| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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Purity: =99.97%
| Targets |
Apoptosis
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|---|---|
| ln Vitro |
Fraxetin (7,8-dihydroxy-6-methoxy coumarin), a coumarin derivative, has been reported to possess antioxidative, anti-inflammatory and neuroprotective effects.[1]
Effect of fraxetin on proliferation and apoptosis in breast cancer cells. The aim of the present study was to examine the effect of fraxetin on proliferation and apoptosis in the MCF-7 breast cancer cell line. Cell proliferation was measused using an MTT assay and 4',6-diamidino-2-phenylindole (DAPI) staining was used to determine shrinkage and condensation. RT-PCR was used to examine the expression of factor-associated suicide (Fas) and Fas ligand (FasL) mRNA, and western blot analysis was used to examine Bax and Bcl-2 protein. MTT showed that the proliferation of MCF-7 cells was significantly inhibited by fraxetin in a dose-dependent manner. Fraxetin also induced significant morphological changes of MCF-7 cells, suggestive of apoptosis, whereas DAPI staining showed that fraxetin caused cell shrinkage and chromatin condensation. RT-PCR showed that the expression of Fas and FasL mRNA was upregulated by fraxetin and the western blot analysis revealed that Bax was upregulated and Bcl-2 was downregulated. In conclusion, fraxetin can inhibit the proliferation of MCF-7 cells, induce apoptosis, upregulate Fas, FasL and Bax, and downregulate Bcl-2 to induce apoptosis. These results support the potential therapeutic role for fraxetin in breast cancer.[2] |
| ln Vivo |
The therapeutic effect of fraxetin on ethanol-induced hepatic fibrosis by enhancing ethanol metabolism, inhibiting oxidative stress and modulating inflammatory mediators in rats. The present study was designed to investigate the possible protective effects of fraxetin against ethanol induced liver fibrosis in rats. Rats were underwent intragastric administration of ethanol (5.0-9.5 g/kg) once a day for 24 weeks. Effect of fraxetin against ethanol induced liver fibrosis was investigated by giving 20 or 50 mg/kg fraxetin. At the end of experiment, the livers were collected for histopathological analyses, protein extraction, and enzymatic activities. Our results indicated that fraxetin significantly corrected ethanol-induced hepatic fibrosis, as evidenced by the decrease in serum ALT and AST, the attenuation of histopathological changes. Fraxetin also expedited ethanol metabolism by enhancing the alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) activities. Besides, fraxetin alleviated lipid peroxidation, enhanced hepatic antioxidant capabilities, inhibited CYP2E1 activity, and reduced the inflammatory mediators, including TNF-α and IL-1β via up-regulation of hemeoxygenase-1 (HO-1) protein. In summary, the hepatoprotection of fraxetin is mostly attributed to its antioxidant capability, alcohol metabolism, and anti-inflammation effect. [1]
Fraxetin exhibits its antioxidant effect through increasing the level of GSH and reducing oxidative damage in a Drosophila melanogaster experimental model. Previous we reported that fraxetin could alleviate carbon tetrachloride induced hepatic fibrosis via inhibition of oxidative stress and inflammation |
| Enzyme Assay |
Estimation of hepatic alcohol metabolizing enzyme activities[1]
Alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) were measured in liver homogenate according to the protocol established in previous study. Fas/FasL mRNA expression by RT-PCR [2] MCF-7 cells were seeded in 6-well plates at a density of 1×104 cells/well. After 24 h, the supernatant was suctioned and the cells were cultured in medium containing 20, 40 or 60 µM of fraxetin for 24 h, the cells were collected and total RNA was extracted according to the instructions of the RNA extraction kit. RNA concentration and purity (A260/A280 >1.8, indicating pure RNA) were determined by UV-Vis spectrophotometer. cDNA was obtained via reverse transcription from mRNA according to the instruction of the reverse transcription kit. The expression of target genes was then detected using cDNA as the template by RT-PCR assay, according to the manufacturer's instructions, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as an internal control. The primer sequences for Fas and FasL are shown in Table I. Amplification conditions were: 95°C for 10 min, 95°C for 15 sec, 60°C for 1 min, with 40 cycles of amplification. The Ct value was automatically calculated using the CFX Manager software, and the relative quantification of gene expression was calculated using the 2−ΔCt method, as per the formula: ΔCt (target gene) = Ct (target gene) - Ct (control gene). |
| Cell Assay |
Cell proliferation inhibition rate [2]
Cell proliferation was measured using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay after cells were treated with fraxetin (both from Sigma, St. Louis, MO, USA). MCF-7 cells were seeded in 96-well plates at a concentration of 1×105/ml and each well contained 100 µl. After 24 h, fraxetin was added to 10, 20, 40 and 60 µM final concentrations. The control group was treated without fraxetin. The cells were incubated for 24 and 48 h at 37°C in 5% CO2 and then culture medium was changed. MTT (10 µl) was added to each well at a final concentration of 5 mg/ml. After incubating for 4 h, the optical density (OD) at 570 nm was measured by a microplate reader. The inhibition rate was calculated as: Inhibition rate (%) = (OD value of control group - OD value of experimental group/OD value of control group) × 100%.[2] Morphological observation [2] MCF-7 cells were treated with 20, 40 and 60 µM of fraxetin for 24 h, and the morphological changes were observed and recorded with an inverted microscope).[2] 4,6-Diamidino-2-phenylindole (DAPI) staining [2] MCF-7 cells were seeded in 6-well plates at a density of 1×104 cells/well. After 24 h, the supernatant was suctioned and the cells were cultured in medium containing 20, 40 or 60 µM of fraxetin for 24 h, then washed with pre-cooled phosphate-buffered saline (PBS) 3 times. DAPI solution (1 µg/ml) was added to each well, the cells were incubated at 37°C for 5 min and washed with pre-cooled PBS again. The cells were observed and images were captured using a fluorescence microscope in the dark.[2] |
| Animal Protocol |
Male SPF-Wistar rats (200 ± 20 g) were used in this study. After a period of 1 week, the rats were divided into five groups with 10 rats per group as follows: one untreated group (GI) and four groups (GII–GV) treated with ethanol gavage once a day. The doses of ethanol were increased gradually according to the method of Zhang et al.: 5.0 g/kg/day from 1 to 4 weeks, 7.0 g/kg/day from 5 to 8 weeks, 9.0 g/kg/day from 9 to 12 weeks, and 9.5 g/kg/day from 13 to 24 weeks. At the same time, the rats were treated with 100 mg/kg/day silymarin (GIII) as positive control, 20 mg/kg/day fraxetin (GIV) and 50 mg/kg/day fraxetin (GV) through gavage. At the end of 24 weeks, all rats were killed and blood samples were collected. Liver samples were obtained from all groups and divided into two parts. One part was stored immediately at −80 °C for future analysis, whereas the other part was fixed in 10% formalin for histopathological examination.
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| ADME/Pharmacokinetics |
Metabolism / Metabolites
Fraxetin has known human metabolites that include (2S,3S,4S,5R)-3,4,5-trihydroxy-6-(8-hydroxy-6-methoxy-2-oxochromen-7-yl)oxyoxane-2-carboxylic acid. |
| References |
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| Additional Infomation |
Fraxetin is a hydroxycoumarin that is 6-methoxycoumarin in which the hydrogens at positions 7 and 8 have been replaced by hydroxy groups. It has a role as an Arabidopsis thaliana metabolite, an antimicrobial agent, an apoptosis inhibitor, an apoptosis inducer, an antioxidant, an anti-inflammatory agent, a hepatoprotective agent, an antibacterial agent and a hypoglycemic agent. It is a hydroxycoumarin and an aromatic ether.
Fraxetin has been reported in Salsola laricifolia, Aesculus turbinata, and other organisms with data available. |
| Molecular Formula |
C10H8O5
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|---|---|
| Molecular Weight |
208.1675
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| Exact Mass |
208.037
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| Elemental Analysis |
C, 57.70; H, 3.87; O, 38.43
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| CAS # |
574-84-5
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| PubChem CID |
5273569
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| Appearance |
Light yellow to yellow solid powder
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| Density |
1.5±0.1 g/cm3
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| Boiling Point |
472.0±45.0 °C at 760 mmHg
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| Melting Point |
230-232°C
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| Flash Point |
196.0±22.2 °C
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| Vapour Pressure |
0.0±1.2 mmHg at 25°C
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| Index of Refraction |
1.648
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| Source |
Fraxinus rhynchophylla Hance
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| LogP |
0.59
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
1
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| Heavy Atom Count |
15
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| Complexity |
288
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O1C(C([H])=C([H])C2=C([H])C(=C(C(=C12)O[H])O[H])OC([H])([H])[H])=O
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| InChi Key |
HAVWRBANWNTOJX-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C10H8O5/c1-14-6-4-5-2-3-7(11)15-10(5)9(13)8(6)12/h2-4,12-13H,1H3
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| Chemical Name |
7,8-dihydroxy-6-methoxychromen-2-one
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| Synonyms |
Fraxetin; 574-84-5; 7,8-Dihydroxy-6-methoxy-2H-chromen-2-one; 7,8-Dihydroxy-6-methoxycoumarin; 7,8-dihydroxy-6-methoxychromen-2-one; 2H-1-Benzopyran-2-one, 7,8-dihydroxy-6-methoxy-; 7,8-Dihydroxy-6-methoxy-chromen-2-one; 7,8-Dihydroxy-6-methoxy-2-benzopyrone;
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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 : ~100 mg/mL (~480.38 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (9.99 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 (9.99 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 (9.99 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 | 4.8038 mL | 24.0188 mL | 48.0377 mL | |
| 5 mM | 0.9608 mL | 4.8038 mL | 9.6075 mL | |
| 10 mM | 0.4804 mL | 2.4019 mL | 4.8038 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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