1. Stem Cell/Wnt
  2. Wnt

IWP-2 

Cat. No.: HY-13912 Purity: 97.40%
Handling Instructions

IWP-2 is an inhibitor of Wnt processing and secretion with IC50 of 27 nM.

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

IWP-2 Chemical Structure

IWP-2 Chemical Structure

CAS No. : 686770-61-6

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10 mg USD 102 In-stock
Stock in Sweden
Estimated Time of Arrival: December 31
50 mg USD 348 In-stock
Stock in Sweden
Estimated Time of Arrival: December 31
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  • Biological Activity

  • Protocol

  • Technical Information

  • Purity & Documentation

  • References

Description

IWP-2 is an inhibitor of Wnt processing and secretion with IC50 of 27 nM.

IC50 & Target

IC50: 27 nM (Wnt)[1]

In Vitro

IWP-2, an inhibitor of WNT processing and secretion. IWP-2 significantly enhances the anti-proliferative effect of LEF. It is also obvious that the combination of LEF and IWP-2 could minimize the expression of β-catenin, c-Myc, Cyclin D1, Bcl2 and Bax to the largest extent compared with single agents[2]. Following treatment in the MKN28 cell line for four days, 10-50 μM IWP-2 significantly suppressed the proliferation of MKN28 cells (P<0.05). In addition, anchor-dependent and anchor-independent colony numbers are significantly decreased following IWP-2 treatment (P<0.05)[3].

In Vivo

To evaluate the efficacy of IWP-2 in vivo, 200 μL each of IWP-2-liposome or free liposome i separately injected into C57BL/6 mice intraperitoneally about 2 h before injection of a similar volume of either blue-dye-filled latex beads or E. coli DH5α. IWP-2 causes significant reduction in the uptake of blue beads as well as E. coli as assessed by CFUs in peritoneal lavage cells within 2 h. In addition, the levels of TNF-α and IL-6 in the lavage fluid of the corresponding mice are reduced by 2-4-fold compared with control values. Interestingly, IWP-2 even induces a considerable increase in secretion of the anti-inflammatory cytokine IL-10[4]. Pretreatment with IWP-2 significantly (P<0.05) abolished SP-induced increase of Wnt3a, p-GSK3β, and β-catenin expressions[5].

References
Preparing Stock Solutions
Concentration Volume Mass 1 mg 5 mg 10 mg
1 mM 2.1432 mL 10.7158 mL 21.4316 mL
5 mM 0.4286 mL 2.1432 mL 4.2863 mL
10 mM 0.2143 mL 1.0716 mL 2.1432 mL
Please refer to the solubility information to select the appropriate solvent.
Cell Assay
[2]

IWP-2 is prepared in DMSO and stored, and then diluted with appropriate medium before use[2].

The human RCC cell lines 786O and Caki-2 (5×103) are seeded into 96-well plates. Cell viability is estimated by MST assay after Caki-2 acells are incubated with ncreasing concentrations of LEF together with 20 μM IWP-2 for 48 h.After treatment, 10 μL MTS is added into each well for 2 h incubation. The absorbance is measured using a model ELX800 Micro Plate Reader at 490 nm. For colony formation assay, Caki-2 cells are trypsinized to single cell suspensions and seeded into fresh 6-well plates at 1000 cells/well. Then cells are incubated with LEF at depicted concentrations for 7 days. Colonies are fixed with absolute methanol for 15 min and then stained with 0.1% crystal violet for 20 min. After washing with PBS three times, the colonies with a diameter over 2 mm are visualized by a digital camera[2]. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Animal Administration
[4][5]

Liposome-IWP2 is prepared with L-α-phosphatidylcholine, octadecylamine, and IWP-2 in a 20:2:0.1 Ratsio (Mice)[4].
IWP-2 is prepared in DMSO and diluted with saline or PBS (Rats)[5].

Mice[4]
About 3-mo-old C57BL/6 mice are housed four to five in a cage at 23°C in a 12-h light/dark cycle. Mice are injected intraperitoneally (i.p.) first with either 200 μL of liposome-IWP2 (LI) or liposome (L) and then after 2 h with 1×108 or 2×108 CFU E. coli in 200 μL of sterile PBS. After 2 h or 24 h mice are killed, and the peritoneal cavity is washed with 5 mL of sterile ice-cold PBS. The peritoneal lavage fluid is centrifuged at 300× g for 5 min, the cell pellet is resuspended in RPMI 1640 complete medium, and the supernatant is used for cytokine assay. For ex vivo experiments, peritoneal phagocytes are isolated as above from normal mice, and equal numbers of cells are plated in medium overnight at 37°C in 5% CO2 before performing further experiments.
Rats[5]
Adult, male, and healthy Wistar rats weighing 220-280 g are used. Rats are randomly divided into 6 groups as follows (n=72, 12 per group): (1) Sham group (Group S), (2) I/R group (Group I/R), (3) I/R+DMSO group (Group DMSO), (4) I/R+IWP group (Group IWP), (5) SP group (Group SP), and (6) SP+Wnt inhibitor IWP-2 group (Group SP+IWP). The hearts are continuously perfused for 120 min in Group S. After 10 min of equilibration, the isolated hearts are continuously perfused for 20 min, then subjected to 30 min of ischemia followed by 60 min of reperfusion in Group I/R; Groups DMSO, IWP, SP and SP+IWP receive 15 min of perfusion with K-H solution containing 0.5 mL/L DMSO, 10 µM IWP (SIGMA-ALDRICH, USA), 2.4 vol% Sevoflurane, 2.4 vol% Sevoflurane+10 µM IWP, respectively, followed by 5 min washout before I/R. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

References
Molecular Weight

466.6

Formula

C₂₂H₁₈N₄O₂S₃

CAS No.

686770-61-6

SMILES

O=C(NC1=NC2=CC=C(C)C=C2S1)CSC3=NC(CCS4)=C4C(N3C5=CC=CC=C5)=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

Solvent & Solubility

DMSO: < 4.7 mg/mL

* "<1 mg/mL" means slightly soluble or insoluble. "≥" means soluble, but saturation unknown.

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IWP-2
Cat. No.:
HY-13912
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