Improving virtual screening of G protein-coupled receptors via ligand-directed modeling

G protein-coupled receptors (GPCRs) play crucial roles in cell physiology and pathophysiology. There is increasing interest in using structural information for virtual screening (VS) of libraries and for structure-based drug design to identify novel agonist or antagonist leads. However, the sparse availability of experimentally determined GPCR/ligand complex structures with diverse ligands impedes the application of structure-based drug design (SBDD) programs directed to identifying new molecules with a select pharmacology. In this study, we apply ligand-directed modeling (LDM) to available GPCR X-ray structures to improve VS performance and selectivity towards molecules of specific pharmacological profile. The described method refines a GPCR binding pocket conformation using a single known ligand for that GPCR. The LDM method is a computationally efficient, iterative workflow consisting of protein sampling and ligand docking. We developed an extensive benchmark comparing LDM-refined binding pockets to GPCR X-ray crystal structures across seven different GPCRs bound to a range of ligands of different chemotypes and pharmacological profiles. LDM-refined models showed improvement in VS performance over origin X-ray crystal structures in 21 out of 24 cases. In all cases, the LDM-refined models had superior performance in enriching for the chemotype of the refinement ligand. This likely contributes to the LDM success in all cases of inhibitor-bound to agonist-bound binding pocket refinement, a key task for GPCR SBDD programs. Indeed, agonist ligands are required for a plethora of GPCRs for therapeutic intervention, however GPCR X-ray structures are mostly restricted to their inactive inhibitor-bound state.Author summary: G protein-coupled receptors (GPCRs) are a major target for drug discovery. These receptors are highly dynamic membrane proteins, and have had limited tractability using with biophysical screens that are widely adopted for globular protein targets. Thus, structure-based virtual screening (SBVS) holds great promise as a complement to physical screening for rational design of novel drugs. Indeed, the increasing number of atomic-detail GPCR X-ray crystal structures has coincided with an increase in prospective SBVS studies that have identified novel compounds. However, experimentally solved GPCR structures do not meet the full demand for SBVS, as the GPCR structural landscape is incomplete, lacking both in coverage of available GPCRs, and diversity in both receptor conformations and the chemistry of co-crystalised ligands. Here we present a novel computational GPCR binding pocket refinement method that can generate predictive GPCR/ligand complexes with improved SBVS performance. This ligand-directed modeling workflow uses parallel processing and efficient algorithms to search the GPCR/ligand conformational space faster and more efficiently than the widely used protein refinement method molecular dynamics. In this study, the resulting models are evaluated both structurally, and in retrospective SBVS. We demonstrate improved performance of refined models over their starting structures in the majority of our test cases.