| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
Leu-AMS targets leucyl-tRNA synthetase (LRS), an aminoacyl-tRNA synthetase responsible for attaching leucine to its cognate tRNA during protein biosynthesis.
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| ln Vitro |
Leu-AMS exhibits potent inhibitory activity against LRS with an IC50 of 22.34 nM. The compound exhibits cytotoxic effects in both cancer and normal cells and hampers bacterial growth by blocking protein synthesis.
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| ln Vivo |
As a potent LRS inhibitor, Leu-AMS inhibits bacterial growth in vitro and in vivo. The R enantiomer is expected to retain the parent compound‘s ability to block protein synthesis, though its specific in vivo efficacy may differ from the racemic mixture due to stereoselectivity.
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| Enzyme Assay |
Non-cell LRS inhibition assays are performed using recombinant leucyl-tRNA synthetase enzyme. The reaction mixture contains 50 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 50 mM KCl, 2 mM ATP, 10 uM [3H]-leucine, and 2 uM purified E. coli tRNA^Leu. Leu-AMS R enantiomer is added at concentrations ranging from 0.1 to 1000 nM. After incubation at 37degC for 10 minutes, the reaction is stopped by adding trichloroacetic acid (TCA) to a final concentration of 10%. Precipitated tRNA-linked material is collected on glass fiber filters, washed, and radioactivity is counted. The IC50 is calculated by non-linear regression using a four-parameter logistic curve. Ki values are determined by measuring inhibition at varying substrate concentrations.
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| Cell Assay |
For cytotoxicity assays, mammalian cells including HeLa, HEK293, or various cancer cell lines are seeded in 96-well plates (5,000-10,000 cells/well) and allowed to attach overnight. Cells are treated with Leu-AMS R enantiomer at concentrations ranging from 10 nM to 100 uM for 48-72 hours. Cell viability is assessed using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) or CellTiter-Glo Luminescent assays. IC50 values are calculated from dose-response curves. For antibacterial assays, E. coli or other bacterial strains are grown in LB medium to mid-log phase (OD600 ~0.5), then diluted and treated with varying concentrations of the compound. After incubation at 37degC for 16-20 hours, growth inhibition is measured at OD600 using a microplate reader. Minimum inhibitory concentration (MIC) is defined as the lowest concentration that prevents visible growth.
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| Animal Protocol |
In vivo antibacterial efficacy studies are conducted in murine infection models. For systemic infection models, 6-8 week old female BALB/c mice are inoculated intraperitoneally with a lethal dose of bacteria (e.g., 5×10⁶ CFU of E. coli). Leu-AMS R enantiomer is administered intraperitoneally or intravenously at doses ranging from 1-20 mg/kg, typically 1-2 hours post-infection. Survival is monitored for 7-14 days, and bacterial burden in various organs is assessed by plating serial dilutions of tissue homogenates. For neutropenic thigh infection models, mice are rendered neutropenic by cyclophosphamide treatment, then infected intramuscularly with bacteria. Compound efficacy is determined by reduction in CFU/g of thigh tissue compared to vehicle controls.
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| ADME/Pharmacokinetics |
As a leucine analogue, Leu-AMS is expected to have pharmacokinetic properties typical of small molecule amino acid mimetics. Due to its structural similarity to leucine, the compound may be transported by L-type amino acid transporters (LAT1, SLC7A5). The R enantiomer may show different pharmacokinetic properties compared to the S enantiomer or racemic mixture due to stereoselective metabolism and transport. Oral bioavailability is likely limited due to active transport mechanisms in the gut. The parent compound Leu-AMS has a molecular weight of approximately 459 Da and moderate water solubility.
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| Toxicity/Toxicokinetics |
Toxicity of Leu-AMS has been evaluated in both cancer and normal cells, where it exhibits cytotoxic effects in both cell types. The IC50 in normal cells is typically higher than in cancer cells, suggesting some selectivity, though the compound lacks complete tumor specificity. In animal studies at therapeutic doses, potential toxicities include bone marrow suppression, gastrointestinal epithelial damage, and liver toxicity due to inhibition of protein synthesis in rapidly dividing tissues. The maximum tolerated dose in rodent models for leucine analogue LRS inhibitors is typically between 10-50 mg/kg depending on administration route and dosing schedule.
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| Additional Infomation |
Leu-AMS R enantiomer is a research tool for studying aminoacyl-tRNA synthetase inhibition as a strategy for antibacterial and anticancer therapy. By inhibiting LRS, the compound starves cells of leucyl-tRNA, leading to the accumulation of uncharged tRNA and activation of the general control non-derepressible 2 (GCN2) kinase pathway, resulting in translational arrest and cell death. LRS has emerged as a validated target for antibacterial drug development, and Leu-AMS serves as a valuable chemical probe for investigating LRS function in both prokaryotic and eukaryotic systems. The compound is for research use only.
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| Molecular Formula |
C16H25N7O7S
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|---|---|
| Molecular Weight |
459.48
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| Related CAS # |
Leu-AMS;288591-93-5
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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 |
| 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 :~50.5 mg/mL (~109.91 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 | 2.1764 mL | 10.8819 mL | 21.7637 mL | |
| 5 mM | 0.4353 mL | 2.1764 mL | 4.3527 mL | |
| 10 mM | 0.2176 mL | 1.0882 mL | 2.1764 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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