Demystifying the Molecular Basis of Pyrazoloquinolinones Recognition at the Extracellular α1+/β3- Interface of the GABA Receptor by Molecular Modeling.

GABA receptors are pentameric ligand-gated ion channels that serve as major inhibitory neurotransmitter receptors in the mammalian brain and the target of numerous clinically relevant drugs interacting with different ligand binding sites. Here, we report an in silico approach to investigate the binding of pyrazoloquinolinones (PQs) that mediate allosteric effects through the extracellular α+/β- interface of GABA receptors. First, we docked a potent prototype of PQs into the α1+/β3- site of a homology model of the human α1β3γ2 subtype of the GABA receptor. Next, for each docking pose, we computationally derived protein-ligand complexes for 18 PQ analogs with known experimental potency. Subsequently, binding energy was calculated for all complexes using the molecular mechanics-generalized Born surface area method. Finally, docking poses were quantitatively assessed in the light of experimental data to derive a binding hypothesis. Collectively, the results indicate that PQs at the α1+/β3- site likely exhibit a common binding mode that can be characterized by a hydrogen bond interaction with β3Q64 and hydrophobic interactions involving residues α1F99, β3Y62, β3M115, α1Y159, and α1Y209. Importantly, our results are in good agreement with the recently resolved cryo-Electron Microscopy structures of the human α1β3γ2 and α1β2γ2 subtypes of GABA receptors.