Napabucasin (BBI608) is an orally available Stat3 and Y cell stemness inhibitor.

For cancer stem cells, spheres are dissociated and plated under cancer stem cell culture conditions on coated 96-well plates. After 72 h of culture, wells are dosed with the indicated compounds. Seventy-two hours or 24 h after dosing, CellTiter-Glo 2.0 is added to each well, and the luminescence is measured as described by the manufacturer. IC50 values are calculated by fitting a four parameter dose–response curve to normalized data using GraphPad Prism software. For bulk cells, cells are plated at 5,000 cells per well on 96-well plates. Twenty-four hours after plating, cells are treated with the indicated compounds. Viability is determined at 72 h as described above.(Only for Reference)

H2O: < 1 mg/mL (insoluble or slightly soluble)

DMSO: 6.88 mg/mL (28.64 mM), Sonication is recommended.

acetonitrile: 0.8 mg/mL (3.33 mM), Sonication is recommended.

Ethanol: < 1 mg/mL (insoluble or slightly soluble)