title: The selectivity and promiscuity of brain-neuroregenerative inhibitors between ROCK1 and ROCK2 isoforms: An integration of SB-QSSR modelling, QM/MM analysis and in vitro kinase assay.
abstract: The Rho-associated kinases (ROCKs) have long been recognized as an attractive therapeutic target for various neurological diseases; selective inhibition of ROCK1 and ROCK2 isoforms would result in distinct biological effects on neurogenesis, neuroplasticity and neuroregeneration after brain surgery and traumatic brain injury. However, the discovery and design of isoform-selective inhibitors remain a great challenge due to the high conservation and similarity between the kinase domains of ROCK1 and ROCK2. Here, a structure-based quantitative structure-selectivity relationship (SB-QSSR) approach was used to correlate experimentally measured selectivity with the difference in inhibitor binding to the two kinase isoforms. The resulting regression models were examined rigorously through both internal cross-validation and external blind validation; a nonlinear predictor was found to have high fitting stability and strong generalization ability, which was then employed to perform virtual screening against a structurally diverse, drug-like compound library. Consequently, five and seven hits were identified as promising candidates of 1-o-2 and 2-o-1 selective inhibitors, respectively, from which seven purchasable compounds were tested in vitro using a standard kinase assay protocol to determine their inhibitory activity against and selectivity between ROCK1 and ROCK2. The structural basis, energetic property and biological implication underlying inhibitor selectivity and promiscuity were also investigated systematically using a hybrid quantum mechanics/molecular mechanics (QM/MM) scheme.
authors: a Department of Neurosurgery , People's Hospital affiliated to Jiangsu University , Zhenjiang , China.
authors: L
authors: L
authors: Zhu
authors: a Department of Neurosurgery , People's Hospital affiliated to Jiangsu University , Zhenjiang , China.
authors: Y
authors: Y
authors: Yang
authors: a Department of Neurosurgery , People's Hospital affiliated to Jiangsu University , Zhenjiang , China.
authors: X
authors: X
authors: Lu
mesh_terms: D001921:Brain / Q000201:enzymology*; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000494:pharmacology*; D020128:Inhibitory Concentration 50; D008962:Models, Theoretical; D011485:Protein Binding; D020033:Protein Isoforms / Q000037:antagonists & inhibitors*; D021281:Quantitative Structure-Activity Relationship; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors*
chemical_list: D004791:Enzyme Inhibitors; D020033:Protein Isoforms; C515851:ROCK1 protein, human; C515852:ROCK2 protein, human; D054460:rho-Associated Kinases
keywords: ROCK1/ROCK2; brain neuroregeneration; inhibitor selectivity and promiscuity; quantitative structure–selectivity relationship