IF images of uninjured, 48 hpi, and 72 hpi mouse airway epithelia wild-type mice treated with vehicle or LGK974 assessing BrdU incorporation. Relative to vehicle-treated mice, LGK974-treated mice exhibited an attenuated SAE with few BrdU+ ABSCs, thereby phenocopying the repair response of K5-CreER;Ctnnb1fl/fl mouse tracheas.
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.
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Description
LGK974 (WNT974) is an orally bioavailable and specific Porcupine (PORCN) inhibitor with an IC50 of 0.1 nM[1].
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].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
LGK974 Related Antibodies
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].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Room temperature in continental US; may vary elsewhere.
Storage
Powder
-20°C
3 years
4°C
2 years
In solvent
-80°C
2 years
-20°C
1 year
Solvent & Solubility
In Vitro:
DMSO : ≥ 32 mg/mL (80.72 mM; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
*"≥" means soluble, but saturation unknown.
Preparing Stock Solutions
ConcentrationSolventMass
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
View the Complete Stock Solution Preparation Table
*Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles. Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.
For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day. The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.
Protocol 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
This protocol yields a suspended solution of 2.5 mg/mL. Suspended solution can be used for oral and intraperitoneal injection.
Taking 1 mL working solution as an example, add 100 μLDMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
Protocol 2
Add each solvent one by one: 10% DMSO 90% Corn Oil
Solubility: ≥ 2.5 mg/mL (6.31 mM); Clear solution
This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown). If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Taking 1 mL working solution as an example, add 100 μLDMSO stock solution (25.0 mg/mL) to 900 μLCorn oil, and mix evenly.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:
Dosage
mg/kg
Animal weight (per animal)
g
Dosing volume (per animal)
μL
Number of animals
Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
%
DMSO+
%
+
%
Tween-80
+
%
Saline
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
The co-solvents required include: DMSO,
. All of co-solvents are available by MedChemExpress (MCE).
, Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Calculation results:
Working solution concentration:
mg/mL
Method for preparing stock solution:
mg
drug dissolved in
μL
DMSO (Stock solution concentration: mg/mL).
The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
Method for preparing in vivo working solution for animal experiments: Take
μL DMSO stock solution, add
μL .
μL , mix evenly, next add
μL Tween 80, mix evenly, then add
μL Saline.
Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution
If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
[1]. Liu J, et al. 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.
[Content Brief]
[2]. Tammela T, et al. A Wnt-producing niche drives proliferative potential and progression in lung adenocarcinoma. Nature. 2017 May 18;545(7654):355-359.
[Content Brief]
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
[1]. Liu J, et al. 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.
[Content Brief]
[2]. Tammela T, et al. A Wnt-producing niche drives proliferative potential and progression in lung adenocarcinoma. Nature. 2017 May 18;545(7654):355-359.
[Content Brief]
[1]. Liu J, et al. 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]. Tammela T, et al. A Wnt-producing niche drives proliferative potential and progression in lung adenocarcinoma. Nature. 2017 May 18;545(7654):355-359.
Complete Stock Solution Preparation Table
*Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles. Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.
Optional Solvent
ConcentrationSolventMass
1 mg
5 mg
10 mg
25 mg
DMSO
1 mM
2.5224 mL
12.6122 mL
25.2245 mL
63.0612 mL
5 mM
0.5045 mL
2.5224 mL
5.0449 mL
12.6122 mL
10 mM
0.2522 mL
1.2612 mL
2.5224 mL
6.3061 mL
15 mM
0.1682 mL
0.8408 mL
1.6816 mL
4.2041 mL
20 mM
0.1261 mL
0.6306 mL
1.2612 mL
3.1531 mL
25 mM
0.1009 mL
0.5045 mL
1.0090 mL
2.5224 mL
30 mM
0.0841 mL
0.4204 mL
0.8408 mL
2.1020 mL
40 mM
0.0631 mL
0.3153 mL
0.6306 mL
1.5765 mL
50 mM
0.0504 mL
0.2522 mL
0.5045 mL
1.2612 mL
60 mM
0.0420 mL
0.2102 mL
0.4204 mL
1.0510 mL
80 mM
0.0315 mL
0.1577 mL
0.3153 mL
0.7883 mL
LGK974 Related Classifications
Help & FAQs
Do most proteins show cross-species activity?
Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.