Upadacitinib

Cell lines

Ba/F3 cells

Preparation Method

Ba/F3 cells were washed and resuspended in Hank’s balanced salt solution at a density of 2x10⁷ cells/mL. Five microliters of cell suspension were added to a 384-well, low-volume, white-walled polystyrene plate that contained 5μL of Upadacitinib. Cells were incubated with compound (final DMSO concentration 0.5%) for 30min at 37℃ before proceeding with pSTAT5 detection.

Reaction Conditions

0, 0.01, 0.1, 0.5, 1, 2μM;

Applications

Upadacitinib exhibited greater than 40-fold selectivity for JAK1 (IC₅₀ = 0.045μM) compared to JAK2 (IC₅₀ = 0.109μM) and demonstrated significant selectivity against JAK3 (approximately 130-fold) and TYK2 (approximately 190-fold).

Animal models

Female Lewis rats

Preparation Method

Arthritis was induced in female Lewis rats (weight, 125 – 150 g)by a single intradermal injection of 0.1mL of microbacterium tuberculosis emulsion into the right hind footpad (Day 0). Rats were dosed as indicated orally by gavage twice a day for 10 days (Day7 – Day17) post immunization with either vehicle or Upadacitinib. To evaluate the severity of arthritis, paw swelling was evaluated with a water displacement plethysmograph every other day up to Day 17. On Day 17, all rats were exsanguinated by cardiac puncture under isolfuorane anesthesia. Left rear paws were scanned using a Micro-CT.Bone volume and density were determined in a 360μm vertical section encompassing the tarsal section of the paw.

Dosage form

0.1, 0.3, 1, 3, 10mg/kg; twice a day for 10 days; p.o.

Applications

Upadacitinib administration improved synovial hypertrophy, inflammation, cartilage damage and bone erosion at the 3 and 10mg/kg dose groups.

References:
[1]Parmentier JM, Voss J, Graff C, et al. In vitro and in vivo characterization of the JAK1 selectivity of upadacitinib (ABT?494). BMC Rheumatol. 2018;2:23.
[2]Young DA, Hegen M, Ma HL, Whitters MJ, Albert LM, Lowe L, Senices M, Wu PW, Sibley B, Leathurby Y, Brown TP, Nickerson-Nutter C, Keith JC Jr, Collins M. Blockade of the interleukin-21/interleukin-21 receptor pathway ameliorates disease in animal models of rheumatoid arthritis. Arthritis Rheum. 2007 Apr;56(4):1152-63. PMID: 17393408.