Ruxolitinib

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency: 

IC50

JAK1 3.3 nM, JAK2 2.8 nM

Off target activity

Potency (off target): 

IC50

Off target protein and potency: 

TYK2 19 nMJAK3 428 nM

Potency assay (off target): 

Ruxolitinib was not active against a panel of 26 additional kinases at ~300 nM.

In cell validation

On target activity in cells

Potency in cells: 

IC50

280 nM

Potency assay (cells): 

Inhibition of substrate phosphorylation in whole blood.

Target engagement assay (cells): 

Indirect: inhibition of protein substrate phosphorylation

Off target activity in cells

Potency assay, off target (cells): 

In various cells, Ruxolitinib did not impact BCR-ABL or KIT signaling pathways.

In vivo validation

Organism: 

Balb/c mice

Dose: 

90 mg/kg

Route of delivery: 

Oral

Organ of Interest

Organ (O): 

spleenblood

Target engagement assay: 

Indirect: inhibition of protein substrate phosphorylation in white blood cells