AS1517499 is a blood-brain barrier permeable inhibitor of STAT6 phosphorylation (IC50= 21 nM).

STAT6:21 nM, RhoA (HBSM cells):21 nM, STAT6 (HBSM cells):21 nM

METHODS: Western Blot was used to detect the expression levels of AS1517499 in human BSM cells treated with 100 nM AS1517499.
RESULTS: Treatment of BSM cells with AS1517499 inhibited IL-13-induced STAT6 phosphorylation and up-regulation of RhoA. [1]

METHODS: To investigate the effect of AS1517499 on the development of antigen-induced BSM hyperreactivity, AS1517499 (10 mg/kg) was administered intraperitoneally to mice 1 hour before each ovalbumin exposure.
RESULTS: AS1517499 completely inhibited the antigenicity induced upregulation of RhoA and BSM hyperreactivity. [1]

Normal human BSM cells (hBSMCs) are maintained in SmBM medium supplemented with 5% fetal bovine serum, 0.5 ng/mL human epidermal growth factor (hEGF), 5 μg/mL insulin, 2 ng/mL human fibroblast growth factor-basic (hFGFb), 50 μg/mL gentamicin, and 50 ng/mL amphotericin B. Cells are maintained at 37°C in a humidified atmosphere (5% CO2), fed every 48 to 72 hours, and passaged when cells reached 90 to 95% confluence. Then the hBSMCs (passages 7-9) are seeded in 6-well plates and 8-well chamber slides at a density of 3,500 cells/cm2 and, when 80 to 85% confluence observed, cells are cultured without serum for 24 hours before addition of recombin is ant human IL-13. AS1517499 (100 nM) or its vehicle (0.3% DMSO) is treated 30 minutes before the addition of IL-13 (100 ng/mL). In some experiments, AS1517499 is treated 0 (co-incubation), 3, or 12 hours after the addition of IL-13. In another series of experiments, a selective Rho-kinase inhibitor Y-27632 (1 μM) or its vehicle (0.3% DMSO) is also applied 15 minutes before the IL-13 application. At the indicated time after the IL-13 treatment, cells are washed with PBS, immediately collected, and disrupted with 1× SDS sample buffer (250 μL/well), and used for Western blot analyses[2] .

Mice[2]