以3个已报道的苯并噻唑类Rho关联含卷曲螺旋蛋白激酶(ROCK)抑制剂(化合物1～3)为研究对象,经分子动力学模拟获得其在ROCK2蛋白结合口袋中的稳定结合构象,通过分子对接结果从氨基酸角度初步揭示了此类抑制剂的结构-活性关系(SAR);然后,对这3个抑制剂进行MM/GBSA结合自由能(ΔGbind)研究,结合自由能计算可知ΔGbind与化合物活性之间具有良好的相关性,且范德华作用能(ΔGVDW)对ΔGbind的贡献最大.通过自由能分解获得了对于高活性抑制剂具有重要影响的关键残基.最后,根据分子对接和自由能研究结果设计并合成了3类新型苯并噻唑类似物(D1～D10).生物学评价结果表明,这10个化合物分别具有11～288 nmol/L(ROCK1)和2～105 nmol/L(ROCK2)的抑制活性.其中,化合物D3～D5在人肝微粒体代谢研究中展现出比已报道化合物更高的代谢稳定性.本研究不仅为高活性ROCK抑制剂的设计提供了理论指导,也为ROCK的应用研究提供了一系列结构新颖的高活性抑制剂.%Rock has been considered to provide a pharmacological strategy for preventing and treating multiple sclerosis, pulmonary hypertension, glaucoma, cardiovascular disease, erectile dysfunction and cancer. With 3 previously reported and benzothiazole-based ROCK inhibitors( 1—3) as the research targets, the structure-activity relationship ( SAR ) was preliminary revealed from amino acid level by molecular docking after obtaining the stable ROCK2-ligand complexes in the binding pocket through molecular dynamic simulations. Then MM/GBSA free energy calculations of compounds 1—3 showed that there was good correlation between binding affinity(ΔGbind ) and inhibitory activities, and van der Waals interaction (ΔGVDW ) contributing toΔGbind most. And the key amino acids with outstanding contribution for high inhibition were obtained through free energy analysis. Finally, 3 series of benzothiazoles( D1—D10) were designed according to the results of molecular docking and free energy calculations. In the biological evaluation, compounds D1—D10 exhibited 2—105 nmol/L IC50 values against ROCK2 and 11—288 nmol/L IC50 values against ROCK1. Compounds D3—D5 exhibited higher metabolic stability than reported compounds 1 and 3 in human liver microsome studies. This work not only gave theoretical guidance for the design of highly potent ROCK, but also offered a series of highly active ROCK inhibitors with intellectual property right for fundamental research and application of ROCK.
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