| Size | Price | Stock | Qty |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| 500mg |
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| 1g |
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| Other Sizes |
Purity: ≥98%
| Targets |
Wnt/β-catenin
MSAB targets β-catenin, a key transcriptional coactivator in the Wnt signaling pathway. Wnt signaling is critical for embryonic development, cell proliferation, and differentiation. In many cancers, this pathway is constitutively activated, leading to the accumulation of β-catenin in the nucleus and the transcription of oncogenes. MSAB binds directly to β-catenin and promotes its degradation, thereby reducing its nuclear levels and inhibiting the transcription of Wnt target genes. |
|---|---|
| ln Vitro |
MSAB (2-10 μM) specifically lowers Wnt-dependent cells' survival while having minimal effect on Wnt-independent cells and healthy human cells [1]. T cell factor (TCF) luciferase reporter activity in HCT116 cells is inhibited by MSAB (0.01-10 μM; 20 hours) [1]. In HEK293T cells, Wnt3a-induced elevations in TOP-Luc activation and active β-catenin levels are inhibited by MSAB (20 hours) [1]. In HCT116 cells, MSAB (0.5-10 μM; 20 hours) lowers the levels of endogenous Wnt target gene mRNA and protein [1]. Protease-dependent MSAB (5 μM) stimulates β-catenin degradation in HCT116 cells for 16 hours [1].
In vitro, MSAB acts as a selective inhibitor of Wnt/β-catenin signaling. It binds to β-catenin, promoting its degradation and specifically downregulating Wnt/β-catenin target genes. This activity has been demonstrated in various cell lines. The compound shows potent anti-tumor effects against Wnt-dependent cancer cells. Its selectivity for the Wnt pathway makes it a valuable tool for studying the role of this signaling cascade in cancer and other diseases. |
| ln Vivo |
In a mouse xenograft model, MSAB (10–20 mg/kg; daily intraperitoneal injection for 2 weeks) reduces the formation of tumors in Wnt-dependent cancer cells [1]. In MMTV-Wnt1 transgenic mice, MSAB (10–20 mg/kg; intraperitoneally given twice daily for two weeks) suppresses tumor growth [1].
In vivo, MSAB has shown potent anti-tumor activity in mouse models. In xenograft mouse models and MMTV-Wnt1 transgenic mice, MSAB (10-20 mg/kg; i.p. daily for 2 weeks) inhibits the growth of Wnt-dependent cancer cells. These findings confirm that targeting β-catenin for degradation is an effective strategy for inhibiting Wnt-driven tumors. The compound's in vivo efficacy highlights its potential as a therapeutic agent for cancers with aberrant Wnt signaling. |
| Enzyme Assay |
High throughput chemical screening was performed as previously described (Raj et al., 2011; Stanton et al., 2009) with modifications. For screening of inhibitors targeting Wnt/β-catenin signaling pathway, HCT116-TOP cells (HCT116 cells stably transfected with TOPFLASH (TCF/LEF1-optimized promoter)-firefly luciferase reporter) were seeded using automated plate filler in 384-well plates and incubated at 37°C overnight. Next day, each small molecule compound was pin-transferred to each plate. Plates were covered with lids and incubated at 37°C for 20 h. The following day, assa y plates were allowed to equilibrate to room temperature for 10 min. Luciferase assay reagent was added to each well, incubated for 15 min, then read using a Perkin Elmer Envision luminometer to quantitate luciferase levels. Compounds that decrease luciferase activity were selected as hits. Luciferase values were normalized to the positive and negative controls to determine a normalized percent inhibition. Based on the normalized luciferase values, compounds showing >50% inhibition in both replicates compared to DMSO control were considered as active compounds. All small molecules were tested in duplicates[1].
In vitro assays for MSAB typically involve studying its effects on β-catenin and Wnt signaling. The compound's ability to bind to β-catenin and promote its degradation can be assessed by Western blotting, where a decrease in β-catenin protein levels is observed. The effect on Wnt signaling is measured using reporter gene assays, such as the TCF/LEF luciferase reporter, which quantifies the transcriptional activity of β-catenin. The expression of Wnt target genes (e.g., AXIN2, MYC, CCND1) is analyzed by RT-PCR. |
| Cell Assay |
Cell viability assay [1]
Cell viability was assayed by Sulforhodamine B based In Vitro Toxicology Assay Kit. Cells were plated in 6-well plates, and after reaching 60-70% confluency, the cells were treated with chemicals at concentrations and durations as indicated in the figures and figure legends. Staining and quantitative analysis were performed according to the manufacturer’s manual. All experiments were performed as duplicates. Luciferase reporter assays[1] To measure transcriptional activity of Wnt, NF-κB, iNOS, or NOTCH, we transiently transfected HCT116 cells or HEK293T cells with TOP-FLASH, FOP-FLASH, NF-κB, iNOS, or NOTCH luciferase reporter, along with an internal Renilla luciferase reporter plasmid as a control (hRL-null). Transfection was performed with lipofectamine 2000 according to the manufacturer’s protocol. Luciferase activity was measured with the Dual Luciferase Reporter Assay System as according to the manufacturer’s manual. The results were normalized to the control Renilla activity. The reported data represent the average of three independent experiments. For cell-based assays, Wnt-dependent cancer cell lines are cultured in standard media. Cells are seeded in multi-well plates and treated with MSAB at various concentrations for a defined period. Following treatment, the effects on β-catenin levels and Wnt signaling are assessed using the methods described above. Cell viability and proliferation are measured using standard assays like MTT or CellTiter-Glo. The effects on cell cycle and apoptosis can also be analyzed. All experiments include appropriate vehicle controls and are performed in triplicate. |
| Animal Protocol |
Animal/Disease Models: Athymic nude mice (5-6 weeks) injected with HCT116, HT115, H23 or H460 cells [1]
Doses: 10, 20 mg/kg Route of Administration: Daily intraperitoneal (ip) injection for 2 weeks Experimental Results: Various mice The size and weight are diminished in Wnt-dependent manner in HCT116, HT115, and H23 tumor types. For xenograft tumor models, cancer cell line HCT116, HT115, H23, or H460 (2x 106) was injected subcutaneously (s.c.) into the flanks of athymic nude mice (NCr nude, 5-6 week old). We also used MMTV-Wnt1 transgenic mice. About 15% of these mice develop mammary tumors between 6 weeks and 3 months of age in MMTV-Wnt1 transgenic mice model. Tumors were allowed to grow to 40 mm3 prior to intraperitoneal injection. One week after cellular inoculation, mice were treated by intraperitoneal injection with vehicle or MSAB (10 or 20 mg/kg) every day for two weeks. Two weeks after intraperitoneal injection, the mice were euthanized and tumor weight was measured. Tumor dimensions were measured, and volume was calculated by length (L) and width (W) using the formula (volume = π/6 x L x W2).[1] In vivo animal studies with MSAB are typically conducted in mouse models of Wnt-dependent cancers. The compound is formulated in a suitable vehicle, such as 10% DMSO, 40% PEG300, 5% Tween 80, and 45% saline. It is administered via intraperitoneal (IP) injection at doses of 10-20 mg/kg daily. Endpoints for efficacy studies include tumor volume measurement, analysis of β-catenin levels and Wnt target gene expression in tumor tissue, and assessment of tumor cell proliferation and apoptosis. All procedures must be conducted in accordance with institutional guidelines. |
| ADME/Pharmacokinetics |
Pharmacokinetic data for MSAB are not extensively reported in public literature. The compound has a molecular weight of 305.35 and is soluble in DMSO. It is typically stored as a powder at -20°C. For in vivo administration, it is formulated in suitable vehicles. A comprehensive ADME profile is essential for interpreting its in vivo efficacy.
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| Toxicity/Toxicokinetics |
Toxicological data for MSAB are primarily derived from in vivo efficacy studies, where it was tolerated at the tested doses. The compound is for research use only and is not for human consumption. Potential toxicity could arise from on-target effects on Wnt signaling, which is important for normal tissue homeostasis. Comprehensive toxicology studies would be required for any future therapeutic development.
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| References | |
| Additional Infomation |
The Wnt/β-catenin signaling pathway plays an important role in tissue homeostasis, and its dysregulation can lead to a variety of human diseases. Abnormal activation of β-catenin is oncogenic and is a key driver in the development and progression of human cancer. Although targeting the oncogenic β-catenin pathway has great potential in cancer treatment, the development of specific inhibitors is still insufficient. We screened small molecule compounds that act on the Wnt/β-catenin signaling pathway using a T-cytokine (TCF)-dependent luciferase reporter system and identified MSAB (methyl 3-{[(4-methylphenyl)sulfonyl]amino}benzoate) as a selective inhibitor of the Wnt/β-catenin signaling pathway. MSAB showed potent antitumor activity against Wnt-dependent cancer cells in vitro and in murine cancer models. MSAB binds to β-catenin, promotes its degradation, and specifically downregulates the expression of Wnt/β-catenin target genes. Our findings may represent an effective therapeutic strategy for cancers dependent on the Wnt/β-catenin signaling pathway. [1]
MSAB is a research-grade chemical probe, and its primary application is in the study of the Wnt/β-catenin signaling pathway. Its ability to bind to β-catenin and promote its degradation makes it a valuable tool for validating this pathway as a therapeutic target in cancer. The compound is not an approved drug and has not entered clinical trials. It is commercially available from various chemical suppliers for research purposes only. |
| Molecular Formula |
C15H15NO4S
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|---|---|
| Molecular Weight |
305.3489
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| Exact Mass |
305.072
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| Elemental Analysis |
C, 59.00; H, 4.95; N, 4.59; O, 20.96; S, 10.50
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| CAS # |
173436-66-3
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| PubChem CID |
1159052
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| Appearance |
Typically exists as white to off-white solids at room temperature
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| Density |
1.3±0.1 g/cm3
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| Boiling Point |
456.5±55.0 °C at 760 mmHg
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| Melting Point |
152-154°C
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| Flash Point |
229.9±31.5 °C
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| Vapour Pressure |
0.0±1.1 mmHg at 25°C
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| Index of Refraction |
1.600
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| LogP |
1.04
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
21
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| Complexity |
449
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC1=CC=C(C=C1)S(=O)(=O)NC2=CC=CC(=C2)C(=O)OC
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| InChi Key |
CVKBYFCJQSPBOI-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C15H15NO4S/c1-11-6-8-14(9-7-11)21(18,19)16-13-5-3-4-12(10-13)15(17)20-2/h3-10,16H,1-2H3
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| Chemical Name |
methyl 3-[(4-methylphenyl)sulfonylamino]benzoate
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| Synonyms |
METHYL 3-(4-METHYLBENZENESULFONAMIDO)BENZOATE; methyl 3-{[(4-methylphenyl)sulfonyl]amino}benzoate; MSAB?; CHEMBL5189621; SCHEMBL16116948; EX-A4298;
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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) |
DMSO : ~250 mg/mL (~818.73 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (6.81 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.08 mg/mL (6.81 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.2749 mL | 16.3747 mL | 32.7493 mL | |
| 5 mM | 0.6550 mL | 3.2749 mL | 6.5499 mL | |
| 10 mM | 0.3275 mL | 1.6375 mL | 3.2749 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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