Structure-based identification of GIRK2-PIP2 modulators among known drugs and metabolites using docking, MM-GBSA, ADMET, and molecular dynamics

G protein-gated inwardly rectifying potassium (GIRK) channels are key regulators of neuronal excitability, making them promising therapeutic targets for central nervous system disorders. Their activation depends on phosphatidylinositol-4,5-bisphosphate (PIP2), which stabilizes the channels open state. A deeper understanding of GIRK-PIP2 interactions could uncover new physiological roles and pave the way for therapies that modulate channel function. This study aimed to advance the targeting of GIRK channels at the PIP2-binding site. Over one million compounds were screened against GIRK2 (PDB ID: 4KFM) using high-throughput virtual screening. A core constraint with a root-mean-square deviation (RMSD) < 2 [A] was applied to assure the accuracy and binding close to PIP2-binding site. The top-scoring ligands were redocked with Glide (SP, XP) and binding free energy was estimated using Molecular Mechanics Generalized Born Surface Area method. The most promising compounds were analyzed for pharmacokinetic/physicochemical properties, followed by molecular dynamics (MD) simulations over 200 ns in membrane bilayer. MD analysis revealed three known compounds (Rosuvastatin, CID: 54365126 and 7304563) as potential competitive GIRK2 modulators, exhibiting stable interactions with residues critical for binding endogenous activators (PIP2, cholesterol), and GIRK-acting drugs. Docking analyses also revealed strong binding to GIRK2 for various metabolites, including leukotrienes, resolvins, acyl-CoAs, and polyphosphates, including adenosine-triphosphate (ATP) and thiamine-triphosphate. Notably, some of the identified compounds can affect similar ion channels, indicating potential cross-reactivity with GIRK2. Furthermore, the binding modes of acyl-CoAs and polyphosphates closely resemble PIP2s hydrophobic and phosphate group engagement. Together, these findings offer promising candidates for experimental validation and therapeutic development. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=153 SRC="FIGDIR/small/653795v1_ufig1.gif" ALT="Figure 1"> View larger version (41K): org.highwire.dtl.DTLVardef@1325e66org.highwire.dtl.DTLVardef@1d3835corg.highwire.dtl.DTLVardef@1550df3org.highwire.dtl.DTLVardef@107007e_HPS_FORMAT_FIGEXP M_FIG C_FIG Abbreviations: ADMET - Absorption, Distribution, Metabolism, Elimination, Toxicity; CID - PubChem Compound Identification; MM-GBSA - Molecular Mechanics with Generalized Born and Surface Area solvation; RMSD - Root-Mean-Square Deviation; SP - Standard Precision; PIP2 - phosphatidylinositol-4,5-bisphosphate; XP - Extra Precision. HIGHLIGHTSO_LIMulti-target screening identifies selective modulators of the GIRK2 channel C_LIO_LILigands target the PIP2-binding site, a novel interface for GIRK2 modulation C_LIO_LIMM-GBSA confirms binding affinity and ligand stability post-docking C_LIO_LIDynamics and bioinformatics predict selectivity and off-target interactions C_LIO_LIStatins and leukotriene-modifying drugs are strong GIRK2 modulator candidates C_LI