GSK269962A (GSK269962A HCl) is a selective ROCK(Rho-associated protein kinase) inhibitor with IC50 values of 1.6 and 4 nM for ROCK1 and ROCK2, respectively.

ROCK2:4 nM, ROCK1:1.6 nM

GSK269962A 完全抑制了由血管紧张素 II 在人类平滑肌中诱导的肌动蛋白应力纤维的形成。这种对肌动蛋白纤维形成的抑制效应从约 1 μM GSK269962A 开始观察到。GSK269962A 在预收缩的大鼠主动脉（组织浴）中诱导了血管舒张，IC50 为 35 nM。由 GSK269962A 诱导的舒张是可逆的。GSK269962A 抑制了 IL-6 mRNA 的转录，并降低了巨噬细胞中 LPS 诱导的 IL-6 和 TNF-α 蛋白产量。

GSK269962A的口服给药在自发性高血压鼠中显著而依剂量减少全身血压。血压降低的效果是急性的且显著的，口服后约2小时观察到血压降低的最大效应。血压的降低伴随着心率的急性、依剂量增加，这可能是由于激活了压力反射机制。使用10 mg/kg剂量的GSK 269962通过ROCK抑制，进而引发VV（排空量）、PVR（排空后残余量）、VT（阈量容积）、VE（排空效率）、ICI（宫缩间期）、BC（膀胱顺应性）、以及VTNVC（诱发NVC的容积阈值）的增加。

The enzyme activity and kinetics of the purified ROCK1(3-543) are determined using scintillation proximity assay. In this assay, purified ROCK1 is incubated with peptide substrate (Biotin-Ahx-AKRRLSSLRA-CONH2), and 33ATP and the subsequent incorporation of 33P into the peptide is quantified by streptavidin bead capture. For IC50 determination, test compounds are dissolved at 10 mM in 100% DMSO, with subsequent serial dilution in 100% DMSO. Compounds are typically assayed over an 11-point dilution range with a concentration in the assay of 10 μM to 0.2 nM in 3-fold dilutions. For dose-response curves, data are normalized and expressed as percentage inhibition using the formula 100×[(U-C1)/(C2-C1)], where U is the unknown value, C1 is the average of the high signal (0%) control wells, and C2 is the average of the low signal (100%) control wells. Curve fitting is performed The results for each compound are recorded as pIC50 values[1].

Human primary smooth muscle cells were serum-starved(cells were grown on coverslips, and at approximately 50% confluence, they were serum-starved overnight) and stimulated with AngII (100 nM) for 2 h. ROCK inhibitors (3 μM for SB-772077-B or GSK269962A) were added 30 min before AngII stimulation, and cells were fixed and stained with rhodamine phalloidin. Confocal images of actin stain were obtained.(Only for Reference)