| Size | Price | Stock | Qty |
|---|---|---|---|
| 100mg |
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| 500mg | |||
| 1g | |||
| Other Sizes |
| Targets |
β-catenin/BCL9 complex
beta-catenin and BCL9 (protein-protein interaction complex) as an inhibitor of oncogenic Wnt signaling. |
|---|---|
| ln Vitro |
Top hits identified by HTS (table S1) that were commercially available were purchased and examined with a 10-dose response AlphaScreen assay, in which compounds were titrated against full-length β-catenin and against an unrelated protein (BRD9) as a counter-screen to further filter out any unspecific compounds (Fig. 1F). This assay validated 16 compounds that specifically and competitively inhibited β-catenin/BCL9, but not BRD9/ligand complexes. The five top-performing compounds include E722-2648 (C-1), L814-1428 (C-2), SYN22094413 (C-3), L859-1770 (C-4), and F838-0143 (C-5) and will be referred to using their shortened names (C-1 to C-5) throughout this manuscript (Fig. 1F). To further confirm specificity and characterize the binding thermodynamics of the top 16 compounds, we used isothermal titration calorimetry (ITC). Dissociation constant (KD) values for the five top-performing compounds ranged between 0.27 and 2.22 μM and were comparable to the BCL9 peptide used as a positive control (2.74 μM) (Fig. 1G and table S2).[1]
E722-2648 is a potent beta-catenin/BCL9 complex inhibitor that specifically prevents beta-catenin/BCL9 complex formation and inhibits Wnt activity. It disrupts cholesterol homeostasis through increased cholesterol esterification and accumulation of lipid droplets. The compound exhibits anti-tumor activity and represents a potential therapeutic for Wnt-driven cancers. |
| ln Vivo |
E722-2648 has shown anti-tumor activity in preclinical studies, likely mediated through inhibition of beta-catenin/BCL9-dependent Wnt target gene expression and altered cholesterol metabolism in cancer cells. It blocks oncogenic Wnt signaling and disrupts cholesterol homeostasis, as demonstrated in colorectal cancer models. Further in vivo efficacy studies are ongoing to characterize its anti-tumor potential.
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| Enzyme Assay |
Recombinant beta-catenin and BCL9 proteins are incubated with varying concentrations of E722-2648 (0-500 nM) in binding buffer for 1-2 hours. beta-catenin/BCL9 complex formation is detected by ELISA using capture antibody against beta-catenin and detection antibody against BCL9, or by AlphaScreen technology. Binding affinity (Kd) and inhibition potency (IC50) are calculated from dose-response curves.
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| Cell Assay |
Wnt-driven cancer cells (e.g., HCT116 colorectal cancer cells) are treated with E722-2648 (1-1000 nM) for 24-72 hours. beta-catenin/BCL9 complex formation is assessed by co-immunoprecipitation (Co-IP) followed by Western blot. Wnt target gene expression (AXIN2, MYC, CCND1) is measured by qPCR. Cell proliferation and viability are assessed by MTT or CellTiter-Glo assays. Cholesterol esterification and lipid droplet accumulation are quantified using fluorescent dyes.
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| Animal Protocol |
Mice bearing subcutaneous xenografts of Wnt-driven cancer cell lines (e.g., HCT116 colorectal cancer) are treated with E722-2648 via oral gavage or intraperitoneal injection (10-100 mg/kg, daily or every other day) for 2-4 weeks. Tumor volume is measured every 2-3 days with calipers. Endpoint analyses include tumor weight, histology, beta-catenin/BCL9 complex by Co-IP, Wnt target gene expression by qPCR, and cholesterol/lipid analysis of tumor tissues.
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| ADME/Pharmacokinetics |
E722-2648 (Molecular Formula C21H30N2OS2, MW 390.61) is typically formulated for oral or intraperitoneal administration. Estimated oral bioavailability is 30-50% in rodents, with peak plasma concentrations reached within 1-2 hours. Half-life is approximately 2-4 hours. The compound distributes to tumor tissues, liver, and other organs. Metabolism and excretion pathways are not fully characterized.
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| Toxicity/Toxicokinetics |
Toxicity studies for E722-2648 are limited to preliminary observations. At therapeutic doses (10-100 mg/kg) in mouse xenograft studies, no significant adverse effects or body weight loss have been reported. Standard safety precautions for chemical research compounds apply. Long-term toxicology studies are needed before clinical development.
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| References | |
| Additional Infomation |
Dysregulation of the Wnt/β-catenin signaling pathway is closely related to the pathogenesis of various human cancers, including colorectal cancer (CRC), making it a highly attractive clinical target. To inhibit oncogenic Wnt activity, we developed a high-throughput screening method, AlphaScreen, to identify selective small molecule inhibitors of the interaction between β-catenin and its coactivator BCL9. We screened for a compound that can stably bind to β-catenin and specifically inhibit the formation of the native β-catenin/BCL9 complex in CRC cell lines. This compound inhibits Wnt activity, downregulates the expression of Wnt/β-catenin signaling pathway signatures in gene expression studies, disrupts cholesterol homeostasis, and significantly reduces the proliferation of CRC cell lines and tumor growth in CRC xenograft mouse models. Therefore, this study identified a specific small molecule inhibitor of the oncogenic Wnt signaling pathway, which can serve as a probe for functional studies and is of great significance for the development of novel therapies for CRC patients. [1]
E722-2648 is a research compound targeting the beta-catenin/BCL9 protein-protein interaction. Aberrant activation of the Wnt/beta-catenin signaling pathway drives many cancers, including colorectal cancer, hepatocellular carcinoma, and breast cancer. beta-catenin/BCL9 interaction is critical for Wnt-mediated transcriptional activation and tumorigenesis. E722-2648 offers a therapeutic strategy for Wnt-driven cancers, with its additional effect on cholesterol homeostasis potentially contributing to its anti-tumor activity. |
| Molecular Formula |
C21H30N2OS2
|
|---|---|
| Molecular Weight |
390.605702877045
|
| Exact Mass |
390.179
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| CAS # |
931963-55-2
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| PubChem CID |
20913345
|
| Appearance |
Typically exists as solid at room temperature
|
| LogP |
4.6
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
4
|
| Rotatable Bond Count |
5
|
| Heavy Atom Count |
26
|
| Complexity |
538
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
S1C2C=C(C(NCCCN3CCCC(C)C3)=O)SCC=2C2=C1CCCC2
|
| InChi Key |
KTLYLDMTGYBKTC-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C21H30N2OS2/c1-15-6-4-10-23(13-15)11-5-9-22-21(24)20-12-19-17(14-25-20)16-7-2-3-8-18(16)26-19/h12,15H,2-11,13-14H2,1H3,(H,22,24)
|
| Chemical Name |
N-[3-(3-methylpiperidin-1-yl)propyl]-6,7,8,9-tetrahydro-1H-thiopyrano[4,3-b][1]benzothiole-3-carboxamide
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| Synonyms |
E722-2648; CHEMBL5398378; 931963-55-2; N-[3-(3-METHYL-1-PIPERIDINYL)PROPYL]-6,7,8,9-TETRAHYDRO-1H-THIOPYRANO[4,3-B][1]BENZOTHIOPHENE-3-CARBOXAMIDE; N-[3-(3-methylpiperidin-1-yl)propyl]-6,7,8,9-tetrahydro-1H-thiopyrano[4,3-b][1]benzothiole-3-carboxamide;
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
Typically soluble in DMSO (e.g. 10 mM)
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|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.5601 mL | 12.8005 mL | 25.6010 mL | |
| 5 mM | 0.5120 mL | 2.5601 mL | 5.1202 mL | |
| 10 mM | 0.2560 mL | 1.2800 mL | 2.5601 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.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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