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: Ram Kumar Mishra, Reshma Alokam, Sarthak Mohan Singhal, Geethasai Srivathsav, Dharamarajan Sriram, Neerja Kaushik-Basu, Dinesh Manvar, Perumal Yogeeswari
• Journal: Journal of chemical information and modeling (J Chem Inf Model) Vol. 54 Issue 10 Pg. 2876-86 (Oct 27 2014) ISSN: 1549-960X [Electronic] United States
• PMID: 25254429 (Publication Type: Journal Article)

Chemical References

• Antineoplastic Agents
• Ligands
• Methylmercury Compounds
• Protein Kinase Inhibitors
• Recombinant Proteins
• Small Molecule Libraries
• 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine
• rho-Associated Kinases
• fasudil

Topics

• 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine (analogs & derivatives, chemistry, pharmacology)
• Antineoplastic Agents (chemistry, pharmacology)
• Binding Sites
• Cell Line, Tumor
• Cell Survival (drug effects)
• Crystallography, X-Ray
• Drug Design
• Hepacivirus (drug effects)
• Hepatocytes (drug effects, enzymology, pathology)
• High-Throughput Screening Assays
• Humans
• Ligands
• Methylmercury Compounds (antagonists & inhibitors, toxicity)
• Molecular Conformation
• Molecular Dynamics Simulation
• Neurons (drug effects, enzymology, pathology)
• Protein Binding
• Protein Kinase Inhibitors (chemistry, pharmacology)
• Recombinant Proteins (chemistry, genetics)
• Small Molecule Libraries (chemistry, pharmacology)
• Structure-Activity Relationship
• Thermodynamics
• User-Computer Interface
• rho-Associated Kinases (antagonists & inhibitors, chemistry, genetics)