Ruxolitinib (INCB018424)

Background

Ruxolitinib dihydrochloride is a specific inhibitor of Janus-associated kinase (JAK1 and JAK2). Ruxolitinib is a small molecular with the formula C17H21N6O4P and a molecular weight of 404. Ruxolitinib phosphate is a cyclopentyl propionitrile derivative competitive with administered ATP and shows inhibitory activity on JAK1 and JAK2. Ruxolitinib inhibits phosphorylation of JAK1 / 2, STAT5, and ERK1 / 2, reducing cell proliferation.

Chemical properties

Physical appearance: A solid

Storage: Store at -20 ° C

M.Wt: 306.37

CAS No: 941678-49-5

Formula: C17H18N6

Synonyms: Ruxolitinib, INCB018424, INCB-018424

Solubility: Insoluble in H2O; ≥15.32 mg / ml in DMSO; ≥17.53 mg / ml in EtOH

Chemical name: (3R) -3-cyclopentyl-3- [4- (7H-pyrrolo [2,3-d] pyrimidin-4-yl) pyrazol-1-yl] propanonitrile

Canonical SMILES: C1CCC (C1) C (CC # N) N2C = C (C = N2) C3 = C4C = CNC4 = NC = N3

Shipping conditions: Evaluation sample solution: ship with blue ice. All other sizes available – ship with RT or blue ice upon request.

General advice: To obtain better solubility, heat the tube to 37 ° C and shake it in the ultrasonic bath for a while. The stock solution can be stored below -20 ° C for several months.

Protocol

Cell experiment: [1]

Cell lines

Primary mononuclear cells isolated from PV patients or normal controls

Preparation method

The solubility of this compound in DMSO is> 10 mM. General advice to obtain a higher concentration: Heat the tube at 37 ° C for 10 minutes and/or shake it in the ultrasound bath for a while. The stock solution can be stored below -20 ° C for several months.

Reaction conditions

IC50: erythroid parents: 407 nM for normal donors, 223 nM for PV donors myeloid parents: 511 nM for normal donors, 444 nM for 14 day PV donors

Applications

The growth of clonogenic erythroid (BFU-E) and myeloid (CFU-M) progenitors were evaluated in colony formation assays in the presence of increasing concentrations of INCB018424. With INCB018424, dose-dependent inhibition of growth of erythroid and myeloid progenitors was observed. The mean IC50 for INCB018424 against erythroid progenitors was 407 nM for normal donors and 223 nM for PV donors. A similar effect was observed in myeloid progenitors (CFU-M), with IC50 values ​​of 511 nM and 444 nM for the control and PV samples, respectively.

Animal experiment: [2]

Animal models

C57BL / 6N mice

Dosage form

Oral administration, 75 mg / kg

Applications

Mice that received 75 mg/kg of ruxolitinib or vehicle 6 hours before and 6 hours after OVA / CpG injection were analyzed for the expression of activation markers in splenic CDs CD11c 1CD81. Lower expression levels of CD40, CD80, CD86 as well as MHC I and II molecules were detected in animals challenged with ruxolitinib. The mice were then administered ruxolitinib or vehicle 6 hours before, as well as 6 hours and 18 hours after priming with OVA / CpG and adoptive transfer of CFSE-labeled OT-I cells. Analysis of transferred CFSE-labeled OT-I T cells revealed reduced proliferation, CD25 expression, and IFN production in ruxolitinib-pretreated mice.

Other notes

Test the solubility of all compounds indoor, and the actual solubility may differ slightly from the theoretical value. This is due to an error in the experimental system and is normal.

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