Evaluation of drug-muscarinic receptor affinities using cell membrane chromatography and radioligand binding assay in guinea pig jejunum membrane

Acta Pharmacol Sin. 2005 Jan;26(1):113-6. doi: 10.1111/j.1745-7254.2005.00015.x.

Abstract

Aim: To study if cell membrane chromatography (CMC) could reflect drug-receptor interaction and evaluate the affinity and competitive binding to muscarinic acetylcholine receptor (mAChR).

Methods: The cell membrane stationary phase (CMSP) was prepared by immobilizing guinea pig jejunum cell membrane on the surface of a silica carrier, and was used for the rapid on-line chromatographic evaluation of ligand binding affinities to mAChR. The affinity to mAChR was also evaluated from radioligand binding assays (RBA) using the same jejunum membrane preparation.

Results: The capacity factor (k') profiles in guinea pig jejunum CMSP were: (-)QNB (15.4)>(+)QNB (11.5)>atropine (5.35)>pirenzepine (5.26)>4-DAMP (4.45)>AF-DX116 (4.18)>pilocarpine (3.93)>acetylcholine (1.31). These results compared with the affinity rank orders obtained from radioligand binding assays indicated that there was a positive correlation (r2= 0.8525, P<0.0001) between both data sets.

Conclusion: The CMC method can be used to evaluate drug-receptor affinities for drug candidates.

MeSH terms

  • Animals
  • Atropine / metabolism
  • Binding, Competitive
  • Cell Membrane / metabolism*
  • Chromatography, Affinity / methods*
  • Drug Interactions
  • Female
  • Guinea Pigs
  • In Vitro Techniques
  • Jejunum / ultrastructure
  • Male
  • Pirenzepine / metabolism
  • Quinuclidinyl Benzilate / metabolism
  • Radioligand Assay
  • Receptors, Muscarinic / metabolism*

Substances

  • Receptors, Muscarinic
  • Pirenzepine
  • Quinuclidinyl Benzilate
  • Atropine