Dihydromyricetin-Ampeloptin-DataSheet-生命科学试剂-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEDihydromyricetinCat. No.: HY-N0112CAS No.: 27200-12-0Synonyms: Ampeloptin; Ampelopsin分式: CHO分量: 320.25作靶点: mTOR; Autophagy作通路: PI3K/Akt/mTOR; Autophagy储存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性数据体外

2、实验 DMSO : 100 mg/mL (312.26 mM)* means soluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制备储备液1 mM 3.1226 mL 15.6128 mL 31.2256 mL5 mM 0.6245 mL 3.1226 mL 6.2451 mL10 mM 0.3123 mL 1.5613 mL 3.1226 mL请根据产品在不同溶剂中的溶解度，选择合适的溶剂配制储备液，并请注意储备液的保存式和期限。体内实验请根据您的实验动物和给药式选择适当的溶解案，配制前请先配制澄

3、清的储备液，再依次添加助溶剂(为保证实验结果的可靠性，体内实验的作液，建议您现现配，当天使；澄清的储备液可以根据储存条件，适当保存；以下溶剂前的百分 指该溶剂在您配制终溶液中的体积占)：1. 请依序添加每种溶剂： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (7.81 mM); Clear solution2. 请依序添加每种溶剂： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.5 mg/mL (7.81 mM); Clear solution1/3 Mast

4、er of Small Molecules 您边的抑制剂师www.MedChemE3. 请依序添加每种溶剂： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (7.81 mM); Clear solutionBIOLOGICAL ACTIVITY物活性 Dihydromyricetin种有效的氢嘧啶酶 (dihydropyrimidinase) 抑制剂，IC50 为 48 M。Dihydromyricetin 可通过抑制 mTOR 信号从激活噬。Dihydromyricetin 抑制 mTOR 复合体 (mTORC1/2)形成。IC50 & Target

5、Dihydropyrimidinase mTORC1 mTORC2 Autophagy48 M (IC50)体外研究 Dihydromyricetin, a flavonol, significantly inhibits the catalytic activities of dihydropyrimidinase toward boththe natural substrate dihydrouracil and xenobiotic substrate 5-propyl-hydantoin. Dihydromyricetin exhibits asignificant inhibitor

6、y effect on the activities of dihydropyrimidinase for both substrates, even more thanMyricetin does. The IC50 values of Dihydromyricetin for dihydropyrimidinase determined from the titrationcurves using Dihydrouracil and 5-propyl-hydantoin are 482 and 402 M, respectively 1. Dihydromyricetin(DHM) sup

7、plementation significantly reverses the increased phosphorylation of mTOR at Ser2448 (p-mTOR)during D-gal administration, which suggests that Dihydromyricetin can activate autophagy through inhibitingmTOR signaling 2.体内研究 Changes in learning and memory capacity in rats administrated normal control g

8、roup, D-gal group, D-gal+Dihydromyricetin (100 mg/kg) group, D-gal+Dihydromyricetin (200 mg/kg) group assessed by morriswater maze (MWM) (n=10 per group). Dihydromyricetin (DHM) treatment significantly shortens the escapelatency when compared with D-gal-induced model group 2.PROTOCOLKinase Assay 1 A

9、 rapid spectrophotometric assay is used to determine the enzymatic activity for hydantoinase, allantoinase,dihydroorotase, and imidase. Dihydrouracil, 5-propyl-hydantoin, and phthalimide are used as substrates.Unless explicitly stated otherwise, Dihydrouracil (2 mM) is used as the substrate in the s

10、tandard assay ofdihydropyrimidinase. Briefly, the decrease in absorbancy at 230, 248, and 298 nm is measured uponhydrolysis of Dihydrouracil, 5-propyl-hydantoin, and Phthalimide as the substrate at 25C, respectively. Tostart the reaction, the purified dihydropyrimidinase (10-70 g) is added to a 2 mL

11、 solution containing thesubstrate and 100 mM Tris-HCl (pH 8.0). Substrate hydrolysis is monitored with a UV/vis spectrophotometer.The extinction coefficient of each substrate is determined experimentally by direct measurement with aspectrophotometer. The extinction coefficients of Dihydrouracil, 5-p

12、ropyl-hydantoin, and Phthalimide are0.683 mM-1cm-1 at 230 nm, 0.0538 mM-1cm-1 at 248 nm, and 3.12 mM-1cm-1 at 298 nm, respectively. Theinitial rates of change are a function of enzyme concentration within the absorbance range of 0.01-0.18 min-1. A unit of activity is defined as the amount of enzyme

13、catalyzing the hydrolysis of 1 mol substrate/min, andthe specific activity is expressed in terms of units of activity per milligram of enzyme. The kinetic parametersKm and Vmax are determined from a non-linear plot by fitting the hydrolyzing rate from individual2/3 Master of Small Molecules 您边的抑制剂师w

14、ww.MedChemEexperiments to the Michaelis-Menten equation 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 2 Hippocampus and cortex tissue samples are homogenized in lysis buffer containing 20 mM Tris (pH 7.5), 135mM NaCl, 2 mM EDTA, 2 mM DTT,

15、 25 mM -glycerophosphate, 2 mM sodium pyrophosphate, 10% glycerol,1% Triton X-100, 1 mM sodium orthovanadate, 10 mM NaF, 10 g/mL aprotinin, 10 g/mL leupeptin, and 1mM PMSF for 30 min on ice and centrifuged at 12000g at 4C for 30 min. The supernatant is collected andprotein quantification is carried

16、out using a BCA kit. The protein samples are boiled in the presence ofsample buffer at 95C for 5 min. The target protein is separated by sodium dodecyl sulfate polyacrylamidegel electrophoresis (SDS), transferred to nitrocellulose membrane, and then probed by correspondingprimary and secondary antib

17、odies. Finally, the target protein is visualized by enhanced chemiluminescence(ECL) reagent exposure to X-ray film 2.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Rats 2Administration 2 Totally 40 male Sprague-Dawley (SD) rats (age: 8 weeks old

18、; body weight: 16020 g) are used. The rats arerandomly divided into four groups including normal control group, D-gal model group, and D-gal combinedwith DHM at the doses of 100 and 200 mg/kg-d groups with 10 rats in each group. All rats are housed at theenvironment with room temperature of 222C and

19、 a dark-light cycle (12 h: 12h), and provided theaccessibility to food and water ad libitum. After adapting to new environment for 1 week, the rats from DHMgroups are administered with DHM dissolved in distilled water at the designated dosages by gavage once aday at 8:00am for 6 consecutive weeks. T

20、he rats from the normal control group are administrated withdistilled water. Except from the normal control group, the rats from other groups are subjected tosubcutaneous injection of D-gal at the dose of 150 mg/kg.d for 6 consecutive weeks. Each administration ofDHM should be 2 h ahead of D-gal inj

21、ection.MCE has not independently confirmed the accuracy of these methods. They are for reference only.户使本产品发表的科研献 Mol Ther. 2019 May 8;27(5):1051-1065. Biochem Biophys Res Commun. 2018 Sep 3;503(1):297-303.See more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Huang CY. Inhibition of a Putative Dihydropyrimi