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

A Metaxas*, Berckel BNM van, PJ Klein, J Verbeek, EC Nash, EJM Kooijman, VA Renjaän, SSV Golla, R Boellaard, JAM Christiaans, AD Windhorst, JE Leysen

*Corresponding author for this work

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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 & Perspectives
Volume7
Issue number1
Number of pages15
ISSN2052-1707
DOIs
Publication statusPublished - Feb 2019

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Methylguanidine
Dizocilpine Maleate
N-Methyl-D-Aspartate Receptors
Memantine
Tritium
Ionophores
Ketamine
Glycine
Glutamic Acid
Ligands

Keywords

  • N
  • NMDA receptor
  • N′-diaryl-N-methylguanidine
  • [ H]GMOM
  • [ H]MK-801
  • binding
  • ion-channel
  • Rats, Wistar
  • Rats
  • Male
  • Receptors, N-Methyl-D-Aspartate/antagonists & inhibitors
  • Carbon Radioisotopes/administration & dosage
  • Dizocilpine Maleate/administration & dosage
  • Animals
  • Inhibitory Concentration 50
  • Guanidines/administration & dosage
  • Positron-Emission Tomography/methods

Cite this

Metaxas, A ; van, Berckel BNM ; Klein, PJ ; Verbeek, J ; Nash, EC ; Kooijman, EJM ; Renjaän, VA ; Golla, SSV ; Boellaard, R ; Christiaans, JAM ; Windhorst, AD ; Leysen, JE. / 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. In: Pharmacology Research & Perspectives. 2019 ; Vol. 7, No. 1.
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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.",
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author = "A Metaxas and van, {Berckel BNM} and PJ Klein and J Verbeek and EC Nash and EJM Kooijman and VA Renja{\"a}n and SSV Golla and R Boellaard and JAM Christiaans and AD Windhorst and JE Leysen",
year = "2019",
month = "2",
doi = "10.1002/prp2.458",
language = "English",
volume = "7",
journal = "Pharmacology Research & Perspectives",
issn = "2052-1707",
publisher = "JohnWiley & Sons Ltd.",
number = "1",

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Metaxas, A, van, BBNM, Klein, PJ, Verbeek, J, Nash, EC, Kooijman, EJM, Renjaän, VA, Golla, SSV, Boellaard, R, Christiaans, JAM, Windhorst, AD & Leysen, JE 2019, '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', Pharmacology Research & Perspectives, vol. 7, no. 1, e00458. https://doi.org/10.1002/prp2.458

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, A; van, Berckel BNM; Klein, PJ; Verbeek, J; Nash, EC; Kooijman, EJM; Renjaän, VA; Golla, SSV; Boellaard, R; Christiaans, JAM; Windhorst, AD; Leysen, JE.

In: Pharmacology Research & Perspectives, Vol. 7, No. 1, e00458, 02.2019.

Research output: Contribution to journalJournal articleResearchpeer-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, A

AU - van, Berckel BNM

AU - Klein, PJ

AU - Verbeek, J

AU - Nash, EC

AU - Kooijman, EJM

AU - Renjaän, VA

AU - Golla, SSV

AU - Boellaard, R

AU - Christiaans, JAM

AU - Windhorst, AD

AU - Leysen, JE

PY - 2019/2

Y1 - 2019/2

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.

KW - N

KW - NMDA receptor

KW - N′-diaryl-N-methylguanidine

KW - [ H]GMOM

KW - [ H]MK-801

KW - binding

KW - ion-channel

KW - Rats, Wistar

KW - Rats

KW - Male

KW - Receptors, N-Methyl-D-Aspartate/antagonists & inhibitors

KW - Carbon Radioisotopes/administration & dosage

KW - Dizocilpine Maleate/administration & dosage

KW - Animals

KW - Inhibitory Concentration 50

KW - Guanidines/administration & dosage

KW - Positron-Emission Tomography/methods

UR - http://europepmc.org/abstract/med/30784206

U2 - 10.1002/prp2.458

DO - 10.1002/prp2.458

M3 - Journal article

VL - 7

JO - Pharmacology Research & Perspectives

JF - Pharmacology Research & Perspectives

SN - 2052-1707

IS - 1

M1 - e00458

ER -