In vivo dose-exposure profiling of 1a was conducted using a small cohort of three male mice was subjected to single ip doses at 10, 30, and 50 mg/kg. Blood was drawn at 0.5 and 3 h, and subsequent LC/MS analysis.

PMA/Iono induced NF-kB luciferase assay was done in HEK293- NF-kB-Luc stable cells HEK293-NF-kB-Luc cells were seeded in 96-well white plates overnight. CID-2858522 was diluted in culture medium and 5 mkL was added to the cells to reach a final concentration 5mkM. 2 h later, 5 mkL PMA/Iono (200 ng/mL) was added (final concentration = 10 ng/mL) and cells were treated for another 16 h. 50 mkL. BriteLite solution (Perkin–Elmer) was added to each well and luminescence was read using a luminometor.