| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
AKR1C enzymes are the primary molecular targets. Coumberone is an isoform-selective substrate for all AKR1C family members (AKR1C1, AKR1C2, AKR1C3, and AKR1C4). It is enzymatically reduced to the fluorescent alcohol coumberol.
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| ln Vitro |
The maximum rates of coumberone (30 μM; 24 hours; HCT116 cells) are observed in AKR1C3 substrates [1]. One can utilize coumberone (5 μM; 24 hours; COS-1 cells) to selectively read out either isoform, AKR1C2 or AKR1C3 optically [2]. For IMR32 cells, coumberone (5 μM; 24 hours) allows for real-time imaging of AKR1C induction [2]. Coumberone did, in fact, boost metabolism (80 hours; IMR-32 cells). Ten times more catalytically efficient than AKR1C2 is cumberone for AKR1C3 in vitro [2].
In cell-free assays, Coumberone exhibits activity as a fluorogenic substrate for AKR1C enzymes. Upon reduction by AKR1C1-4 in the presence of NADPH cofactor, Coumberone is converted to coumberol, generating a strong fluorescence turn-on signal. It is used to measure AKR1C enzymatic activity. |
| ln Vivo |
There is no specific in vivo activity data available for this compound. As a fluorogenic probe for AKR1C activity, it can be used in live cells to monitor AKR1C induction, substrate specificity, and inhibitor efficacy in real time.
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| Enzyme Assay |
A standard protocol involves incubating Coumberone (e.g., 10-50 uM) with varying concentrations of purified AKR1C enzyme (e.g., AKR1C1, AKR1C2, AKR1C3, or AKR1C4) and the cofactor NADPH (e.g., 100-200 uM) in an assay buffer (e.g., 100 mM potassium phosphate, pH 7.4) at 37degC. The increase in fluorescence over time (e.g., Ex/Em: typically around 360/460 nm) is measured with a fluorescence spectrophotometer.
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| Cell Assay |
Immunofluorescence[2]
Cell Types: IMR32 cells Tested Concentrations: 5 μM Incubation Duration: 24 hrs (hours) Experimental Results: Enabled real-time imaging of AKR1C induction. A general protocol for cellular experiments involves seeding cells that express AKR1C enzymes (e.g., liver cells or cancer cell lines) in a 96-well plate and incubating them with Coumberone (e.g., 10-50 uM) in culture medium for 1-4 hours at 37degC. After incubation, the fluorescence in the medium or cell lysate is measured. A specific AKR1C inhibitor (e.g., medroxyprogesterone acetate, 1-10 uM) can be used as a control to confirm specificity. |
| Animal Protocol |
There is no standard in vivo protocol for this compound. A general protocol for studying AKR1C activity in animals would involve intravenous or intraperitoneal injection of Coumberone (e.g., 1-20 mg/kg) into a mouse model, followed by measurement of coumberol accumulation in tissues (e.g., liver, kidney) by fluorescence imaging or HPLC analysis.
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| ADME/Pharmacokinetics |
General PK properties for this small molecule probe are not applicable, as it is not a drug. For in vivo use, Coumberone would be formulated in DMSO:PEG:water (e.g., 10:40:50) or in saline with co-solvents.
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| Toxicity/Toxicokinetics |
General toxicity for fluorogenic probes is considered low. No specific toxicity studies are reported for Coumberone. Standard safety precautions for handling laboratory chemicals should be observed.
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| References |
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| Additional Infomation |
Coumberone is a research tool for studying aldo-keto reductase 1C enzymes, which are important in steroid hormone metabolism, prostaglandin synthesis, and are drug targets for cancer and inflammatory diseases. Coumberone can be used to screen for AKR1C inhibitors in high-throughput assays. This product is for research use only.
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| Molecular Formula |
C22H19NO3
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|---|---|
| Molecular Weight |
345.39
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| Exact Mass |
345.136
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| CAS # |
878019-47-7
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| PubChem CID |
11588246
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
3.787
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
26
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| Complexity |
621
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1CC2=CC3=C(C4=C2N(C1)CCC4)OC(=O)C=C3C(=O)C5=CC=CC=C5
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| InChi Key |
DGJJSADMDWUKOJ-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C22H19NO3/c24-19-13-17(21(25)14-6-2-1-3-7-14)18-12-15-8-4-10-23-11-5-9-16(20(15)23)22(18)26-19/h1-3,6-7,12-13H,4-5,8-11H2
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| Chemical Name |
6-benzoyl-3-oxa-13-azatetracyclo[7.7.1.02,7.013,17]heptadeca-1(17),2(7),5,8-tetraen-4-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 |
| 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: 16.67 mg/mL (48.26 mM )
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1.67 mg/mL (4.84 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 16.7 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.8953 mL | 14.4764 mL | 28.9528 mL | |
| 5 mM | 0.5791 mL | 2.8953 mL | 5.7906 mL | |
| 10 mM | 0.2895 mL | 1.4476 mL | 2.8953 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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