IWP-O1 is a novel and potent porcn (acyltransferase Porcupine) inhibitor with an EC50 value of 80 pM in a cultured cell reporter assay of Wnt signaling. It effectively suppresses the phosphorylation of Dvl2/3 in HeLa cells. The acyltransferase Porcupine (Porcn) is essential for the secretion of Wnt proteins which contribute to embryonic development, tissue regeneration, and tumorigenesis. Additionally, IWP-O1 has significantly improved metabolic stability over our previously reported Porcn inhibitors.

Physicochemical Properties

Molecular Formula	C26H20N6O
Molecular Weight	432.47660446167
CAS #	2074607-48-8
Related CAS #	2074607-48-8
PubChem CID	134154594
Appearance	Light yellow to yellow solid powder
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	6
Heavy Atom Count	33
Complexity	614
Defined Atom Stereocenter Count	0
SMILES	O=C(CN1C(C2C=CC=CC=2)=C(C2C=CN=CC=2)N=N1)NC1C=CC(=CN=1)C1C=CC=CC=1
Chemical Name	5-Phenyl-N-(5-phenylpyridin-2-yl)-4-(pyridin-4-yl)-1H-1,2,3-triazol-1-acetamide
Synonyms	IWP-O1; IWPO1; IWP O1
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 Note: This product requires protection from light (avoid light exposure) during transportation and storage.
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

Targets	IWP-O1 (17) is 2.5 times more active than the experimental medication LGK974 and suppresses Wnt signaling in L-Wnt-STF cells with an EC50 value of 80 pM [1].
ln Vitro	IWP-O1 (17) is 2.5 times more active than the experimental medication LGK974 and suppresses Wnt signaling in L-Wnt-STF cells with an EC50 value of 80 pM [1]. IWP-O1 inhibited Wnt signaling in L-Wnt-STF cells with an EC₅₀ of 80 pM, which is 2.5 times more potent than the investigational Porcn inhibitor LGK974 (EC₅₀ = 0.2 nM)[1] IWP-O1 suppressed phosphorylation of Dishevelled (Dvl2/3) and low-density lipoprotein receptor-related protein 6 (LRP6) in HeLa cells, indicating inhibition of both β-catenin-dependent and -independent Wnt signaling pathways[1]
Cell Assay	Wnt signaling inhibition was assessed using L-Wnt-STF cells, which stably express a Wnt-responsive luciferase reporter. Cells were treated with compounds, and luciferase activity was measured to determine EC₅₀ values[1] Western blot analysis was performed in HeLa cells treated with IWP-O1 or LGK974 for 24 hours. Cell lysates were probed with antibodies against phosphorylated Dvl2/3, total Dvl2/3, phosphorylated LRP6, total LRP6, and actin as a loading control[1]
ADME/Pharmacokinetics	IWP-O1 exhibited significantly improved metabolic stability compared to earlier Porcn inhibitors. In murine liver S9 fractions, IWP-O1 remained stable with no degradation observed after 4 hours of incubation[1] In murine plasma, IWP-O1 had a half-life of 130 minutes[1]
References	[1]. Development of a triazole class of highly potent Porcn inhibitors. Bioorg Med Chem Lett. 2016 Dec 15;26(24):5891-5895.
Additional Infomation	IWP-O1 belongs to a triazole class of Porcn inhibitors designed based on a general scaffold identified from four earlier inhibitor classes[1] The compound functions by preventing the secretion of Wnt proteins, thereby inhibiting Wnt signaling pathways involved in embryogenesis, tissue homeostasis, and tumorigenesis[1] IWP-O1 features a 1,2,3-triazole core, which contributes to its high metabolic stability, making it suitable for in vivo model studies in mice[1]

Solubility Data

Solubility (In Vitro)

DMSO:≥ mg/mL Water:N/A Ethanol:N/A

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (4.81 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (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 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.08 mg/mL (4.81 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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.3122 mL	11.5612 mL	23.1225 mL
	5 mM	0.4624 mL	2.3122 mL	4.6245 mL
	10 mM	0.2312 mL	1.1561 mL	2.3122 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.