GW-1100-DataSheet-生命科学试剂-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEGW-1100Cat. No.: HY-50691CAS No.: 306974-70-9分式: CHFNOS分量: 520.58作靶点: GPR40作通路: GPCR/G Protein储存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性数据体外实验 DMSO : 50 mg/mL (96.05 mM; Need ultrasonic)H2O : 90% c

2、orn oilSolubility: 2.5 mg/mL (4.80 mM); Clear solutionBIOLOGICAL ACTIVITY1/3 Master of Small Molecules 您边的抑制剂师www.MedChemE物活性 GW-1100选择性的 GPR40 拮抗剂，pIC50为6.9。IC50 & Target pIC50: 6.9 (GPR40) 1体外研究 GW-1100 (GW1100) dose dependently inhibits GPR40-mediated Ca2+ elevations stimulated by GW9508 andlinol

3、eic acid (pIC50 values of 5.990.03 and 5.990.06, respectively). GW-1100 at a concentration of 1 Mproduces a significant rightward shift in the concentration-response curve to GW9508 (pEC50=7.170.08 inthe absence and pEC50=6.790.09 in the presence of 1 M GW-1100; P50 response 2. GW-1100 (GW1100)reduc

4、es FFAR1 ligand-induced intracellular calcium in CHO-K1/bFFAR1 cells and neutrophils. CHO-K1/bFFAR1 cells are incubated for 15 min with 10 M GW1100 or vehicle (0.1% DMSO) and then stimulatedwith vehicle, oleic acid, linoleic acid or GW9508. GW-1100 significantly reduces the increase in intracellular

5、calcium induced by 300 M oleic acid (AUC(60-150 s), p(60-150 s), p(60-150 s), p 3.体内研究 The intracerebroventricular injection of DHA (50 g) and GW9508 (1.0 g), a GPR40-selective agonist,significantly reduces mechanical allodynia and thermal hyperalgesia at day 7, but not at day 1, after CFAinjection.

6、 These effects are inhibited by intracerebroventricular pretreatment with GW-1100 (10 g), a GPR40antagonist 4.PROTOCOLCell Assay 3 CHO-K1/bFFAR1 or CHO-K1/pcDNA3.1 cells (2106 cells/2 mL) are loaded with 2.5 M Fura-2AMfluorescent indicator dye in recording buffer (10 mM HEPES, 140 mM NaCl, 2 mM CaCl

7、2, 21 mM MgCl2, 25mM KCl, 10 mM glucose, pH 7.4) for 30 min, washed three times with recording buffer, and returned to theincubator for 10 min. Cells are incubated with different concentrations of propionic acid (1, 10 and 30 mM),oleic acid (0-500 M), linoleic acid (0-200 M), GW9508 (0-100 M), ionom

8、ycin (2 M), thapsigargin (2 M)or vehicle (0.1% DMSO). The fatty acid concentrations used in all experiments are in the range ofconcentrations of healthy and peripartum cows. In another set of experiments, cells are incubated with either10 M GW-1100 for 15 min, 2 M U73122 for 3 min or vehicle (0.1% D

9、MSO) for 15 min and then stimulatedwith either 300 M oleic acid, 100 M linoleic acid or 10 M GW9508. Cellular fluorescence (Ca2+) ismeasured at 509 nm emission with 340/380 nm dual wavelength excitation using a LS55 spectrofluorimeter.Cuvette temperatures are maintained at 37C with constant stirring

10、 3.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice 4Administration 4 Male ddY mice (age, 4 weeks) are housed in cages at 23-24C with a 12-h light-dark cycle (lights from 8 amto 8 pm) and food and waterad libitum. DHA (50 g/mouse), the select

11、ive GPR40-agonist GW9508 (1.0-25g/mouse) and the GPR40 antagonist GW1100 (1-10 g/mouse) are dissolved in 1% DMSO and the solutionis diluted with saline before von Frey testing (1% DMSO final concentration). The doses of GW9508 arechosen based upon our previous publication, whereas GW-1100 is selecte

12、d on the basis of previous reportsand our preliminary experiments. Under a non-anesthetized state, DHA and GW9508 are administered viathe intracerebroventricular (i.c.v.) route 10 min before CFA injection, and GW1100 is administered via thei.c.v. route 10 min before GW9508 injection. Flavopiridol (5

13、 and 15 nmol/mouse), a cyclin-dependent kinaseinhibitor, is administered by i.c.v. injection into the left lateral ventricle of the mice twice a day (at 9:00 and2/3 Master of Small Molecules 您边的抑制剂师www.MedChemE19:00) after CFA treatment.MCE has not independently confirmed the accuracy of these metho

14、ds. They are for reference only. J Endocrinol. 2018 Nov 1. pii: JOE-18-0432.R1. Sci Rep. 2017 Jun 28;7(1):4351. Am J Pathol. 2015 Jan;185(1):185-96. J Agric Food Chem. 2019 Feb 20. RSC Adv. 2019 May.See more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Stoddart LA, et al. Uncovering

15、the pharmacology of the G protein-coupled receptor GPR40: high apparent constitutive activity inguanosine 5-O-(3-35Sthio)triphosphate binding studies reflects binding of an endogenous agonist. Mol Pharmacol. 2007 Apr;71(2. Briscoe CP, et al. Pharmacological regulation of insulin secretion in MIN6 ce

16、lls through the fatty acid receptor GPR40: identification ofagonist and antagonist small molecules. Br J Pharmacol. 2006 Jul;148(5):619-28.3. Manosalva C, et al. Cloning, identification and functional characterization of bovine free fatty acid receptor-1 (FFAR1/GPR40) inneutrophils. PLoS One. 2015 Mar 19;10(3):e0119715.4. Nakamoto K, et al. Hypothalamic GPR40 signaling activated by free long chain fatty acids suppresses CFA-induced inflammatory