title: Discovery of a novel ROCK2 inhibitor with anti-migration effects via docking and high-content drug screening.
abstract: Rho-associated protein kinase (ROCK) mediated the reorganization of the actin cytoskeleton and has been implicated in the spread and metastatic process of cancer. In this study, structure-based high-throughput virtual screening was used to identify candidate compounds targeting ROCK2 from a chemical library. Moreover, high-content screening based on neurite outgrowth of SH-SY5Y cells (a human neuroblastoma cell line) was used for accelerating the identification of compounds with characteristics of ROCK2 inhibitors. The effects of bioactive ROCK2 inhibitor candidates were further validated using other bioassays including cell migration and wound healing in SH-SY5Y cells. Through the combined virtual and high-content drug screening, the compound 1,3-benzodioxol-5-yl[1-(5-isoquinolinylmethyl)-3-piperidinyl]-methanone (BIPM) was identified as a novel and potent ROCK2 inhibitor. Exposure of SH-SY5Y cells to BIPM led to significant changes in neurite length, cell migration and actin stress fibers. Further experiments demonstrated that BIPM was able to significantly inhibit phosphorylation of cofilin, a regulatory protein of actin cytoskeleton. These results suggest that BIPM could be considered as a promising scaffold for the further development of ROCK2 inhibitors for anti-cancer metastasis.
authors.affiliation: State Key Laboratory of Quality Research in Chinese Medicine, and Institute of Chinese Medical Sciences, University of Macau, Macao, China. simonlee@umac.mo.
authors.forename: Cheong-Meng
authors.initials: CM
authors.lastname: Chong
authors.affiliation: State Key Laboratory of Quality Research in Chinese Medicine, and Institute of Chinese Medical Sciences, University of Macau, Macao, China. simonlee@umac.mo.
authors.forename: Man-Teng
authors.initials: MT
authors.lastname: Kou
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, Zhejiang 310058, China. tingjunhou@zju.edu.cn tingjunhou@hotmail.com.
authors.forename: Peichen
authors.initials: P
authors.lastname: Pan
authors.affiliation: State Key Laboratory of Quality Research in Chinese Medicine, and Institute of Chinese Medical Sciences, University of Macau, Macao, China. simonlee@umac.mo.
authors.forename: Hefeng
authors.initials: H
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Quality Research in Chinese Medicine, and Institute of Chinese Medical Sciences, University of Macau, Macao, China. simonlee@umac.mo.
authors.forename: Nana
authors.initials: N
authors.lastname: Ai
authors.affiliation: State Key Laboratory of Quality Research in Chinese Medicine, and Institute of Chinese Medical Sciences, University of Macau, Macao, China. simonlee@umac.mo.
authors.forename: Chuwen
authors.initials: C
authors.lastname: Li
authors.affiliation: State Key Laboratory of Quality Research in Chinese Medicine, and Institute of Chinese Medical Sciences, University of Macau, Macao, China. simonlee@umac.mo.
authors.forename: Hai-Jing
authors.initials: HJ
authors.lastname: Zhong
authors.affiliation: State Key Laboratory of Quality Research in Chinese Medicine, and Institute of Chinese Medical Sciences, University of Macau, Macao, China. simonlee@umac.mo.
authors.forename: Chung-Hang
authors.initials: CH
authors.lastname: Leung
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, Zhejiang 310058, China. tingjunhou@zju.edu.cn tingjunhou@hotmail.com and Institute of Functional Nano & Soft Materials (FUNSOM) and Jiangsu Key Laboratory for Carbon-Based Functional Materials & Devices, Soochow University, Suzhou, Jiangsu 215123, China.
authors.forename: Tingjun
authors.initials: T
authors.lastname: Hou
authors.affiliation: State Key Laboratory of Quality Research in Chinese Medicine, and Institute of Chinese Medical Sciences, University of Macau, Macao, China. simonlee@umac.mo.
authors.forename: Simon Ming-Yuen
authors.initials: SM
authors.lastname: Lee
mesh_terms: D001665:Binding Sites; D015415:Biomarkers; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D003198:Computer Simulation; D055808:Drug Discovery; D004354:Drug Screening Assays, Antitumor; D004789:Enzyme Activation / Q000187:drug effects; D057166:High-Throughput Screening Assays; D006801:Humans; D008968:Molecular Conformation; D062105:Molecular Docking Simulation; D056004:Molecular Dynamics Simulation; D015394:Molecular Structure; D011485:Protein Binding; D047428:Protein Kinase Inhibitors / Q000737:chemistry* / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects; D054852:Small Molecule Libraries; D057188:Workflow; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism
chemical_list: D015415:Biomarkers; D047428:Protein Kinase Inhibitors; D054852:Small Molecule Libraries; C515852:ROCK2 protein, human; D054460:rho-Associated Kinases