IN SILICO CHARACTERIZATION OF AMINO-PYRAZOLE DERIVATIVES AS MULTI-TARGET LIGANDS TO MODULATE ALZHEIMER’S DISEASE AND RELATED TAUOPATHIES VIA GSK-3BETA, FYN, AND DYRK1A INHIBITION

This study aims to investigate the molecular basis of target engagement for a series of amino-pyrazole-based multikinase inhibitors designed to modulate Alzheimer’s disease and tauopathies. Our team performed a comprehensive in silico analysis to evaluate how structural modifications impact the binding affinity and selectivity against Glycogen Synthase Kinase-3beta (GSK-3beta), FYN proto-oncogene, and Dual-Specificity Tyrosine-Phosphorylation-Regulated Kinase 1A (DYRK1A).
Computational docking simulations were executed using MolSoft ICM-Pro. This in silico approach allowed for the identification of critical hydrogen-bond (HB) networks within the highly conserved ATP-binding pockets of the three kinases. Specifically, docking predicted that the amino-pyrazole moiety forms essential HBs with the hinge region residues (e.g., Asp133 in GSK-3beta and Glu239/Leu241 in DYRK1A). Furthermore, our analysis identified that the removal of the cyclopropyl substituent led to a loss of hydrophobic interactions with gatekeeper residues like Phe238 in DYRK1A, explaining significant drops in potency observed in experimental assays.
The docking poses were subsequently compared with high-resolution X-ray crystallographic structures, which confirmed our predicted binding modes. In particular, docking analysis highlighted the role of the meta-carbamoyl group of the lead compound in establishing additional stabilizing interactions with Asn292 and Asp307 in DYRK1A. These in silico findings elucidate the molecular determinants required for balanced triple-kinase inhibition, providing a rational framework for the further refinement of disease-modifying agents for tau-related neurodegeneration. These findings, corroborated by experimental validation, elucidate the molecular determinants required for balanced triple-kinase inhibition.