WAY 316606

SKU: orb1302834

Description

WAY 316606 is a small molecule inhibitor of secreted frizzled-related protein 1 (sFRP-1), which antagonizes Wnt signaling. This compound is used in research to probe Wnt pathway biology in both cellular and animal models, particularly in studies of bone formation and osteoporosis.

Research Area

Signal Transduction, Stem Cell & Developmental Biology

Key Properties

• CAS Number: 915759-45-4
• MW: 448.48
• Purity: >99.99% (May vary between batches)
• Formula: C₁₈H₁₉F₃N₂O₄S₂
• SMILES: FC(F)(F)c1ccc(cc1S(=O)(=O)NC1CCNCC1)S(=O)(=O)c1ccccc1
• Target: Wnt/beta-catenin
• Solubility: DMSO:50 mg/mL (111.49 mM)

Bioactivity

• Target IC50:
  SFRP1:0.5 μM
• In Vivo:
  WAY-316606 effectively promotes bone formation, as demonstrated in neonatal murine calvarial assays, with the capability to increase total bone area by up to 60% in a dose-dependent manner, achieving an EC50 of approximately 1 nM. This compound exhibits favorable aqueous solubility, displays moderate to low inhibition of cytochrome P450 isozymes (3A4, 2D6, 2C9), and maintains good stability in both rat and human liver microsomes (t1/2>60 min for each species). Moreover, in female Sprague-Dawley rats, WAY-316606 shows high plasma clearance (77 mL/min/kg, surpassing hepatic blood flow) following a single intravenous bolus dose (2 mg/kg), leading to a swift reduction in plasma drug levels regardless of administration route .
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• In Vitro:
  The EC50 of WAY-316606 for Wnt-Luciferase Activity from U2-OS Cells is 0.65 μM. WAY-316606 binds to the secreted frizzled-related protein (sFRP)-1 inhibitor with a KD of 0.08 μM and inhibits sFRP-1 with an EC50 of 0.65 μM. WAY-316606 also binds to sFRP-2, albeit over 10 times weaker with a KD of 1 μM. Using a fluorescence polarization binding assay that employs a fluorescent probe compound and purified human sFRP-1 protein in a competitive-binding format, the IC50 for WAY-316606 is 0.5 μM .
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• Cell Research:
  U2OS bone cells are infected with recombinant adenovirus 5 (Ad5) WNT3 at a multiplicity of infection (MOI) of 2, followed by infection with Ad5-sFRP-1 and Ad5-16xTCF-luciferase, each at an MOI of 10. Four hours after infection, the cells are frozen in sterile cryogenic vials at a cell density of 9×106 cells/mL and stored in a 150°C freezer. For the assay, a vial of frozen cells is thawed, and the cells are resuspended in plating medium [phenol red-free RPMI 1640 medium containing 5% fetal calf serum, 2 mM GlutaMAX-l, and 1% (v/v) penicillin-streptomycin] to a final cell density of 1.5×105 cells/mL. The resuspended cells are then plated in 96-well tissue culture treated plates at a volume of 100 μL of cell suspension/well (i.e., 1.5×104 cells/well). The plates are incubated at 37°C inside a 5% CO2/ 95% humidified air incubator for 5 h or until the cells have attached and started to spread. Prior to the addition of WAY-316606, the medium is replaced with 50 μL/well of phenol red-free RPMI 1640 containing 10% fetal calf serum, 2 mM GlutaMAX-l, and 1% (v/v) penicillin-streptomycin. WAY-316606, or vehicle (typically DMSO), diluted in phenol red-free RPMI 1640 containing 2 mM GlutaMAX-l, and l % (v/v) penicillin-streptomycin are then added to the wells in replicates of 4 wells/dilution and the plates are incubated at 37°C overnight. Dose response experiments are performed with the compounds in 2-fold serial dilutions from 10000 4.9 nM. After the overnight incubation, the cells are washed twice with 150 uL/well of PBS w/o calcium or magnesium and lysed with 50 μL/well of 1× cell culture lysis reagent on a shaker at room temperature for 30 min. Aliquots of the cell lysates (30 μL) are transferred to 96-well luminometer plates, and the luciferase activity is measured in a MicroLumat PLUS luminometer using 100 μL/well of a luciferase substrate.
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Storage & Handling

• Storage: -20°C
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Alternative Names

βcatenin, SARP-2, Secreted frizzled related protein 1, sFRP-1, WAY316606, WAY-316606, WAY 316606, Wnt, Wnt/b-catenin, Wnt/betacatenin, Wnt/β-catenin, bcatenin, betacatenin, beta-catenin, FrzA, Inhibitor, inhibit

Compound Identifiers

PubChem CID

16727102