Structure-Based Virtual Screening and Biological Evaluation of a Calpain Inhibitor for Prevention of Selenite-Induced Cataractogenesis in an in Vitro System

• Published in: Journal of chemical information and modeling 2015-08, Vol.55 (8), p.1686-1697
• Main Authors: Muralidharan, Arumugam Ramachandran, Selvaraj, Chandrabose, Singh, Sanjeev Kumar, Sheu, Joen-Rong, Thomas, Philip A, Geraldine, Pitchairaj
• Format: Article
• Language: English
• Subjects: Animals; Antioxidants; Calcium; Calcium - metabolism; Calpain - antagonists & inhibitors; Calpain - chemistry; Calpain - metabolism; Cataract - metabolism; Cataract - pathology; Cataract - prevention & control; Cell Line; Cells; Crystallography, X-Ray; Cysteine Proteinase Inhibitors - chemistry; Cysteine Proteinase Inhibitors - therapeutic use; Drug Discovery; Humans; Lens, Crystalline - drug effects; Lens, Crystalline - metabolism; Lens, Crystalline - pathology; Models, Molecular; Proteases; Protein Conformation; Rats, Wistar; Rodents; Selenious Acid; Tissues
• ISSN: 1549-9596; 1549-960X
• DOI: 10.1021/acs.jcim.5b00092

Calpains belong to the family of calcium-dependent, structurally related intracellular cysteine proteases that exhibit significant functions in evolution of different types of cataracts in human as well as animal models. Application of calpain inhibitors generated through a virtual screening workflow may provide new avenues for the prevention of cataractogenesis. Hence, in the current study, compounds were first screened for potent calpain inhibitory activity by employing a structure-based approach, and the screening results were then validated through biological experiments in rat lenses. A hit compound, HTS08688, was obtained by structure-based virtual screening. A micromolar concentration of HTS08688 was found to prevent in vitro cataractogenesis in isolated Wistar rat lenses, while maintaining the antioxidant and calcium concentrations at near normal levels. Inhibition of superoxide anion generation, as observed through cytochemical localization studies, and maintenance of structural integrity, as demonstrated by histological analysis of lenticular tissue, also suggested that HTS08688 can ameliorate the cataractous condition induced by selenite in an in vitro rodent model. A cell proliferation assay was performed; the IC 50 value of the screened calpain inhibitor, HTS08688, against human lenticular epithelial cells-b3 was found to be 177 μM/mL. This combined theoretical and experimental approach has demonstrated a potent lead compound, HTS08688, that exhibits putative anticataractogenic activity by virtue of its potential to inhibit calpain.