Inhibition of in vivo [3H]MK-801 binding by NMDA receptor open channel blockers and GluN2B antagonists in rats and mice

Fernandes, Alda; Wojcik, Trevor; Baireddy, Praveena; Pieschl, Rick L.; Newton, Amy; Tian, Yuan; Yang, Hong; Bristow, Linda J.; Li, Yuwen

doi:10.1016/j.ejphar.2015.08.044

Cited by 19 publications

(22 citation statements)

References 32 publications

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“…Specifically, the conformational change produced by binding to the GluN2B allosteric site closes channels containing the GluN2B subunit, prevents [ 3 H]MK-801 from accessing its binding site in the channel pore, and reduces [ 3 H]MK-801 binding. Previous studies have shown that GluN2B NAMs maximally inhibit [ 3 H]MK-801 binding by ∼60%, suggesting that GluN2B-containing NMDA receptors represent 60% of the total NMDA receptor population labeled in vivo (Fernandes et al, 2015). In a similar manner, treatment with BMS-986163 also inhibited [ 3 H]MK-801 binding in rats, confirming functional NMDA receptor inhibition in vivo.…”

Section: Discussionsupporting

confidence: 59%

“…The in vivo characterization of BMS-986169 and the phosphate prodrug BMS-986163 focused on 1) confirmation of occupancy and functional inhibition of GluN2B receptors, 2) testing in assays of antidepressant drug-like effects and side effects, and 3) testing on translational measures that can be investigated in healthy humans. GluN2B occupancy in rodents was measured by ex vivo [ 3 H]Ro 25-6981 binding (Fernandes et al, 2015) and showed a dose-dependent increase that fully saturated at doses $3 mg/kg. Occupancy was tightly correlated with exposure, and occupancy/exposure relationships were similar after i.v.…”

Section: Discussionmentioning

confidence: 99%

“…dosing with BMS-986169 or BMS-986163, confirming rapid and effective cleavage of the prodrug. Functional inhibition of GluN2B receptors was demonstrated by inhibition of in vivo [ 3 H]MK-801 binding (Fernandes et al, 2015). [ 3 H]MK-801 is a radiotracer that labels all NMDA receptor subtypes present in an open conformational state.…”

Section: Discussionmentioning

confidence: 99%

“…[ 3 H]MK-801 is a radiotracer that labels all NMDA receptor subtypes present in an open conformational state. The assay can be used to show both direct binding site competition following treatment with other NMDA receptor open-channel blockers and binding inhibition due to the functional effects of agents acting at other modulatory sites (Murray et al, 2000;Fernandes et al, 2015). Specifically, the conformational change produced by binding to the GluN2B allosteric site closes channels containing the GluN2B subunit, prevents [ 3 H]MK-801 from accessing its binding site in the channel pore, and reduces [ 3 H]MK-801 binding.…”

Section: Discussionmentioning

confidence: 99%

“…In Vivo [ 3 H]MK-801 Binding in Rats. Studies to determine inhibition of in vivo [ 3 H]MK-801 binding were conducted at BMS using methods previously described (Fernandes et al, 2015). For the doseresponse study, rats with previously implanted jugular vein catheters were randomly assigned (n 5 4/group) to treatment with either vehicle (0.9% saline, 2 ml/kg, i.v.…”

Section: Methodsmentioning

confidence: 99%

See 4 more Smart Citations

Preclinical Characterization of (R)-3-((3S,4S)-3-fluoro-4-(4-hydroxyphenyl)piperidin-1-yl)-1-(4-methylbenzyl)pyrrolidin-2-one (BMS-986169), a Novel, Intravenous, Glutamate N-Methyl-d-Aspartate 2B Receptor Negative Allosteric Modulator with Potential in Major Depressive Disorder

Bristow

Gulia

Weed

et al. 2017

J Pharmacol Exp Ther

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()-3-((3S,4S)-3-fluoro-4-(4-hydroxyphenyl)piperidin-1-yl)-1-(4-methylbenzyl)pyrrolidin-2-one (BMS-986169) and the phosphate prodrug 4-((3,4)-3-fluoro-1-((R)-1-(4-methylbenzyl)-2-oxopyrrolidin-3-yl)piperidin-4-yl)phenyl dihydrogen phosphate (BMS-986163) were identified from a drug discovery effort focused on the development of novel, intravenous glutamate -methyl-d-aspartate 2B receptor (GluN2B) negative allosteric modulators (NAMs) for treatment-resistant depression (TRD). BMS-986169 showed high binding affinity for the GluN2B subunit allosteric modulatory site (K = 4.03-6.3 nM) and selectively inhibited GluN2B receptor function in Xenopus oocytes expressing human -methyl-d-aspartate receptor subtypes (IC = 24.1 nM). BMS-986169 weakly inhibited human ether-a-go-go-related gene channel activity (IC = 28.4 M) and had negligible activity in an assay panel containing 40 additional pharmacological targets. Intravenous administration of BMS-986169 or BMS-986163 dose-dependently increased GluN2B receptor occupancy and inhibited in vivo [3H](+)-5-methyl-10,11-dihydro-5H-dibenzo[]cyclohepten-5,10-imine ([3H]MK-801) binding, confirming target engagement and effective cleavage of the prodrug. BMS-986169 reduced immobility in the mouse forced swim test, an effect similar to intravenous ketamine treatment. Decreased novelty suppressed feeding latency, and increased ex vivo hippocampal long-term potentiation was also seen 24 hours after acute BMS-986163 or BMS-986169 administration. BMS-986169 did not produce ketamine-like hyperlocomotion or abnormal behaviors in mice or cynomolgus monkeys but did produce a transient working memory impairment in monkeys that was closely related to plasma exposure. Finally, BMS-986163 produced robust changes in the quantitative electroencephalogram power band distribution, a translational measure that can be used to assess pharmacodynamic activity in healthy humans. Due to the poor aqueous solubility of BMS-986169, BMS-986163 was selected as the lead GluN2B NAM candidate for further evaluation as a novel intravenous agent for TRD.

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Section: Discussionsupporting

confidence: 59%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 3 more Smart Citations

Preclinical Characterization of (R)-3-((3S,4S)-3-fluoro-4-(4-hydroxyphenyl)piperidin-1-yl)-1-(4-methylbenzyl)pyrrolidin-2-one (BMS-986169), a Novel, Intravenous, Glutamate N-Methyl-d-Aspartate 2B Receptor Negative Allosteric Modulator with Potential in Major Depressive Disorder

Bristow

Gulia

Weed

et al. 2017

J Pharmacol Exp Ther

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Exploring the overlapping binding sites of ifenprodil and EVT‐101 in GluN2B‐containing NMDA receptors using novel chicken embryo forebrain cultures and molecular modeling

Fjelldal

Freyd

Evenseth

et al. 2019

Pharmacology Res & Perspec

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N‐methyl‐ d ‐aspartate receptors ( NMDAR ) are widely expressed in the brain. GluN2B subunit‐containing NMDAR s has recently attracted significant attention as potential pharmacological targets, with emphasis on the functional properties of allosteric antagonists. We used primary cultures from chicken embryo forebrain (E10), expressing native GluN2B‐containing NMDA receptors as a novel model system. Comparing the inhibition of calcium influx by well‐known GluN2B subunit‐specific allosteric antagonists, the following rank order of potency was found: EVT ‐101 ( EC 50 22 ± 8 nmol/L) > Ro 25‐6981 ( EC 50 60 ± 30 nmol/L) > ifenprodil ( EC 50 100 ± 40 nmol/L) > eliprodil ( EC 50 1300 ± 700 nmol/L), similar to previous observations in rat cortical cultures and cell lines overexpressing chimeric receptors. The less explored Ro 04‐5595 had an EC 50 of 186 ± 32 nmol/L. Venturing to explain the differences in potency, binding properties were further studied by in silico docking and molecular dynamics simulations using x‐ray crystal structures of GluN1/GluN2B amino terminal domain. We found that Ro 04‐5595 was predicted to bind the recently discovered EVT ‐101 binding site, not the ifenprodil‐binding site. The EVT ‐101 binding pocket appears to accommodate more structurally different ligands than the ifenprodil‐binding site, and contains residues essential in ligand interactions necessary for calcium influx inhibition. For the ifenprodil site, the less effective antagonist (eliprodil) fails to interact with key residues, while in the EVT ‐101 pocket, difference in potency might be explained by differences in ligand‐receptor interaction patterns.

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MK801-induced locomotor activity in preweanling and adolescent male and female rats: role of the dopamine and serotonin systems

et al. 2020

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Inhibition of in vivo [3H]MK-801 binding by NMDA receptor open channel blockers and GluN2B antagonists in rats and mice

Cited by 19 publications

References 32 publications

Preclinical Characterization of (R)-3-((3S,4S)-3-fluoro-4-(4-hydroxyphenyl)piperidin-1-yl)-1-(4-methylbenzyl)pyrrolidin-2-one (BMS-986169), a Novel, Intravenous, Glutamate N-Methyl-d-Aspartate 2B Receptor Negative Allosteric Modulator with Potential in Major Depressive Disorder

Preclinical Characterization of (R)-3-((3S,4S)-3-fluoro-4-(4-hydroxyphenyl)piperidin-1-yl)-1-(4-methylbenzyl)pyrrolidin-2-one (BMS-986169), a Novel, Intravenous, Glutamate N-Methyl-d-Aspartate 2B Receptor Negative Allosteric Modulator with Potential in Major Depressive Disorder

Exploring the overlapping binding sites of ifenprodil and EVT‐101 in GluN2B‐containing NMDA receptors using novel chicken embryo forebrain cultures and molecular modeling

MK801-induced locomotor activity in preweanling and adolescent male and female rats: role of the dopamine and serotonin systems

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