LGK-974 (formerly known as WNT-974; LGK-974 and NVP-LGK974) is a novel potent, orally bioavailable and specific/selective PORCN (Porcupine) inhibitor with potential anticancer activity. It inhibits Wnt signaling with an IC50 of 0.4 nM in TM3 cells. LGK974 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. LGK-974 is being investigated for the treatment of cancers driven by the Wnt pathway in a Wnt ligand-dependent manner.

Physicochemical Properties

Molecular Formula	C23H20N6O
Molecular Weight	396.44
Exact Mass	396.169
CAS #	1243244-14-5
Related CAS #	2375595-49-4 (2HCl);1243244-14-5;
PubChem CID	46926973
Appearance	White to light brown solid powder
Density	1.3±0.1 g/cm3
Boiling Point	650.9±55.0 °C at 760 mmHg
Flash Point	347.5±31.5 °C
Vapour Pressure	0.0±1.9 mmHg at 25°C
Index of Refraction	1.652
LogP	2
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	5
Heavy Atom Count	30
Complexity	560
Defined Atom Stereocenter Count	0
InChi Key	XXYGTCZJJLTAGH-UHFFFAOYSA-N
InChi Code	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)
Chemical Name	2-(2',3-dimethyl-[2,4'-bipyridin]-5-yl)-N-(5-(pyrazin-2-yl)pyridin-2-yl)acetamide
Synonyms	NVP-LGK974; NVPLGK974; NVP LGK974; LGK974; LGK-974; LGK 974; WNT974; WNT 974; WNT-974
HS Tariff Code	2934.99.9001
Storage	Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

ln Vitro	LGK974 efficiently displaces [3H]-GNF-1331 in PORCN radioligand binding assays, with an IC50 of 1 nM. In HN30 cells, LGK974 efficiently lowers Wnt-dependent AXIN2 mRNA levels with an IC50 of 0.3 nM[1].
ln Vivo	The medication LGK974, which targets the Wnt-specific acyltransferase Porcupine. Strongly suppressing Wnt signaling, well-tolerated, and highly effective in rodent tumor models is LGK974. Rats without tumors were used in toxicology tests at doses of 3 and 20 mg/kg. LGK974 was well tolerated at an effective dose of 3 mg/kg per day for 14 days, and no aberrant histopathological abnormalities were observed in tissues that depend on Wnt, such as the stomach, intestines, or skin. Rats given very high dosages (20 mg/kg per day) for 14 days showed loss of intestinal epithelial cells, which is consistent with the idea that Wnt is necessary for intestinal tissue homeostasis [1].
Animal Protocol	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)
References	[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.
Additional Infomation	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.

Solubility Data

Solubility (In Vitro)

DMSO: 79 mg/mL (199.3 mM) Water:<1 mg/mL Ethanol:<1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: 2.5 mg/mL (6.31 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with heating and sonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (6.31 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 3: 2% DMSO+corn oil:5 mg/mL  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.5224 mL	12.6122 mL	25.2245 mL
	5 mM	0.5045 mL	2.5224 mL	5.0449 mL
	10 mM	0.2522 mL	1.2612 mL	2.5224 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.