Ligand based pharmacophore modelling and integrated computational approaches in the quest for small molecule inhibitors against hCA IX

Abstract

Carbonic anhydrase IX is an important biomarker to fight hypoxic tumours in both initial and metastatic stages of many forms of cancer. Overexpression of hCA IX in the hypoxic environment, has an active role in pH maintenance and makes the hCA IX a better target for the inhibitors targeting specific types of cancer stages. Being a member of the carbonic anhydrase family and having sixteen isoforms, it is important to have a selective inhibition of hCA IX to limit the disruption in the biological and metabolic pathways where other isoforms of hCA are localised and to avoid the other toxicity and adverse effects we try to find selective hCA IX inhibitors from a natural derivative. In the process of finding selective hCA inhibitors we developed a pharmacophore model based on existing inhibitors with IC₅₀ values of less than 50 nm, which is then validated with the external decoy set and used for database searching followed by virtual screening to identify the hits based on the pharmacophore fit score and RMSD. Molecular docking studies were performed to identify protein ligand interaction and molecular dynamics simulation studies to analyse the stability of the complex and DFT studies were carried out. The initial screening yielded 43 hits with the RMSD value less than 1, which when subjected to docking exhibited very good interaction with key residues ZN301, HIS94, HIS96 and HIS119. The top 4 compounds in the molecular dynamics simulation studies for 100 ns provided useful insights on the stability of the complex and the DFT studies confirmed the energy variation between HOMO and LUMO is within an acceptable range. An average binding score of −7.8 Kcal mol⁻¹ for the lead compounds and high stability margin in the dynamics study concludes that these lead compounds demonstrated outstanding potential for hCA IX inhibitory action theoretically and that further experimental studies for selective inhibition are inevitable.