| Size | Price | Stock | Qty |
|---|---|---|---|
| 1g |
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| Other Sizes |
| Targets |
Benzarone acts primarily as an allosteric inhibitor of the EYA family of phosphatases (specifically EYA1, EYA2, and EYA3) . By binding to a site distinct from the active pocket, it disrupts the functional complex formation between EYA and its transcriptional partners (such as SIX5 and p300), thereby inhibiting the expression of downstream target genes involved in cell proliferation and migration . Furthermore, Benzarone is a potent inhibitor of the Human Uric Acid Transporter 1 (URAT1) , with an IC₅₀ of 2.8 μM in oocyte models, which explains its historical role in modulating serum uric acid levels .
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| ln Vitro |
In vitro, Benzarone demonstrates significant inhibitory effects on URAT1 with an IC₅₀ of 2.8 μM . In cancer research, a derivative of Benzarone (DS-1-38) potently inhibits the growth of Sonic Hedgehog (SHH) subgroup medulloblastoma cells, an effect linked to EYA antagonism . Studies on cellular metabolism show that low concentrations (0.03 mM) of Benzarone stimulate the incorporation of acetate and palmitate into lipids in cultured human arterial smooth muscle cells, while higher concentrations (>0.2 mM) suppress lipid labeling and protein biosynthesis . Additionally, it inhibits SAG-induced GLI-luciferase activity in SL2 cells, indicating suppression of the Hedgehog signaling pathway .
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| ln Vivo |
In vivo studies have shown that Benzarone can influence serum lipid profiles. In hypertensive rats fed a cholesterol-rich diet, administration of Benzarone resulted in a 10% reduction in body weight, a relative increase in serum HDL, and decreases in LDL and VLDL, without altering total cholesterol or triglycerides . In the context of cancer biology, the Benzarone derivative DS-1-38 demonstrates excellent brain penetrance and significantly prolongs the lifespan of genetically engineered mouse models of SHH medulloblastoma, confirming its in vivo efficacy against this tumor type . Furthermore, Benzarone has been shown to lower the oxygen consumption of the rat portal vein while blocking spontaneous phasic activity .
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| Enzyme Assay |
Methodology for EYA Binding Affinity (SPR or ITC): To assess direct binding of Benzarone to EYA proteins, Surface Plasmon Resonance (SPR) can be employed. Recombinant EYA protein is immobilized on a sensor chip. Varying concentrations of Benzarone (e.g., 0.1–100 μM) are flowed over the chip, and binding responses are measured in real-time to calculate the dissociation constant (Kd). Alternatively, Isothermal Titration Calorimetry (ITC) can be used to directly measure the heat change upon binding, providing thermodynamic parameters such as stoichiometry (n), enthalpy (ΔH), and entropy (ΔS).
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| Cell Assay |
Methodology (EYA Transactivation / Western Blot Assay): Human cells (e.g., HCEC-1CT or MB21 cells) are seeded in 6-well plates and cultured overnight . Upon reaching 70-80% confluency, cells are treated with Benzarone at various concentrations (e.g., 0, 10, 20, 30 µM) or vehicle control (DMSO) for 24-48 hours . For mechanistic studies, cells are lysed for immunoprecipitation using anti-EYA3-conjugated agarose to pull down protein complexes, followed by Western blotting with antibodies against EYA3, p300, and SIX5 to assess complex disruption . For viability assays (e.g., in Daoy or MB21 cells), cells are treated with compounds (10 µM) replenished every 2-3 days, and viable cells are counted after 5-7 days using trypan blue exclusion or CellTiter-Glo assays .
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| Animal Protocol |
Methodology (Rat Pharmacokinetics and Metabolism): Male Sprague-Dawley rats (weighing ~200-250g) are typically used. For oral administration, a suspension of Benzarone is given via gavage at a dose of 2 mg/kg . For intravenous studies, a solution is administered via the tail vein. Blood samples are collected at predetermined time points (e.g., 0, 0.5, 1, 2, 4, 6, 8, 12, 24 hours post-dose). To study biliary excretion, bile ducts are cannulated in anesthetized animals to collect bile for up to 24 hours . Plasma, urine, feces, and bile samples are analyzed for total radioactivity (if using 14C-labeled compound) and metabolite profiling via HPLC to determine absorption, excretion patterns, and metabolic pathways .
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| ADME/Pharmacokinetics |
Benzarone is rapidly absorbed following oral administration in humans, with peak plasma concentrations of total radioactivity reached between 1-2 hours . It demonstrates extensive protein binding, with over 99% of the compound bound to human plasma proteins in vitro . The pharmacokinetics show significant species differences. In humans, unchanged Benzarone is not detectable in plasma (>99% of circulating radioactivity represents metabolites), and the drug is primarily eliminated via urinary excretion (73% of the dose) . Conversely, in rats and dogs, fecal excretion predominates (>80%), largely due to substantial biliary excretion (up to 72% in rats), indicating enterohepatic circulation .
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| Toxicity/Toxicokinetics |
The toxicity profile of Benzarone is linked to its effects on mitochondrial function. In rat hepatocytes, Benzarone acts as a mitochondrial uncoupler and reduces the mitochondrial membrane potential at higher concentrations (≥20 µM for membrane potential; ≥50 µM for uncoupling), leading to decreased State 3 respiration and an impaired Respiratory Control Ratio (RCR) . It also inhibits mitochondrial fatty acid β-oxidation (IC₅₀ of 34 µM for ketone body synthesis) and induces oxidative stress in human HepG2 cells starting at 10 µM . Chronic exposure (8 hours) at high concentrations (100 µM) induces late apoptosis and cytochrome c release . According to GHS classification from various notifications, Benzarone is labeled as a Warning substance: suspected of damaging fertility or the unborn child (H361) and very toxic to aquatic life with long-lasting effects (H410) .
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| References | |
| Additional Infomation |
Benzarone is a member of 1-benzofuran.
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| Molecular Formula |
C17H14O3
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|---|---|
| Molecular Weight |
266.2913
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| Exact Mass |
266.094
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| Elemental Analysis |
C, 76.68; H, 5.30; O, 18.02
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| CAS # |
1477-19-6
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| PubChem CID |
255968
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| Appearance |
Brown to gray solid powder
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| Density |
1.234g/cm3
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| Boiling Point |
473.6ºC at 760mmHg
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| Melting Point |
124.3°
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| Flash Point |
240.2ºC
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| Vapour Pressure |
1.36E-09mmHg at 25°C
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| Index of Refraction |
1.638
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| LogP |
3.931
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
20
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| Complexity |
346
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CCC1=C(C2=CC=CC=C2O1)C(=O)C3=CC=C(C=C3)O
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| InChi Key |
RFRXIWQYSOIBDI-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C17H14O3/c1-2-14-16(13-5-3-4-6-15(13)20-14)17(19)11-7-9-12(18)10-8-11/h3-10,18H,2H2,1H3
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| Chemical Name |
(2-ethyl-1-benzofuran-3-yl)-(4-hydroxyphenyl)methanone
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| Synonyms |
Benzarone; 1477-19-6; (2-Ethylbenzofuran-3-yl)(4-hydroxyphenyl)methanone; Benzaron; Methanone, (2-ethyl-3-benzofuranyl)(4-hydroxyphenyl)-;
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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 (~375.5 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (9.39 mM) (saturation unknown) in 10% DMSO + 40% PEG300 +5% Tween-80 + 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 25.0 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.7553 mL | 18.7765 mL | 37.5530 mL | |
| 5 mM | 0.7511 mL | 3.7553 mL | 7.5106 mL | |
| 10 mM | 0.3755 mL | 1.8777 mL | 3.7553 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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