Inhibitory activities

IC50 values for TG101348 were determined commercially using the InVitrogen kinase profiling service for a 223 kinase screen that included JAK2 and JAK2V617F or Carna Biosciences for the screen of all Janus kinase family members including JAK1 and Tyk2. ATP concentration was set to approximately the Km value for each kinase.

Cell lines

Human erythroleukemic cell line (HEL) with JAK2V617F mutation; Ba/F3 cells

Preparation method

Soluble in DMSO > 10 mM. General tips for obtaining a higher concentration: Please warm the tube at 37℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months.

Reaction Conditions

0.1, 0.3, 1, 3, and 10 μM; 72 h.

Applications

In HEL and Ba/F3 cells with JAK2V617F mutation, TG101348 (SAR302503) inhibits cell proliferation with IC50 values of 300 nM and 420 nM, respectively. TG101348 also reduces STAT5 phosphorylation and induces apoptosis in a dose-dependent way.

Animal models

C57Bl/6 mice; C57BL/6 mice intravenously injected with whole bone marrow expressing JAK2V617F.

Dosage form

30, 100, and 200 mg/kg; 0.5, 1, 3, 5, 7, and 24 hr postdose; administrated orally.60 mg/kg, 120 mg/kg; oral gavage twice daily; from day 28 on for 42 days.

Applications

In C57Bl/6 mice, maximum plasma concentrations (Cmax) of TG101348 are 0.68, 3.58, and 4.28 μM at 3 h postdose at doses 30, 100, and 200 mg/kg, respectively. In mice with polycythemia vera induced by JAK2V617F, TG101348 reduces hematocrit by 5.1% and 17.9% at 60 mg/kg and 120 mg/kg, respectively. Also, TG101348 dose-dependently inhibits splenomegaly and polycythemia.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

References:

[1]. Wernig G, Kharas MG, Okabe R, et al. Efficacy of TG101348, a selective JAK2 inhibitor, in treatment of a murine model of JAK2V617F-induced polycythemia vera. Cancer Cell, 2008, 13(4): 311-320.