| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg |
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| Targets |
EAAT2
WAY-213613 hydrochloride specifically targets the excitatory amino acid transporter 2 (EAAT2, also known as GLT-1). It acts as a potent, selective, non-substrate (non-transportable) reuptake inhibitor with an IC50 of 85 nM.. It shows selectivity for EAAT2 over EAAT1 (IC50 = 5 microM) and EAAT3 (IC50 = 3.8 microM), with approximately 50-100 fold selectivity.. |
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| ln Vitro |
With IC50 values of 5004 nM, 85 nM, and 3787 nM, respectively, WAY-213613 hydrochloride (0-100 μM) exhibits inhibitory activity against human EAAT1, EAAT2, and EAAT3 isoforms[1]. With Ki values of 15 nM, 41 nM, and 55 nM, WAY-213613 hydrochloride (3,30,300 nM) inhibits synaptosome L-[3H] glutamate absorption [1]. When EAAT1-, EAAT2-, or EAAT3 is injected into oocytes, WAY-213613 hydrochloride (0-100 μM) blocks glutamate-induced currents in a concentration-dependent manner (IC50 values: 48, 0.13, and 4.0 μM, respectively)[1]. With strong selectivity for ionophilic receptors and EAAT2, WAY-213613 hydrochloride (0.5-50 μM) exerts inhibiting effects on NMDA stimulation responses [1].
In vitro, WAY-213613 hydrochloride is a potent EAAT2 inhibitor. It demonstrates an IC50 of 85 nM for EAAT2, indicating a high binding affinity to this specific glutamate transporter.. Furthermore, it exhibits selectivity for EAAT2 over EAAT3 (IC50 = 3.8 microM) and EAAT1 (IC50 = 5 microM), confirming it as a selective inhibitor for the predominant CNS glutamate transporter.. Its mechanism is as a non-substrate inhibitor, meaning it blocks the transporter without being translocated. |
| ln Vivo |
Specific in vivo activity data for WAY-213613 hydrochloride has not been detailed in the provided search results. As a potent and selective EAAT2 inhibitor, it is hypothesized to produce a robust increase in extracellular glutamate levels in the brain. This makes it a valuable tool for studying conditions associated with glutamate dysregulation, such as epilepsy, stroke, and excitotoxic neurodegeneration. It is frequently used in preclinical models to investigate glutamate-mediated mechanisms..
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| Enzyme Assay |
The standard in vitro protocol for assessing EAAT inhibition is a radiolabeled substrate uptake assay using cells expressing the specific EAAT isoforms. HEK293 cells stably expressing human EAAT2 are seeded in 24-well plates. Upon reaching confluency, the cells are pre-incubated with varying concentrations of WAY-213613 hydrochloride (0.01-10,000 nM) for 10 minutes at 37degC in assay buffer. Then, [3H]-L-glutamate (a radiolabeled substrate, e.g., 50 nM) is added to the cells and incubated for 5-10 minutes. The uptake reaction is terminated by washing the cells three times with ice-cold PBS. The cells are lysed with 0.2 N NaOH, and the radioactivity in the lysate is measured using a liquid scintillation counter. The IC50 of 85 nM is calculated from the inhibition curve using the Cheng-Prusoff equation.
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| Cell Assay |
The in vitro cellular assay for WAY-213613 hydrochloride involves the use of primary rat cortical astrocytes or human induced pluripotent stem cell (iPSC)-derived astrocytes, which endogenously express high levels of EAAT2. Cells are seeded in 96-well black-walled plates and cultured for 7-14 days to ensure high EAAT2 expression. The medium is replaced with a sodium-containing assay buffer. A fluorescent glutamate indicator (e.g., a FRET-based glutamate sensor or a dye such as Glutamate-Labeled Aspartate, ASP-4) is added to the cells. The cells are then pre-treated with varying concentrations of WAY-213613 hydrochloride (0.01-10,000 nM) for 10 minutes. Glutamate uptake is initiated by adding 10 uM L-glutamate. The decrease in fluorescence, reflecting the clearance of glutamate from the extracellular medium, is monitored in real-time using a fluorescence plate reader. The IC50 is determined from the dose-response curve.
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| Animal Protocol |
An in vivo protocol for WAY-213613 hydrochloride would involve the use of a microdialysis probe implanted into the hippocampus or striatum of an anesthetized rat. After a stabilization period, artificial cerebrospinal fluid (aCSF) containing the compound (10-100 uM) is perfused through the probe. Dialysate samples are collected at regular intervals (every 10 minutes) and analyzed for glutamate concentration using high-performance liquid chromatography (HPLC) with fluorescence detection (e.g., using o-phthalaldehyde derivatization). The increase in extracellular glutamate levels is used as a pharmacodynamic marker to confirm EAAT2 inhibition. Parallel experiments can assess behavioral endpoints such as seizure activity (EEG recording) or motor coordination (rotarod) to study the consequences of elevated glutamate.
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| ADME/Pharmacokinetics |
Detailed pharmacokinetic data for WAY-213613 hydrochloride is not provided. As a small molecule with a molecular weight of 468.74, it is likely designed for oral administration, though its charged hydrochloride salt may influence absorption. It is soluble in water and DMSO. To evaluate its CNS penetration, a standard PK study involves intravenous (IV) administration (1 mg/kg) and intraperitoneal (IP) administration (10 mg/kg) in mice, followed by serial plasma and brain tissue collection. The concentration of the compound in both matrices is analyzed by LC-MS/MS to calculate brain-to-plasma ratio (Kp), clearance (Cl), and half-life (T1/2).
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| Toxicity/Toxicokinetics |
Specific toxicological data for WAY-213613 hydrochloride is not available. As an EAAT2 inhibitor that blocks glutamate reuptake, the primary safety concern in vivo is the overstimulation of glutamate receptors (excitotoxicity), which could lead to neuronal death and seizures. Standard safety assessment would involve an acute toxicity study in mice to determine the maximum tolerated dose (MTD) and to observe for overt signs of CNS toxicity, such as seizures, tremors, and mortality. Histopathological examination of brain tissue would be conducted to assess for neuronal necrosis. An in vitro hERG (human ether-à-go-go-related gene) channel assay would also be performed to assess the risk of cardiac QT prolongation.
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| References |
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| Additional Infomation |
WAY-213613 hydrochloride is a research-grade chemical and is not approved for clinical use. It is a potent and selective tool for studying EAAT2/GLT-1 function in the central nervous system. Because EAAT2 is responsible for 90-95% of glutamate uptake in the forebrain, this compound is critical for understanding the role of glutamate transporters in synaptic transmission, plasticity, and neurotoxicity. It is widely used in preclinical models of epilepsy, amyotrophic lateral sclerosis (ALS), stroke, and traumatic brain injury to investigate the pathophysiology of glutamate excitotoxicity. It is for research use only.
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| Molecular Formula |
C16H14BRCLF2N2O4
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|---|---|
| Molecular Weight |
451.65
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| Exact Mass |
449.979
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| CAS # |
2450268-84-3
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| Related CAS # |
WAY-213613;868359-05-1
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| PubChem CID |
139290995
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
26
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| Complexity |
477
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| Defined Atom Stereocenter Count |
1
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| SMILES |
C1(=CC(Br)=C(C=C1F)OC1C=CC(NC(=O)C[C@@H](C(=O)O)N)=CC=1)F.Cl
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| InChi Key |
IBCWXGJQAQXHPZ-ZOWNYOTGSA-N
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| InChi Code |
InChI=1S/C16H13BrF2N2O4.ClH/c17-10-5-11(18)12(19)6-14(10)25-9-3-1-8(2-4-9)21-15(22)7-13(20)16(23)24;/h1-6,13H,7,20H2,(H,21,22)(H,23,24);1H/t13-;/m0./s1
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| Chemical Name |
(2S)-2-amino-4-[4-(2-bromo-4,5-difluorophenoxy)anilino]-4-oxobutanoic acid;hydrochloride
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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, 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) |
DMSO: 50 mg/mL (110.71 mM)
H2O: < 0.1 mg/mL |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.54 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% 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 25.0 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. Solubility in Formulation 2: ≥ 2.5 mg/mL (5.54 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 25.0 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. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (5.54 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.2141 mL | 11.0705 mL | 22.1410 mL | |
| 5 mM | 0.4428 mL | 2.2141 mL | 4.4282 mL | |
| 10 mM | 0.2214 mL | 1.1071 mL | 2.2141 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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