20.500.12556/DiRROS-23663 Water-based pharmacophore modeling in kinase inhibitor design a case study on fyn and lyn protein kinases Water-based pharmacophore modeling is an emerging approach in inhibitor design that leverages the dynamics of explicit water molecules within ligand-free, water-filled binding sites to derive 3D pharmacophores for virtual screening. In this study, we assess the potential of this strategy through a case study targeting the ATP binding sites of Fyn and Lyn protein kinases─members of the Src family that have been less explored in anticancer drug discovery compared to other family members. Molecular dynamics simulations of multiple kinase structures were used to generate and validate several water-derived pharmacophores, which were subsequently employed to screen chemically diverse libraries of compounds. Two active compounds were identified in biochemical assays: a flavonoid-like molecule with low-micromolar inhibitory activity and a weaker inhibitor from the library of nature-inspired synthetic compounds. Structural analysis via molecular docking and simulations revealed that key predicted interactions, particularly with the hinge region and the ATP binding pocket, were retained in the bound states of these hits. However, interactions with more flexible regions, such as the N-terminal lobe and activation loop, were less consistently captured. These findings outline both the strengths and challenges of using water-based pharmacophores: while effective at modeling conserved core interactions, they may miss peripheral contacts governed by protein flexibility. Incorporating ligand information where available may help address this challenge. Overall, water-based pharmacophore modeling presents a promising ligand-independent strategy for identifying novel chemotypes and exploring undercharged chemical and conformational space in kinases as well as other therapeutically relevant targets. drug discovery inhibitors ligands peptides proteins pharmacophores odkrivanje zdravil inhibitorji ligandi peptidi proteini farmakofori true false true Angleški jezik Slovenski jezik © 2025 The Authors. Published by American Chemical Society Neznano 2025-09-23 11:53:58 2025-09-23 12:02:25 2025-10-24 03:54:46 0000-00-00 00:00:00 2025 0 0 Nasl. z nasl. zaslona; Opis vira z dne 1. 9. 2025; str. 9747−9761 iss. 18 Vol. 65 2025 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 2025-01-01 66.011:615 1549-960X 10.1021/acs.jcim.5c01478 247223299 https://pubs.acs.org/doi/10.1021/acs.jcim.5c01478 1 3b093cc6-9863-11f0-9bb7-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=35035 RAZ_Ljubic_Martin_2025.pdf RAZ_Ljubic_Martin_2025.pdf 1 5E57DEB6B69CE0450EEC3296DF0C14BC c6e30cff3f33361545ed342cfaa0e138fc7831e88a95bd1d929f931c1191108c c465ca13-9876-11f0-9bb7-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=35041 Kemijski inštitut 0 0 0