1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEAM095 free acidCat. No.: HY-16040CAS No.: 1228690-36-5分式: CHNO分量: 456.49作靶點: LPL 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)體外實驗 DMSO : 67.3 mg/mL (147.43 mM; Need ultra

2、sonic and warming)Mass Solvent1 mg 5 mg 10 mg Concentration制備儲備液1 mM 2.1906 mL 10.9531 mL 21.9063 mL5 mM 0.4381 mL 2.1906 mL 4.3813 mL10 mM 0.2191 mL 1.0953 mL 2.1906 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。體內(nèi)實驗 請根據(jù)您的實驗動物和給藥式選擇適當(dāng)?shù)娜芙獍福渲魄罢埾扰渲瞥吻宓膬湟海僖来翁砑又軇?為保證實驗結(jié)果的可靠性，體內(nèi)實驗的作液，建議您現(xiàn)現(xiàn)配，當(dāng)天使；澄清的儲備液

3、可以根據(jù)儲存條件，適當(dāng)保存；以下溶劑前的百分 指該溶劑在您配制終溶液中的體積占)：1. 請依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 2.25 mg/mL (4.93 mM); Clear solutionBIOLOGICAL ACTIVITY物活性 AM095 free acid種有效的 LPA1 受體拮抗劑，對重 組和 LPA1 的 IC50 值分別為 0.98 和 0.73 M。1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEIC50 & Target IC50: 0.98 M (human LP

4、A1), 0.73 M (mouse LPA1)體外研究 AM095 inhibits the LPA-induced calcium flux of CHO cells stably transfected with human or mouse LPA1.The IC50 for AM095 antagonism of LPA-induced calcium flux of human or mouse LPA1-transfected CHOcells is 0.025 and 0.023 M, respectively 1. AM095 reduces LPA-induced vaso

5、relaxation by appr 90% at 10M as compared to vehicle control 2. AM095 inhibits LPA-driven chemotaxis of CHO cells overexpressingmouse LPA1 (IC50=778 nM) and human A2058 melanoma cells (IC50=233 nM) 3.體內(nèi)研究 Pharmacological antagonism of LPA1 with AM095 significantly attenuates bleomycin-induced dermal

6、 fibrosis1. AM095 has high oral bioavailability and a moderate half-life and is well tolerated at the doses tested inrats and dogs after oral and intravenous dosing. AM095 dose-dependently reduces LPA-stimulated histaminerelease. AM095 attenuates bleomycin-induced increases in collagen, protein, and

7、 inflammatory cellinfiltration in bronchalveolar lavage fluid. AM095 decreases kidney fibrosis in a mouse unilateral ureteralobstruction model 3.PROTOCOLKinase Assay 3 Assays are conducted using both hLPA1/CHO and mLPA1/CHO cells. A cell pellet of hLPA1/CHO ormLPA1/CHO cells is resuspended in appr 2

8、0 mL of ice-cold membrane buffer containing 10 mM HEPES, pH7.4, 1 mM dithiothreitol, and protease inhibitors. Cells are sonicated, and the cell lysate is centrifuged at 2000rpm for 10 min at 4C. The supernatant is further centrifuged at 25,000 rpm for 70 min at 4C. The membranepellet is resuspended

9、in 5 mL of ice-cold membrane buffer and homogenized using a Potter-Elvehjem tissuegrinder. Final protein concentration is determined using the Bradford Protein Assay Kit. Known amounts ofAM095 (diluted in dimethyl sulfoxide) or vehicle (dimethyl sulfoxide) are added to 25 to 40 g of hLPA1/CHOor mLPA

10、1/CHO membranes and 0.1 nM 35S-GTPS in buffer (50 mM HEPES, 0.1 mM NaCl, 10 mMMgCl2, 50 g/mL saponin, pH 7.5) containing 0.2% fatty acid-free human serum albumin and 5 M GDP. Totest for LPA1 antagonist activity, the ability of AM095 to inhibit GTPS binding stimulated by 900 nM LPA(18:1) is measured.

11、 Alternatively, to test for agonist effects, the ability of AM095 to stimulate GTPS bindingin the absence of LPA is measured. Reactions are incubated for 30 min at 30C, before harvestingmembranes onto glass filter binding plates and washing three times with cold buffer containing 50 mMHEPES, pH 7.4,

12、 100 mM NaCl, 10 mM MgCl2 using a Brandel 96-tip cell harvester. Plates are dried and thencpm are evaluated by using a Packard TopCount NXT microplate scintillation counter.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice underwent UUO or sha

13、m surgery to the left kidney. In brief, a longitudinal, upper left incision isAdministration 3 performed to expose the left kidney. The renal artery is located and a 6/0 silk thread is passed between theartery and the ureter. The thread is looped around the ureter and knotted three times insuring fu

14、ll ligation ofureter. The kidney is returned to abdomen, the abdominal muscle is sutured, and the skin is closed withstaples. The contralateral (right) kidney served as an uninjured control. AM095 (30 mg/kg) or vehicle (water)is given 1 to 4 h before UUO and then b.i.d. thereafter by oral gavage. Af

15、ter 8 days, mice are euthanizedusing inhaled CO2, and the kidneys are harvested and cut in half for histopathological and biochemicalanalysis of fibrosis. To assess fibrosis, half of the kidney is fixed in 10% neutral buffered formalin and stainedusing Massons trichrome. The other half of the kidney

16、 is frozen at 80C for subsequent biochemical2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEanalysis of collagen content.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 Biomol Ther (Seoul). 2017 Mar 1;25(2):194-201. Biomol Ther (Seoul). 201

17、4 Feb;22(2):129-35. Biochem Biophys Res Commun. 2015 May 29;461(2):378-82.See more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Castelino FV, et al. Amelioration of dermal fibrosis by genetic deletion or pharmacologic antagonism of lysophosphatidic acid receptor 1in a mouse model of

18、scleroderma. Arthritis Rheum. 2011 May;63(5):1405-15.2. Ruisanchez E, et al. Lysophosphatidic acid induces vasodilation mediated by LPA1 receptors, phospholipase C, and endothelial nitricoxide synthase. FASEB J. 2014 Feb;28(2):880-90.3. Swaney, J. S., et al. Pharmacokinetic and pharmacodynamic characterization of an oral lysophosphatidic acid type 1 receptor-selectiveantagonist. Jo