X‐ray crystallography and free energy calculations reveal the binding mechanism of A2A adenosine receptor antagonists

DOI: 10.1002/anie.202003788

Authors: Willem Jespers (Uppsala University) , Gregory Verdon (Sosei Heptares) , Jhonny Azuaje (Universidade de Santiago de Compostela) , Maria Majellaro (Universidade de Santiago de Compostela) , Henrik Keränen (Uppsala University) , Xerardo García‐mera (Universidade de Santiago de Compostela) , Miles Congreve (Sosei Heptares) , Francesca Deflorian (Sosei Heptares) , Chris De Graaf (Sosei Heptares) , Andrei Zhukov (Sosei Heptares) , Andrew S. Dore (Sosei Heptares) , Jonathan S. Mason (Sosei Heptares) , Johan Åqvist (Uppsala University) , Robert M. Cooke (Sosei Heptares) , Eddy Sotelo (Universidade de Santiago de Compostela) , Hugo Gutiérrez‐de‐terán (Uppsala University)
Co-authored by industrial partner: Yes

Type: Journal Paper
Journal: Angewandte Chemie International Edition

State: Published (Approved)
Published: July 2020
Diamond Proposal Number(s): 17185

Abstract: We present a robust protocol based on iterations of free energy perturbation (FEP) calculations, chemical synthesis, biophysical mapping and X‐ray crystallography to reveal the binding mode of an antagonist series to the A2A adenosine receptor (AR). Eight A2AAR binding site mutations from biophysical mapping experiments were initially analyzed with sidechain FEP simulations, performed on alternate binding modes. The results distinctively supported one binding mode, which was subsequently used to design new chromone derivatives. Their affinities for the A2AAR were experimentally determined and investigated through a cycle of ligand‐FEP calculations, validating the binding orientation of the different chemical substituents proposed. Subsequent X‐ray crystallography of the A2AAR with a low and a high affinity chromone derivative confirmed the predicted binding orientation. The new molecules and structures here reported were driven by free energy calculations, and provide new insights on antagonist binding to the A2AAR, an emerging target in immuno‐oncology.

Journal Keywords: adenosine receptors; biophysical mapping (BPM); free energy perturbation (FEP); G protein-coupled receptor (GPCR)

Subject Areas: Chemistry, Biology and Bio-materials


Instruments: I24-Microfocus Macromolecular Crystallography

Documents:
anie.202003788.pdf

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