CKG012 is an inhibitor of the Wnt/β-catenin signaling pathway that inhibits the release of HMGB1 and the transcription of β-catenin. It exhibits attenuating activities against cecal ligation and puncture (CLP)-induced sepsis and multiple myeloma cancer [1] [2].

CGK012 (0-20 μM) reduces excessive vascular permeability in HUVECs by inhibiting LPS-induced acetylation of HMGB1 and expression of SIRT1, thereby suppressing the release of HMGB1 and the expression of pathogen-associated molecules such as TLR2/4, without affecting cell viability [1]. It improves inflammatory responses by decreasing adhesion and migration of inflammatory immune cells, and by inhibiting the production of pro-inflammatory cytokines IL-6, TNF-α, and transcription factors NF-kB and ERK1/2 [1]. Moreover, CGK012 (0-20 μM) promotes β-catenin phosphorylation and degradation, suppressing β-catenin-dependent gene expression, and inhibiting the proliferation of multiple myeloma cells RPMI-8226 with an IC50 of 5.08 μM [2]. In cell viability assays [1] using HUVECs at concentrations of 5-100 μM over 48 hours, CGK012 showed no impact on cell viability. Western blot analysis [1] in HUVECs, HEK293-FL reporter, and RPMI-8226 cells at 0-20 μM for 12 hours demonstrated reduced levels of SIRT1 and acetylated HMGB1, as well as decreased β-catenin in HEK293-FL reporter and RPMI-8226. Cell migration assays [1] with HUVECs at 0-20 μM for 6 hours showed inhibition of neutrophil migration through HUVEC monolayers.

CGK012 (0.05-0.53 mg/kg, intravenously, double dose) suppresses HMGB1 release and immune cell migration, thereby enhancing vascular cell stability and survival rates in C57BL/6 mice with sepsis induced by cecal ligation and puncture [1]. Animal Model: CLP-induced sepsis in C57BL/6 mice [1]. Dosage: 0.26-0.53 mg/kg. Administration: double doses, 12 or 50 hours post-surgery. Result: Decreased HMGB1 expression and increased survival rate.