title: Design of novel rho kinase inhibitors using energy based pharmacophore modeling, shape-based screening, in silico virtual screening, and biological evaluation.
abstract: Rho-associated protein kinase (ROCK) plays a key role in regulating a variety of cellular processes, and dysregulation of ROCK signaling or expression is implicated in numerous diseases and infections. ROCK proteins have therefore emerged as validated targets for therapeutic intervention in various pathophysiological conditions such as diabetes-related complications or hepatitis C-associated pathogenesis. In this study, we report on the design and identification of novel ROCK inhibitors utilizing energy based pharmacophores and shape-based approaches. The most potent compound 8 exhibited an IC50 value of 1.5 μM against ROCK kinase activity and inhibited methymercury-induced neurotoxicity of IMR-32 cells at GI50 value of 0.27 μM. Notably, differential scanning fluorometric analysis revealed that ROCK protein complexed with compound 8 with enhanced stability relative to Fasudil, a validated nanomolar range ROCK inhibitor. Furthermore, all compounds exhibited ≥96 μM CC50 (50% cytotoxicity) in Huh7 hepatoma cells, while 6 compounds displayed anti-HCV activity in HCV replicon cells. The identified lead thus constitutes a prototypical molecule for further optimization and development as anti-ROCK inhibitor.
authors.affiliation: Computer-Aided Drug Design Lab, Department of Pharmacy, Birla Institute of Technology & Science-Pilani, Hyderabad Campus , Jawahar Nagar, Hyderabad-500078, Andhra Pradesh, India.
authors.forename: Ram Kumar
authors.initials: RK
authors.lastname: Mishra
authors.forename: Reshma
authors.initials: R
authors.lastname: Alokam
authors.forename: Sarthak Mohan
authors.initials: SM
authors.lastname: Singhal
authors.forename: Geethasai
authors.initials: G
authors.lastname: Srivathsav
authors.forename: Dharamarajan
authors.initials: D
authors.lastname: Sriram
authors.forename: Neerja
authors.initials: N
authors.lastname: Kaushik-Basu
authors.forename: Dinesh
authors.initials: D
authors.lastname: Manvar
authors.forename: Perumal
authors.initials: P
authors.lastname: Yogeeswari
mesh_terms: D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives / Q000737:chemistry / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000737:chemistry* / Q000494:pharmacology; D001665:Binding Sites; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D018360:Crystallography, X-Ray; D015195:Drug Design; D016174:Hepacivirus / Q000187:drug effects; D022781:Hepatocytes / Q000187:drug effects / Q000201:enzymology / Q000473:pathology; D057166:High-Throughput Screening Assays; D006801:Humans; D008024:Ligands; D008767:Methylmercury Compounds / Q000037:antagonists & inhibitors / Q000633:toxicity; D008968:Molecular Conformation; D056004:Molecular Dynamics Simulation; D009474:Neurons / Q000187:drug effects / Q000201:enzymology / Q000473:pathology; D011485:Protein Binding; D047428:Protein Kinase Inhibitors / Q000737:chemistry* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D054852:Small Molecule Libraries / Q000737:chemistry* / Q000494:pharmacology; D013329:Structure-Activity Relationship; D013816:Thermodynamics; D014584:User-Computer Interface; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics
chemical_list: D000970:Antineoplastic Agents; D008024:Ligands; D008767:Methylmercury Compounds; D047428:Protein Kinase Inhibitors; D011994:Recombinant Proteins; D054852:Small Molecule Libraries; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D054460:rho-Associated Kinases; C049347:fasudil