Stemolecule™ Wnt Inhibitor IWP-3

Stemolecule Wnt Inhibitor IWP-3 was identified in a high throughput screen for antagonists of the Wnt/β-catenin pathway. Wnt Inhibitor IWP-3 prevents palmitylation of Wnt proteins by Porcupine (Porcn), a membrane-bound O-acyltransferase, thereby blocking Wnt secretion and activity. It also blocks phosphorylation of the Lrp6 receptor and accumulation of both Dvl2 and β-catenin¹. The inhibition of the Wnt pathway through the use of IWP-3 has also been shown to promote the formation of cardiomyocytes from human embryonic stem cell-derived mesoderm cells².

Stemgent and the Stemolecule brand name are trademarks of REPROCELL Inc., Japan.

Product Name: Stemolecule Wnt Inhibitor IWP-3

Catalog Number: 04-0035

Size: 2 mg

Alternate Name(s): 2-(3-(4-fluorophenyl)-3,4,6,7-tetrahydro-4-oxothieno[3,2-d]pyrimidin-2-ylthio)-N-(6-methylbenzo[d]thiazol-2-yl)acetamide

Chemical Formula: C₂₂H₁₇FN₄O₂S₃

Molecular Weight: 484.59

CAS Number: 687561-60-0

Purity: Greater than 99 % by HPLC analysis

Formulation: Light yellow solid

Solubility: This molecule is soluble in DMSO at 100 mM.

Reconstitution: For a 10 mM concentrated stock solution of Wnt Inhibitor IWP-3, reconstitute the compound by adding 412.7 µL of DMSO to the entire contents of the vial. If precipitate is observed, warm the solution to 37 °C for 2 to 5 minutes. For cell culture, prewarm the medium prior to adding reconstituted compound. Note: For most cell culture applications, the total DMSO concentration should be kept below 0.5%.

Storage and Stability: Store powder at 4 °C protected from light. Information about the stability of Stemolecules in solution is largely not available. As a general guideline, we recommend that stock solution be freshly made and stored in aliquots at −20 °C, protected from light. The effect of storage of stock solutions should be verified for each application.

Quality Control: The purity of Wnt Inhibitor IWP-3 was determined by HPLC analysis. The accurate mass was determined by mass spectrometry. Cellular toxicity of Wnt Inhibitor IWP-3 was tested on mouse embryonic stem cells.

Specification Sheets:

• 04-0035 Specifications Sheet

Safety Data Sheets:

• 04-0035 Safety Data Sheet

1. Chen, B., Dodge, M.E., Tang, W., Lu, J., Ma, Z., Fan, C.W., Wei, S., Hao, W., Kilgore, J., Williams, N.S., Roth, M.G., Amatruda, J.F., Chen, C., and Lum, L. Small molecule-mediated disruption of Wnt-dependent signaling in tissue regeneration and cancer. Nat Chem Biol 5: 100-107 (2009).
2. Willems, E., Spiering, S., Davidovics, H., Lanier, M., Xia, Z., Dawson, M., Cashman, J., and Mercola, M. Small-molecule inhibitors of the Wnt pathway potently promote cardiomyocytes from human embryonic stem cell-derived mesoderm. Circ Res 109: 360-364 (2011).

Additional Publications

• Kitano Y; Nishimura S; Kato TM; Ueda A; Takigawa K; Umekaga M; Nomura M; Kawakami A; Ogawa H; Xu H; Hotta A; Takasu N; Tsukahara M. Generation of hypoimmunogenic induced pluripotent stem cells by CRISPR-Cas9 system and detailed evaluation for clinical application. Molecular Therap: Methods Clinical Develop 26:15 (2022).
• Tsujisaka T; Hatani T; Okubo C; Ito R; Kimura A; Narita M; Chonabayashi K ;Funakoshi S; Lucena-Cacace A; Toyoda T; Osafune K; Kimura T; Saito H; Yoshida Y.. Purification of human iPSC-derived cells at large scale using microRNA switch and magnetic-activated cell sorting. Stem Cell Rep 17:1772 (2022).
• Okamoto R; Goto I; Nishimura Y; Kobayashi I; Hashizume R; Yoshida Y; Ito R; Kobayashi Y; Nishikawa M; Ali Y; Saito S; Tanaka T; Sawa Y; Ito M; Dohi K. Gap junction protein beta 4 plays an important role in cardiac function in humans, rodents, and zebrafish. PLos One :240129 (2020).
• Matsura K; Wada M; Sakaguchi K; Matsuhashi Y; Shimizu T. Adequate taylor couette flow-mediated shear stress is useful for dissociating human iPS cell-derived cell aggregates. Regen Therap in press:https://doi.org/10.1016/j.reth.2019.04.006 (2019).
• Ito E; Miyagawa S; Takeda A; Kawamura A; Harada A; Iseoka H; Yajima S; Sougawa N; Michizuki-Oda N; Yasuda S; Sato Y; Sawa Y. Tumorigenicity assay essential for facilitating safety studies of hiPSC-derived cardiomyocytes for clinical application. Sci Rep 9:1881 (2019).
• Lee H; Haller C; Manneville C; Doll T; Fruh I; Gubser Keller C; Richards SM; Ibig-Rehm Y; Patoor M; Goette M; Bouchez LC; Mueller M. Identification of Small Molecules Which Induce Skeletal Muscle Differentiation in Embryonic Stem Cells via Activation of the Wnt and Inhibition of Smad2/3 and Sonic Hedgehog Pathways. Stem Cells 34:299-310 (2016).
• Funakoshi S; Miki K; Takaki T; Okubo C; hatani T; Chonabayashi K; Nishikawa M; Takei I; Oishi A; Narita M; Hoshijima M; Kimura T; Yamanaka S; Yoshida Y. Enhanced engraftment, proliferation, and therapeutic potential in heart using optimized human iPSC-derived cardiomyocytes. Sci Rep 6:19111 (2016).
• Cheesboro J. Mechanism of segmentation in the American cockroach, Perplaneta americana. Ph.D. Thesis, University of Sussex (2012).