The First Potent and Selective Inhibitors of the Glycine Transporter Type 2

Caulfield, Wilson; Collie, Iain T.; Dickins, Rachel S.; Epemolu, Ola; McGuire, Ross; Hill, David R.; Mcvey, Gillian; Morphy, J. Richard; Rankovic, Zoran; Sundaram, Hardy

doi:10.1021/jm0011272

Cited by 91 publications

(90 citation statements)

References 15 publications

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“…Rabbit and rat antibodies against the GLYT2 N terminus have been previously characterized (9,31). The GLYT2 inhibitor ORG25543 was generously donated by Dr. Zoran Rankovic, Organon, Scotland, United Kingdom (32). The GLYT1 inhibitor N-[3-(4-fluorophenyl)-3-(4-phenylphenoxy)-propyl]sarcosine (NFPS) was a gift of Dr. Jesús Benavides (Sanofi-Aventis, Vitry sur Seine, France).…”

Section: Methodsmentioning

confidence: 99%

P2Y Purinergic Regulation of the Glycine Neurotransmitter Transporters

Jiménez

Zafra

Pérez-Sen

et al. 2011

Journal of Biological Chemistry

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The sodium-and chloride-coupled glycine neurotransmitter transporters (GLYTs) control the availability of glycine at glycine-mediated synapses. The mainly glial GLYT1 is the key regulator of the glycine levels in glycinergic and glutamatergic pathways, whereas the neuronal GLYT2 is involved in the recycling of synaptic glycine from the inhibitory synaptic cleft. In this study, we report that stimulation of P2Y purinergic receptors with 2-methylthioadenosine 5 -diphosphate in rat brainstem/spinal cord primary neuronal cultures and adult rat synaptosomes leads to the inhibition of GLYT2 and the stimulation of GLYT1 by a paracrine regulation. These effects are mainly mediated by the ADP-preferring subtypes P2Y 1 and P2Y 13 because the effects are partially reversed by the specific antagonists N 6 -methyl-2 -deoxyadenosine-3 ,5 -bisphosphate and pyridoxal-5 -phosphate-6-azo(2-chloro-5-nitrophenyl)-2,4-disulfonate and are totally blocked by suramin. P2Y 12 receptor is additionally involved in GLYT1 stimulation. Using pharmacological approaches and siRNAmediated protein knockdown methodology, we elucidate the molecular mechanisms of GLYT regulation. Modulation takes place through a signaling cascade involving phospholipase C activation, inositol 1,4,5-trisphosphate production, intracellular Ca 2؉ mobilization, protein kinase C stimulation, nitric oxide formation, cyclic guanosine monophosphate production, and protein kinase G-I (PKG-I) activation. GLYT1 and GLYT2 are differentially sensitive to NO/cGMP/PKG-I both in brain-derived preparations and in heterologous systems expressing the recombinant transporters and P2Y 1 receptor. Sensitivity to 2-methylthioadenosine 5 -diphosphate by GLYT1 and GLYT2 was abolished by small interfering RNA (siRNA)-mediated knockdown of nitric-oxide synthase. Our data may help define the role of GLYTs in nociception and pain sensitization.

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Section: Methodsmentioning

confidence: 99%

P2Y Purinergic Regulation of the Glycine Neurotransmitter Transporters

Jiménez

Zafra

Pérez-Sen

et al. 2011

Journal of Biological Chemistry

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“…In particular, selective inhibitors of glycine transporters GLYT1 and GLYT2, the former largely glial and the latter mostly present on glycinergic neurons (Zafra et al 1995a,b;Poyatos et al 1997), have become available (Atkinson et al 2001;Caulfield et al 2001;Herdon et al 2001); these compounds have opened the possibility to labelling selectively glycinergic nerve endings with radioactive glycine and subsequently investigating the mechanisms of glycine release. Taking advantage of this opportunity, we recently studied glycine release from synaptosomes of spinal cord, the region in which the density of glycinergic terminals is the highest and in which glycinergic transmission has been best established (Luccini and Raiteri 2007).…”

Section: +mentioning

confidence: 99%

Glycinergic nerve endings in hippocampus and spinal cord release glycine by different mechanisms in response to identical depolarizing stimuli

Luccini¹,

Romei²,

Raiteri³

2008

Journal of Neurochemistry

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Studies on hippocampal glycine release are extremely rare. We here investigated release from mouse hippocampus glycinergic terminals selectively pre‐labelled with [3H]glycine through transporters of the GLYT2 type. Purified synaptosomes were incubated with [3H]glycine in the presence of the GLYT1 blocker NFPS to abolish uptake (∼ 30%) through GLYT1. The non‐GLYT1‐mediated uptake was entirely sensitive to the GLYT2 blocker Org25543. Depolarization during superfusion with high‐K+ (15–50 mmol/L) provoked overflows totally dependent on external Ca2+, whereas in the spinal cord the 35 or 50 mmol/L KCl‐evoked overflow (higher than that in hippocampus) was only partly dependent on extraterminal Ca2+. In the hippocampus, the Ca2+‐dependent 4‐aminopyridine (1 mmol/L)‐evoked overflow was five‐fold lower than that in spinal cord. The component of the 10 μmol/L veratridine‐induced overflow dependent on external Ca2+ was higher in the hippocampus than that in spinal cord, although the total overflow in the hippocampus was only half of that in the spinal cord. Part of the veratridine‐evoked hippocampal overflow occurred by GLYT2 reversal and part by bafilomycin A1‐sensitive exocytosis dependent on cytosolic Ca2+ generated through the mitochondrial Na+/Ca2+ exchanger. As glycine sites on NMDA receptors are normally not saturated, understanding mechanisms of glycine release should facilitate pharmacological modulation of NMDA receptor function.

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“…VVZ-149 showed a lower binding affinity to GlyT2, with an IC 50 of 0.86 μM, compared with the previously published GlyT2 inhibitors, Org-25543 and ALX-1393, which presented IC 50 values within the nanomolar range. 11,15,23 Those potent compounds had undesirable dose-limiting toxicity such as tremor and stereotypy caused by complete blockade of GlyT2 or coinhibition of GlyT1. 11 VVZ-149 is considered less likely to exhibit toxicity because of GlyT2 inhibition, and severe toxicity because of VVZ-149 has not been observed in animal studies.…”

Section: Discussionmentioning

confidence: 99%

“…Urine was collected 0-6, 6-12, 12-24, and 24-32 hours after the initiation of dosing. In the MAD study, serial blood samples were collected using heparinized tubes 0, 0.5, 1, 2, 4, 4.25, 4.5, 5, 6, 7, and 8 hours after initiation of the administration of the first and fifth doses and 0, 0.5, 1, 2, 4, 4.25, 4.5, 5,6,7,8,15,20,28, and 40 hours after initiation of the administration of the sixth dose. Urine was collected 0-8 hours after initiation of the administration of the first and fifth doses and 0-8, 8-16, 16-24, and 24-40 hours after initiation of the administration of the sixth dose.…”

Section: Blood and Urine Collectionmentioning

confidence: 99%

“…11 Targeting multiple analgesic pathways could induce greater synergistic effects than targeting a single pathway. [12][13][14][15] For example, coinhibition of GlyT2 and serotonin 2A (5HT 2 A) receptors using selective antagonists (ORG-25543 and MDL11,939, respectively) exert synergistic antinociceptive effects in a rat spinal nerve ligation model. 16 The 5HT 2 A receptor belongs to the serotonin receptor family and is mainly expressed in nociceptive sensory neurons.…”

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confidence: 99%

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Safety, Tolerability, and Pharmacokinetic Characteristics of a Novel Nonopioid Analgesic, VVZ‐149 Injections in Healthy Volunteers: A First‐in‐Class, First‐in‐Human Study

Lee

Kim

et al. 2017

The Journal of Clinical Pharma

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VVZ-149, a dual antagonist of GlyT2 and 5HT 2 A receptors, is an investigational analgesic with a novel mechanism of action that is currently under earlystage clinical development as an injectable agent for the treatment of postoperative pain. Here, the safety, tolerability, and pharmacokinetics of VVZ-149 injections in healthy male volunteers were explored in a randomized, double-blind, single-and multiple-ascending-dose (SAD and MAD, respectively), placebo-controlled clinical study. Subjects randomly received a 4-hour intravenous infusion of 0.25-8 mg/kg VVZ-149 or placebo in the SAD study (n = 46) or a 4-hour intravenous infusion of 4-7 mg/kg VVZ-149 or placebo twice daily for 3 days in the MAD study (n = 20). Serial blood and urine samples were collected for the pharmacokinetic analysis of VVZ-149 and its active metabolite (VVZ-368). Noncompartmental and compartmental pharmacokinetic analyses were performed. Various dosing scenarios were simulated to identify the adequate dosing regimen for the subsequent trials. Plasma exposure to VVZ-149 and VVZ-368 showed a dose-proportional increase. VVZ-149 did not accumulate in the plasma, whereas the plasma concentration of VVZ-368 increased by 1.23-to 2.49-fold after the fifth and sixth doses, respectively, in the MAD trial. Among the simulated dosing regimens, a loading dose followed by a maintenance dose was found to be an optimal dosing regimen, yielding the effective concentration estimated from animal studies in rat models of neuropathic or inflammatory pain. Single-or multiple-dose administration of VVZ-149 was generally well tolerated. These results showed that 0.5-8 mg/kg VVZ-149 exhibited linear pharmacokinetic characteristics and can be safely administered in further clinical studies.

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The First Potent and Selective Inhibitors of the Glycine Transporter Type 2

Cited by 91 publications

References 15 publications

P2Y Purinergic Regulation of the Glycine Neurotransmitter Transporters

P2Y Purinergic Regulation of the Glycine Neurotransmitter Transporters

Glycinergic nerve endings in hippocampus and spinal cord release glycine by different mechanisms in response to identical depolarizing stimuli

Safety, Tolerability, and Pharmacokinetic Characteristics of a Novel Nonopioid Analgesic, VVZ‐149 Injections in Healthy Volunteers: A First‐in‐Class, First‐in‐Human Study

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