ZW4864 is a selective inhibitor of the β-catenin/B-Cell Lymphoma 9 protein (β-catenin/BCL9 PPI) interaction, demonstrating oral activity. It inhibits β-catenin/BCL9 PPI with a K_i value of 0.76 μM and an IC_50 value of 0.87 μM.

β-catenin-BCL9:0.76 μM (Ki), β-catenin-BCL9:0.87 μM (IC50)

ZW4864, at concentrations ranging from 10 to 40 μM over periods of 24 to 72 hours, has been demonstrated to modulate β-catenin signaling pathways selectively in various cell lines, including SW480, MDA-MB-231, MDA-MB-468, and MCF10A. Specifically, it lowers the levels of Axin2 and cyclin D1 proteins in SW480 and MDA-MD-231 cells, demonstrating a suppression of β-catenin target gene transcription without impacting the expression of the housekeeper gene HPRT. Furthermore, ZW4864 is effective in inducing rapid apoptosis in triple-negative breast cancer cells that exhibit hyperactive β-catenin signaling, leaving normal MCF10A mammary epithelial cells unaffected. Its mechanism of action includes binding to β-catenin, disrupting its interaction with B-cell lymphoma 9 (BCL9) but not with E-cadherin, thus inhibiting β-catenin signaling pathway activation, downregulating oncogenic target genes, and reducing cancer cell invasiveness. This compound also significantly diminishes TOPFlash luciferase activity in a dose-dependent manner in β-catenin-expressing cells, indicating a targeted suppression of β-catenin transactivation. These findings, validated through Western Blot Analysis, Apoptosis Analysis, and RT-PCR, underscore ZW4864's potential as a selective inhibitor of β-catenin signaling in cancer research.

ZW4864, administered orally at a dosage of 20 mg/kg, demonstrates favorable pharmacokinetic properties, achieving an oral bioavailability (F) of 83%[1]. At a higher dosage of 90 mg/kg, the compound is observed to alter tumor growth in mice[1]. Furthermore, ZW4864 effectively inhibits β-catenin target gene expression within a patient-derived xenograft mouse model. These studies were conducted using C57BL/6 mice for pharmacokinetic analysis at 20 mg/kg dosage and general mice population at 90 mg/kg dosage, showcasing the compound's potential in moderating tumor progression and exhibiting excellent pharmacokinetic profiles[1].