The in vivo efficacy of JW74 is assessed using SW480 cell xenografts, employing relatively high doses of 150 or 300 mg/kg due to its rapid degradation observed in both human liver microsome analysis and pharmacokinetic studies. Mass spectrometry confirms the presence of JW74 in tumors and plasma, with tumor concentrations ranging between 4.2 to 72.1 μmol/kg for the 150 mg/kg dose, 1.9 to 11.1 μmol/kg for the 300 mg/kg dose, and a plasma concentration of 2.8 μM for both dosages^[1].

JW74 is noted for diminishing canonical Wnt signaling activity in the ST-Luc assay, achieving an IC50 of 790 nM^[1].

The treatment of U2OS cells with 10 μM JW74 for 72 hours results in a cellular viability drop to 80% compared to DMSO-treated cells. Additionally, flow cytometric analysis, after 48 hours of treatment with either DMSO or 10 μM JW74, reveals a reduction in Ki-67 expression from 97.5% in DMSO-treated cells to 86.7% in JW74-treated cells^[2].