1. Stem Cell/Wnt
  2. Porcupine

LGK974 

Cat. No.: HY-17545 Purity: 98.97%
Handling Instructions

LGK974 is a potent and specific Porcupine (PORCN) inhibitor with an IC50 of 0.1 nM.

For research use only. We do not sell to patients.

LGK974 Chemical Structure

LGK974 Chemical Structure

CAS No. : 1243244-14-5

Size Price Stock Quantity
Free Sample (0.5-1 mg)   Apply now  
10 mM * 1 mL in DMSO USD 132 In-stock
Estimated Time of Arrival: December 31
5 mg USD 120 In-stock
Estimated Time of Arrival: December 31
10 mg USD 204 In-stock
Estimated Time of Arrival: December 31
50 mg USD 720 In-stock
Estimated Time of Arrival: December 31
100 mg USD 1128 In-stock
Estimated Time of Arrival: December 31
200 mg   Get quote  
500 mg   Get quote  

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    LGK974 purchased from MCE. Usage Cited in: Exp Cell Res. 2016 Jul 15;345(2):206-17.

    Inhibition of PORCN abolishes Runx2 induction stimulated by β-GP. VSMCs are pretreated with LGK974 (1nM) for 30 min, then incubated with β-GP (10mM) for 24h, and the cell lysates are immunoblotted with antibodies against Runx2 and β-actin, respectively.
    • Biological Activity

    • Protocol

    • Technical Information

    • Purity & Documentation

    • References

    Description

    LGK974 is a potent and specific Porcupine (PORCN) inhibitor with an IC50 of 0.1 nM.

    IC50 & Target

    Porcupine[1]

    In Vitro

    LGK974 effectively displaces [3H]-GNF-1331 with an IC50 of 1 nM in the PORCN radioligand binding assay. LGK974 potently reduces Wnt-dependent AXIN2 mRNA levels in HN30 cells with an IC50 of 0.3 nM[1].

    In Vivo

    LGK974, a drug that targets Porcupine, a Wnt-specific acyltransferase. LGK974 potently inhibits Wnt signaling, has strong efficacy in rodent tumor models, and is well-tolerated. Toxicology studies are performed on nontumor bearing rats at 3 and 20 mg/kg. At the efficacious dose of 3 mg/kg per day for 14 d, LGK974 is well-tolerated without abnormal histopathological findings in Wnt-dependent tissues, including the intestine, stomach, and skin. When rats are administrated a very high dose of 20 mg/kg per day for 14 d, loss of intestinal epithelium is observed, consistent with the concept that Wnt is required for intestinal tissue homeostasis[1].

    Clinical Trial
    Solvent & Solubility
    In Vitro: 

    DMSO : ≥ 32 mg/mL (80.72 mM)

    *"≥" means soluble, but saturation unknown.

    Preparing
    Stock Solutions
    Concentration Solvent Mass 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
    *Please refer to the solubility information to select the appropriate solvent.
    In Vivo:
    • 1.

      Add each solvent one by one:  10% DMSO    90% (20% SBE-β-CD in saline)

      Solubility: 2.5 mg/mL (6.31 mM); Suspended solution; Need ultrasonic and warming

    • 2.

      Add each solvent one by one:  10% DMSO    90% corn oil

      Solubility: ≥ 2.5 mg/mL (6.31 mM); Clear solution

    References
    Kinase Assay
    [1]

    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].

    MCE has not independently confirmed the accuracy of these methods. They are for reference only.

    Cell Assay
    [1]

    HN30 cells and UMSCC cells are used. For TaqMan assay, 2×106 cells per well are plated into six-well cell culture plates and treated with or without LGK974 in amultipoint dose-response. RNA samples are collected after 48 h. For colony formation assays, 2×103 cells per well are plated into six-well cell culture plates with or without compound treatment. Cells are stained with crystal violet 1 wk later[1].

    MCE has not independently confirmed the accuracy of these methods. They are for reference only.

    Animal Administration
    [1]

    Mice and Rats[1]
    Nude mice (or nude rats) bearing the mouse mammary tumor virus-Wnt1, HN30, or SNU1076 tumors are randomized according to tumor volume. LGK974 is formulated in 10% (vol/vol) citrate buffer (pH 2.8)/90% (vol/vol) citrate buffer (pH 3.0) or 0.5% MC/0.5% Tween 80 and administered by oral gavage at a dosing volume of 10 μL/g animal body weight. Body weight is monitored daily, and tumor sizes are assessed three times per week after the tumors are palpable. Tumor sizes are determined by using caliper measurements. Tumor volumes are calculated with a formula (length×width×height)/2. The plasma concentrations and exposures of LGK974 in the tumor-bearing nude mice (n=2 per dosing group) are determined on day 14. Blood samples (50 μL) are collected by serial retroorbital sampling at 1, 3, 7, 16, and 24 h postdose. The blood samples are centrifuged, and plasma is separated and frozen until analysis by liquid chromatography/MS/MS. For tolerability studies, LGK974 is administrated to nontumor-bearing Wistar rats one time per day by oral gavage at 3 or 20 mg/kg per day. Necropsies are performed at the end of the study. Tissues are fixed in 10% (vol/vol) neutralbuffered formalin, sectioned, and subjected to H&E staining.

    MCE has not independently confirmed the accuracy of these methods. They are for reference only.

    References
    Molecular Weight

    396.44

    Formula

    C₂₃H₂₀N₆O

    CAS No.

    1243244-14-5

    SMILES

    O=C(NC1=NC=C(C2=NC=CN=C2)C=C1)CC3=CN=C(C4=CC(C)=NC=C4)C(C)=C3

    Storage
    Powder -20°C 3 years
      4°C 2 years
    In solvent -80°C 6 months
      -20°C 1 month
    Shipping

    Room temperature in continental US; may vary elsewhere

    Purity: 98.97%

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    Product Name:
    LGK974
    Cat. No.:
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    Cat. No.: HY-17545