1. Stem Cell/Wnt
  2. sFRP-1

WAY 316606 

Cat. No.: HY-10858 Purity: 99.61%
Handling Instructions

WAY 316606 is an inhibitor of the secreted protein sFRP-1, an endogenous antagonist of the secreted glycoprotein Wnt. The affinity of WAY-316606 for sFRP-1 is determined using the FP binding assay with IC50 of 0.5 μM.

For research use only. We do not sell to patients.

WAY 316606 Chemical Structure

WAY 316606 Chemical Structure

CAS No. : 915759-45-4

Size Price Stock Quantity
10 mM * 1 mL in DMSO USD 404 In-stock
Estimated Time of Arrival: December 31
5 mg USD 367 In-stock
Estimated Time of Arrival: December 31
10 mg USD 552 In-stock
Estimated Time of Arrival: December 31
50 mg USD 1234 In-stock
Estimated Time of Arrival: December 31
100 mg USD 1974 In-stock
Estimated Time of Arrival: December 31
200 mg   Get quote  
500 mg   Get quote  

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Customer Review

    WAY 316606 purchased from MCE. Usage Cited in: Int J Dev Neurosci. 2018 May;66:24-32. 

    Whole-mount anti-5-HT (black) immunostaining of embryos treated with DMSO, WAY-316606, Sfrp1 protein, Sfrp1 plus WAY-316606, or Shh in utero electroporation together with WAY-316606 injection. Black arrows indicate misdirected axons.

    WAY 316606 purchased from MCE. Usage Cited in: Int J Dev Neurosci. 2018 May;66:24-32. 

    Quantification of the percentage of normal and misdirected 5-HT axons for the different groups.
    • Biological Activity

    • Protocol

    • Technical Information

    • Purity & Documentation

    • References

    Description

    WAY 316606 is an inhibitor of the secreted protein sFRP-1, an endogenous antagonist of the secreted glycoprotein Wnt. The affinity of WAY-316606 for sFRP-1 is determined using the FP binding assay with IC50 of 0.5 μM.

    IC50 & Target

    IC50: 0.5 μM (sFRP-1)[1]

    In Vitro

    The EC50 of WAY-316606 for Wnt-Luciferase Activity from U2-OS Cells is 0.65 μM[1]. WAY-316606 binds to 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[2].

    In Vivo

    WAY-316606 increases bone formation when tested in a neonatal murine calvarial assay. WAY-316606 increases total bone area up to 60% in a dose-dependent manner with an EC50 of about 1 nM. WAY-316606 has good aqueous solubility, moderate to low inhibition of cytochrome p450 isozymes (3A4, 2D6, 2C9) and good stability in rat and human liver microsomes (t1/2>60 min in each species). In female Sprague-Dawley rats, WAY-316606 exhibits high plasma clearance (77 mL/min/kg, greater than hepatic blood flow) following a single intravenous bolus dose (2 mg/kg), which results in a rapid decline of drug exposure in the plasma despite the route of administration[2].

    Solvent & Solubility
    In Vitro: 

    10 mM in DMSO

    Preparing
    Stock Solutions
    Concentration Solvent Mass 1 mg 5 mg 10 mg
    1 mM 2.2298 mL 11.1488 mL 22.2975 mL
    5 mM 0.4460 mL 2.2298 mL 4.4595 mL
    10 mM 0.2230 mL 1.1149 mL 2.2298 mL
    *Please refer to the solubility information to select the appropriate solvent.
    References
    Kinase Assay
    [2]

    WAY-316606 binding to purified sFRP is determined by spectroscopy methods. The sFRP-1 or -2 stock solutions are diluted to 1 μM in a buffered solution and the initial fluorescence is measured. Increasing concentrations of WAY-316606 (0 to 50 μM) are added to the protein in the cuvette and incubated for 5 min prior to assessing fluorescence intensity using a Fluoromax-2 fluorometer. In control experiments, the DMSO (vehicle control)-matched buffer solution is used. Fluorescence spectra are scanned in the ratio mode (S/R, signal/reference) to compensate for variations in lamp output as a function of wavelength. Fluorescence changes are fitted to a quadratic equation to obtain apparent dissociation constants[2].

    MCE has not independently confirmed the accuracy of these methods. They are for reference only.

    Cell Assay
    [1]

    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 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. WAY-316606, or vehicle (typically DMSO), diluted in phenol red-free RPMI 1640 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 luciferase substrate.

    MCE has not independently confirmed the accuracy of these methods. They are for reference only.

    References
    Molecular Weight

    448.48

    Formula

    C₁₈H₁₉F₃N₂O₄S₂

    CAS No.

    915759-45-4

    SMILES

    O=S(C1=CC(S(=O)(C2=CC=CC=C2)=O)=CC=C1C(F)(F)F)(NC3CCNCC3)=O

    Storage
    Powder -20°C 3 years
      4°C 2 years
    In solvent -80°C 6 months
      -20°C 1 month
    Shipping

    Room temperature in continental US; may vary elsewhere

    Purity: 99.61%

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    Product Name:
    WAY 316606
    Cat. No.:
    HY-10858
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    WAY 316606

    Cat. No.: HY-10858