Binding kinetics redefine the antagonist pharmacology of the corticotropin-releasing factor type 1 receptor

J Pharmacol Exp Ther. 2012 May;341(2):518-31. doi: 10.1124/jpet.111.188714. Epub 2012 Feb 22.


Corticotropin-releasing factor (CRF) receptor antagonists are under preclinical and clinical investigation for stress-related disorders. In this study the impact of receptor-ligand binding kinetics on CRF₁ receptor antagonist pharmacology was investigated by measuring the association rate constant (k₁), dissociation rate constant (k₋₁), and kinetically derived affinity at 37°C. Three aspects of antagonist pharmacology were reevaluated: comparative binding activity of advanced compounds, in vivo efficacy, and structure-activity relationships. Twelve lead compounds, with little previously noted difference of affinity, varied substantially in their kinetic binding activity with a 510-fold range of kinetically derived affinity (k₋₁/k₁), 170-fold range of k₋₁, and 13-fold range of k₁. The k₋₁ values indicated previous affinity measurements were not close to equilibrium, resulting in compression of the measured affinity range. Dissociation was exceptionally slow for three ligands (k₋₁ t(1/2) of 1.6-7.2 h at 37°C). Differences of binding behavior were consistent with in vivo pharmacodynamics (suppression of adrenocorticotropin in adrenalectomized rats). Ligand concentration-effect relationships correlated with their kinetically derived affinity. Two ligands that dissociated slowly (53 and 130 min) produced prolonged suppression, whereas only transient suppression was observed with a more rapidly dissociating ligand (16 min). Investigating the structure-activity relationship indicated exceptionally low values of k₁, approaching 100,000-fold less than the diffusion-limited rate. Retrospective interpretation of medicinal chemistry indicates optimizing specific elements of chemical structure overcame kinetic barriers in the association pathway, for example, constraint of the pendant aromatic orthogonal to the ligand core. Collectively, these findings demonstrate receptor binding kinetics provide new dimensions for understanding and potentially advancing the pharmacology of CRF₁ receptor antagonists.

MeSH terms

  • Animals
  • Binding, Competitive
  • HEK293 Cells
  • Humans
  • Kinetics
  • Ligands
  • Protein Binding
  • Radioligand Assay / methods
  • Rats
  • Receptors, Corticotropin-Releasing Hormone / antagonists & inhibitors*
  • Receptors, Corticotropin-Releasing Hormone / chemistry
  • Receptors, Corticotropin-Releasing Hormone / metabolism*
  • Retrospective Studies
  • Structure-Activity Relationship


  • Ligands
  • Receptors, Corticotropin-Releasing Hormone
  • CRF receptor type 1