| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
Ki: 11 μM (EAAT1), 14 μM (EAAT3), 19 μM (EAAT2)[1]
L-threo-3-Hydroxyaspartic acid specifically targets the excitatory amino acid transporters (EAATs), which are responsible for clearing the neurotransmitter glutamate from the synaptic cleft. It is a potent EAAT inhibitor. It exhibits competitive inhibitory activity against EAAT1, EAAT2, and EAAT3, with Ki values of 11 microM, 19 microM, and 14 microM, respectively. It is also a non-transportable inhibitor of EAAT5.. |
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| ln Vitro |
In vitro, L-threo-3-Hydroxyaspartic acid is a potent EAAT inhibitor. It shows inhibition constants (Ki) of 11 microM for EAAT1, 19 microM for EAAT2, and 14 microM for EAAT3 in HEK293 cell lines expressing these transporters.. These values indicate a strong binding affinity for the transporter, blocking the reuptake of glutamate. It is a non-transportable inhibitor, meaning it competes for the binding site but is not moved across the membrane, leading to sustained blockade..
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| ln Vivo |
Specific in vivo activity data for L-threo-3-Hydroxyaspartic acid has not been detailed in the provided search results. As an EAAT inhibitor, it would be expected to increase extracellular glutamate levels in the brain following administration. This could be studied in animal models of neurological diseases where glutamate excitotoxicity plays a role, such as epilepsy or stroke. By blocking glutamate reuptake, it can be used as a tool to study the behavioral and physiological consequences of elevated synaptic glutamate.
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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 EAAT1, EAAT2, or EAAT3 are seeded in 24-well plates. After reaching confluency, the cells are pre-incubated with varying concentrations of L-threo-3-Hydroxyaspartic acid (0.1-1000 uM) for 10 minutes at 37degC. 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 Ki values are calculated from the inhibition curves using the Cheng-Prusoff equation..
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| Cell Assay |
The in vitro cellular assay for L-threo-3-Hydroxyaspartic acid involves the use of primary neuronal cultures or cell lines that express EAATs. Rat cortical astrocytes or neurons are seeded in 96-well plates and cultured for 7-14 days. The assay medium is replaced with HEPES-buffered salt solution containing 1 microM of the fluorescent glutamate indicator (e.g., Glutamate Sensor or a fluorescently labeled aspartate analog). The cells are pretreated with varying concentrations (0.1-1000 uM) of L-threo-3-Hydroxyaspartic acid for 10 minutes. Glutamate uptake is initiated by adding 10 microM 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 inhibition of glutamate uptake is calculated as a percentage of the control, and the Ki is determined from the dose-response curve.
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| Animal Protocol |
An in vivo protocol for L-threo-3-Hydroxyaspartic acid 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 (0.1-1 mM) is perfused through the probe. Dialysate samples are collected at regular intervals (every 10-20 minutes) and analyzed for glutamate concentration using high-performance liquid chromatography (HPLC) with fluorescence detection. The increase in extracellular glutamate levels is used to confirm the pharmacological activity of the EAAT inhibitor. Additional behavioral endpoints, such as seizure activity (electroencephalogram, EEG) or the induction of neuronal damage, can be assessed.
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| ADME/Pharmacokinetics |
Detailed pharmacokinetic data for L-threo-3-Hydroxyaspartic acid is not available. As a charged amino acid analog (molecular weight 149.1 g/mol), it is expected to have very poor oral bioavailability and a short plasma half-life due to rapid metabolism and renal excretion.. It does not readily cross the blood-brain barrier (BBB). Therefore, for in vivo CNS studies, it is typically administered by intracerebroventricular (ICV) injection or by direct infusion into the brain via microdialysis or osmotic minipumps to achieve sufficient concentrations in the brain parenchyma.
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| Toxicity/Toxicokinetics |
Specific toxicological data for L-threo-3-Hydroxyaspartic acid is not available. As an inhibitor of glutamate reuptake, the primary safety concern is the overstimulation of glutamate receptors (excitotoxicity) leading to neuronal cell death. High concentrations of the compound in the brain would cause a rapid and sustained increase in extracellular glutamate, which could trigger seizures and neurodegeneration. Standard safety assessment would involve monitoring for seizure activity (clinical observation and EEG) and histopathological analysis of brain tissue to assess neuronal damage (e.g., silver staining or Fluoro-Jade C staining).
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| References |
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| Additional Infomation |
(3S)-3-hydroxy-L-aspartic acid is the (3S)-diastereomer of 3-hydroxy-L-aspartic acid and is a metabolite. It is the conjugate acid of (3S)-3-hydroxy-L-aspartic acid (2-) and (3S)-3-hydroxy-L-aspartic acid (1-), and also the enantiomer of (3R)-3-hydroxy-D-aspartic acid. (3S)-3-hydroxy-L-aspartic acid is a metabolite found or produced in Saccharomyces cerevisiae.
L-threo-3-Hydroxyaspartic acid is a research-grade chemical and is not approved for clinical use. It is the most potent of the substrate inhibitors for EAATs. Its unique property as a "non-transportable" inhibitor makes it a valuable pharmacological tool to distinguish the effects of blocking the transporter from the effects of substrate translocation. This compound is used extensively in neurobiology research to study the role of EAATs in glutamate homeostasis, synaptic transmission, and the pathophysiology of neurological disorders such as epilepsy, stroke, amyotrophic lateral sclerosis (ALS), and Alzheimer's disease. |
| Molecular Formula |
C4H7NO5
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|---|---|
| Molecular Weight |
149.10
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| Exact Mass |
149.032
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| CAS # |
7298-99-9
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| Related CAS # |
DL-threo-3-Hydroxyaspartic acid;4294-45-5
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| PubChem CID |
443239
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| Appearance |
White to off-white solid powder
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| Density |
1.738 g/cm3
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| Boiling Point |
368.7ºCat 760 mmHg
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| Flash Point |
176.8ºC
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| LogP |
-4.4
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
10
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| Complexity |
156
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| Defined Atom Stereocenter Count |
2
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| SMILES |
[C@H]([C@@H](C(=O)O)O)(C(=O)O)N
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| InChi Key |
YYLQUHNPNCGKJQ-LWMBPPNESA-N
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| InChi Code |
InChI=1S/C4H7NO5/c5-1(3(7)8)2(6)4(9)10/h1-2,6H,5H2,(H,7,8)(H,9,10)/t1-,2-/m0/s1
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| Chemical Name |
(2S,3S)-2-amino-3-hydroxybutanedioic acid
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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 |
| 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 | 6.7069 mL | 33.5345 mL | 67.0691 mL | |
| 5 mM | 1.3414 mL | 6.7069 mL | 13.4138 mL | |
| 10 mM | 0.6707 mL | 3.3535 mL | 6.7069 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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