Toward G protein-coupled receptor structure-based drug design using X-ray lasers

Rational structure-based drug design (SBDD) relies on the availability of a large number of co-crystal structures to map the ligand-binding pocket of the target protein and use this information for lead-compound optimization via an iterative process. While SBDD has proven successful for many drug-discovery projects, its application to G protein-coupled receptors (GPCRs) has been limited owing to extreme difficulties with their crystallization. Here, a method is presented for the rapid determination of multiple co-crystal structures for a target GPCR in complex with various ligands, taking advantage of the serial femtosecond crystallography approach, which obviates the need for large crystals and requires only submilligram quantities of purified protein. The method was applied to the human β2-adrenergic receptor, resulting in eight room-temperature co-crystal structures with six different ligands, including previously unreported structures with carvedilol and propranolol. The generality of the proposed method was tested with three other receptors. This approach has the potential to enable SBDD for GPCRs and other difficult-to-crystallize membrane proteins.

Download Full-text

Recent Advances in Structure-Based Drug Design Targeting Class A G Protein-Coupled Receptors Utilizing Crystal Structures and Computational Simulations

Journal of Medicinal Chemistry ◽

10.1021/acs.jmedchem.6b01453 ◽

2017 ◽

Vol 61 (1) ◽

pp. 1-46 ◽

Cited By ~ 39

Author(s):

Yoonji Lee ◽

Shaherin Basith ◽

Sun Choi

Keyword(s):

Drug Design ◽

Crystal Structures ◽

G Protein ◽

G Protein Coupled Receptors ◽

Computational Simulations ◽

Structure Based Drug Design ◽

Class A ◽

Recent Advances ◽

G Protein Coupled

Download Full-text

Structure-Based Drug Design for G Protein-Coupled Receptors

Progress in Medicinal Chemistry ◽

10.1016/b978-0-444-63380-4.00001-9 ◽

2014 ◽

pp. 1-63 ◽

Cited By ~ 50

Author(s):

Miles Congreve ◽

João M. Dias ◽

Fiona H. Marshall

Keyword(s):

Drug Design ◽

G Protein ◽

G Protein Coupled Receptors ◽

Structure Based Drug Design ◽

G Protein Coupled

Download Full-text

Progress in Structure Based Drug Design for G Protein-Coupled Receptors

Journal of Medicinal Chemistry ◽

10.1021/jm200371q ◽

2011 ◽

Vol 54 (13) ◽

pp. 4283-4311 ◽

Cited By ~ 174

Author(s):

Miles Congreve ◽

Christopher J. Langmead ◽

Jonathan S. Mason ◽

Fiona H. Marshall

Keyword(s):

Drug Design ◽

G Protein ◽

G Protein Coupled Receptors ◽

Structure Based Drug Design ◽

G Protein Coupled

Download Full-text

Stabilised G protein-coupled receptors in structure-based drug design: a case study with adenosine A2A receptor

MedChemComm ◽

10.1039/c2md20164j ◽

2013 ◽

Vol 4 (1) ◽

pp. 52-67 ◽

Cited By ~ 18

Author(s):

Stephen P. Andrews ◽

Benjamin Tehan

Keyword(s):

Parkinson’S Disease ◽

Drug Design ◽

G Protein ◽

G Protein Coupled Receptors ◽

Adenosine A2a Receptor ◽

Structure Based Drug Design ◽

A2a Receptor ◽

Adenosine A2a ◽

G Protein Coupled

The first example of structure-based drug design with stabilised GPCRs has enabled the identification of a preclinical candidate for the treatment of Parkinson's disease.

Download Full-text

Modelling altered signalling of G-protein coupled receptors in inflamed environment to advance drug design

10.1101/2021.10.20.465073 ◽

2021 ◽

Author(s):

Sourav Ray ◽

Arne Thies ◽

Vikram Sunkara ◽

Hanna Wulkow ◽

M. Özgür Celik ◽

...

Keyword(s):

Drug Design ◽

G Protein ◽

Binding Pocket ◽

Reaction Rates ◽

G Protein Coupled Receptors ◽

Lower Probability ◽

Design Synthesis ◽

Channel Inhibition ◽

G Protein Coupled

We previously reported the successful design, synthesis and testing of the prototype opioid painkiller NFEPP that does not elicit adverse side effects. Uniquely, this design was based on mathematical modelling of extracellular interactions between G-protein coupled receptors (GPCRs) and ligands, recognizing that GPCRs function differently under pathological versus healthy conditions. We now present a novel stochastic model of GPCR function that includes intracellular dissociation of G-protein subunits and modulation of plasma membrane calcium channels associated with parameters of inflamed and healthy tissue (pH, radicals). The model is validated against in vitro experimental data for NFEPP and fentanyl ligands at different pH values. We found markedly reduced calcium channel inhibition induced by NFEPP at normal pH compared to lower pH, in contrast to the effect of fentanyl, and enhanced constitutive G-protein activation but lower probability of ligand binding with increasing radical concentrations. By means of molecular dynamics simulations, we also assessed qualitative changes of reaction rates due to additional disulfide bridges inside the GPCR binding pocket. The results suggest that, compared to radicals, low pH is a more important determinant of overall GPCR function in an inflamed environment. Future drug design efforts should take this into account.

Download Full-text

Structure‐Based Drug Design for G Protein‐Coupled Receptors

Burger's Medicinal Chemistry and Drug Discovery ◽

10.1002/0471266949.bmc269 ◽

2021 ◽

pp. 1-59

Author(s):

Miles Congreve ◽

John A. Christopher ◽

Chris Graaf

Keyword(s):

Drug Design ◽

G Protein ◽

G Protein Coupled Receptors ◽

Structure Based Drug Design ◽

G Protein Coupled

Download Full-text

Engineering of Challenging G Protein-Coupled Receptors for Structure Determination and Biophysical Studies

Molecules ◽

10.3390/molecules26051465 ◽

2021 ◽

Vol 26 (5) ◽

pp. 1465

Author(s):

Yann Waltenspühl ◽

Janosch Ehrenmann ◽

Christoph Klenk ◽

Andreas Plückthun

Keyword(s):

High Resolution ◽

Drug Design ◽

G Protein ◽

Drug Targets ◽

Native Mass Spectrometry ◽

G Protein Coupled Receptors ◽

Resonance Spectroscopy ◽

Structure Based Drug Design ◽

Biophysical Studies ◽

G Protein Coupled

Membrane proteins such as G protein-coupled receptors (GPCRs) exert fundamental biological functions and are involved in a multitude of physiological responses, making these receptors ideal drug targets. Drug discovery programs targeting GPCRs have been greatly facilitated by the emergence of high-resolution structures and the resulting opportunities to identify new chemical entities through structure-based drug design. To enable the determination of high-resolution structures of GPCRs, most receptors have to be engineered to overcome intrinsic hurdles such as their poor stability and low expression levels. In recent years, multiple engineering approaches have been developed to specifically address the technical difficulties of working with GPCRs, which are now beginning to make more challenging receptors accessible to detailed studies. Importantly, successfully engineered GPCRs are not only valuable in X-ray crystallography, but further enable biophysical studies with nuclear magnetic resonance spectroscopy, surface plasmon resonance, native mass spectrometry, and fluorescence anisotropy measurements, all of which are important for the detailed mechanistic understanding, which is the prerequisite for successful drug design. Here, we summarize engineering strategies based on directed evolution to reduce workload and enable biophysical experiments of particularly challenging GPCRs.

Download Full-text