Coumberone is a metabolic fluorescent probe with isoform selectivity for all AKR1C isoforms. All four members of the AKR1C family can reduce coumberone to its fluorescent alcohol coumberol. Coumberone may be utilized in AKR1C research.

Physicochemical Properties

Molecular Formula	C22H19NO3
Molecular Weight	345.39
Exact Mass	345.136
CAS #	878019-47-7
PubChem CID	11588246
Appearance	Light yellow to yellow solid powder
LogP	3.787
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	2
Heavy Atom Count	26
Complexity	621
Defined Atom Stereocenter Count	0
InChi Key	DGJJSADMDWUKOJ-UHFFFAOYSA-N
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
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
HS Tariff Code	2934.99.9001
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)

Biological Activity

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].
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.
References	[1]. A novel fluorometric assay for aldo-keto reductase 1C3 predicts metabolic activation of the nitrogen mustard prodrug PR-104A in human leukaemia cells. Biochem Pharmacol. 2014;88(1):36-45. [2]. Imaging induction of cytoprotective enzymes in intact human cells: coumberone, a metabolic reporter for human AKR1C enzymes reveals activation by panaxytriol, an active component of red ginseng. J Am Chem Soc. 2008;130(43):14123-14128.

Solubility Data

Solubility (In Vitro)

DMSO: 16.67 mg/mL (48.26 mM )

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.