G007-LK is a selective inhibitor of TNKS1 and TNKS2, with IC50s of 46 nM and 25 nM, respectively.

TNKS2:25 nM, TNKS1:46 nM

G007-LK demonstrated anti-tumor efficacy in xenograft and genetically engineered CRC models. In the COLO-320DM model, it reduced levels of tankyrases 1 and 2, stabilized AXIN1 and AXIN2, and decreased β-catenin levels. G007-LK treatment increased the expression of KRT20 and TM4SF4 in COLO-320DM tumors. Administered at 20 mg/kg twice daily, G007-LK achieved a 61% inhibition of tumor growth. Additionally, it reduced Wnt/β-catenin signaling and cell proliferation in normal intestinal tissue.

G007-LK inhibits Wnt/β-catenin signaling by preventing poly(ADP-ribosyl)ation-dependent AXIN degradation, thus promoting β-catenin destabilization. In cell culture, it completely blocks ligand-driven Wnt/β-catenin signaling and demonstrates inhibition of approximately 50% in APC mutation-driven signaling across most CRC cell lines. G007-LK reduces mitotic COLO-320DM cells from 24% to 12% and S-phase HCT-15 cells from 28% to 18% at a concentration of 0.2 μM. It also inhibits colony formation in CRC cell lines COLO-320DM and SW403. Furthermore, G007-LK suppresses organ growth with an IC50 of 80 nM.

TNKS1 and TNKS2 in vitro biochemical assays: G007-LK inhibitory activity at various doses (duplicates) is tested twice by TNKS1, TNSK2 Chemiluminescent Assay Kits, and the luminescence is measured.

Cell lines: APC-mutant CRC cell lines COLO-320DM. Concentrations: ~0.2 μM. Method: For colony formation assays,cells are seeded at 500 cells/well in 2 mL medium.Cell line in triplicate wells is treated with either 0.06% DMSO or compound in 0.06% DMSO and incubated for up to 17 days or until colonies became sufficiently large to quantify.Colonies are stained with 200 μL of 12 mM 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide to each well for 1 h,and colony numbers are quantitated with a GelCount scanner at 1200 dpi resolution.

Animal Models: Human APC –mutant CRC xenograft COLO-320DM. Formulation: 15% DMSO,17.5% Cremophor EL,8.75% ethanol,8.75% Miglyol 810N,50% PBS. Dosages: 20 mg/kg. Administration: intraperitoneal injection twice daily