Biochanin-A-4-Methylgenistein-DataSheet-生命科學(xué)試劑-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEBiochanin ACat. No.: HY-14595CAS No.: 491-80-5Synonyms: 4-Methylgenistein; Olmelin分式: CHO分量: 284.26作靶點(diǎn): FAAH作通路: Metabolic Enzyme/Protease; Neuronal Signaling儲存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month

2、溶解性數(shù)據(jù)體外實(shí)驗 DMSO : 51 mg/mL (179.41 mM)* means soluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制備儲備液1 mM 3.5179 mL 17.5895 mL 35.1791 mL5 mM 0.7036 mL 3.5179 mL 7.0358 mL10 mM 0.3518 mL 1.7590 mL 3.5179 mL請根據(jù)產(chǎn)品在不同溶劑中的溶解度，選擇合適的溶劑配制儲備液，并請注意儲備液的保存式和期限。BIOLOGICAL ACTIVITY物活性 Bioch

3、anin A種天然存在的脂 酸酰胺解酶 (FAAH) 抑制劑，抑制FAAH，作于，F(xiàn)AAH，IC50 分別為 1.8，1.4 和 2.4 M。IC50 & Target IC50: 1.8 M (mouse FAAH), 1.4 M (rat FAAH), 2.4 M (human FAAH) 11/3 Master of Small Molecules 您邊的抑制劑師www.MedChemE體外研究 Biochanin A inhibits the hydrolysis of 0.5 M AEA by mouse, rat and human FAAH with IC50 s of 1.8,

4、 1.4 and2.4 M respectively. FAAH is inhibited by Biochanin A with a pIC50 value of 6.210.02, corresponding to anIC50 value of 0.62 M. Biochanin A produces significant inhibition of the URB597-sensitive tritium retentionat high nanomolar-low micromolar concentrations. Experiments are run with human F

5、AAH and 0.5 M3HAEA with assay conditions giving these higher utilization rates, the activity is still inhibited by BiochaninA, Genistein, Formononetin and Daidzein in the low micromolar range (IC50s of 6.0, 8.4, 12 and 30 M,respectively) 1.體內(nèi)研究 Biochanin A is tested at doses of 30, 100 and 300 g. Th

6、e highest dose also reduced formalin-induced ERKphosphorylation in a manner antagonized by AM251. Thus, Biochanin A behaved like URB597 after localadministration to the paw. In anaesthetized mice, URB597 (30 g i.pl.) and Biochanin A (100 g i.pl.) bothinhibit the spinal phosphorylation of extracellul

7、ar signal-regulated kinase produced by the intraplantarinjection of formalin. The effects of both compounds are significantly reduced by the CB1 receptorantagonist/inverse agonist AM251 (30 g i.pl.). Biochanin A (15 mg/k i.v.) does not increase brain AEAconcentrations, but produces a modest potentia

8、tion of the effects of 10 mg/kg i.v. AEA in the tetrad test.Biochanin A (15 mg/kg i.v.) is without effects on its own, but significantly potentiates the effects of AEA (10mg/kg i.v.) 1.PROTOCOLKinase Assay 1 For experiments with FAAH, rat liver homogenates, mouse brain homogenates and membranes from

9、 COS7cells transfected with the human enzyme are used. Frozen (80C) livers from adult C57BL/6 mice andfrozen brains (minus cerebella) from adult Wistar or Sprague-Dawley rats are thawed and homogenized in 20mM HEPES, 1 mM MgCl2, pH 7. The homogenates are centrifuged at 35000g for 20 min at 4C. After

10、resuspension in buffer followed by recentrifugation and a second resuspension in buffer, the pellets areincubated at 37C for 15 min. This incubation is undertaken in order to hydrolyse all endogenous FAAHsubstrates. The homogenates are then centrifuged as above, recentrifuged and resuspended in 50 m

11、M Tris-HCl buffer, pH 7.4, containing 1 mM EDTA and 3 mM MgCl2. The homogenates are then frozen at 80C inaliquots until used for assay. FAAH is assayed in the homogenates and in the COS7 cell membranes using0.5 M (unless otherwise stated) 3HAEA labelled in the ethanolamine part of the molecule. Blan

12、k values areobtained by the use of buffer rather than homogenate. In the experiments comparing effects of Biochanin Aupon FAAH and FAAH-2, the same assay is used but with 16 nM 3Holeoylethanolamide (3HOEA) assubstrate and with an incubation phase at room temperature. The choice of OEA rather than AE

13、A for FAAH-2is motivated by the relative rates of hydrolysis: OEA is metabolized four times faster than AEA by FAAH-2,whereas for FAAH the rate of hydrolysis of OEA is about a third of that for AEA. When 0.5 M 3HAEA isused as substrate, assay conditions for rat brain and mouse liver are chosen so th

14、at 3HAEA is metabolizedin all cases. For 16 nM 3HOEA, a limited supply of an expensive ligand meant that optimization is notpossible, and the amount of substrate utilized is higher (341 and 0.50.1% for FAAH and its correspondingmock-transfected, respectively; 402 and 210.4 for FAAH-2 and its corresp

15、onding mock-transfectedrespectively) 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice 12/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEAdministration 1 ICR mice are used for the behavioural tests measuring spontaneous activity (over a 10 m

16、in testing period),rectal temperature, ring immobility (over a 5 min testing period) and nociceptive threshold (tail flick tests).AEA and Biochanin A are dissolved in a vehicle consisting of ethanol, Emulphor-620 and physiological salinein a ratio of 1:1:18 v/v, and administered i.v. to the animals

17、via the tail vein (injection volume 10 L/g bodyweight). The degree of antinociception is expressed as percentage of maximum possible effect (%MPE),defined as (test-control time)/(10-control time)100.MCE has not independently confirmed the accuracy of these methods. They are for reference only.REFERENCES1. Thors L, et al. Biochanin A, a naturally occurring inhibitor of fatty a