Characterization of the subtype selectivity of the allosteric modulator heptane-1,7-bis-(dimethyl-3'-phthalimidopropyl) ammonium bromide (c7/3-phth) at cloned muscarinic acetylcholine receptors

Arthur Christopoulos, Jennifer L. Sorman, Fred Mitchelson, Esam E. El-Fakahany

Research output: Contribution to journalArticleResearchpeer-review

Abstract

The present study investigated the interaction between the muscarinic acetylcholine receptor (mAChR) allosteric modulator heptane-1,7-bis-(dimethyl-3'-phthalimidopropyl) ammonium bromide (C7/3-phth) and the orthosteric antagonist [3H]N-methylscopolamine ([3H]NMS) at the five cloned human mAChRs expressed in Chinese hamster ovary cells. Equilibrium binding studies, using two different concentrations of radioligand, showed the interaction between C7/3-phth and [3H]NMS to be characterized by different degrees of negative cooperativity, depending on the receptor subtype. The modulator exhibited the highest affinity (85 nM) for the unoccupied M2 receptor and the lowest affinity for the unoccupied M5 receptor, the latter being approximately 100-fold lower. In contrast, the highest degree of negative cooperativity was observed at the M5 receptor, whereas lowest negative cooperativity was found at the M1 and M4 receptors. Non-equilibrium dissociation kinetic studies also confirmed the allosteric properties of C7/3-phth at all five mAChRs and yielded independent estimates of the modulator affinity for the occupied receptor. The latter estimates showed good agreement with those calculated using parameter values determined from the equilibrium experiments. The present results extend previous findings that C7/3-phth is a potent allosteric modulator at mAChRs, particularly the M2 subtype, and also highlight the effects of cooperativity on apparent drug-receptor subtype selectivity. Copyright (C) 1999 Elsevier Science Inc.

Original languageEnglish
Pages (from-to)171-179
Number of pages9
JournalBiochemical Pharmacology
Volume57
Issue number2
DOIs
Publication statusPublished - 15 Jan 1999

Keywords

  • Allosteric modulation
  • Cooperativity
  • Dissociation kinetics
  • Equilibrium binding
  • Muscarinic receptors

Cite this

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title = "Characterization of the subtype selectivity of the allosteric modulator heptane-1,7-bis-(dimethyl-3'-phthalimidopropyl) ammonium bromide (c7/3-phth) at cloned muscarinic acetylcholine receptors",
abstract = "The present study investigated the interaction between the muscarinic acetylcholine receptor (mAChR) allosteric modulator heptane-1,7-bis-(dimethyl-3'-phthalimidopropyl) ammonium bromide (C7/3-phth) and the orthosteric antagonist [3H]N-methylscopolamine ([3H]NMS) at the five cloned human mAChRs expressed in Chinese hamster ovary cells. Equilibrium binding studies, using two different concentrations of radioligand, showed the interaction between C7/3-phth and [3H]NMS to be characterized by different degrees of negative cooperativity, depending on the receptor subtype. The modulator exhibited the highest affinity (85 nM) for the unoccupied M2 receptor and the lowest affinity for the unoccupied M5 receptor, the latter being approximately 100-fold lower. In contrast, the highest degree of negative cooperativity was observed at the M5 receptor, whereas lowest negative cooperativity was found at the M1 and M4 receptors. Non-equilibrium dissociation kinetic studies also confirmed the allosteric properties of C7/3-phth at all five mAChRs and yielded independent estimates of the modulator affinity for the occupied receptor. The latter estimates showed good agreement with those calculated using parameter values determined from the equilibrium experiments. The present results extend previous findings that C7/3-phth is a potent allosteric modulator at mAChRs, particularly the M2 subtype, and also highlight the effects of cooperativity on apparent drug-receptor subtype selectivity. Copyright (C) 1999 Elsevier Science Inc.",
keywords = "Allosteric modulation, Cooperativity, Dissociation kinetics, Equilibrium binding, Muscarinic receptors",
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Characterization of the subtype selectivity of the allosteric modulator heptane-1,7-bis-(dimethyl-3'-phthalimidopropyl) ammonium bromide (c7/3-phth) at cloned muscarinic acetylcholine receptors. / Christopoulos, Arthur; Sorman, Jennifer L.; Mitchelson, Fred; El-Fakahany, Esam E.

In: Biochemical Pharmacology, Vol. 57, No. 2, 15.01.1999, p. 171-179.

Research output: Contribution to journalArticleResearchpeer-review

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