title: High-throughput screening against ~6.1 million structurally diverse, lead-like compounds to discover novel ROCK inhibitors for cerebral injury recovery.
abstract: Rho-associated protein kinase (ROCK) has been recognized as an attractive therapeutic target to promote neurogenesis, neuroregeneration, and neurorecovery after cerebral injury. Here, a high-throughput screening protocol was described to discover novel ROCK inhibitors from a large chemical library containing ~6.1 million structurally diverse, lead-like compounds. The protocol employed empirical rules such as ADMET evaluation and chemical similarity analysis to exclude those of drug-unlike candidates, and then molecular docking and binding affinity predictions were performed to suggest few promising candidates with high scores. Consequently, five compounds were successfully identified to have satisfactory activity profile with IC50 values at nanomolar level. In order to elucidate the molecular mechanism of inhibitor binding to target, the complex structures of ROCK kinase domain with the five identified compounds were modeled and examined in detail. A number of polar chemical forces such as hydrogen bonds and cation-π interactions as well as nonpolar contacts such as π-π stacking and hydrophobic forces were revealed at the complex interface, conferring high affinity and strong specificity to inhibitor binding. In addition, several key residues around the kinase active site, including Val90, Lys105, Asn203, and Phe368, were found to play an important role in binding.
authors.affiliation: Department of Nursing, People's Hospital Affiliated to Jiangsu University, Zhenjiang, 212002, China. haolinggong@126.com.
authors.forename: Haoling
authors.initials: H
authors.lastname: Gong
authors.affiliation: Department of Neurosurgery, People's Hospital Affiliated to Jiangsu University, Zhenjiang, 212002, China.
authors.forename: Zhicheng
authors.initials: Z
authors.lastname: Yuan
authors.affiliation: Department of Neurosurgery, People's Hospital Affiliated to Jiangsu University, Zhenjiang, 212002, China.
authors.forename: Liping
authors.initials: L
authors.lastname: Zhan
mesh_terms: D000818:Animals; D001930:Brain Injuries / Q000188:drug therapy*; D018938:Caco-2 Cells; D020134:Catalytic Domain; D004285:Dogs; D004353:Drug Evaluation, Preclinical; D057166:High-Throughput Screening Assays; D006801:Humans; D061985:Madin Darby Canine Kidney Cells; D062105:Molecular Docking Simulation; D056004:Molecular Dynamics Simulation; D047428:Protein Kinase Inhibitors / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology* / Q000627:therapeutic use; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism
chemical_list: D047428:Protein Kinase Inhibitors; D054460:rho-Associated Kinases
keywords: Blood-brain barrier; CNS; Cerebral injury; High-throughput screening; Neuroregeneration; Rho-associated protein kinase