Design of a True Bivalent Ligand with Picomolar Binding Affinity for a G Protein-Coupled Receptor Homodimer

Overview

Journal J Med Chem

Publisher American Chemical Society

Specialty Chemistry

Date 2018 Sep 27

PMID 30257092

Citations 15

Authors

Daniel Pulido

Veronica Casado-Anguera

Laura Perez-Benito

Estefania Moreno

Arnau Cordomi

Laura Lopez

Antoni Cortes

Sergi Ferre

Leonardo Pardo

Vicent Casado

Miriam Royo

Affiliations

Soon will be listed here.

Abstract

Bivalent ligands have emerged as chemical tools to study G protein-coupled receptor dimers. Using a combination of computational, chemical, and biochemical tools, here we describe the design of bivalent ligand 13 with high affinity ( K = 21 pM) for the dopamine D receptor (DR) homodimer. Bivalent ligand 13 enhances the binding affinity relative to monovalent compound 15 by 37-fold, indicating simultaneous binding at both protomers. Using synthetic peptides with amino acid sequences of transmembrane (TM) domains of DR, we provide evidence that TM6 forms the interface of the homodimer. Notably, the disturber peptide TAT-TM6 decreased the binding of bivalent ligand 13 by 52-fold and had no effect on monovalent compound 15, confirming the DR homodimer through TM6 ex vivo. In conclusion, by using a versatile multivalent chemical platform, we have developed a precise strategy to generate a true bivalent ligand that simultaneously targets both orthosteric sites of the DR homodimer.

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References

Bonaventura J, Navarro G, Casado-Anguera V, Azdad K, Rea W, Moreno E . Allosteric interactions between agonists and antagonists within the adenosine A2A receptor-dopamine D2 receptor heterotetramer. Proc Natl Acad Sci U S A. 2015; 112(27):E3609-18. PMC: 4500251. DOI: 10.1073/pnas.1507704112. View

Guitart X, Navarro G, Moreno E, Yano H, Cai N, Sanchez-Soto M . Functional selectivity of allosteric interactions within G protein-coupled receptor oligomers: the dopamine D1-D3 receptor heterotetramer. Mol Pharmacol. 2014; 86(4):417-29. PMC: 4164978. DOI: 10.1124/mol.114.093096. View

Hubner H, Schellhorn T, Gienger M, Schaab C, Kaindl J, Leeb L . Structure-guided development of heterodimer-selective GPCR ligands. Nat Commun. 2016; 7:12298. PMC: 4963535. DOI: 10.1038/ncomms12298. View

Manglik A, Kruse A, Kobilka T, Thian F, Mathiesen J, Sunahara R . Crystal structure of the µ-opioid receptor bound to a morphinan antagonist. Nature. 2012; 485(7398):321-6. PMC: 3523197. DOI: 10.1038/nature10954. View

Santos R, Ursu O, Gaulton A, Bento A, Donadi R, Bologa C . A comprehensive map of molecular drug targets. Nat Rev Drug Discov. 2016; 16(1):19-34. PMC: 6314433. DOI: 10.1038/nrd.2016.230. View

Berque-Bestel I, Lezoualch F, Jockers R . Bivalent ligands as specific pharmacological tools for G protein-coupled receptor dimers. Curr Drug Discov Technol. 2008; 5(4):312-8. DOI: 10.2174/157016308786733591. View

Jorg M, May L, Mak F, Lee K, Miller N, Scammells P . Synthesis and pharmacological evaluation of dual acting ligands targeting the adenosine A2A and dopamine D2 receptors for the potential treatment of Parkinson's disease. J Med Chem. 2014; 58(2):718-38. DOI: 10.1021/jm501254d. View

Kaczor A, Jorg M, Capuano B . The dopamine D2 receptor dimer and its interaction with homobivalent antagonists: homology modeling, docking and molecular dynamics. J Mol Model. 2016; 22(9):203. PMC: 5023759. DOI: 10.1007/s00894-016-3065-2. View

Orru M, Bakesova J, Brugarolas M, Quiroz C, Beaumont V, Goldberg S . Striatal pre- and postsynaptic profile of adenosine A(2A) receptor antagonists. PLoS One. 2011; 6(1):e16088. PMC: 3019225. DOI: 10.1371/journal.pone.0016088. View

10.

Daniels D, Lenard N, Etienne C, Law P, Roerig S, Portoghese P . Opioid-induced tolerance and dependence in mice is modulated by the distance between pharmacophores in a bivalent ligand series. Proc Natl Acad Sci U S A. 2005; 102(52):19208-13. PMC: 1323165. DOI: 10.1073/pnas.0506627102. View

11.

Casado V, Cortes A, Ciruela F, Mallol J, Ferre S, Lluis C . Old and new ways to calculate the affinity of agonists and antagonists interacting with G-protein-coupled monomeric and dimeric receptors: the receptor-dimer cooperativity index. Pharmacol Ther. 2007; 116(3):343-54. DOI: 10.1016/j.pharmthera.2007.05.010. View

12.

Ferre S, Casado V, Devi L, Filizola M, Jockers R, Lohse M . G protein-coupled receptor oligomerization revisited: functional and pharmacological perspectives. Pharmacol Rev. 2014; 66(2):413-34. PMC: 3973609. DOI: 10.1124/pr.113.008052. View

13.

Mohr K, Schmitz J, Schrage R, Trankle C, Holzgrabe U . Molecular alliance-from orthosteric and allosteric ligands to dualsteric/bitopic agonists at G protein coupled receptors. Angew Chem Int Ed Engl. 2012; 52(2):508-16. DOI: 10.1002/anie.201205315. View

14.

Moreno E, Quiroz C, Rea W, Cai N, Mallol J, Cortes A . Functional μ-Opioid-Galanin Receptor Heteromers in the Ventral Tegmental Area. J Neurosci. 2016; 37(5):1176-1186. PMC: 5296795. DOI: 10.1523/JNEUROSCI.2442-16.2016. View

15.

Stuart A, Fiser A, Sanchez R, Melo F, Sali A . Comparative protein structure modeling of genes and genomes. Annu Rev Biophys Biomol Struct. 2000; 29:291-325. DOI: 10.1146/annurev.biophys.29.1.291. View

16.

Busnelli M, Kleinau G, Muttenthaler M, Stoev S, Manning M, Bibic L . Design and Characterization of Superpotent Bivalent Ligands Targeting Oxytocin Receptor Dimers via a Channel-Like Structure. J Med Chem. 2016; 59(15):7152-66. DOI: 10.1021/acs.jmedchem.6b00564. View

17.

Pronk S, Pall S, Schulz R, Larsson P, Bjelkmar P, Apostolov R . GROMACS 4.5: a high-throughput and highly parallel open source molecular simulation toolkit. Bioinformatics. 2013; 29(7):845-54. PMC: 3605599. DOI: 10.1093/bioinformatics/btt055. View

18.

Gaitonde S, Gonzalez-Maeso J . Contribution of heteromerization to G protein-coupled receptor function. Curr Opin Pharmacol. 2016; 32:23-31. PMC: 5395348. DOI: 10.1016/j.coph.2016.10.006. View

19.

Kern A, Albarran-Zeckler R, Walsh H, Smith R . Apo-ghrelin receptor forms heteromers with DRD2 in hypothalamic neurons and is essential for anorexigenic effects of DRD2 agonism. Neuron. 2012; 73(2):317-32. PMC: 3269786. DOI: 10.1016/j.neuron.2011.10.038. View

20.

Guo W, Shi L, Javitch J . The fourth transmembrane segment forms the interface of the dopamine D2 receptor homodimer. J Biol Chem. 2002; 278(7):4385-8. DOI: 10.1074/jbc.C200679200. View