| Size | Price | Stock | Qty |
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| 250mg |
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| 500mg |
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| 1g |
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| 2g |
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| 5g | |||
| Other Sizes |
Purity: ≥98%
| Targets |
PI3K
Esculetin acts through multiple molecular targets and pathways. Its primary recognized activity is the inhibition of lipoxygenases (LOs), specifically 5-LO and 12-LO. This inhibition is likely due to its ability to bind iron, as it also inhibits other iron-containing enzymes. Esculetin is a potent inhibitor of 5-lipoxygenase, with reported IC50 values of 4 µM, 1.46 µM for the formation of 5-HETE, and 0.4 µM in some studies. It inhibits 12-LO with an IC50 of 2.5 µM. Esculetin also inhibits the histone demethylase JARID1B (IC50 = 4.6 µM) and the lysine-specific demethylase KDM5B. Furthermore, esculetin directly targets β-catenin, suppressing the Wnt-β-catenin pathway. It also inhibits the PI3K/Akt pathway, which is involved in platelet-derived growth factor (PDGF)-induced airway smooth muscle cell phenotype switching. |
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| ln Vitro |
Esculetin decreases cell proliferation by inducing G1 phase cell cycle arrest, which is associated with the down-regulation of cyclin D1/CDK4 and cyclin E/CDK2 complexes by the activation of p27KIP[2]. Esculetin significantly reduces HCC cell proliferation, with an IC50 value of 2.24 mM, in a concentration- and time-dependent manner. In SMMC-7721 cells, it blocks the cell cycle at the S phase and induces apoptosis with a notable increase in caspase-3 and caspase-9 activity, but it has no effect on caspase-8 activity. Esculetin treatment causes the mitochondrial membrane potential to collapse both in vitro and in vivo, along with an uptick in the transcriptional gene Bax and a decrease in the translational gene Bcl-2 expression. Esculetin inhibits the growth of hepatocellular carcinoma cells both in vitro and in vivo, and its mechanisms involve the start of a mitochondrial-mediated, caspase-dependent apoptosis pathway[3].
Esculetin demonstrates significant in vitro biological activities. It is a potent antioxidant that protects cells from oxidative stress-induced damage by scavenging reactive oxygen species (ROS). Esculetin has been shown to inhibit the proliferation of human T cells stimulated by mitogens in a dose-dependent manner. In cancer research, esculetin suppresses the proliferation of human colon cancer cells by directly targeting β-catenin and promotes TRAIL-induced apoptosis in cancer cells. It also inhibits the production of matrix metalloproteinases (MMPs) in cartilage explants. Additionally, esculetin exhibits antimicrobial activity against various bacteria, including B. cereus, S. aureus, and E. coli. In vitro, esculetin shows a concentration-dependent relaxant effect on isolated guinea-pig tracheal chains, with an EC50 of 60 µM. |
| ln Vivo |
Esculetin significantly decreases tumor growth in mice bearing Hepa1-6 cells. With increasing doses of esculetin, tumor weight is decreased by 20.33, 40.37, and 55.42%. It's important to note that this animal study found no evidence of esculetin's toxicity[3]. At a dose of 5 mg/kg, esculetin exhibits intestinal anti-inflammatory activity in a rat experimental model of inflammatory bowel disease brought on by trinitrobenzenesulfonic acid[4].
Esculetin has demonstrated efficacy in various in vivo models. It reduces croton oil-induced ear edema in mice, indicating anti-inflammatory activity. It also inhibits acetylcholine-induced writhing in mice with an ED50 of 69 mg/kg, suggesting analgesic properties. In a rat model of colon carcinogenesis induced by 1,2-dimethylhydrazine, esculetin inhibited the formation of aberrant crypt foci and tumors. Oral administration of a prodrug of esculetin, CPA-926, suppressed cartilage destruction in a rabbit model of experimental osteoarthritis. Treatment with esculetin also suppressed tumor growth in vivo. |
| Enzyme Assay |
In vitro enzyme assays for esculetin typically measure its ability to inhibit lipoxygenase activity. For 5-LO inhibition, the enzyme is incubated with its substrate, arachidonic acid, in the presence of increasing concentrations of esculetin. The formation of 5-HETE, a 5-lipoxygenase product, is then measured using chromatographic or radiometric methods. The compound's IC50 for inhibiting 5-HETE formation is determined from dose-response curves. Esculetin inhibits 5-LO noncompetitively. Similar assays can be performed to assess 12-LO inhibition. Its ability to inhibit JARID1B can be evaluated using histone demethylase activity assays.
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| Cell Assay |
Tests are performed in 96-well plates. For mature adipocytes, cells are seeded (5000 cells/well), grown to confluency, induced to differentiate, and grown to maturation. Preadipocytes are cultured overnight before treatment at a seeding density of 2500 cells per well. For 6, 12, 24, or 48 hours, cells are either incubated with DMSO or esculetin at progressively higher concentrations. The seeding density for the post-confluent preadipocytes is 2500 cells/well, and the cells are grown to confluency prior to treatment. For Days 0 to 2, 2 to 4, and 4 to 6 of adipogenesis, esculletin (100 or 200 M) is added to the induction medium in a carrier containing 0.01% DMSO. Every two days, medium is changed. Cells are treated with 20 μL of the MTS assay solution per well before being washed three times with DMEM/10% FBS before the assay for cell viability. 100 μL of DMEM/10% FBS medium is then added to each well. 25 μL of 10% sodium dodecyl sulfate is then added to each well after cells have been incubated at 37 °C for one hour. To calculate the formazan concentration, which is inversely proportional to the number of live cells, a plate reader is used to measure the absorbance at 490 nm.
Cell-based assays for esculetin involve treating various cell lines with the compound and measuring its effects on cell proliferation, apoptosis, and signaling pathways. For example, human T cells can be treated with esculetin to assess its inhibitory effect on mitogen-induced proliferation. Cancer cell lines can be used to study its effects on cell viability, apoptosis (e.g., TRAIL-induced), and the Wnt-β-catenin pathway. The compound's antioxidant activity can be assessed by measuring its ability to protect cells from H2O2-induced damage in fibroblast cell lines. |
| Animal Protocol |
C57BL/6J mice implanted with Hepa1-6 cells
200, 400, or 700 mg/kg/day i.p In vivo animal studies for esculetin are conducted in various disease models. In the croton oil-induced ear edema model in mice, esculetin is typically applied topically, and its anti-inflammatory effect is assessed by measuring ear swelling. In the mouse writhing test, esculetin is administered orally or intraperitoneally to evaluate its analgesic activity. In carcinogenesis models, such as the DMH-induced colon cancer model in rats, esculetin is administered orally, and the number of aberrant crypt foci and tumors is assessed. |
| ADME/Pharmacokinetics |
Esculetin (CAS 305-01-1) has a molecular weight of 178.14 g/mol. It is soluble in DMSO (32 mg/mL) but insoluble in water and ethanol. For storage, the powder should be kept at -20°C, where it is stable for up to 3 years. In solution, it can be stored at -20°C for up to 6 months.
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| Toxicity/Toxicokinetics |
Esculetin is generally considered to have low toxicity. It has been shown to have no significant cytotoxic effect on normal murine macrophages. However, as with any research compound, appropriate safety precautions should be taken when handling it.
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| References | |
| Additional Infomation |
Esculetin is a hydroxycoumarin, a derivative of umbelliferone, in which the hydrogen atom at position 6 is replaced by a hydroxyl group. It is used in ultraviolet (UV) absorbers. Esculetin possesses antioxidant, UV filtering, and plant metabolic activity. It has been reported to be found in Caragana frutex, Sonchus fruticosus, and other organisms with relevant data.
Esculetin (6,7-dihydroxycoumarin) is a naturally occurring coumarin with a wide range of biological activities. It is a potent, selective, and non-competitive inhibitor of 5-lipoxygenase and 12-lipoxygenase. Its diverse pharmacological properties, including anti-inflammatory, antioxidant, anti-tumor, and immunomodulatory effects, make it a valuable research tool for studying various diseases. Esculetin is not approved for therapeutic use and is intended for research purposes only. |
| Molecular Formula |
C9H6O4
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|---|---|
| Molecular Weight |
178.14154
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| Exact Mass |
178.026
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| Elemental Analysis |
C, 60.68; H, 3.40; O, 35.92
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| CAS # |
305-01-1
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| Related CAS # |
305-01-1
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| PubChem CID |
5281416
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| Appearance |
Yellow to brown solid powder
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| Density |
1.6±0.1 g/cm3
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| Boiling Point |
469.7±45.0 °C at 760 mmHg
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| Melting Point |
271-273 °C(lit.)
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| Flash Point |
201.5±22.2 °C
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| Vapour Pressure |
0.0±1.2 mmHg at 25°C
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| Index of Refraction |
1.689
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| LogP |
0.98
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
0
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| Heavy Atom Count |
13
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| Complexity |
248
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O=C1C=CC2=C(O1)C=C(O)C(O)=C2
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| InChi Key |
ILEDWLMCKZNDJK-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C9H6O4/c10-6-3-5-1-2-9(12)13-8(5)4-7(6)11/h1-4,10-11H
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| Chemical Name |
6,7-dihydroxychromen-2-one
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| Synonyms |
CCRIS 7065; CCRIS-7065; CCRIS7065; Cichorigenin; NSC 26428; Asculetine; NSC-26428; NSC26428
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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: 35~125 mg/mL (196.5~701.7 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (11.68 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 (11.68 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. 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 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (11.68 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 | 5.6136 mL | 28.0678 mL | 56.1356 mL | |
| 5 mM | 1.1227 mL | 5.6136 mL | 11.2271 mL | |
| 10 mM | 0.5614 mL | 2.8068 mL | 5.6136 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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