Binding characterization of N-(2-chloro-5-thiomethylphenyl)-N′-(3-[ 3 H] 3 methoxy phenyl)-N′-methylguanidine ([ 3 H]GMOM), a non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist

Athanasios Metaxas, Bart N. M. van Berckel, Pieter J. Klein, Joost Verbeek, Emily C. Nash, Esther J. M. Kooijman, V. ronique A. Renjaän, Sandeep S. V. Golla, Ronald Boellaard, Johannes A. M. Christiaans, Albert D. Windhorst, Josée E. Leysen

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Labeled with carbon-11, N-(2-chloro-5-thiomethylphenyl)-N′-(3-methoxyphenyl)-N′-methylguanidine ([ 11 C]GMOM) is currently the only positron emission tomography (PET) tracer that has shown selectivity for the ion-channel site of N-methyl-D-aspartate (NMDA) receptors in human imaging studies. The present study reports on the selectivity profile and in vitro binding properties of GMOM. The compound was screened on a panel of 80 targets, and labeled with tritium ([ 3 H]GMOM). The binding properties of [ 3 H]GMOM were compared to those of the reference ion-channel ligand [ 3 H](+)-dizocilpine maleate ([ 3 H]MK-801), in a set of concentration-response, homologous and heterologous inhibition, and association kinetics assays, performed with repeatedly washed rat forebrain preparations. GMOM was at least 70-fold more selective for NMDA receptors compared to all other targets examined. In homologous inhibition and concentration-response assays, the binding of [ 3 H]GMOM was regulated by NMDA receptor agonists, albeit in a less prominent manner compared to [ 3 H]MK-801. Scatchard transformation of homologous inhibition data produced concave upward curves for [ 3 H]GMOM and [ 3 H]MK-801. The radioligands showed bi-exponential association kinetics in the presence of 100 μmol L −1 l-glutamate/30 μmol L −1 glycine. [ 3 H]GMOM (3 nmol L −1 and 10 nmol L −1 ) was inhibited with dual affinity by (+)-MK-801, (R,S)-ketamine and memantine, in both presence and absence of agonists. [ 3 H]MK-801 (2 nmol L −1 ) was inhibited in a monophasic manner by GMOM under baseline and combined agonist conditions, with an IC 50 value of ~19 nmol L −1 . The non-linear Scatchard plots, biphasic inhibition by open channel blockers, and bi-exponential kinetics of [ 3 H]GMOM indicate a complex mechanism of interaction with the NMDA receptor ionophore. The implications for quantifying the PET signal of [ 11 C]GMOM are discussed.
Original languageEnglish
Article numbere00458
JournalPharmacology Research and Perspectives
Volume7
Issue number1
DOIs
Publication statusPublished - 19 Feb 2019

Cite this

@article{212b257e1fc749c6a9767b4f044ca42d,
title = "Binding characterization of N-(2-chloro-5-thiomethylphenyl)-N′-(3-[ 3 H] 3 methoxy phenyl)-N′-methylguanidine ([ 3 H]GMOM), a non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist",
abstract = "Labeled with carbon-11, N-(2-chloro-5-thiomethylphenyl)-N′-(3-methoxyphenyl)-N′-methylguanidine ([ 11 C]GMOM) is currently the only positron emission tomography (PET) tracer that has shown selectivity for the ion-channel site of N-methyl-D-aspartate (NMDA) receptors in human imaging studies. The present study reports on the selectivity profile and in vitro binding properties of GMOM. The compound was screened on a panel of 80 targets, and labeled with tritium ([ 3 H]GMOM). The binding properties of [ 3 H]GMOM were compared to those of the reference ion-channel ligand [ 3 H](+)-dizocilpine maleate ([ 3 H]MK-801), in a set of concentration-response, homologous and heterologous inhibition, and association kinetics assays, performed with repeatedly washed rat forebrain preparations. GMOM was at least 70-fold more selective for NMDA receptors compared to all other targets examined. In homologous inhibition and concentration-response assays, the binding of [ 3 H]GMOM was regulated by NMDA receptor agonists, albeit in a less prominent manner compared to [ 3 H]MK-801. Scatchard transformation of homologous inhibition data produced concave upward curves for [ 3 H]GMOM and [ 3 H]MK-801. The radioligands showed bi-exponential association kinetics in the presence of 100 μmol L −1 l-glutamate/30 μmol L −1 glycine. [ 3 H]GMOM (3 nmol L −1 and 10 nmol L −1 ) was inhibited with dual affinity by (+)-MK-801, (R,S)-ketamine and memantine, in both presence and absence of agonists. [ 3 H]MK-801 (2 nmol L −1 ) was inhibited in a monophasic manner by GMOM under baseline and combined agonist conditions, with an IC 50 value of ~19 nmol L −1 . The non-linear Scatchard plots, biphasic inhibition by open channel blockers, and bi-exponential kinetics of [ 3 H]GMOM indicate a complex mechanism of interaction with the NMDA receptor ionophore. The implications for quantifying the PET signal of [ 11 C]GMOM are discussed.",
author = "Athanasios Metaxas and {van Berckel}, {Bart N. M.} and Klein, {Pieter J.} and Joost Verbeek and Nash, {Emily C.} and Kooijman, {Esther J. M.} and Renja{\"a}n, {V. ronique A.} and Golla, {Sandeep S. V.} and Ronald Boellaard and Christiaans, {Johannes A. M.} and Windhorst, {Albert D.} and Leysen, {Jos{\'e}e E.}",
note = "{\circledC} 2019 The Authors. Pharmacology Research & Perspectives published by John Wiley & Sons Ltd, British Pharmacological Society and American Society for Pharmacology and Experimental Therapeutics.",
year = "2019",
month = "2",
day = "19",
doi = "10.1002/prp2.458",
language = "English",
volume = "7",
journal = "Pharmacology Research and Perspectives",
issn = "2052-1707",
publisher = "Wiley-Blackwell Publishing Ltd",
number = "1",

}

Binding characterization of N-(2-chloro-5-thiomethylphenyl)-N′-(3-[ 3 H] 3 methoxy phenyl)-N′-methylguanidine ([ 3 H]GMOM), a non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist. / Metaxas, Athanasios; van Berckel, Bart N. M.; Klein, Pieter J.; Verbeek, Joost; Nash, Emily C.; Kooijman, Esther J. M.; Renjaän, V. ronique A.; Golla, Sandeep S. V.; Boellaard, Ronald; Christiaans, Johannes A. M.; Windhorst, Albert D.; Leysen, Josée E.

In: Pharmacology Research and Perspectives, Vol. 7, No. 1, e00458, 19.02.2019.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Binding characterization of N-(2-chloro-5-thiomethylphenyl)-N′-(3-[ 3 H] 3 methoxy phenyl)-N′-methylguanidine ([ 3 H]GMOM), a non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist

AU - Metaxas, Athanasios

AU - van Berckel, Bart N. M.

AU - Klein, Pieter J.

AU - Verbeek, Joost

AU - Nash, Emily C.

AU - Kooijman, Esther J. M.

AU - Renjaän, V. ronique A.

AU - Golla, Sandeep S. V.

AU - Boellaard, Ronald

AU - Christiaans, Johannes A. M.

AU - Windhorst, Albert D.

AU - Leysen, Josée E.

N1 - © 2019 The Authors. Pharmacology Research & Perspectives published by John Wiley & Sons Ltd, British Pharmacological Society and American Society for Pharmacology and Experimental Therapeutics.

PY - 2019/2/19

Y1 - 2019/2/19

N2 - Labeled with carbon-11, N-(2-chloro-5-thiomethylphenyl)-N′-(3-methoxyphenyl)-N′-methylguanidine ([ 11 C]GMOM) is currently the only positron emission tomography (PET) tracer that has shown selectivity for the ion-channel site of N-methyl-D-aspartate (NMDA) receptors in human imaging studies. The present study reports on the selectivity profile and in vitro binding properties of GMOM. The compound was screened on a panel of 80 targets, and labeled with tritium ([ 3 H]GMOM). The binding properties of [ 3 H]GMOM were compared to those of the reference ion-channel ligand [ 3 H](+)-dizocilpine maleate ([ 3 H]MK-801), in a set of concentration-response, homologous and heterologous inhibition, and association kinetics assays, performed with repeatedly washed rat forebrain preparations. GMOM was at least 70-fold more selective for NMDA receptors compared to all other targets examined. In homologous inhibition and concentration-response assays, the binding of [ 3 H]GMOM was regulated by NMDA receptor agonists, albeit in a less prominent manner compared to [ 3 H]MK-801. Scatchard transformation of homologous inhibition data produced concave upward curves for [ 3 H]GMOM and [ 3 H]MK-801. The radioligands showed bi-exponential association kinetics in the presence of 100 μmol L −1 l-glutamate/30 μmol L −1 glycine. [ 3 H]GMOM (3 nmol L −1 and 10 nmol L −1 ) was inhibited with dual affinity by (+)-MK-801, (R,S)-ketamine and memantine, in both presence and absence of agonists. [ 3 H]MK-801 (2 nmol L −1 ) was inhibited in a monophasic manner by GMOM under baseline and combined agonist conditions, with an IC 50 value of ~19 nmol L −1 . The non-linear Scatchard plots, biphasic inhibition by open channel blockers, and bi-exponential kinetics of [ 3 H]GMOM indicate a complex mechanism of interaction with the NMDA receptor ionophore. The implications for quantifying the PET signal of [ 11 C]GMOM are discussed.

AB - Labeled with carbon-11, N-(2-chloro-5-thiomethylphenyl)-N′-(3-methoxyphenyl)-N′-methylguanidine ([ 11 C]GMOM) is currently the only positron emission tomography (PET) tracer that has shown selectivity for the ion-channel site of N-methyl-D-aspartate (NMDA) receptors in human imaging studies. The present study reports on the selectivity profile and in vitro binding properties of GMOM. The compound was screened on a panel of 80 targets, and labeled with tritium ([ 3 H]GMOM). The binding properties of [ 3 H]GMOM were compared to those of the reference ion-channel ligand [ 3 H](+)-dizocilpine maleate ([ 3 H]MK-801), in a set of concentration-response, homologous and heterologous inhibition, and association kinetics assays, performed with repeatedly washed rat forebrain preparations. GMOM was at least 70-fold more selective for NMDA receptors compared to all other targets examined. In homologous inhibition and concentration-response assays, the binding of [ 3 H]GMOM was regulated by NMDA receptor agonists, albeit in a less prominent manner compared to [ 3 H]MK-801. Scatchard transformation of homologous inhibition data produced concave upward curves for [ 3 H]GMOM and [ 3 H]MK-801. The radioligands showed bi-exponential association kinetics in the presence of 100 μmol L −1 l-glutamate/30 μmol L −1 glycine. [ 3 H]GMOM (3 nmol L −1 and 10 nmol L −1 ) was inhibited with dual affinity by (+)-MK-801, (R,S)-ketamine and memantine, in both presence and absence of agonists. [ 3 H]MK-801 (2 nmol L −1 ) was inhibited in a monophasic manner by GMOM under baseline and combined agonist conditions, with an IC 50 value of ~19 nmol L −1 . The non-linear Scatchard plots, biphasic inhibition by open channel blockers, and bi-exponential kinetics of [ 3 H]GMOM indicate a complex mechanism of interaction with the NMDA receptor ionophore. The implications for quantifying the PET signal of [ 11 C]GMOM are discussed.

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UR - https://www.ncbi.nlm.nih.gov/pubmed/30784206

U2 - 10.1002/prp2.458

DO - 10.1002/prp2.458

M3 - Article

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JO - Pharmacology Research and Perspectives

JF - Pharmacology Research and Perspectives

SN - 2052-1707

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