产品描述

G007-LK是一种有效的选择性tankyrase抑制剂，对TNKS1/2的IC50分别为46 nM 和 25 nM。

靶点（IC50 & Targe）

PARP

体外研究

G007-LK reduces Wnt/β-catenin signaling by preventing poly(ADP-ribosyl)ation-dependent AXIN degradation, thereby promoting β-catenin destabilization. G007-LK completely blocks ligand-driven Wnt/β-catenin signaling in cell culture and display approximately 50% inhibition of APC mutation–driven signaling in most CRC cell lines. G007-LK (0.2 μM) reduces the number of COLO-320DM cells in mitosis from 24% to 12% and decreases HCT-15 cells in S-phase from 28% to 18%. G007-LK suppresses colony formation in CRC lines COLO-320DM and SW403. G007-LK suppresses organoids growth with IC50 of 80 nM. [2]

体内研究

G007-LK exhibits antitumor efficacy in xenograft and genetically engineered CRC models. In the COLO-320DM model, G007-LK reduces tankyrases 1 and tankyrases 2 protein levels, stabilizes AXIN1 and AXIN2, and decreases β-catenin level. Wnt/β-catenin signaling is clearly inhibited in a dose-dependent manner in the efficacy study tumors as indicated by reduced expression of β-catenin–activated genes NKD1, APCDD1, and TNFRSF19 (TROY), as well as increased expression of β-catenin–repressed gene CLIC3. G007-LK treatment increases expression of KRT20 and TM4SF4 in COLO-320DM tumors. G007-LK (20 mg/kg twice daily) achieves 61% tumor growth inhibition. G007-LK reduces Wnt/β-catenin signaling and cell proliferation in normal intestine. [2]

激酶实验

TNKS1 and TNKS2 in vitro biochemical assays:

G007-LK inhibitory activity at various doses (duplicates) is tested twice against TNKS1, TNSK2 Chemiluminescent Assay Kits, and the luminescence is measured on a GloMax Luminometer.

细胞实验

Cell lines: APC-mutant CRC cell lines COLO-320DM

Concentrations: ~0.2 μM

Incubation Time: 7 to 13 days

Method: For colony formation assays, cells are seeded at 500 cells/well in 2 mL of 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, changing medium and compound every third day. Colonies are stained by adding 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.

(Only for Reference)

动物实验

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

(Only for Reference)

参考文献

[1] Voronkov A, et al. J Med Chem, 2013, 56(7), 3012-3023.

[2] Lau T, et al. Cancer Res, 2013, 73(10), 3132-3144.