| Size | Price | Stock | Qty |
|---|---|---|---|
| 250mg |
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| 500mg |
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| Other Sizes |
| Targets |
1. 4-Methylesculetin inhibits the nuclear factor-kappa B (NF-κB) signaling pathway, as evidenced by reduced phosphorylation of NF-κB p65 protein in LPS-stimulated RAW264.7 cells and DSS-induced colonic tissues [1]
2. 4-Methylesculetin suppresses the mitogen-activated protein kinase (MAPK) signaling pathway, including decreased phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2), c-Jun N-terminal kinase (JNK), and p38 MAPK in inflamed cells and tissues [1] 3. 4-Methylesculetin downregulates the expression of pro-inflammatory mediators, such as interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and interleukin-1 beta (IL-1β), at the mRNA and protein levels [1] |
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| ln Vitro |
1. In LPS-stimulated RAW264.7 murine macrophage cells: 4-Methylesculetin (at concentrations of 10, 20, and 40 μM) dose-dependently reduced the mRNA expression of IL-6 and TNF-α. At 40 μM, it decreased IL-6 mRNA level by approximately 60% and TNF-α mRNA level by approximately 50% compared to the LPS-only group. It also inhibited the phosphorylation of NF-κB p65, ERK1/2, JNK, and p38 MAPK proteins [1]
2. In Caco-2 human intestinal epithelial cells: 4-Methylesculetin (20 and 40 μM) increased the expression of tight junction protein zonula occludens-1 (ZO-1) and enhanced the transepithelial electrical resistance (TEER) value, which indicated improved intestinal barrier function. It also reduced LPS-induced increase in paracellular permeability [1] |
| ln Vivo |
In C57BL/6 mice with dextran sulfate sodium (DSS)-induced ulcerative colitis:
1. Oral administration of 4-Methylesculetin (100 mg/kg body weight, once daily for 7 days) significantly attenuated DSS-induced weight loss (from ~20% weight loss in DSS group to ~8% in drug-treated group), reduced disease activity index (DAI) score (from ~8 in DSS group to ~3 in drug-treated group), and restored colon length (from ~4 cm in DSS group to ~6 cm, close to normal group) [1] 2. 4-Methylesculetin ameliorated colonic histopathological damage: it reduced the degree of mucosal erosion, inflammatory cell infiltration, and crypt destruction, with the histopathological score decreasing from ~7 in DSS group to ~2 in drug-treated group [1] 3. 4-Methylesculetin downregulated the protein expression of TNF-α, IL-6, and IL-1β in colonic tissues, and inhibited the phosphorylation of NF-κB p65, ERK1/2, JNK, and p38 MAPK in colonic epithelial cells [1] |
| Enzyme Assay |
1. Western Blot assay for protein phosphorylation and expression:
- Cell or colonic tissue samples were lysed to extract total protein, and protein concentration was determined using a protein assay kit. - Equal amounts of protein (30-50 μg per lane) were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene difluoride (PVDF) membranes. - Membranes were blocked with non-fat milk for 1 hour at room temperature, then incubated with primary antibodies against target proteins (e.g., p-NF-κB p65, NF-κB p65, p-ERK1/2, ERK1/2, ZO-1, TNF-α) overnight at 4°C. - After washing with TBST buffer, membranes were incubated with horseradish peroxidase (HRP)-conjugated secondary antibodies for 1 hour at room temperature. - Protein bands were visualized using an enhanced chemiluminescence (ECL) detection system, and band intensity was quantified using image analysis software [1] 2. Quantitative real-time polymerase chain reaction (qRT-PCR) assay for mRNA expression: - Total RNA was extracted from cells or tissues using an RNA extraction kit, and RNA purity and concentration were measured. - Complementary DNA (cDNA) was synthesized from total RNA using a reverse transcription kit. - qRT-PCR was performed using specific primers for IL-6, TNF-α, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH, internal control) in a PCR system. - The relative mRNA expression level was calculated using the 2^(-ΔΔCt) method [1] |
| Cell Assay |
1. RAW264.7 macrophage cell assay (inflammation model):
- Cells were seeded in 6-well plates at a density of 5×10^5 cells per well and cultured overnight. - Cells were pretreated with 4-Methylesculetin (10, 20, 40 μM) for 1 hour, then stimulated with LPS (1 μg/mL) for 24 hours. - After incubation, cells were collected for total protein extraction (for Western Blot) or total RNA extraction (for qRT-PCR) to detect inflammatory mediators and signaling pathway proteins [1] 2. Caco-2 intestinal epithelial cell assay (barrier function model): - Cells were seeded in transwell inserts at a density of 1×10^5 cells per insert and cultured for 21 days to form a confluent monolayer. - The monolayer was treated with 4-Methylesculetin (20, 40 μM) for 1 hour, then stimulated with LPS (1 μg/mL) for 24 hours. - TEER value was measured using a volt-ohmmeter to evaluate barrier integrity. Cells were also collected for Western Blot to detect ZO-1 protein expression [1] |
| Animal Protocol |
1. Animal grouping and colitis induction:
- Male C57BL/6 mice (6-8 weeks old) were randomly divided into three groups: normal control group, DSS-induced colitis group, and 4-Methylesculetin-treated colitis group (n=6 per group). - Colitis was induced by providing mice with drinking water containing 3.5% (w/v) DSS for 7 consecutive days; the normal group received regular drinking water [1] 2. Drug administration: - 4-Methylesculetin was dissolved in 0.5% carboxymethyl cellulose sodium (CMC-Na) solution. - The drug-treated group received oral gavage of 4-Methylesculetin at a dose of 100 mg/kg body weight once daily for 7 days, starting from the first day of DSS administration. - The normal group and DSS group received oral gavage of an equal volume of 0.5% CMC-Na solution [1] 3. Sample collection and detection: - Mice were weighed daily to record weight changes, and DAI score was evaluated daily based on weight loss, stool consistency, and rectal bleeding. - On day 8, mice were euthanized. Colons were isolated, measured for length, and divided into two parts: one part was fixed in formalin for histopathological analysis, and the other part was stored at -80°C for protein and RNA extraction [1] |
| References | |
| Additional Infomation |
6,7-Dihydroxy-4-methylcoumarin is a hydroxycoumarin, which is the product of 4-methylcoumarin with hydroxyl groups at the 3 and 4 positions. It is an inhibitor of hyaluronic acid synthesis. In addition, it is also used as a fluorescence sensor to monitor the consumption of boric acid in the Suzuki coupling reaction; its fluorescence can be easily detected by the naked eye using a standard 365 nm UV lamp. It has the effects of inhibiting hyaluronic acid synthesis, anti-oxidation and anti-inflammation. 1. 4-Methylesculin is a natural coumarin derivative with potential anti-inflammatory activity. Its mechanism of improving DSS-induced intestinal inflammation mainly involves two aspects: inhibiting the activation of NF-κB and MAPK signaling pathways, thereby reducing the production of pro-inflammatory cytokines; and enhancing intestinal barrier function by upregulating the expression of tight junction proteins [1]. 2. This study confirmed that 4-methylesculin has a protective effect against DSS-induced ulcerative colitis in mice, suggesting that it may become a candidate drug for the treatment of inflammatory bowel disease (IBD) [1].
|
| Molecular Formula |
C10H8O4
|
|---|---|
| Molecular Weight |
192.1681
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| Exact Mass |
192.042
|
| CAS # |
529-84-0
|
| PubChem CID |
5319502
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| Appearance |
Solid powder
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| Density |
1.5±0.1 g/cm3
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| Boiling Point |
455.5±45.0 °C at 760 mmHg
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| Melting Point |
274-276 °C(lit.)
|
| Flash Point |
190.8±22.2 °C
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| Vapour Pressure |
0.0±1.2 mmHg at 25°C
|
| Index of Refraction |
1.652
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| LogP |
2.08
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| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
0
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| Heavy Atom Count |
14
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| Complexity |
284
|
| Defined Atom Stereocenter Count |
0
|
| InChi Key |
KVOJTUXGYQVLAJ-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C10H8O4/c1-5-2-10(13)14-9-4-8(12)7(11)3-6(5)9/h2-4,11-12H,1H3
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| Chemical Name |
6,7-dihydroxy-4-methylchromen-2-one
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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 Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
DMSO : ~50 mg/mL (~260.19 mM)
H2O : < 0.1 mg/mL |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (13.01 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 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 2: ≥ 2.08 mg/mL (10.82 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. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (10.82 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 5.2037 mL | 26.0186 mL | 52.0373 mL | |
| 5 mM | 1.0407 mL | 5.2037 mL | 10.4075 mL | |
| 10 mM | 0.5204 mL | 2.6019 mL | 5.2037 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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