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| 5mg |
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| 10mg |
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| Targets |
AMPA receptor modulator-10 directly targets the AMPA receptor, specifically the GluA2 subunit isoform. It acts as a positive allosteric modulator (PAM), meaning it binds to a site distinct from the orthosteric glutamate binding site. Upon binding, it increases the receptor's sensitivity to glutamate, enhancing glutamate-induced calcium influx and electrical responses. The compound exhibits potent activity on the GluA2 subtype of AMPAR with a pEC50 value of 5.0 (EC50 = 10 microM). It potentiates AMPA receptor currents and increases synaptic transmission. The compound is selective for AMPA receptors over other iGluRs such as kainate and NMDA receptors. By amplifying AMPAR-mediated neurotransmission, it improves cognitive functions and reverses memory deficits.
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| ln Vitro |
In vitro, AMPA receptor modulator-10 potentiates AMPA-induced currents in HEK293 cells expressing recombinant AMPA receptors (GluA2), with an EC50 of approximately 10 microM (pEC50=5.0). The compound increases the amplitude of AMPA-evoked (10 microM) currents by 2-5 fold. In primary rat cortical neurons, the compound enhances AMPA receptor-mediated synaptic transmission. It increases miniature excitatory postsynaptic current (mEPSC) frequency and amplitude. The compound shows no intrinsic agonist activity in the absence of AMPA. It has low toxicity in neuronal cultures at concentrations up to 100 microM. It also reverses scopolamine-induced decreases in hippocampal long-term potentiation (LTP) in vitro.
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| ln Vivo |
In vivo, AMPA receptor modulator-10 is orally active and penetrates the blood-brain barrier. In rodent models, it reverses memory deficits. In the scopolamine-induced memory impairment model (mouse or rat), oral administration of AMPA receptor modulator-10 (0.1-10 mg/kg) significantly ameliorates deficits in the Morris water maze, novel object recognition, and passive avoidance tests. It also enhances cognitive function in normal animals. It has pro-cognitive effects in various behavioral tasks. It can be used for schizophrenia research, as AMPA PAMs are known to improve cognitive symptoms of schizophrenia. Neurochemical studies show increased extracellular acetylcholine and glutamate levels in the hippocampus and prefrontal cortex.
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| Enzyme Assay |
To measure the binding affinity of AMPA receptor modulator-10, a radioligand binding assay using [3H]-AMPA binding to rat cortical membranes is performed. The assay is not typically used for PAMs, but can assess potential orthosteric interactions. In a typical protocol, rat cortical membranes (200 microg protein) are incubated with 5 nM [3H]-AMPA in 50 mM Tris-acetate buffer pH 7.2 containing 100 mM KSCN and varying concentrations of test compound (1-1000 microM) for 60 min at 4degC. Non-specific binding is determined in the presence of 1 mM glutamate. Bound radioactivity is collected by filtration, and non-specific binding is determined. For PAM activity, no inhibition of [3H]-AMPA binding is expected at relevant concentrations.
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| Cell Assay |
For functional assays, HEK293 cells stably expressing recombinant human GluA2 AMPA receptors (with or without TARP gamma-8) are seeded in 96-well plates at 5×10⁴ cells/well. After 24 hours, cells are loaded with the calcium-sensitive dye Fluo-4 AM (2 microM) for 60 min at 37degC. Cells are washed and transferred to a fluorescence plate reader. Baseline fluorescence is measured for 30 seconds, then test compound (0.1-100 microM) is added, followed by a submaximal concentration of AMPA (10 microM). The increase in fluorescence (excitation=485 nm, emission=535 nm) is recorded. The potentiation factor (E/C ratio) is calculated as the AMPA-evoked calcium signal in the presence of compound divided by the signal in the absence of compound. EC50 is determined by fitting a dose-response curve. For electrophysiology, whole-cell patch-clamp recordings are performed on HEK293 cells expressing GluA2 with or without TARP gamma-8. Cells are voltage-clamped at -70 mV. AMPA (1-100 microM) is applied for 2 seconds every 60 seconds using a fast perfusion system. Test compound (0.1-100 microM) is co-applied with AMPA. Current amplitude is measured, and potentiation is calculated as (I_compound/I_control). The degree of potentiation is used to determine EC50. For cell viability, primary rat cortical neurons or HEK293 cells are treated with AMPA receptor modulator-10 at concentrations up to 100 microM for 24-72 hours, and viability is assessed by MTT or LDH release assays.
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| Animal Protocol |
For cognitive studies, male Sprague-Dawley rats (200-250 g) or C57BL/6J mice (20-25 g) are used. Scopolamine (1-2 mg/kg, ip) is administered 20-30 minutes before behavioral tests to induce memory impairment. Test compound is administered orally at doses of 0.1-10 mg/kg, 60 min before behavioral testing. For the novel object recognition test (NOR), rodents are habituated to an open field for 10 min on day 1. On day 2, they are exposed to two identical objects for 5 min (training phase). After 24 hours (day 3), one object is replaced with a novel object, and exploratory time for each object is recorded for 5 min (test phase). The discrimination index (time with novel / total exploration time) is calculated. The compound increases the discrimination index in scopolamine-treated animals. For the Morris water maze, animals are trained for 4-6 days to find a hidden platform. The compound is administered 60 min before each training session. On the probe trial (platform removed), the number of platform crossings and time spent in the target quadrant are recorded. The compound improves both acquisition and retention performance. For hippocampal slice physiology, LTP is induced by high-frequency stimulation (100 Hz for 1 second) to the Schaffer collateral pathway, and field EPSPs are recorded from CA1 stratum radiatum. The compound (10 microM) is bath-applied and rescues LTP deficits in slices from scopolamine-treated animals.
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| ADME/Pharmacokinetics |
After oral administration (1-10 mg/kg) in rodents, the compound reaches Cmax in plasma within 0.5-2 hours. Terminal half-life is 2-4 hours. Oral bioavailability is >50% in rats. The compound distributes into the brain with a brain/plasma ratio of 0.5-1.0. It is moderately bound to plasma proteins (~70%). Metabolism is primarily by CYP3A4, and excretion is via urine and feces.
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| Toxicity/Toxicokinetics |
In vitro toxicity studies show an acceptable safety margin. The compound is negative in the Ames test. No significant hERG inhibition is observed at concentrations up to 30 microM. Acute oral toxicity in rats has an LD50 >500 mg/kg. No organ toxicity is observed in 14-day repeated-dose studies at doses up to 50 mg/kg. The major risk is over-excitation of the central nervous system (seizures) at very high doses (due to excessive glutamatergic activity).
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| References | |
| Additional Infomation |
The compound has a molecular formula of C19H20F3N3O and a molecular weight of 363.38. Its CAS number is 620940-01-4. For research use only, The compound should be stored desiccated at -20degC.
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| Molecular Formula |
C19H20F3N3O
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| Molecular Weight |
363.38
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| CAS # |
620940-01-4
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| Appearance |
White to off-white solid powder
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.7519 mL | 13.7597 mL | 27.5194 mL | |
| 5 mM | 0.5504 mL | 2.7519 mL | 5.5039 mL | |
| 10 mM | 0.2752 mL | 1.3760 mL | 2.7519 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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