LGK974 (WNT974)

Catalog No.S7143 Batch:S714305

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Technical Data

Formula

C23H20N6O
Molecular Weight 396.44 CAS No. 1243244-14-5
Solubility (25°C)* In vitro DMSO 79 mg/mL (199.27 mM)
Water Insoluble
Ethanol Insoluble
In Vivo (Add solvents to the product individually and in order.)
Homogeneous suspension
CMC-NA
≥5mg/ml Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

Biological Activity

Description LGK974 (WNT974) is a potent and specific PORCN inhibitor that inhibits Wnt signalling with an IC50 of 0.4 nM in TM3 cells, and is currently in Phase 1.
Targets
Porcn
(TM3 cells)
In vitro In the PORCN radioligand binding assay, LGK974 (WNT974) effectively displaces [3H]-GNF-1331 with an IC50 of 1 nM and shows no major cytotoxicity in cells up to 20 µM. It shows comparable inhibitory activities against all tested Wnts with IC50 ranging from 0.05 to 2.4 nM, which is consistent with the genetic loss of PORCN phenotype. This compound specifically inhibits the growth of three RNF43-mutant cell lines, HPAF-II, PaTu 8988S, and Capan-2.
In Vivo In a murine MMTV-Wnt1 tumour model and a human head and neck squamous cell carcinoma model (HN30), LGK974 (WNT974) (3 mg/kg) inhibits Wnt signalling in vivo and induces tumour regression without significant body weight loss in the mice. This compound (5 mg/kg, p.o., BID) also inhibits tumour growth of RNF43-mutant pancreatic tumours (HPAF-II and Capan-2) in vivo.
Features Orally bioavailable Porcupine-specific inhibitor that has been tested in Phase I clinical trials for treatment of malignancies dependent on Wnt ligands.

Protocol (from reference)

Cell Assay:

[2]
  • Cell Lines

    HPAF-II, PaTu 8988S, and Capan-2 cells

  • Concentrations

    ~1 μM

  • Incubation Time

    3 days

  • Method

    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 (WNT974). 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 humidified atmosphere containing 5% CO2. Cells are fixed with final 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.
Animal Study:

[1]
  • Animal Models

    A murine MMTV-Wnt1 tumor model and a human head and neck squamous cell carcinoma model (HN30)

  • Dosages

    ~3 mg/kg daily

  • Administration

    Oral gavage

References

  • https://pubmed.ncbi.nlm.nih.gov/24277854/
  • https://pubmed.ncbi.nlm.nih.gov/23847203/

Customer Product Validation

Nestin silencing inhibits the Wnt/β-catenin signaling in breast CSC. Nestin-si and Nestin-c breast CSC were cultured for 48 h, and the relative levels of target proteins and phosphorylation were determined by western blotting. Data are presented as the mean ± SD of the levels of target proteins vs. the β-actin control, or the phosphorylated vs. the total form, from six separate experiments. In addition, Nestinhigh in the presence or absence of LGK974 (1 nM), Nestinlow in the presence or absence of SB216763 (5 uM), and control CSC were tested for their proliferation via mammosphere formation assays. Quantitative analysis of the formed mammospheres. *P <0.05, vs. the Nestin-c or control, except for specifically indicated. CSC, cancer stem cell; SD, standard deviation.

Data from [ Breast Cancer Res , 2014 , 16(4), 408 ]

(c) Immunohistochemical images of intestinal sections from E-Rspo2 and R26-rtTA/P-Rspo3 mice treated with either DMSO or LGK974. BrdU was stained for proliferation. Scale bars, 100 μm.

Data from [ , , Nat Commun, 2017, 8:15945 ]

AP1/Jun noncanonical Wnt activity measured in the presence of LGK974 (porcupine inhibitor) and JNK-IN-8 (JNK inhibitor) (both 1 μM). F: Aldosterone secretion during the same experiment (n=4). Results are expressed in mean SEM, and P values show significance between treatments and baseline control.

Data from [ , , J Clin Endocrinol Metab, 2015, 100(6): E836-44 ]

(A) Body weight changes over the study by treatment group (LGK low: LGK974 at 3 mg/kg/day; LGK high: LGK974 at 6 mg/kg/kg/day; C59: Wnt-C59). (B) Femur length at necropsy. (C) Total body bone mineral density (BMD) at necropsy. (D) Spine BMD at necropsy.

Data from [ , , J Endocrinol, 2018, 238(1):13-23 ]

Sellecks LGK974 (WNT974) Has Been Cited by 94 Publications

CD36 enrichment in HER2-positive mesenchymal stem cells drives therapy refractoriness in breast cancer [ J Exp Clin Cancer Res, 2025, 44(1):19] PubMed: 39833955
Wnt signaling activation induces CTCF binding and loop formation at cis-regulatory elements of target genes [ Genome Res, 2025, 10.1101/gr.279684.124] PubMed: 40550689
Tamoxifen induces protection against manganese toxicity by REST upregulation via the ER-α/Wnt/β-catenin pathway in neuronal cells [ J Biol Chem, 2025, 301(6):108529] PubMed: 40280417
Developmental signals control chromosome segregation fidelity during pluripotency and neurogenesis by modulating replicative stress [ Nat Commun, 2024, 15(1):7404] PubMed: 39191776
Loss of ARID3A perturbs intestinal epithelial proliferation-differentiation ratio and regeneration [ J Exp Med, 2024, 221(10)e20232279] PubMed: 39150450
Physiological regulation of neuronal Wnt activity is essential for TDP-43 localization and function [ EMBO J, 2024, 43(16):3388-3413] PubMed: 38918634
GPA33 expression in colorectal cancer can be induced by WNT inhibition and targeted by cellular therapy [ Oncogene, 2024, ] PubMed: 39472498
Unbalancing cAMP and Ras/MAPK pathways as a therapeutic strategy for cutaneous neurofibromas [ JCI Insight, 2024, 9(3)e168826] PubMed: 38175707
Higher overall survival rates of oral squamous cell carcinoma treated with metronomic neoadjuvant chemotherapy [ Am J Cancer Res, 2024, 14(3):1033-1051] PubMed: 38590400
Porcupine inhibitors LGK-974 and ETC-159 inhibit Wnt/β-catenin signaling and result in inhibition of the fibrosis [ Toxicol In Vitro, 2024, 104:105986] PubMed: 39647516

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

NOT FOR HUMAN, VETERINARY DIAGNOSTIC OR THERAPEUTIC USE.