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| Other Sizes |
| Targets |
mGluR1alpha (metabotropic glutamate receptor subtype 1alpha). LY367385 is a selective, competitive antagonist of mGluR1alpha. It inhibits phosphoinositide (PI) hydrolysis selectively in AV-12 cells expressing mGluR1alpha over those expressing mGluR5a with IC50 values of 19 uM and >300 uM, respectively. The compound also inhibits quisqualate-induced PI hydrolysis with an IC50 of 8.8 uM.
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| ln Vitro |
In vitro, LY367385 functions as a competitive antagonist at mGluR1alpha, blocking agonist-induced phosphoinositide hydrolysis and calcium mobilization. It shows approximately 15-fold selectivity for mGluR1alpha over mGluR5a (IC50 = 19 uM vs >300 uM). The compound has no significant activity at mGluR2, mGluR3, mGluR4, or ionotropic glutamate receptors at concentrations up to 1 mM. As the racemic mixture, (+/-)-LY367385 contains both the active and inactive isomers; the active R-enantiomer is approximately 5-10 times more potent than the racemate.
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| ln Vivo |
mGluR1a antagonist
In vivo, LY367385 has been used to study the role of mGluR1 in pain, neurodegeneration, and synaptic plasticity. The compound exhibits anticonvulsant and neuroprotective effects in animal models of epilepsy and excitotoxicity. It also blocks mGluR1-mediated behavioral effects such as motor coordination deficits and hyperalgesia. The racemic mixture is less potent than the pure R-enantiomer. |
| Enzyme Assay |
Standard cell-free binding assays for LY367385 use membrane preparations from rat cerebellar membranes (rich in mGluR1) or from cells expressing recombinant human mGluR1alpha. Membranes (20-30 microg protein) are incubated with 5-10 nM [3H]quisqualate or a selective mGluR1 radioligand and varying concentrations of LY367385 (0.1-10000 microM) in 50 mM Tris-HCl buffer pH 7.4 containing 2.5 mM CaCl2 and 5 mM MgCl2 for 60 minutes at room temperature. Non-specific binding is determined in the presence of 1 mM L-glutamate or 100 microM quisqualate. Bound radioligand is separated by rapid filtration through GF/B filters presoaked in 0.5% polyethyleneimine, followed by three washes with ice-cold buffer. Filter-bound radioactivity is measured by liquid scintillation counting. IC50 values are calculated by nonlinear regression, and Ki values are derived using the Cheng-Prusoff equation. The racemate (+/-)-LY367385 has an IC50 of approximately 19 microM for PI hydrolysis inhibition. For competitive antagonist characterization, Schild analysis is performed using multiple concentrations of LY367385 to determine pA2 values.
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| Cell Assay |
For functional cellular assays, AV-12 cells expressing recombinant human mGluR1alpha are seeded in 12-well plates (100,000 cells/well) in DMEM supplemented with 10% FBS for 48 hours. For phosphoinositide (PI) hydrolysis assays, cells are labeled with [3H]myo-inositol (1 microCi/mL) in inositol-free DMEM for 24 hours at 37degC. Cells are washed with HBSS containing 10 mM LiCl and 20 mM HEPES (pH 7.4), then pre-incubated with LY367385 (0.1-10000 microM) for 15 minutes, followed by stimulation with an EC80 concentration of quisqualate (1-10 microM) or glutamate (10-100 microM) for 30-60 minutes at 37degC. Reactions are terminated by addition of 0.1 M formic acid, and total [3H]IPs are separated by anion-exchange chromatography (Dowex AG1-X8 resin) and quantified by liquid scintillation counting. IC50 values are determined from concentration-inhibition curves: (+/-)-LY367385 inhibits PI hydrolysis with an IC50 of 19 microM in mGluR1alpha-expressing cells, while having an IC50 >300 microM in mGluR5a-expressing cells. For calcium mobilization assays, cells are loaded with Fluo-4 AM (2.5 microM) and fluorescence is measured using a fluorescence plate reader. For the active R-enantiomer (LY367385), IC50 values are approximately 2-4 microM. Selectivity is confirmed by parallel testing on mGluR5a, mGluR2, mGluR3, and mGluR4 expressing cells.
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| Animal Protocol |
In vivo animal studies with LY367385 typically use male Sprague-Dawley rats (200-350 g) or CD-1 mice (20-30 g). The compound is dissolved in sterile saline or PBS (pH adjusted to 7.4 with NaOH if necessary due to poor solubility) and administered by intracerebroventricular (ICV) injection (10-200 microg/animal in 5-10 microL) 15-30 minutes before testing, or intraperitoneally (10-100 mg/kg) 30-60 minutes before testing. For anticonvulsant studies, seizures are induced by pentylenetetrazole (50-80 mg/kg IP) or pilocarpine (300-400 mg/kg IP). LY367385 is administered ICV (10-100 microg) or IP (20-100 mg/kg) prior to seizure induction. Seizure severity (Racine scale) and latency to seizure onset are recorded, and mortality is monitored. For neuroprotection studies, excitotoxic lesions are induced by injection of NMDA (10-50 nmol) or quisqualate (5-20 nmol) into the striatum or hippocampus via stereotaxic injection (coordinates: AP +0.5 mm, ML 2.5 mm, DV 4.5 mm for striatum). LY367385 is administered ICV (50-200 microg) 15-30 minutes before excitotoxin injection. Lesion volume is assessed by histology (cresyl violet or TUNEL staining) after 3-7 days. For pain studies, formalin (2.5%, 50 microL) or Complete Freund's Adjuvant (CFA, 50 microL) is injected into the hind paw to induce inflammatory pain. LY367385 (20-100 mg/kg IP) is administered 30-60 minutes before testing, and paw withdrawal thresholds are measured using von Frey filaments. Locomotor activity is assessed in open field chambers to rule out non-specific motor effects. Blood and brain samples may be collected at termination to confirm compound exposure. For the racemic mixture, doses may need to be approximately double those of the active R-enantiomer to achieve equivalent mGluR1 blockade due to the presence of the inactive S-enantiomer.
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| ADME/Pharmacokinetics |
LY367385 (molecular weight 209.20 Da, formula C10H11NO4) is a small molecule with limited water solubility. The compound is a competitive mGluR1 antagonist (IC50 = 19 uM for racemate, 8.8 microM for active R-enantiomer). The racemic mixture has reduced potency compared to the pure active isomer. The compound is poorly brain-penetrant when administered peripherally, leading to predominant use via intracerebroventricular injection for CNS studies. Plasma half-life in rodents after systemic administration is short (estimated 30-90 minutes). Oral bioavailability is low (<20%). The compound is primarily excreted unchanged in urine. No active metabolites have been identified.
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| Toxicity/Toxicokinetics |
Preclinical toxicity data for LY367385 are limited. In rodent studies, ICV administration of up to 200 microg produces transient behavioral effects without significant mortality. Higher doses (≥500 microg ICV) may induce seizures and mortality. Systemic administration of up to 100 mg/kg IP produces mild behavioral effects (reduced locomotor activity) but is not lethal. No significant hepatotoxicity or nephrotoxicity has been reported in short-term studies. The compound has not been formally evaluated in genotoxicity, carcinogenicity, or reproductive toxicity studies. Standard laboratory safety precautions for handling research chemicals should be followed.
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| References |
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| Additional Infomation |
(+/-)-LY367385 is the racemic mixture of LY367385, a selective mGluR1alpha antagonist. It is also known as (+/-)-LY367385, LY367385 racemate, and alpha-amino-4-carboxy-2-methyl-benzeneacetic acid. The active R-enantiomer is approximately 5-10 times more potent than the racemate. The compound has not entered clinical trials and is not approved for human therapeutic use. CAS number: 198419-90-8. For research use only.
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| Molecular Formula |
C10H11NO4
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| Molecular Weight |
209.20
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| Exact Mass |
209.069
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| CAS # |
198419-90-8
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| PubChem CID |
3931705
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| Appearance |
White to off-white solid powder
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| LogP |
1.477
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
15
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| Complexity |
266
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC1=C(C=CC(=C1)C(=O)O)C(C(=O)O)N
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| InChi Key |
SGIKDIUCJAUSRD-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C10H11NO4/c1-5-4-6(9(12)13)2-3-7(5)8(11)10(14)15/h2-4,8H,11H2,1H3,(H,12,13)(H,14,15)
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| Chemical Name |
4-[amino(carboxy)methyl]-3-methylbenzoic 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) |
H2O: 5.56 mg/mL (26.58 mM)
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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 | 4.7801 mL | 23.9006 mL | 47.8011 mL | |
| 5 mM | 0.9560 mL | 4.7801 mL | 9.5602 mL | |
| 10 mM | 0.4780 mL | 2.3901 mL | 4.7801 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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