AVE-0991 - Angiotensin Receptor Agonist - 生命科學(xué)試劑 - MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEAVE 0991Cat. No.: HY-15778CAS No.: 304462-19-9分式: CHNOS分量: 580.72作靶點(diǎn): Angiotensin Receptor作通路: GPCR/G Protein儲存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實(shí)驗(yàn) DMSO : 100 mg/mL (172.20 mM)H2O : 40% P

2、EG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (4.31 mM); Clear solution2. 請依序添加每種溶劑： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.5 mg/mL (4.31 mM); Suspended solution; Need ultrasonic1/4 Master of Small Molecules 您邊的抑制劑師www.MedChemE3. 請依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (4

3、.31 mM); Clear solutionBIOLOGICAL ACTIVITY物活性 AVE 0991多肽，有服活性的管緊張素-(1-7)受體激動(dòng)劑，IC50 為21 nM。IC50 & Target IC50: 2135 nM (Ang-(1-7) receptor) 1體外研究 AVE 0991 is a nonpeptide compound that evokes effects similar to Ang-(1-7) on the endothelium. AVE 0991and unlabeled Ang-(1-7) compete for high-affinity bi

4、nding of 125I-Ang-(1-7) to bovine aortic endothelial cellmembranes with IC50s of 2135 and 220280 nM, respectively. Peak concentrations of NO and O2- releaseby AVE 0991 sodium salt and Ang-(1-7) (both 10 M) are not significantly different (NO: 29520 and 27025nM; O2-: 182 and 204 nM). However, the rel

5、eased amount of bioactive NO is 5 times higher for AVE0991 in comparison to Ang-(1-7) 1.體內(nèi)研究 AVE 0991 (0.58 nmol/g) produces a significant decrease of water diuresis in WT mice compared withvehicle-treated animals (0.060.03 mL versus 0.270.05; n=9 for each group; P2O versus 681.1165.8mOsm/KgH2O in v

6、ehicle-treated mice; PMas abolishes the antidiuretic effect of AVE 0991 during waterloading (0.370.10 mL n=9 versus 0.270.03 mL n=11 in AVE 0991-treated mice). As observed withC57BL/6 mice, administration of AVE 0991 (0.58 nmol/g) in water-loaded Swiss mice also produces asignificant decrease of the

7、 urinary volume compared with vehicle-treated animals (0.130.05 mL n=16versus 0.510.04 mL n=40; P 2. One week of treatment with AVE-0991 produces a significant decrease inperfusion pressure (56.550.86 vs. 68.730.69 mmHg in vehicle-treated rats) and an increase in systolictension (11.400.05 vs. 9.840

8、.15 g in vehicle-treated rats), rate of tension rise (+dT/dt; 184.300.50 vs.155.201.97 g/s in vehicle-treated rats), rate of tension fall (dT/dt; 179.601.39 vs. 150.802.42 g/s invehicle-treated rats). A slight increase in heart rate (HR) is also observed (220.400.71 vs. 214.200.74beats/min in vehicl

9、e-treated rats 3.PROTOCOLKinase Assay 1 Binding of 125I-Ang-(1-7) is performed. Briefly, 100 g of membranes from primary cultured bovine aorticendothelial cells (BAECs, passage 1) are incubated in a total volume of 200 L for 45 minutes at 25C inHEPES-buffered saline (10 mM HEPES, 0.1 M NaCl, 5 mM Mg

10、Cl2) containing 0.2% BSA and proteaseinhibitor cocktail Complete (Boehringer Mannheim). Saturable binding of 125I-Ang-(1-7) is calculated bysubtracting nonspecific binding (40% to 50%), determined in the presence of 10 M unlabeled Ang-(1-7) fromtotal binding. Competition experiments with increasing

11、concentrations of AVE 0991 and unlabeled Ang-(1-7)are performed in the presence of 10 nM 125I-Ang-(1-7). Assays are terminated by vacuum filtration (15mm Hg) over Durapore filters (0.65 m, Opak 96-well plates) presoaked with 1% BSA. The filters are washed3 times with each 100 L of PBS (50 mM, NaHPO4

12、 and 0.15 M NaCl, pH 7.2). Radioactivity on dried filters isquantified with a gamma counter 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.2/4 Master of Small Molecules 您邊的抑制劑師www.MedChemECell Assay 1 Monkey kidney (COS) cells and Chinese hamster ova

13、ry (CHO) cells are stably transfected withrat Mas cDNA driven by a cytomegalovirus promoter and selected by neomycin. 125I-Ang-(1-7) (0.510-9 mol/L) is incubated in 24-well plates for 60 minutes at 4C in 0.3 mL of serum-free medium (DMEM)supplemented with 0.2% BSA, 0.005% bacitracin, 0.1 mol/L PMSF,

14、 and 0.5 mol/L orthophenanthrolinewith Mas-transfected COS cells in the presence or absence of AVE 0991 (AVE, 10-10 to -5 mol/L). After 2ishes with ice-cold serum-free DMEM, cells are disrupted with 0.1% Triton X-100. Bound radioactivity ismeasured in a gamma counter. Binding of rhodamine-Ang-(1-7)

15、in Mas-transfected CHO cells is performedunder similar conditions using 210-9 mol/L rhodamine-labeled-Ang-(1-7) in the presence or absence of AVE(10-6 mol/L), CV11974 (10-6 mol/L), or PD123319 (10-6 mol/L). NSB is determined in the presence of 10-6mol/L Ang-(1-7) 1.MCE has not independently confirme

16、d the accuracy of these methods. They are for reference only.Animal Mice 2Administration 23 Swiss male mice, Mas-KO (Mas-/-) male mice on the pure genetic background C57BL/6, and WT C57BL/6control mice (Mas+/+) are used. Water diuresis is induced by intraperitoneal water injection (0.05 mL/g ofbody

17、weight BW) in conscious mice. Drugs are administered in the same injection with water load atprefixed volumes (0.01 mL/g BW). In the first set of experiments, WT mice (C57BL/6, control group) or Mas-KO mice are treated with: (1) 0.58 nmol/g AVE 0991 (n=9, control; n=11, Mas-KO mice); or (2) vehicle

18、forAVE 0991 (10 M KOH, 0.01 mL/g; n=9, control; n=9, Mas-KO). In the second set, Swiss mice are treatedwith: (1) vehicle (n=36); (2) 0.58 nmol/g AVE 0991 (n=16); (3) 46 pmol/g Ang-(1-7) antagonist A-779 (n=4);(4) 2 nmol/g losartan or valsartan (n=5); (5) 2 nmol/g AT2 receptor antagonists PD123319 or

19、 PD123177(n=9); (6) AVE 0991 combined with A-779; (7) AVE 0991 combined with losartan or valsartan (n=4 for each);(8) or AVE 0991combined with PD123319 (n=5) or PD123177 (n=4). The urinary volume is measured for 60minutes after water loading, and urine samples are obtained to determine the osmolalit

20、y. The dose of AVE0991is based in preliminary experiments performed in Swiss mice.Rats 3Male Wistar rats weighting 250-300 g are used. Rats are treated either with AVE-0991 (1 mg/kg, n=9) orvehicle (0.9% NaCl, n=11) administered orally by gavage. At the end of the 7 day period of AVE-0991treatment,

21、the animals are decapitated 10-15 min after intraperitoneal injection of 400 IU of heparin. After thethorax is opened, the heart is carefully dissected, removed from the thoracic cavity, and placed in a platecontaining ice-cold Krebs-Ringer solution (KRS) to attenuate any potential cardiac damage du

22、ring dissectionof aorta artery.MCE has not independently confirmed the accuracy of these methods. They are for reference only.戶使本產(chǎn)品發(fā)表的科研獻(xiàn) Blood. 2015 Jan 22;125(4):710-9. Redox Biol. 2019 Jan;20:75-86. Diabetes. 2017 Aug;66(8):2201-2212. Aging (Albany NY). 2018 Apr 17;10(4):645-657. PLoS One. 2015 Nov 5;10(11):e0142087.See more customer validations on HYPERLINK / www.MedChemE3/4 Master of Small Molecules 您邊的抑制劑師www.MedChemEREFERENCES1. Wiemer G, et al. AVE 0991, a nonpeptide mimic of the effects of angiotensin-(1-7) on the endothelium. Hyperten