5-HT2C Receptor Structures Reveal the Structural Basis of GPCR Polypharmacology

Drugs frequently require interactions with multiple targets-via a process known as polypharmacology-to achieve their therapeutic actions. Currently, drugs targeting several serotonin receptors, including the 5-HT2C receptor, are useful for treating obesity, drug abuse, and schizophrenia. The competing challenges of developing selective 5-HT2C receptor ligands or creating drugs with a defined polypharmacological profile, especially aimed at G protein-coupled receptors (GPCRs), remain extremely difficult. Here, we solved two structures of the 5-HT2C receptor in complex with the highly promiscuous agonist ergotamine and the 5-HT2A-C receptor-selective inverse agonist ritanserin at resolutions of 3.0 Å and 2.7 Å, respectively. We analyzed their respective binding poses to provide mechanistic insights into their receptor recognition and opposing pharmacological actions. This study investigates the structural basis of polypharmacology at canonical GPCRs and illustrates how understanding characteristic patterns of ligand-receptor interaction and activation may ultimately facilitate drug design at multiple GPCRs.

[1]  Hualiang Jiang,et al.  Structural Basis for Molecular Recognition at Serotonin Receptors , 2013, Science.

[2]  J. Hagan,et al.  Functional characterisation of the human cloned 5‐HT7 receptor (long form); antagonist profile of SB‐258719 , 1998, British journal of pharmacology.

[3]  R. Abagyan,et al.  Biased probability Monte Carlo conformational searches and electrostatic calculations for peptides and proteins. , 1994, Journal of molecular biology.

[4]  H. Hamm,et al.  Conformational flexibility and structural dynamics in GPCR-mediated G protein activation: a perspective. , 2013, Journal of molecular biology.

[5]  R. Abagyan,et al.  Conserved binding mode of human beta2 adrenergic receptor inverse agonists and antagonist revealed by X-ray crystallography. , 2010, Journal of the American Chemical Society.

[6]  B. Roth,et al.  Antagonist Functional Selectivity: 5-HT2A Serotonin Receptor Antagonists Differentially Regulate 5-HT2A Receptor Protein Level In Vivo , 2011, Journal of Pharmacology and Experimental Therapeutics.

[7]  R. Rodriguiz,et al.  5-HT2C Agonists Modulate Schizophrenia-Like Behaviors in Mice , 2017, Neuropsychopharmacology.

[8]  S. Rasmussen,et al.  Crystal Structure of the β2Adrenergic Receptor-Gs protein complex , 2011, Nature.

[9]  Bryan L Roth,et al.  Structure and function of serotonin G protein-coupled receptors. , 2015, Pharmacology & therapeutics.

[10]  J. D. den Boer,et al.  Ritanserin as add‐on medication to neuroleptic therapy for patients with chronic or subchronic schizophrenia , 2000, Human psychopharmacology.

[11]  Randy J. Read,et al.  Acta Crystallographica Section D Biological , 2003 .

[12]  D. E. Nichols,et al.  Crystal Structure of an LSD-Bound Human Serotonin Receptor , 2017, Cell.

[13]  David E. Nichols,et al.  Psychedelics , 2016, Pharmacological Reviews.

[14]  B. Roth.  Drugs and valvular heart disease. , 2007, The New England journal of medicine.

[15]  R. Stevens,et al.  Microscale fluorescent thermal stability assay for membrane proteins. , 2008, Structure.

[16]  M. Holahan,et al.  Aripiprazole, A Drug that Displays Partial Agonism and Functional 
Selectivity , 2017, Current Neuropharmacology.

[17]  Clemens Vonrhein,et al.  Exploiting structure similarity in refinement: automated NCS and target-structure restraints in BUSTER , 2012, Acta crystallographica. Section D, Biological crystallography.

[18]  T. K. Harden,et al.  Quantification of isozyme-specific activation of phospholipase C-beta2 by Rac GTPases and phospholipase C-epsilon by Rho GTPases in an intact cell assay system. , 2006, Methods in enzymology.

[19]  George Papadatos,et al.  The ChEMBL bioactivity database: an update , 2013, Nucleic Acids Res..

[20]  Collaborative Computational,et al.  The CCP4 suite: programs for protein crystallography. , 1994, Acta crystallographica. Section D, Biological crystallography.

[21]  R. Mailman,et al.  Aripiprazole, A Novel Atypical Antipsychotic Drug with a Unique and Robust Pharmacology , 2003, Neuropsychopharmacology.

[22]  Stephen L Garland,et al.  A ligand's view of target similarity: chemogenomic binding site-directed techniques for drug discovery. , 2011, Current topics in medicinal chemistry.

[23]  Henry Lin,et al.  Structure-based discovery of opioid analgesics with reduced side effects , 2016, Nature.

[24]  V. Cherezov,et al.  Crystallizing membrane proteins using lipidic mesophases , 2009, Nature Protocols.

[25]  Brian M. Smith,et al.  Lorcaserin, a Novel Selective Human 5-Hydroxytryptamine2C Agonist: in Vitro and in Vivo Pharmacological Characterization , 2008, Journal of Pharmacology and Experimental Therapeutics.

[26]  Arthur Christopoulos,et al.  Crystal structures of the M1 and M4 muscarinic acetylcholine receptors , 2016, Nature.

[27]  Anat Levit,et al.  D4 dopamine receptor high-resolution structures enable the discovery of selective agonists , 2017, Science.

[28]  P. Emsley,et al.  Features and development of Coot , 2010, Acta crystallographica. Section D, Biological crystallography.

[29]  G. Barnea,et al.  The genetic design of signaling cascades to record receptor activation , 2008, Proceedings of the National Academy of Sciences.

[30]  Bryan L. Roth,et al.  Parallel Functional Activity Profiling Reveals Valvulopathogens Are Potent 5-Hydroxytryptamine2B Receptor Agonists: Implications for Drug Safety Assessment , 2009, Molecular Pharmacology.

[31]  Albert C. Pan,et al.  Structure and Dynamics of the M3 Muscarinic Acetylcholine Receptor , 2012, Nature.

[32]  E. sanders-Bush,et al.  Constitutively active 5-hydroxytryptamine2C receptors reveal novel inverse agonist activity of receptor ligands. , 1994, The Journal of biological chemistry.

[33]  Yang I Li,et al.  An Expanded View of Complex Traits: From Polygenic to Omnigenic , 2017, Cell.

[34]  R. Stevens,et al.  Structural Features for Functional Selectivity at Serotonin Receptors , 2013, Science.

[35]  Shuxing Zhang,et al.  Computational polypharmacology: a new paradigm for drug discovery , 2017, Expert opinion on drug discovery.

[36]  Michael J. Keiser,et al.  The Presynaptic Component of the Serotonergic System is Required for Clozapine's Efficacy , 2011, Neuropsychopharmacology.

[37]  B. Roth,et al.  Magic shotguns versus magic bullets: selectively non-selective drugs for mood disorders and schizophrenia , 2004, Nature Reviews Drug Discovery.

[38]  F. Wurm,et al.  Transfecting mammalian cells: optimization of critical parameters affecting calcium-phosphate precipitate formation. , 1996, Nucleic acids research.

[39]  Stefano Costanzi,et al.  Computational studies to predict or explain G protein coupled receptor polypharmacology. , 2014, Trends in pharmacological sciences.

[40]  Gert Vriend,et al.  GPCRDB information system for G protein-coupled receptors , 2003, Nucleic Acids Res..

[41]  Randy J. Read,et al.  Phaser crystallographic software , 2007, Journal of applied crystallography.

[42]  Gebhard F. X. Schertler,et al.  Two distinct conformations of helix 6 observed in antagonist-bound structures of a β1-adrenergic receptor , 2011, Proceedings of the National Academy of Sciences.

[43]  Maria F. Sassano,et al.  Automated design of ligands to polypharmacological profiles , 2012, Nature.

[44]  J. Palacios,et al.  A short history of the 5-HT2C receptor: from the choroid plexus to depression, obesity and addiction treatment , 2017, Psychopharmacology.

[45]  R. Stevens,et al.  How Ligands Illuminate GPCR Molecular Pharmacology , 2017, Cell.

[46]  J. Ballesteros,et al.  [19] Integrated methods for the construction of three-dimensional models and computational probing of structure-function relations in G protein-coupled receptors , 1995 .

[47]  R. Stevens,et al.  High-resolution crystal structure of an engineered human beta2-adrenergic G protein-coupled receptor. , 2007, Science.

[48]  John P. Overington,et al.  Comprehensive characterization of the Published Kinase Inhibitor Set , 2016, Nature Biotechnology.

[49]  B. Roth,et al.  Design and Discovery of Functionally Selective Serotonin 2C (5-HT2C) Receptor Agonists. , 2016, Journal of medicinal chemistry.

[50]  Maria F. Sassano,et al.  PRESTO-TANGO: an open-source resource for interrogation of the druggable human GPCR-ome , 2015, Nature Structural &Molecular Biology.

[51]  B. Roth,et al.  Lorcaserin and pimavanserin: emerging selectivity of serotonin receptor subtype-targeted drugs. , 2013, The Journal of clinical investigation.

[52]  Vadim Cherezov,et al.  Serial Femtosecond Crystallography of G Protein-Coupled Receptors. , 2018, Annual review of biophysics.

[53]  J. M. Kennedy,et al.  Standard binding and functional assays related to medications development division testing for potential cocaine and opiate narcotic treatment medications. , 1998, NIDA research monograph.

[54]  Bas Vroling,et al.  GPCRdb: an information system for G protein-coupled receptors , 2015, Nucleic Acids Res..

[55]  M. Babu,et al.  Molecular signatures of G-protein-coupled receptors , 2013, Nature.

[56]  R. Stevens,et al.  Rastering strategy for screening and centring of microcrystal samples of human membrane proteins with a sub-10 µm size X-ray synchrotron beam , 2009, Journal of The Royal Society Interface.