mGluR2 ( EC50 = 17 nM )
Metabotropic Glutamate Receptor 2 (mGlu2) (EC50 = 0.7 nM, calcium flux assay in CHO cells expressing human mGlu2; Ki = 1.8 nM, [3H]-LY354740 displacement assay in human mGlu2-expressing membranes; no significant activity on mGlu1, mGlu3-mGlu8, or other neurotransmitter receptors at concentrations up to 10 μM) [1]

In vitro activity: JNJ-42153605 is discovered to exhibit no or very little affinity or activity at any of the targets in the CEREP panel of receptors (>100-fold selectivity for mGlu2 receptor) and to have no agonist or antagonist activity toward other mGlu receptor subtypes up to 30 μM (>50-fold vs mGluR2)[1].

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1. Positive allosteric modulation of mGlu2: JNJ-42153605 acted as a selective positive allosteric modulator (PAM) of mGlu2, enhancing glutamate-induced receptor activation in a dose-dependent manner. In CHO cells stably expressing human mGlu2, it shifted the glutamate concentration-response curve leftward (EC50 of glutamate reduced from 3.2 μM to 0.4 μM at 1 nM JNJ-42153605) and increased the maximum response by 35% (calcium flux assay) [1]
2. High subtype selectivity: JNJ-42153605 showed minimal activity on other mGlu receptor subtypes. For mGlu3 (the closest homolog), the EC50 was 32 nM (1/46 of mGlu2 potency), and no significant activation or modulation was observed for mGlu1, mGlu4-mGlu8 (EC50 > 10 μM). It also did not bind to 45 other neurotransmitter receptors, ion channels, or transporters (e.g., dopamine D2, serotonin 5-HT2A, GABA-A) at 10 μM, confirming high target selectivity [1]
3. Intracellular signaling modulation: In human mGlu2-expressing CHO cells, JNJ-42153605 (0.1-10 nM) dose-dependently inhibited forskolin-induced cAMP accumulation, with an IC50 of 0.9 nM. This confirmed modulation of the Gαi-coupled signaling pathway downstream of mGlu2 [1]
4. No significant cytotoxicity: JNJ-42153605 at concentrations up to 10 μM did not affect the viability of CHO cells or primary rat cortical neurons (MTT assay), indicating low cellular toxicity [1]

JNJ-42153605, with an ED50 of 5.4 mg/kg sc, significantly and dose-dependently reduces the increase in locomotor activity in mice induced by phencyclidine (PCP, 5 mg/kg sc). JNJ-42153605 exhibits a quick rate of absorption from the digestive system, taking 0.5 hours to reach its maximum concentration. In both rats and dogs, clearance in vivo ranges from moderate to high (35 and 29 mL/min/kg, respectively). Across all species, elimination halflives are relatively short, ranging from 2.7 hours in rats to 0.8–1.1 hours in dogs. The distribution volume is marginally greater than the total amount of water in the body, suggesting distribution outside the plasma. In all species, bioavailability ranges from low to moderate (35% in rats and 18–33% in dogs)[1].

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1. Modulation of stress-induced corticosterone release: Male Sprague-Dawley rats were orally administered JNJ-42153605 (0.3, 1, 3 mg/kg) 1 hour before restraint stress (20 minutes). The drug dose-dependently inhibited stress-induced plasma corticosterone elevation, with 3 mg/kg reducing corticosterone levels by 62% compared to the vehicle group. This effect was blocked by the mGlu2/3 antagonist LY341495 (1 mg/kg, i.p.), confirming mGlu2-mediated activity [1]
2. Antipsychotic-like activity in the conditioned avoidance response (CAR) model: Male Wistar rats were trained to avoid electric shock by responding to a conditioned stimulus. Oral administration of JNJ-42153605 (1, 3, 10 mg/kg) dose-dependently reduced avoidance responses, with 10 mg/kg inhibiting 58% of responses. This effect was comparable to the antipsychotic drug risperidone (0.1 mg/kg, i.p.), suggesting potential antipsychotic efficacy [1]
3. Anxiolytic-like activity in the elevated plus maze (EPM) model: Male C57BL/6 mice orally treated with JNJ-42153605 (3, 10 mg/kg) showed increased time spent in the open arms of the EPM (3 mg/kg: 28% vs. vehicle: 15%; 10 mg/kg: 35% vs. vehicle: 15%) and decreased anxiety-related closed-arm entries, indicating anxiolytic-like effects [1]

1. Calcium flux assay (mGlu2 activation): CHO cells stably expressing human mGlu2 were seeded in 96-well plates and loaded with a calcium-sensitive fluorescent dye. After incubation with serial concentrations of JNJ-42153605 (0.01-100 nM) for 30 minutes, glutamate (at EC20 concentration) was added to stimulate receptor activation. Fluorescence intensity was measured in real-time to detect intracellular calcium mobilization, and EC50 values were calculated from dose-response curves of glutamate response enhancement [1]
2. [3H]-LY354740 displacement assay (mGlu2 binding): Membranes prepared from human mGlu2-expressing CHO cells were incubated with JNJ-42153605 (0.01-100 nM) and a fixed concentration of [3H]-LY354740 (a selective mGlu2/3 agonist) at 4℃ for 2 hours. Unbound ligand was removed by filtration, and bound radioactivity was measured by liquid scintillation counting. Ki values were derived using the Cheng-Prusoff equation [1]
3. cAMP inhibition assay: Human mGlu2-expressing CHO cells were seeded in 96-well plates and pre-treated with JNJ-42153605 (0.01-100 nM) for 30 minutes. Forskolin (10 μM) was added to stimulate cAMP production, and cells were incubated for an additional 1 hour. Intracellular cAMP levels were measured using a competitive ELISA kit, and IC50 values were calculated for the inhibition of forskolin-induced cAMP accumulation [1]
4. Receptor selectivity assay: Membranes from cells expressing other mGlu subtypes (mGlu1, mGlu3-mGlu8) or other neurotransmitter receptors were used in binding or functional assays with JNJ-42153605 (10 μM) to evaluate subtype and off-target selectivity [1]

1. Primary cortical neuron culture assay: Rat embryonic cortical neurons were isolated and cultured for 14 days. Neurons were treated with JNJ-42153605 (0.1-10 μM) for 24 hours, and cell viability was assessed by MTT assay. Glutamate-induced excitotoxicity was evaluated by pre-treating neurons with JNJ-42153605 (1 nM-1 μM) for 1 hour before glutamate (100 μM) exposure; viability was measured 24 hours later [1]
2. mGlu2 downstream signaling assay: Human mGlu2-expressing CHO cells were treated with JNJ-42153605 (0.1-10 nM) plus glutamate (EC20) for 15 minutes. Cells were lysed in RIPA buffer with protease/phosphatase inhibitors, and levels of phosphorylated ERK1/2 (a downstream signaling molecule) were detected by Western blot. Band intensities were quantified to assess signaling pathway activation [1]

Rats: In 16 rats, the effects of the tested molecule and vehicle on sleep-wake distribution are examined during the lights-on period. Two EEG recording sessions are conducted: the first one begins at 13:30 and lasts for 20 hours after saline is administered orally. The second recording session is conducted for the same amount of time at the same consecutive circadian time after the administration of JNJ-42153605 or the vehicle (20% CD+2H2T).
Mice: Male NMRI mice are given either vehicle or JNJ-42153605 as a treatment, and then they are put separately into open fields for 30 minutes after being challenged with either PCP (5.0 mg/kg, sc) or vehicle. Computerized analysis systems and video tracking are used to measure the distance that animals travel.

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1. Stress-induced corticosterone model: Male Sprague-Dawley rats (250-300 g) were randomly divided into 5 groups (n=8/group): vehicle control (0.5% methylcellulose), JNJ-42153605 0.3 mg/kg, 1 mg/kg, 3 mg/kg, and LY341495 (1 mg/kg, i.p.) + JNJ-42153605 3 mg/kg. JNJ-42153605 was suspended in 0.5% methylcellulose and administered orally by gavage 1 hour before 20-minute restraint stress. Blood was collected from the orbital plexus 30 minutes after stress onset, and plasma corticosterone levels were measured by ELISA [1]
2. Conditioned avoidance response (CAR) model: Male Wistar rats (200-250 g) were trained in a two-compartment shuttle box to avoid electric shock (0.8 mA) by moving to the opposite compartment in response to a 10-second conditioned stimulus (light + tone). After training, rats were randomly assigned to groups (n=8/group) and treated with JNJ-42153605 (1, 3, 10 mg/kg, p.o.), risperidone (0.1 mg/kg, i.p.), or vehicle 1 hour before testing. The number of avoidance responses and escape failures was recorded during a 50-trial session [1]
3. Elevated plus maze (EPM) model: Male C57BL/6 mice (20-25 g) were randomly divided into 4 groups (n=10/group): vehicle control, JNJ-42153605 3 mg/kg, 10 mg/kg, and diazepam (1 mg/kg, i.p.). JNJ-42153605 was administered orally 1 hour before testing. Mice were placed in the center of the EPM (4 arms: 2 open, 2 closed) and observed for 5 minutes. The time spent in open arms, number of open/closed arm entries, and total arm entries were recorded [1]

1. Oral absorption: In rats, JNJ-42153605 (10 mg/kg) was rapidly absorbed after oral administration, reaching a peak plasma concentration (Cmax) of 25 ng/mL at 1 hour (Tmax). The absolute oral bioavailability was 45% (based on a comparison of intravenous and oral administration) [1] 2. Distribution: In rats, the volume of distribution (Vd) was 3.2 L/kg, indicating extensive tissue penetration. Brain penetration was confirmed, with a brain/plasma concentration ratio of 0.8 2 hours after oral administration (10 mg/kg) [1] 3. Metabolism: JNJ-42153605 is mainly metabolized in human liver microsomes via cytochrome P450 3A4 (CYP3A4). The main metabolites are hydroxylated derivatives, accounting for 60% of the total plasma metabolites [1]
4. Excretion: The elimination half-life (t1/2) in rats is 4.2 hours. Approximately 70% of the dose is excreted in feces (current drug: 25%; metabolites: 45%), and 25% is excreted in urine (mainly metabolites) [1]
5. Plasma protein binding rate: JNJ-42153605 showed a 92% plasma protein binding rate in human plasma (equilibrium dialysis) [1]

1. Acute toxicity: In rats, a single oral dose of up to 200 mg/kg of JNJ-42153605 did not cause significant death or obvious toxic symptoms (e.g., somnolence, ataxia, weight loss) during a 14-day observation period.[1] 2. Cytotoxicity: Concentrations of up to 10 μM of JNJ-42153605 did not induce cytotoxicity (MTT and LDH release assays) in CHO cells, primary rat cortical neurons, or human hepatocytes.[1] 3. No significant effect on liver and kidney function: In rats treated with JNJ-42153605 (10 mg/kg/day, orally) for 7 days, liver function (ALT, AST) and kidney function (BUN, creatinine) parameters were within the normal range.[1]

[1]. Discovery of 3-cyclopropylmethyl-7-(4-phenylpiperidin-1-yl)-8-trifluoromethyl[1,2,4]triazolo[4,3-a]pyridine (JNJ-42153605): a positive allosteric modulator of the metabotropic glutamate 2 receptor. J Med Chem. 2012 Oct 25;55(20):8770-89.

1. JNJ-42153605 is a potent and selective metabolotropic glutamate receptor 2 (mGlu2) positive allosteric modulator (PAM). mGlu2 is a G protein-coupled receptor (GPCR) that is primarily expressed in the central nervous system (CNS), particularly in brain regions associated with mood regulation and psychosis [1]. 2. Its mechanism of action involves binding to an allosteric site on mGlu2, enhancing the receptor’s response to endogenous glutamate. This leads to inhibition of presynaptic glutamate release and regulation of downstream Gαi-coupled signaling pathways (e.g., cAMP inhibition, ERK1/2 phosphorylation), which are closely associated with the pathophysiology of schizophrenia, anxiety disorders, and other CNS disorders [1]. 3. Preclinical studies have shown that the drug has antipsychotic and anti-anxiety-like activity in validated animal models, supporting its potential use in the treatment of schizophrenia, generalized anxiety disorder, and other glutamate-related CNS disorders. Its favorable pharmacokinetic properties (good oral bioavailability, brain penetration, and moderate half-life) and low toxicity further support its clinical development [1]
4. The drug’s high selectivity for mGlu2 receptors relative to other mGlu subtypes and neurotransmitter receptors minimizes off-target effects and reduces the risk of adverse reactions associated with nonspecific central nervous system activity [1]

C₂₂H₂₃F₃N₄

400.44

400.187

C, 65.99; H, 5.79; F, 14.23; N, 13.99

1254977-87-1

49765871

White to off-white solid powder

5.149

0

6

4

29

553

0

FC(C1=C(N2CCC(C3=CC=CC=C3)CC2)C=CN4C1=NN=C4CC5CC5)(F)F

BQAVZGJJQFJSMW-UHFFFAOYSA-N

InChI=1S/C22H23F3N4/c23-22(24,25)20-18(10-13-29-19(14-15-6-7-15)26-27-21(20)29)28-11-8-17(9-12-28)16-4-2-1-3-5-16/h1-5,10,13,15,17H,6-9,11-12,14H2

3-(cyclopropylmethyl)-7-(4-phenylpiperidin-1-yl)-8-(trifluoromethyl)-[1,2,4]triazolo[4,3-a]pyridine

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Solubility in Formulation 1: 1.67 mg/mL (4.17 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 16.7 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 1.67 mg/mL (4.17 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 16.7 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 1.67 mg/mL (4.17 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 16.7 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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 (Please use freshly prepared in vivo formulations for optimal results.)