Opioid peptide receptor studies. 14. Stereochemistry determines agonist efficacy and intrinsic efficacy in the [(35)S]GTP-gamma-S functional binding assay

Synapse. 2001 Jan;39(1):64-9. doi: 10.1002/1098-2396(20010101)39:1<64::AID-SYN9>3.0.CO;2-J.


Previous data obtained with the cloned rat mu opioid receptor demonstrated that stereochemistry affects the four parameters of the ligand-receptor interaction: potency (ED(50)), efficacy (maximal stimulation), intrinsic efficacy (effect as a function of receptor occupation), and binding affinity. This study evaluated the activities of structurally diverse opioid receptor ligands in the [(35)S]GTP-gamma-S binding assay, comparing the relationship between receptor binding, activation, efficacy, and intrinsic efficacy. The data, obtained with cloned rat mu receptors, demonstrated that an analgetic, (-)-5-m-hydroxyphenyl-2-methylmorphan (NIH8508), and its (+)-isomer (NIH8509), behave as partial agonists, but had different intrinsic efficacy in the [(35)S]GTP-gamma-S binding assay. Replacement of the methyl group with the phenethyl group on the piperidine nitrogen of NIH8508 and NIH8509 [(1R,5S)-AH019 and (1S, 5R)-AH019] increased affinity for the mu receptor and eliminated any agonist effect, supporting the hypothesis that certain structural features make these compounds antagonists. These study also show that all of the fully efficacious mu agonists studied here had high levels of intrinsic efficacy, producing a 50% response at about 10% receptor occupancy. Comparison of the binding K(i) in competitively inhibiting [(125)I]IOXY binding to the functional K(i) for opioid antagonists [K(i)(IOXY)/K(i)(GTP-gamma-S)] provides more detailed evidence that the [(35)S]GTP-gamma-S binding assay can be used to reliably determine apparent functional antagonist K(i) values in addition to agonist ED(50), efficacy and intrinsic efficacy.

MeSH terms

  • Analgesics, Opioid / chemistry*
  • Analgesics, Opioid / pharmacokinetics*
  • Animals
  • Binding Sites / drug effects*
  • Binding Sites / physiology
  • Guanosine 5'-O-(3-Thiotriphosphate) / pharmacokinetics
  • Mice
  • Molecular Conformation*
  • Morphinans / metabolism
  • Morphinans / pharmacology*
  • Radioligand Assay / statistics & numerical data
  • Receptors, Opioid, mu / agonists*
  • Receptors, Opioid, mu / metabolism*
  • Sulfur Radioisotopes
  • Tumor Cells, Cultured


  • AH019
  • Analgesics, Opioid
  • Morphinans
  • Receptors, Opioid, mu
  • Sulfur Radioisotopes
  • Guanosine 5'-O-(3-Thiotriphosphate)