LGK974 是一种有效的、特异性的、具有口服活性的Porcupine(PORCN) 抑制剂(IC50:0.1 nM)。

LGK-974 is an effective and selective PORCN inhibitor and inhibits Wnt signaling (IC50: 0.4 nM) in TM3 cells. It has been used in trials studying the treatment of Metastatic Colorectal Cancer and Squamous Cell Carcinoma, Head And Neck.

在小鼠MMTV-WNT1肿瘤模型和人的头、颈部鳞状细胞癌模型（hn30）中,LGK-974（3 mg/kg）对Wnt信号通路有抑制作用,其可诱导肿瘤退化且不影响小鼠体重.LGK-974（5 mg/kg,2次/天,p.o.）还可抑制RNF43-突变胰腺肿瘤(HPAF-II 和Capan-2)的生长

LGK-974对测试的Wnts均有抑制作用（IC50：0.05-2.4 nM）。 在PORCN放射配体结合测定中,LGK-974可有效取代[3H]GNF -1331（IC50：1 nM）,且20 μM时细胞毒性也较小。LGK-974可特异抑制RNF43突变细胞株HPAF-II、PaTu 8988S和 Capan-2的生长。

Radioligand binding assay: using the aforementioned membrane preps, filtration binding assays are performed. To reduce nonspecific binding, 96-well filtration plates are precoated as suggested by the manufacturer with 0.1% BSA and then washed four times with 0.1% BSA. Membrane preps (50 μg total protein) are incubated in polypropylene 96-well plates with 6.6 nM 3H-GNF-1331 in the presence or absence of a testing compound in binding buffer (50 mM Tris, pH 7.5, 5 mM MgCl2, 1 mM EDTA, 0.1% BSA) plus EDTA-free protease inhibitor mixture in a final volume of 150 μL for 3 h at room temperature. Binding reaction mixtures are then transferred to the precoated 96-well filtration plates, filtered, and washed using a 96-pin FilterMate Harvester. Radioactive signals are obtained using a Microplate Scintillation Counter TopCount. Curve fitting is performed using Prism[1].

Cells are plated in growth medium in a 96-well plate at a density of 6,000–12,000 cells per well and treated with DMSO or 1 μM LGK974. After 3 d, the cells are treated with fresh growth medium containing 20 μM EdU, which is included in the Click-iT EdU Alexa Fluor 488 HCS assay kit, and the plate was incubated for 2 h at 37 °C in a humidi?ed atmosphere containing 5% CO2. Cells are ?xed with ?nal 4% (mass/vol) paraformaldehyde for 30 min, washed with PBS, permeabilized, and stained with 50 μg/mL Hoechst in PBS for 30 min. After wash, the cells are proceeded to EdU detection according to the instruction of Click-iT EdU assay kit. Triplet wells are performed for each condition. (Only for Reference)

1243244-14-5

C23H20N6O

396.454

LGK 974;WNT974;LGK-974;NVP-LGK974

DMSO:73 mg/mL (184.1 mM)
Ethanol:<1 mgml
Powder: -20°C for 3 years
In solvent: -80°C for 2 years