| Size | Price | Stock | Qty |
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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 |
Ethyl Caffeate primarily targets the transcription factor NF-κB by preventing its binding to DNA. At the molecular level, its effects are mediated through the inhibition of NF-κB-DNA complex formation. The compound does not affect upstream signaling components including IκBα phosphorylation/degradation, NF-κB (p65) translocation to the nucleus, or the activation of mitogen-activated protein kinases (SAPK/JNK, p38, p42/44). The IC₅₀ for inhibiting LPS-induced NO production in RAW 264.7 macrophages is 5.5 μg/mL. [1]
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| ln Vitro |
Ethyl Caffeate significantly inhibits LPS-induced NO production in RAW 264.7 macrophages in a dose-dependent manner, with an IC₅₀ of 5.5 μg/mL. At 2 μg/mL, NO production was inhibited by 35% compared to LPS-only treated cells. [1]
The compound inhibits iNOS mRNA expression; at 1 μg/mL, a 56% reduction in iNOS mRNA was observed, and at 2 μg/mL, iNOS mRNA levels were similar to vehicle control. At the protein level, 0.5 μg/mL ethyl caffeate reduced iNOS protein to approximately 30% of that in LPS-only treated cells. [1] Ethyl Caffeate suppresses LPS-induced COX-2 protein expression; at 5 μg/mL, COX-2 expression was reduced to 45% of LPS-only control. It also markedly suppresses PGE₂ production; at 1 μg/mL, PGE₂ production was significantly inhibited, and at 2-5 μg/mL, total inhibition of PGE₂ was observed. [1] In TPA-treated MCF-7 cells, ethyl caffeate (10-100 μg/mL) downregulates cox-2 transcriptional activity in a dose-dependent manner, reducing full-length cox-2 promoter activity by 72% (at 10 μg/mL), 57% (at 50 μg/mL), and 48% (at 100 μg/mL). [1] Ethyl Caffeate does not affect LPS-induced MAPK phosphorylation (SAPK/JNK, p38, p42/44) in macrophages at concentrations up to 10 μg/mL. It also does not affect IκBα phosphorylation and degradation, nor NF-κB p65 translocation to the nucleus. [1] Using an in vitro EMSA binding assay, ethyl caffeate (10-20 μg/mL) dose-dependently inhibits NF-κB binding to DNA, with complete inhibition at 20 μg/mL. This inhibition is completely reversed by 50 μM DTT. [1] Structure-activity relationship studies show that both the catechol moiety and the α,β-unsaturated ester group are essential for NF-κB DNA binding inhibition. Ethyl caffeate (containing both) is most effective; ethyl 3,4-dihydroxyhydrocinnamate (containing catechol only) shows weaker inhibition; ethyl cinnamate (containing α,β-unsaturated ester only) shows no inhibition. [1] Ethyl caffeine has an IC50 of 5.5 μg/ml and strongly suppresses the generation of nitric oxide (NO) caused by lipopolysaccharide (LPS)[1]. |
| ln Vivo |
In a mouse skin immunohistochemical study, topical application of Ethyl Caffeate significantly inhibits TPA-induced COX-2 expression in the epidermal layer in a dose-dependent manner. At 1 mg/200 μL/site (24 mM), ethyl caffeate showed comparable inhibition to 1 mg/200 μL/site celecoxib (13 mM). At 2 mg/200 μL/site (48 mM), ethyl caffeate was more effective than 10 mg/200 μL/site celecoxib (131 mM) in inhibiting TPA-induced COX-2 expression. [1]
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| Enzyme Assay |
An electrophoretic mobility shift assay (EMSA) was performed to assess NF-κB binding to DNA. Nuclear extracts (4 μg) from LPS-stimulated RAW 264.7 cells were incubated with biotin end-labeled, 22-mer double-stranded NF-κB oligonucleotide in binding buffer containing 10% glycerol, 100 mM KCl, 1.5 mM MgCl₂, and 0.3 mM EDTA for 20 minutes at room temperature. For in vitro binding assays, nuclear extracts from LPS-stimulated cells were treated with different concentrations of test compounds (including Ethyl Caffeate at concentrations such as 10 and 20 μg/mL) at 37°C for 30 minutes before EMSA analysis. DNA-protein complexes were separated on a native 5-10% gradient polyacrylamide gel. Specificity was confirmed by supershift with anti-p65 antibody and competition with unlabeled NF-κB oligonucleotide. [1]
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| Cell Assay |
RAW 264.7 murine macrophages were cultured in DMEM with 10% FBS. For NO production assays, cells (2×10⁵ cells/well) were pretreated with Ethyl Caffeate (0.1-20 μg/mL) for 1 hour, then stimulated with LPS (1 μg/mL) for 24 hours. Nitrite accumulation was measured by Griess reaction (100 μL supernatant mixed with 100 μL Griess reagent, absorbance at 540 nm). Cell viability was determined by MTT assay to exclude cytotoxicity. [1]
For RT-PCR, cells (3×10⁶/well) were treated with ethyl caffeate (0.5-5 μg/mL) for 1 hour, then LPS (1 μg/mL) for 6 hours. Total RNA was isolated using TRIZOL, and iNOS mRNA was amplified with specific primers. GA3PDH was used as internal control. [1] For Western blot, cells were treated with ethyl caffeate for 1 hour, then LPS (1 μg/mL) for 18 hours (iNOS, COX-2) or 30 minutes (MAPK) or 11 minutes (IκBα). Total, cytosolic, or nuclear proteins (20 μg) were separated by SDS-PAGE, transferred to PVDF membranes, and detected with specific antibodies against iNOS, COX-2, phospho-MAPKs, IκBα, p65 NF-κB, PARP, and α-tubulin. [1] For COX-2 promoter assay, MCF-7 cells (1.2×10⁵/well) were co-transfected with pCOX-2-Luc and pRL-TK-Luc using LipofectAMINE. After recovery, cells were treated with TPA (50 ng/mL) alone or with ethyl caffeate (10-100 μg/mL) for 6 hours. Firefly and Renilla luciferase activities were measured using dual luciferase assay. [1] For PGE₂ measurement, RAW 264.7 cells were pretreated with 500 μM aspirin for 3 hours (to inactivate COX-1), washed, then treated with ethyl caffeate (0.1-20 μg/mL) for 1 hour, followed by LPS (1 μg/mL) for 16 hours. PGE₂ in culture media was measured by ACE competitive enzyme immunoassay. [1] |
| Animal Protocol |
Female ICR mice were used. Dorsal skin was shaven. Mice were treated topically with Ethyl Caffeate (1 mg/200 μL/site or 2 mg/200 μL/site) or celecoxib (1 mg/200 μL/site or 10 mg/200 μL/site) for 30 minutes, then treated with TPA (10 nmol/200 μL/site) for 4 hours. After treatment, mice were killed by cervical dislocation. Skin tissues were formalin-fixed, paraffin-embedded, sectioned (4 μm), and deparaffinized. For antigen retrieval, sections were heated in 10 mM citrate buffer (pH 6.0) for 10 minutes. Endogenous peroxidase was blocked with 3% hydrogen peroxide in methanol for 15 minutes. Sections were incubated with polyclonal COX-2 antibody (1:500 dilution) for 1-2 hours at room temperature, then developed using HPR EnVision system with DAB staining, and counterstained with Mayer's hematoxylin. [1]
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| Toxicity/Toxicokinetics |
Ethyl Caffeate was found to have little or no cytotoxicity to RAW 264.7 macrophages at concentrations of 10 μg/mL or below, as determined by MTT assay. At 5 μg/mL, >50% of cells remained viable. [1]
In MCF-7 cells, no cytotoxic effects were observed with ethyl caffeate treatment at concentrations of 10-100 μg/mL over 6 hours as determined by MTT assay. [1] |
| References | |
| Additional Infomation |
Ethyl trans-caffeic acid is an ethyl ester formed by the condensation of the carboxyl group of trans-caffeic acid with ethanol. It possesses anti-inflammatory and antitumor activities. It is an alkyl caffeic acid ester and ethyl ester, functionally related to trans-caffeic acid. Ethyl caffeic acid has been reported in perilla, Himalayan sasanqua, and several other organisms with relevant data.
Ethyl Caffeate was isolated from the ethyl acetate fraction of Bidens pilosa using bioactivity-guided fractionation. Its structure was elucidated by IR, NMR, and EI-MS. Molecular weight: 208 [M]⁺. Melting point: 147-149°C. [1] The proposed mechanism of action is an "oxidation-inhibition" mechanism: the catechol moiety is oxidized to α-benzoquinone, which then reacts with cysteine sulfhydryl groups of the p65 subunit of NF-κB via Michael-type addition, preventing DNA binding. The α,β-unsaturated ester group provides additional reaction centers for covalent interaction with p65. [1] The compound does not affect IL-10 promoter activity, suggesting specificity for COX-2 regulation. [1] |
| Molecular Formula |
C11H12O4
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| Molecular Weight |
208.2106
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| Exact Mass |
208.073
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| CAS # |
102-37-4
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| Related CAS # |
Ethyl trans-caffeate;66648-50-8
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| PubChem CID |
5317238
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| Appearance |
Light yellow to yellow solid powder
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| Density |
1.3±0.1 g/cm3
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| Boiling Point |
377.0±32.0 °C at 760 mmHg
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| Flash Point |
148.4±18.6 °C
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| Vapour Pressure |
0.0±0.9 mmHg at 25°C
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| Index of Refraction |
1.612
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| LogP |
1.72
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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 |
4
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| Heavy Atom Count |
15
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| Complexity |
237
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CCOC(=O)/C=C/C1=CC(=C(C=C1)O)O
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| InChi Key |
WDKYDMULARNCIS-GQCTYLIASA-N
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| InChi Code |
InChI=1S/C11H12O4/c1-2-15-11(14)6-4-8-3-5-9(12)10(13)7-8/h3-7,12-13H,2H2,1H3/b6-4+
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| Chemical Name |
ethyl (E)-3-(3,4-dihydroxyphenyl)prop-2-enoate
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| Synonyms |
Ethyl caffeate; 102-37-4; ethyl trans-caffeate; Ethyl 3,4-dihydroxycinnamate; Caffeic acid ethyl ester;
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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)
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| Solubility (In Vitro) |
DMSO : ~250 mg/mL (~1200.71 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 | 4.8028 mL | 24.0142 mL | 48.0284 mL | |
| 5 mM | 0.9606 mL | 4.8028 mL | 9.6057 mL | |
| 10 mM | 0.4803 mL | 2.4014 mL | 4.8028 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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