瑞舒地尔盐酸二水合物 TQ0319

Product Introduction
Bioactivity


英文名: Ripasudil

描述: Ripasudil (Ripasudil hydrochloride dihydrate) 是一种 ROCK 特异性抑制剂，能够抑制 ROCK1和 ROCK2的活性，IC50值分别为 51 和 19 nM。

细胞实验: Trabecular meshwork (TM) cells are plated on 6 well plates at a density of 1?×?10^4 cells per well in DMEM containing 10% FBS. Following overnight culture, when cells have reached semiconfluence, 1 or 10?μM of Ripasudil, 10?μM of Y-27632, or 10?μM of fasudil are added to culture wells. PBS is used as a control vehicle. After 60?min, drug solutions are removed and replaced with DMEM containing 10% FBS. Cells are observed by phase-contrast microscopy and photographed 60?min after drug application and 2?h after drug removal. For immunohistochemistry, TM cells are plated on gelatin-coated 8 well chamber slides at a density of 1?×?10^4 cells per well in DMEM containing 10% FBS. After overnight culture, when cells reach semiconfluence, cells are incubated in Ripasudil at 1 or 10?μM, Y-27632 at 10?μM, or fasudil at 10?μM for 60?min. PBS is used as a control vehicle. Drug solutions are removed and replaced with DMEM containing 10% FBS after 2?h. Cells are fixed with 4% paraformaldehyde in PBS for 15?min then washed with cytoskeletal buffer (10?mM MES, 150?mM NaCl, 5?mM EGTA, 5?mM MgCl2, 5?mM glucose, pH 6.1) and serum buffer (10% FBS in PBS). Cells are permeabilized with 0.5% Triton X-100 in PBS for 12?min at room temperature and blocked with serum buffer for at least 2?h at 4°C. Filamentous actin (F-actin) is labeled with 0.05?mg/mL Phalloidin-TRITC for 1?h at room temperature. After washing with PBS, cells are mounted with a commercial mounting medium containing DAPI and observed using a fluorescence microscope. The exposure to take images for F-actin and DAPI are 0.1 and 0.05?sec, respectively [2].

激酶实验: ROCK 1 (0.75 ng/mL) and ROCK 2 (0.5 ng/mL) are incubated with various concentrations of Ripasudil, Y-27632, or HA-1077 at 25°C for 90 min in 50 mM Tris-HCl buffer (pH 7.5) containing 100 mM KCl, 10 mM MgCl2, 0.1 mM EGTA, 30 mM Long S6 Kinase Substrate peptide, and 1 mM ATP in a total volume of 40 mL. PKACa, PKC, and CaMKIIa are also incubated with various concentrations of Ripasudil, Y-27632, or HA-1077. PKACa (0.0625 ng/mL) is incubated at 25°C for 30 min in 40 mM Tris-HCl buffer (pH 7.5) containing 20 mM MgCl2, 1 mg/ mL BSA, 5 mM Kemptide peptide substrate, and 1 mM ATP in a total volume of 40 mL. PKC (0.025 ng/mL) is incubated at 25°C for 80 min in 20 mM Tris-HCl buffer (pH 7.5) containing 20 mM MgCl2, 0.4 mM CaCl2, 0.1 mg/mL BSA, 0.25 mM EGTA, 25 ng/mL phosphatidylserine, 2.5 ng/mL diacylglycerol, 0.0075% Triton-X-100, 25 mM DTT, 10 mM Neurogranin (28-43) peptide substrate, and 1 mM ATP in a total volume of 40 mL. CaMKIIa (0.025 ng/mL) is incubated at 25°C for 90 min in 50 mM Tris-HCl buffer (pH 7.5) containing 10 mM MgCl2, 2 mM CaCl2, 0.04 mg/mL BSA, 16 mg/mL purified calmodulin from bovine testis, 500 mM DTT, 50 mM Autocamitide 2, and 1 mM ATP in a total volume of 40 mL. After incubation, 40 mL of KinaseGlo Luminescent Kinase Assay solution is added, and allowed to remain at 25°C for 10 min, and Relative Light Units (RLU) are measured using a luminometer. The RLU without test compound is set as 100% (Control value), and that without enzyme and compound is set as 0% (Normal value). The reaction rate (% of control) is then calculated from the RLU with addition of each concentration of test compounds, and the 50% inhibitory concentrations (IC50) are determined by logistic regression analysis using SAS [1].

动物实验: In the rabbit experiments, 50 mL of vehicle or Ripasudil at concentrations of 0.0625%, 0.125%, 0.25, or 0.5% is instilled into one eye. Intraocular pressure (IOP) is measured in both eyes before and 0.5, 1, 2, 3, 4, and 5 h after instillation. The contralateral eye is not treated. Animals are administered all concentrations of Ripasudil assigned using the Latin square method with intervals of at least 2 d. In the monkey experiments, 20 mL of Ripasudil at concentrations of 0.1%, 0.2%, or 0.4%, and latanoprost at a concentration of 0.005% are instilled into one eye. IOP is measured in both eyes before and 1, 2, 4, 6, and 8 h after instillation. The contralateral eye is not treated. Animals are arranged to receive all formulations with intervals of at least 1 week using the Latin square method. The IOPs are compared with the results for the instillation side at pre-dose and at each time point after the instillation of Ripasudil and are compared with both eyes at each time point.

体外活性: Ripasudil 对CaMKIIα、PKACα和PKC的抑制活性较弱（IC50分别为370 nM、2.1 μM和27 μM）[1]。在培养的小梁细胞（TM细胞）中，Ripasudil（1、10 μM）可引起细胞骨架的变化，包括细胞收缩和圆化以及肌动蛋白束的减少。此外，Ripasudil（5 μM）显著降低Schlemm's管内皮（SCE）细胞单层的跨内皮电阻（TEER），并增加FITC-右旋糖酐的通透性[2]。

体内活性: Ripasudil在浓度依赖的方式下降低猴眼内0.1%至0.4%之间以及兔眼内0.0625%至0.5%之间的眼内压[1]。Ripasudil（1 mg/kg，p.o. 每日一次）在神经压迫(NC)后对视网膜神经节细胞(RGCs)表现出神经保护效果。此外，Ripasudil还能抑制小鼠轴突损伤引起的氧化应激。Ripasudil在NC损伤后抑制RGCs中依时间产生的ROS[3]。

存储条件: Powder: -20°C for 3 years | In solvent: -80°C for 1 year

溶解度: H2O : 45 mg/mL (113.67 mM)


关键字: inhibit | K115 | Ripasudil | Ripasudil Hydrochloride | Rho-kinase | Rho-associated kinase | ROCK | ROK | Ripasudil hydrochloride Dihydrate | Inhibitor | K 115 | Rho-associated protein kinase

相关产品: Narciclasine | SAR407899 hydrochloride | GSK269962A | RKI1313 | RKI-1447 | GKI-1 HCl | Y-33075 dihydrochloride | WAY-656935 | GSK429286A | CRT0066854 hydrochloride

相关库: Anti-Aging Compound Library | Drug Repurposing Compound Library | Wnt/Hedgehog/Notch Compound Library | FDA-Approved Kinase Inhibitor Library | Anti-Cancer Drug Library | Inhibitor Library | Anti-Cancer Clinical Compound Library | Kinase Inhibitor Library | Anti-Cancer Approved Drug Library | TGF-beta/Smad Compound Library

瑞舒地尔盐酸二水合物 TQ0319信息由TargetMol中国为您提供，如您想了解更多关于瑞舒地尔盐酸二水合物 TQ0319报价、型号、参数等信息，欢迎来电或留言咨询。

注：该产品未在中华人民共和国食品药品监督管理部门申请医疗器械注册和备案，不可用于临床诊断或治疗等相关用途