RS-127445-MT-500-DataSheet-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemERS 127445Cat. No.: HY-15419CAS No.: 199864-86-3Synonyms: MT 500分式: CHClFN分量: 317.79作靶點: 5-HT Receptor作通路: GPCR/G Protein; Neuronal Signaling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 : 3

2、1 mg/mL (97.55 mM)* means soluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制備儲備液1 mM 3.1467 mL 15.7337 mL 31.4673 mL5 mM 0.6293 mL 3.1467 mL 6.2935 mL10 mM 0.3147 mL 1.5734 mL 3.1467 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。BIOLOGICAL ACTIVITY物活性 RS 127445種選擇性的 5-HT2B 受體

3、拮抗劑，pKi 為 9.5。IC50 & Target 5-HT1A Receptor 5-HT1B/D Receptor 5-HT3 Receptor 5-HT5 Receptor5.5 (pKi) 6 (pKi) 6 (pKi) 6 (pKi)1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemE5-HT6 Receptor 5-HT2A Receptor 5-HT2C Receptor 5-HT2B Receptor98% of RS-127445 is protein-bound 2.PROTOCOLKinase Assay 1 CHO-K1

4、 cells expressing human 5-HT2A, 5-HT2B or 5-HT2C receptors are harvested using 2 mM EDTA inphosphate buffered saline. Cell membranes are prepared by four cycles of homogenization (Brinkman P10disrupter) and centrifugation (48,000g for 15 min). As previously described, each assay is established so as

5、to achieve steady state conditions and to optimize specific binding. For the 5-HT2A receptor, membranesfrom 1106 cells are incubated with 0.2 nM 3H-ketanserin at 32C for 60 min. Nonspecific binding isdetermined using 10 M methysergide. For the 5-HT2B receptor, membranes from 1.5106 cells areincubate

6、d with 0.2 nM 3H-5-HT at 4C for 120 min. Nonspecific binding is determined using 10 M 5-HT.For the 5-HT2C receptor, membranes from 3105 cells are incubated with 0.5 nM 3H-mesulergine at 32Cfor 60 min. Nonspecific binding is determined using 10 M methysergide. Assays are terminated by vacuumfiltratio

7、n through glass fibre filters (GF/B) which had been pretreated with 0.1% polyethyleneimine. Total andbound radioactivity is determined by liquid scintillation counting. Greater than 90% specific binding is2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEachieved in each of these assays.The selectivi

8、ty of RS-127445 for 5-HT2B receptors is further examined bytesting the compound for affinity at over 100 additional ion channel or receptor binding sites. These studiesare conducted by Cerep using standard protocols 1.MCE has not independently confirmed the accuracy of these methods. They are for re

9、ference only.Cell Assay 1 HEK-293 cells expressing the human 5-HT2B receptor are incubated with 3H-myoinositol (1.67 Ci/mL) in162 cm2 flasks overnight at 37C in an inositol free Hams F12 medium containing 10% dialyzed foetalbovine serum. The cells are harvested, washed five times with phosphate buff

10、ered saline and resuspendedin inositol free Hams F12 media at density of approximately 3106 cells/mL. RS-127445 (10 M) is initiallydissolved in 10% (v/v) DMSO with 90% inositol free Hams F12 medium. Subsequent dilutions are made withinositol free Hams F12 medium. 5-HT is dissolved in inositol free H

11、ams F12 medium containing 100 mM LiCland 1 mM ascorbate. RS-127445, vehicle or other antagonists are pre-incubated with 240 L of cellsuspension at 37C for 20 min. The reactions are initiated by addition of 5-HT. Sixty minutes later, thereactions are terminated by adding 50 L of ice-cold 20% perchlor

12、ic acid, chilled in an ice-water bath for 10min and then neutralized with 160 L of 1 N KOH. Each sample is diluted with 2 ml of 50 mM Tris-HCl, pH7.4 at room temperature. The aqueous portion (2.2 mL) is transferred onto Dowex AG1X8 columns (1 ml, 1 :1, w/v) which had been washed with 5 ml of distill

13、ed water. The columns are then washed with 18 ml ofdistilled water and the inositol phosphates are eluted with 3 ml of 1 N HCl. The eluted radioactivity isdetermined by liquid scintillation spectroscopy using a Packard 1900CA analyzer 1.MCE has not independently confirmed the accuracy of these metho

14、ds. They are for reference only.Animal Rats 1Administration 12 Male Sprague-Dawley rats (200 g) are used. To compare the plasma kinetics of RS-127445 followingdifferent routes of administration, 90 rats are distributed into three treatment groups of 30 rats each. A singledose of RS-127445 (5 mg/kg)

15、dissolved (2.5 mg/mL) in ethanol:propylene glycol : water (10 : 50 : 40, v:v:v),is administered to each rat. At 0.08, 0.25, 0.5, 1, 2, 4, 8 and 24 h after dosing, the rats are anaesthetized andblood samples are collected by cardiac puncture.Mice 2Adult male C57BL/6J mice (25-30 g) are used. The effe

16、cts of RS-127445 (1 nM-10 M, single concentrationper tissue, 15 min contact time) or vehicle (5 or 50 L DMSO) are expressed as the percentage change inamplitude compared with the mean amplitude of four pre-drug, post-EFS contractile responses. The resultsare analysed using a two-sample equal varianc

17、e t-test.MCE has not independently confirmed the accuracy of these methods. They are for reference only.REFERENCES1. Bonhaus DW, et al. RS-127445: a selective, high affinity, orally bioavailable 5-HT2B receptor antagonist. Br J Pharmacol. 1999Jul;127(5):1075-82.2. Bassil AK, et al. Inhibition of colonic motility and defecation by RS-127445 suggests an involvement of the 5-HT2B receptor in rodent