IWP-2

Biological Activity

IWP-2 is an inhibitor of Wnt production. IWP-2 inactivates Porcupine, a membrane-bound O-acyltransferase responsible for palmitoylating Wnt proteins, which is essential for their signaling ability and secretion. IWP-2 selectively inhibits palmitoylation of Wnt, and blocks Wnt-dependent phosphorylation of Lrp6 receptor and Dvl2, and β-catenin accumulation. IWP-2 is useful in both suppressing embryonic stem cell self-renewal and decreasing cancer cell proliferation, migration and invasion.

Product Citations

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  Stem Cell Reports. 2021 Jun 8;16(6):1568-1583.

  Photobiomodulation therapy for hair regeneration: A synergetic activation of β-CATENIN in hair follicle stem cells by ROS and paracrine WNTs
  IWP-2 purchased from AbMole

• 

  Oncol Rep. 2020 Aug;44(2):577-588.

  TNF‑α promotes the malignant transformation of intestinal stem cells through the NF‑κB and Wnt/β‑catenin signaling pathways
  IWP-2 purchased from AbMole

Customer Product Validations & Biological Datas

	Source	ONCOLOGY REPORTS (2020 Aug). Figure 6. IWP-2 (Abmole Bioscience, US)
	Method	western blot
	Cell Lines	NCM460s cells
	Concentrations	1, 10, 100 µM
	Incubation Time	3 h
	Results	Notably, IWP-2 also counteracted TNF-α-induced activation of the NF-κB pathway in NCM460s cells, including increased phosphorylation of IKKα/β and IκBα, decreased protein levels of IκBα and nuclear translocation of p65

	Source	Front Immunol (2018). Figure 6. IWP-2
	Method	injected intravenously
	Cell Lines	C57BL/6 mice
	Concentrations	20 mg/kg
	Incubation Time	4.5 h
	Results	This view is supported by reduced percentages of IFN-γ+ liver CD3+NK1.1+ cells in mice treated with IWP-2, a small molecule inhibitor of the acyltransferase Porcupine, which plays a central role in Wnt secretion

Protocol

Cell Experiment
Cell lines	MKN28 cell line
Preparation method	Cell proliferation assay CellTiter 96 Aqueous Non-Radioactive Cell Proliferation Assay kit (Promega Corporation, Madison, WI, USA) was used. Transwell migration and invasion assay Haptotaxis chambers (8-μm pore size, Corning Costar, Cambridge, MA, USA) without or with 5 mg/ml matrigel (Sigma Aldrich) were used for migration and invasion assays, respectively. The experiments were repeated three times independently.
Concentrations	0,5,10,20,50µM
Incubation time	5 days

Animal Experiment
Animal models	six month-old zebrafish model
Formulation	0.1% DMSO
Dosages	10 μM
Administration	in aquarium water supplemented with 10 μM IWR-1 or in 0.1% DMSO as a control

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

Species	Mouse	Rat	Rabbit	Guinea pig	Hamster	Dog
Weight (kg)	0.02	0.15	1.8	0.4	0.08	10
Body Surface Area (m2)	0.007	0.025	0.15	0.05	0.02	0.5
Km factor	3	6	12	8	5	20

Animal A (mg/kg) = Animal B (mg/kg) multiplied by	Animal B Km
	Animal A Km

For example, to modify the dose of Compound A used for a mouse (20 mg/kg) to a dose based on the BSA for a rat, multiply 20 mg/kg by the K_m factor for a mouse and then divide by the K_m factor for a rat. This calculation results in a rat equivalent dose for Compound A of 10 mg/kg.

Chemical Information

References

Discovery of Inhibitor of Wnt Production 2 (IWP-2) and Related Compounds As Selective ATP-Competitive Inhibitors of Casein Kinase 1 (CK1) δ/ε
Balbina Garcia-Reyes, et al. J Med Chem. 2018 May 10;61(9):4087-4102. PMID: 29630366.

The Wingless homolog Wnt5a stimulates phagocytosis but not bacterial killing
George Maiti, et al. Proc Natl Acad Sci U S A. 2012 Oct 9;109(41):16600-5. PMID: 23012420.