罗氟司特作用机制 - Medchemexpress - MCE中国

1、Product Data SheetRoflumilastCat. No.: HY-15455CAS No.: 162401-32-3分式: CHClFNO分量: 403.21作靶点: Phosphodiesterase (PDE); RSV作通路: Metabolic Enzyme/Protease; Anti-infection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 (124.00 mM)H2O : 0.1 mg/mL (insoluble

2、)* means soluble, but saturation unknown.SolventMass1 mg 5 mg 10 mgConcentration制备储备液1 mM 2.4801 mL 12.4005 mL 24.8010 mL5 mM 0.4960 mL 2.4801 mL 4.9602 mL10 mM 0.2480 mL 1.2400 mL 2.4801 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；旦配成溶液，请分装保存，避免反复冻融造成的产品失效。储备液的保存式和期限：-80C, 6 months; -20C, 1 month。-80C 储存时，请在 6 个

3、内使，-20C 储存时，请在 1 个内使。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。以下溶解案都请先按照 In Vitro 式配制澄清的储备液，再依次添加助溶剂：为保证实验结果的可靠性，澄 的储备液可以根据储存条件，适当保存；体内实验的作液，建议您现现配，当天 使； 以下溶剂前显的百分 指该溶剂在您配制终溶液中的体积占；如在配制过程中出现沉淀、析出现象，可 以通过加热和/或超声的式助溶1. 请依序添加每种溶剂： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (6.20 mM); Clear solution此案可获得 2.5 mg/mL (6

4、.20 mM，饱和度未知) 的澄 溶液，此案不适于实验周 期在半个以上的实验。以 1 mL 作液为例，取 100 L 25.0 mg/mL 的澄 DMSO 储备液加到 900 L 油中，混合均匀。Page 1 of 2 www.MedChemEBIOLOGICAL ACTIVITY物活性 Roflumilast种选择性的 PDE4 抑制剂，作于 PDE4A1，PDEA4，PDEB1 和 PDEB2，IC50 分别为 0.7，0.9，0.7和 0.2 nM。IC & Target IC50: 0.7 nM (PDE4A1), 0.9 nM (PDE4A4), 0.7 nM (PDE4B1), 0

5、.2 nM (PDE4B2)1体外研究 Roflumilast does not affect PDE enzymes apart from PDE4, and is a subnanomolar inhibitor of most PDE4 splicingvariants tested. It showed no PDE4 subtype selectivity apart from PDE4C (4C1, IC50=3 nM; 4C2, IC50=4.3 nM), which isinhibited with a slightly lower potency2. Roflumilast

6、is a potent and selective PDE4 inhibitor. Roflumilast is amonoselective PDE4 inhibitor since it does not affect other PDE isoenzymes, including PDE1, PDE2, PDE3, and PDE5up to 10,000-fold higher concentrations. Roflumilast inhibits human neutrophil functions. Roflumilast inhibits TNFsynthesis in mon

7、ocyte-derived dendritic cells. Rolfumilast inhibits proliferation and cytokine synthesis in CD4+ T cells.Proliferation is inhibited to a maximum of about 60% by Roflumilast with a potency (IC30) of 7 nM3.体内研究 Animal studies with Roflumilast demonstrated that it reduced the accumulation of neutrophil

8、s in bronchoalveolarlavage fluid following short-term exposure of guinea pigs, mice or rats to tobacco smoke, and following exposure ofrats to a combination of tobacco smoke and bacterial lipopolysaccharide, and abolished the lung parenchymal influxof inflammatory cells seen in rats exposed to tobac

9、co smoke for 7 months2. Roflumilast blocks COPD progression inpIgR/ mice. For these studies, 9-month-old WT or pIgR/ mice are treated daily by oral gavage with 100 g ofRoflumilast (5 g/g) or vehicle (4% methylcellulose, 1.3% PEG400) for 3 months and lungs are harvested at 12 monthsof age. Unlike pIg

10、R/ mice treated with vehicle, mice treated with Roflumilast had no progression of small airwaywall remodelling after starting treatment. Strikingly, 12-month-old pIgR/ mice treated with Roflumilast had reducedindices of emphysema compared with 9-month-old pIgR/ mice, indicating that Roflumilast not

11、only blocksprogression of emphysema in this model but apparently facilitates some resolution of the emphysematousdestruction of lung parenchyma4.PROTOCOLKinase Assay 3 PDE activity is determined with some modifications. The assay mixture contain 50 mM Tris (pH 7.4), 5 mM MgCl2, 0.5M cAMP or cGMP, an

12、d 3HcAMP or 3HcGMP (about 30,000 cpm/assay), the indicated concentration of theinhibitor and an aliquot of the enzyme solution at a final assay volume of 200 L. Stock solutions of the compoundsare diluted 1:100 (v/v) in the Tris buffer mentioned above; appropriate dilutions are prepared in 1% (v/v)

13、DMSO/Trisbuffer, which are diluted 1:2 (v/v) in the assays to obtain the desired final concentrations of the inhibitors at a DMSOconcentration of 0.5% (v/v). DMSO itself affected none of the PDE activities. After preincubation for 5 min at 37C, thereaction is started by the addition of substrate (cA

14、MP or cGMP) and the assays are incubated for further 15 min at37C. Then 50 L of 0.2 N HCl is added to stop the reaction and the assays are left on ice for about 10 min. Followingincubation with 25 g of 5-nucleotidase (Crotalus atrox snake venom) for 10 min at 37C, the assays are loaded onQAE Sephade

15、x A-25 (1 mL of bed volume in Poly-Prep chromatography columns). The columns are eluted with 2 mLof 30 mM ammonium formate (pH 6.0) and the eluate is counted for radioactivity. Results are corrected for blankvalues (measured in the presence of denatured protein) that are below 5% of total radioactiv

16、ity. The amount of cyclicnucleotides hydrolyzed did not exceed 30% of the original substrate concentration3.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice4Administration 4 WT or pIgR/ mice are used. For studies using Roflumilast, 200 L of 0

17、.5 mg/mL suspension of Roflumilast or vehicle(4% methylcellulose, 1.3% PEG400 and 5 g drug per mg animal weight) is administered by oral gavage once daily, 5days a week for the duration of treatment. Mice are treated daily by oral gavage with 100 g of Roflumilast (5 g/g)or vehicle (4% methylcellulos

18、e, 1.3% PEG400) for 3 months and lungs are harvested at 12 months of age.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Page 2 of 3 www.MedChemE户使本产品发表的科研献 Int Urol Nephrol. 2018 Nov 24. Int Urol Nephrol. 2017 Oct;49(10):1723-1730.See more customer val

19、idations on HYPERLINK www.MedChemE www.MedChemEREFERENCES1. Hatzelmann A, et al. The preclinical pharmacology of roflumilast-a selective, oral phosphodiesterase 4 inhibitor in development for chronic obstructivepulmonary disease. Pulm Pharmacol Ther. 2010 Aug;23(4):235-56.2. Rabe KF. Update on roflumilast, a phosphodiesterase 4 inhibitor for the treatment of chronic obstructive pulmonary disease. Br J Pharmacol. 2011May;163(1):53-67.3. Hatzelmann A, et al. Anti-inflammatory and immunomodulatory pote