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. Phase 3.
Targets	JAK1	JAK2
IC50	3.3 nM	2.8 nM [1]
In vitro	INCB018424 potently and selectively inhibits JAK2V617F-mediated signaling and proliferation in Ba/F3 cells and HEL cells. INCB018424 markedly increases apoptosis in a dose dependent manner in Ba/F3 cells. INCB018424 (64 nM) results in a doubling of cells with depolarized mitochondria in Ba/F3 cells. INCB018424 inhibits proliferating of erythroid progenitors from normal donors and polycythemia vera patients with IC50 of 407 nM and 223 nM, respectively. INCB018424 demonstrates remarkable potency against erythroid colony formation with IC50 of 67nM. [1]
In vivo	INCB018424 (180 mg/kg, orally, twice a day) results in survive rate of greater than 90% by day 22 in a JAK2V617F-driven mouse model. INCB018424 (180 mg/kg, orally, twice a day) markedly reduces splenomegaly and circulating levels of inflammatory cytokines, and preferentially eliminated neoplastic cells, resulting in significantly prolonged survival without myelosuppressive or immunosuppressive effects in a JAK2V617F-driven mouse model. [1] The primary end point is reached in 41.9% of patients in the Ruxolitinib group as compared with 0.7% in the placebo group in the double-blind trial of myelofibrosis. Ruxolitinib results in maintaining of reduction in spleen volume and improvement of 50% or more in the total symptom score. [2] A total of 28% of the patients in the Ruxolitinib (15 mg twice daily) group has at least a 35% reduction in spleen volume at week 48 in patients with myelofibrosis, as compared with 0% in the group receiving the best available therapy. The mean palpable spleen length has decreased by 56% with Ruxolitinib but has increased by 4% with the best available therapy at week 48. Patients in the ruxolitinib group has an improvement in overall quality-of-life measures and a reduction in symptoms associated with myelofibrosis. [3]
Features

Protocol(Only for Reference)

Kinase Assay: ^[1]

Binding assay

Recombinant proteins are expressed using Sf21 cells and baculovirus vectors and purified with affinity chromatography. JAK kinase assays use a homogeneous time-resolved fluorescence assay with the peptide substrate (-EQEDEPEGDYFEWLE). Each enzyme reaction is carried out with Ruxolitinib or control, JAK enzyme, 500 nM peptide, adenosine triphosphate (ATP; 1mM), and 2% dimethyl sulfoxide (DMSO) for 1 hour. The 50% inhibitory concentration (IC50) is calculated as INCB018424 concentration required for inhibition of 50% of the fluorescent signal.

Cell Assay: ^[1]

Cell lines	Ba/F3 and HEL cells
Concentrations	3 μM
Incubation Time	48 hours
Method	Cells are seeded at 2 × 103/well of white bottom 96-well plates, treated with INCB018424 from DMSO stocks (0.2% final DMSO concentration), and incubated for 48 hours at 37 ℃ with 5% CO2. Viability is measured by cellular ATP determination using the Cell-Titer Glo luciferase reagent or viable cell counting. Values are transformed to percent inhibition relative to vehicle control, and IC50 curves are fitted according to nonlinear regression analysis of the data using PRISM GraphPad.

Animal Study: ^[1]

Animal Models	JAK2V617F-driven mouse model
Dosages	180 mg/kg
Administration	Oral gavage

1

References

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

NCT Number	Recruitment	Conditions	Sponsor /Collaborators	Start Date	Phases
NCT01969838	Recruiting	Primary Myelofibrosis|Post-Polycythemia Vera Myelofibrosis|Post-Essential Thrombocythemia Myelofibrosis	Gilead Sciences	2013-10	Phase 3
NCT01981850	Recruiting	Primary Myelofibrosis|Post-Polycythemia Vera Myelofibrosis|Post-Essential Thrombocythemia Myelofibrosis	Promedior, Inc.	2013-10	Phase 2
NCT01965119	Not yet recruiting	Relapsed or Refractory Hodgkin Lymphoma|Primary Mediastinal Large B-cell Lymphoma	Samsung Medical Center	2013-11
NCT01895842	Not yet recruiting	Leukemia	M.D. Anderson Cancer Center|Incyte Corporation	2014-02	Phase 1
NCT02015208	Not yet recruiting	Chronic Lymphocytic Leukemia	Sunnybrook Health Sciences Centre|Novartis	2014-02	Phase 1|Phase 2

Chemical Information

Download Ruxolitinib (INCB018424) SDF

Chemical Name	(3R)-3-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-cyclopentylpropanenitrile

Preparing Stock Solutions

Stock Solution (1ml DMSO)	1mM	10mM	20mM	30mM
Mass(mg)	0.30637	3.0637	6.1274	-

Customer Reviews (1)

Click to enlarge	Rating
	Source	Yong Weon Yi Georgetown University. Ruxolitinib (INCB018424) purchased from Selleck
	Method	Western Blot
	Cell Lines	HS578T cells
	Concentrations	0-3 μM
	Incubation Time	18 h
	Results	INCB018424 treatment resulted in a reduction of STAT3 phosphorylation in a concentration-dependent manner.

Product Citations (7)

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  Age-associated inflammation inhibits epidermal stem cell function. [Doles J, et al. Genes Dev 2013;26(19):2144-53]

  
  PubMed: 22972935


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  Inhibition of JAKs in macrophages increases lipopolysaccharide-induced cytokine production by blocking IL-10-mediated feedback. [Pattison MJ, et al. J Immunol 2012;189(6):2784-92]

  
  PubMed: 22904308


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  Type I IFN Inhibits Innate IL-10 Production in Macrophages through Histone Deacetylase 11 by Downregulating MicroRNA-145. [Lin L, et al. J Immunol 2013;191(7):3896-904]

  
  PubMed: 23980205