Naunyn-Schmiedeberg's Archives of Pharmacology

, Volume 388, Issue 1, pp 51–65

Structure-bias relationships for fenoterol stereoisomers in six molecular and cellular assays at the β2-adrenoceptor

  • Michael T. Reinartz
  • Solveig Kälble
  • Timo Littmann
  • Takeaki Ozawa
  • Stefan Dove
  • Volkhard Kaever
  • Irving W. Wainer
  • Roland Seifert
Original Article

Abstract

Functional selectivity is well established as an underlying concept of ligand-specific signaling via G protein-coupled receptors (GPCRs). Functionally, selective drugs could show greater therapeutic efficacy and fewer adverse effects. Dual coupling of the β2-adrenoceptor (β2AR) triggers a signal transduction via Gsα and Giα proteins. Here, we examined 12 fenoterol stereoisomers in six molecular and cellular assays. Using β2AR-Gsα and β2AR-Giα fusion proteins, (R,S’)- and (S,S’)-isomers of 4′-methoxy-1-naphthyl-fenoterol were identified as biased ligands with preference for Gs. G protein-independent signaling via β-arrestin-2 was disfavored by these ligands. Isolated human neutrophils constituted an ex vivo model of β2AR signaling and demonstrated functional selectivity through the dissociation of cAMP accumulation and the inhibition of formyl peptide-stimulated production of reactive oxygen species. Ligand bias was calculated using an operational model of agonism and revealed that the fenoterol scaffold constitutes a promising lead structure for the development of Gs-biased β2AR agonists.

Keywords

Functional selectivity β2-Adrenergic receptor Biased ligand Bias quantification Structure-bias relationships Fenoterol 

Abbreviations

7TMR

Seven-transmembrane domain receptor

AC

Adenylyl cyclase

β2AR

β2-Adrenoceptor

β-arr-2

β-Arrestin type 2

BSA

Bovine serum albumin

DHA

Dihydroalprenolol

DMEM

Dulbecco’s modified Eagle’s medium

ECL

Extracellular loop

EDTA

Ethylenediaminetetraacetic acid

EPI

Epinephrine (adrenaline)

fMLF

N-formyl-l-methionyl-l-leucyl-l-phenylalanine

Giα

Inhibitory Gα protein

Gsα

Stimulatory Gα protein

GPCR

G protein-coupled receptor

GTPase

GTP-hydrolyzing activity

ISO

Isoproterenol

PBS

Phosphate-buffered saline

PMSF

Phenylmethanesulfonylfluoride

RGS4

Regulatory protein of G protein signaling, type 4

ROS

Reactive oxygen species

Sf9

Clonal isolate of Spodoptera frugiperda ovary cells

TM

Transmembrane domain

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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Michael T. Reinartz
    • 1
  • Solveig Kälble
    • 1
  • Timo Littmann
    • 1
  • Takeaki Ozawa
    • 2
  • Stefan Dove
    • 3
  • Volkhard Kaever
    • 1
    • 4
  • Irving W. Wainer
    • 5
  • Roland Seifert
    • 1
  1. 1.Institute for PharmacologyHannover Medical SchoolHannoverGermany
  2. 2.Department of Chemistry, School of ScienceThe University of TokyoTokyoJapan
  3. 3.Department of Pharmaceutical and Medicinal Chemistry IIUniversity of RegensburgRegensburgGermany
  4. 4.Core Unit MetabolomicsHannover Medical SchoolHannoverGermany
  5. 5.Laboratory of Clinical Investigation, Biomedical Research Center, National Institute on AgingNational Institutes of HealthBaltimoreUSA

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