LGK974 (WNT974; LGK974)   中文名称: 2',3-二甲基-N-[5-(2-吡嗪基)-2-吡啶基]-[2,4'-联吡啶]-5-乙酰胺

Porcupine (PORCN, IC₅₀ = 0.1 nM), a membrane-bound O-acyltransferase essential for Wnt ligand palmitoylation and secretion [1]

LGK974 在稳定表达Wnt响应荧光素酶报告基因（STF3A）的HEK293细胞中抑制Wnt信号通路，EC₅₀为0.4 nM。100 nM浓度处理可使Wnt3a诱导的荧光素酶活性降低>95%。 [1]

该化合物在体外阻断Wnt配体棕榈酰化，经10 nM LGK974处理24小时的PORCN过表达细胞中，Wnt3a蛋白的棕榈酸盐掺入完全消失。 [1]

在肺腺癌细胞系（如H2030-BrM3）中，LGK974（1 μM）通过抑制自分泌Wnt产生，72小时后使肿瘤细胞增殖减少>80%。Wnt通路活性通过Axin2 mRNA下调（qPCR）评估。 [2]
LGK974 在 PORCN 放射性配体结合测定中有效取代 [3H]-GNF-1331，IC50 为 1 nM。在 HN30 细胞中，LGK974 有效降低 Wnt 依赖性 AXIN2 mRNA 水平，IC50 为 0.3 nM[1]。

在MMTV-Wnt1转基因小鼠（自发性乳腺肿瘤）中，口服LGK974（3 mg/kg/天）诱导5/10小鼠肿瘤完全消退，21天后肿瘤生长抑制率达98%（对比溶剂组）。肿瘤内Wnt靶基因（Axin2, Lgr5）表达下降>90%。 [1]

在肺腺癌患者来源异种移植（PDX）模型中，口服LGK974（20 mg/kg/天）治疗4周后肿瘤生长减少70%。此疗效与肿瘤基质中Wnt配体活性降低相关（通过Axin2原位杂交证实）。 [2]
药物 LGK974，针对 Wnt 特异性酰基转移酶 Porcupine。 LGK974 强烈抑制 Wnt 信号传导，耐受性良好，并且在啮齿动物肿瘤模型中非常有效。没有肿瘤的大鼠以3和20mg/kg的剂量用于毒理学试验。 LGK974 在每天 3 mg/kg 的有效剂量下持续 14 天具有良好的耐受性，并且在依赖 Wnt 的组织（例如胃、肠或皮肤）中没有观察到异常的组织病理学异常。连续 14 天给予非常高剂量（每天 20 mg/kg）的大鼠表现出肠上皮细胞的损失，这与 Wnt 对于肠组织稳态是必需的这一观点是一致的 [1]。

采用体外棕榈酰化实验检测PORCN酶活性。将重组PORCN与Wnt3a肽段底物、³H-棕榈酰辅酶A及LGK974（0.01–100 nM）共孵育，通过闪烁计数法定量放射性棕榈酸盐掺入以计算IC₅₀。 [1]

STF3A-HEK293细胞（Wnt报告基因）经LGK974（0.1–100 nM）处理24小时（±Wnt3a条件培养基），测定荧光素酶活性以计算EC₅₀。 [1]

增殖实验中，肺腺癌细胞（如H2030-BrM3）经LGK974（0.001–10 μM）处理72小时后，采用CellTiter-Glo检测细胞活性。通过Axin2和LEF1的qPCR验证Wnt通路抑制。 [2]

Dissolved in 010% (vol/vol) citrate buffer (pH 2.8)/90% (vol/vol) citrate buffer (pH 3.0) or 0.5% MC/0.5% Tween 80; 3 mg/kg; Oral gavage
A murine MMTV-Wnt1 tumor model and a human head and neck squamous cell carcinoma model (HN30)

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MMTV-Wnt1 mice received oral LGK974 (3 mg/kg or 10 mg/kg) suspended in 0.5% methylcellulose/0.5% Tween-80, administered daily by gavage. Tumor volume was measured by caliper. [1]

Lung adenocarcinoma PDX models were established in NSG mice. Mice were orally dosed with LGK974 (20 mg/kg in 0.5% methylcellulose/0.5% Tween-80) daily for 28 days. Tumor growth was monitored by caliper. [2]

[1]. Targeting Wnt-driven cancer through the inhibition of Porcupine by LGK974. Proc Natl Acad Sci U S A. 2013 Dec 10;110(50):20224-9.

[2]. A Wnt-producing niche drives proliferative potential and progression in lung adenocarcinoma. Nature. 2017 May 18;545(7654):355-359.

LGK974 is a first-in-class PORCN inhibitor that blocks Wnt ligand secretion, thereby suppressing Wnt/β-catenin signaling in Wnt-addicted cancers. [1]

In lung adenocarcinoma, tumor progression is driven by Wnt-producing stromal niches. LGK974 abrogates this paracrine signaling, demonstrating therapeutic potential against stromal-dependent malignancies. [2]
LGK974 is a carboxamide, the structure of which is that of acetamide substituted on carbon by a 2',3-dimethyl-2,4'-bipyridin-5-yl group and on nitrogen by a 5-(pyrazin-2-yl)pyridin-2-yl group. It is a highly potent, selective and orally bioavailable Porcupine inhibitor (a Wnt signalling inhibitor). It has a role as a Wnt signalling inhibitor. It is a member of bipyridines, a member of pyrazines, a member of pyridines and a secondary carboxamide. It is functionally related to an acetamide.
WNT974 has been used in trials studying the treatment of Metastatic Colorectal Cancer and Squamous Cell Carcinoma, Head And Neck.
Porcupine Inhibitor WNT974 is an orally available inhibitor of porcupine (PORCN), with potential antineoplastic activity. Upon oral administration, WNT974 binds to and inhibits PORCN in the endoplasmic reticulum (ER), which blocks post-translational acylation of Wnt ligands and inhibits their secretion. This prevents the activation of Wnt ligands, interferes with Wnt-mediated signaling, and inhibits cell growth in Wnt-driven tumors. Porcupine, a membrane-bound O-acyltransferase (MBOAT), is required for the palmitoylation of Wnt ligands, and plays a key role in Wnt ligand secretion and activity. Wnt signaling is dysregulated in a variety of cancers.

C23H20N6O

396.4445

396.169

C, 69.68; H, 5.08; N, 21.20; O, 4.04

1243244-14-5

2375595-49-4 (2HCl);1243244-14-5;

46926973

White to light brown solid powder

1.3±0.1 g/cm3

650.9±55.0 °C at 760 mmHg

347.5±31.5 °C

0.0±1.9 mmHg at 25°C

1.652

2

97.04

1

6

5

30

560

0

O=C(C([H])([H])C1=C([H])N=C(C2C([H])=C([H])N=C(C([H])([H])[H])C=2[H])C(C([H])([H])[H])=C1[H])N([H])C1C([H])=C([H])C(C2C([H])=NC([H])=C([H])N=2)=C([H])N=1

XXYGTCZJJLTAGH-UHFFFAOYSA-N

InChI=1S/C23H20N6O/c1-15-9-17(12-28-23(15)18-5-6-25-16(2)10-18)11-22(30)29-21-4-3-19(13-27-21)20-14-24-7-8-26-20/h3-10,12-14H,11H2,1-2H3,(H,27,29,30)

2-(2',3-dimethyl-[2,4'-bipyridin]-5-yl)-N-(5-(pyrazin-2-yl)pyridin-2-yl)acetamide

NVP-LGK974; NVPLGK974; 1243244-14-5; 2-(2',3-Dimethyl-[2,4'-bipyridin]-5-yl)-N-(5-(pyrazin-2-yl)pyridin-2-yl)acetamide; CHEBI:78030; NVP LGK974; LGK974; LGK-974; LGK 974; WNT974; WNT 974; WNT-974

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

配方 1 中的溶解度: 2.5 mg/mL (6.31 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 悬浮液； 超声和加热处理
例如，若需制备1 mL的工作液，可将100 μL 25.0 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 2 中的溶解度: ≥ 2.5 mg/mL (6.31 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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配方 3 中的溶解度: 2% DMSO+corn oil:5 mg/mL

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。