| Size | Price | Stock | Qty |
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| 500mg |
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| Other Sizes |
| Targets |
eIF4A3-IN-8 targets eukaryotic initiation factor 4A3 (eIF4A3), an ATP-dependent RNA helicase that is a core component of the exon junction complex (EJC). The compound acts as a selective ATP-competitive inhibitor, binding to the ATP-binding site of eIF4A3 and preventing its helicase activity. eIF4A3 is essential for the formation and function of the EJC, which is deposited on mRNA during splicing and plays critical roles in mRNA export, translation, and nonsense-mediated decay. By inhibiting eIF4A3, eIF4A3-IN-8 disrupts EJC function and allows researchers to study the specific roles of this complex in gene regulation. The compound is used as a chemical probe to investigate the detailed function of eIF4A3 and EJC in cellular processes.
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| ln Vitro |
In vitro, eIF4A3-IN-8 acts as a selective ATP-competitive inhibitor of eIF4A3 helicase activity. The compound inhibits the ATPase and helicase activities of eIF4A3 by competing with ATP for binding to the enzyme's active site. This inhibition disrupts the function of the exon junction complex (EJC) in mRNA processing. The compound's selectivity for eIF4A3 over other helicases makes it a valuable tool for studying the specific functions of this protein. eIF4A3-IN-8 can be used in cell-free assays to measure eIF4A3 ATPase activity using malachite green or coupled enzyme assays. The compound's inhibitory activity is concentration-dependent, and IC50 values can be determined by measuring ATP hydrolysis or RNA unwinding. The compound is also used to study the role of eIF4A3 in mRNA splicing and export.
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| ln Vivo |
eIF4A3-IN-8 is not used as a therapeutic agent but rather as a chemical probe for research purposes. The compound is used in cell biology and gene regulation research to study the functions of eIF4A3 and the exon junction complex (EJC). By inhibiting eIF4A3 activity, the compound allows researchers to investigate the role of the EJC in mRNA splicing, export, translation, and nonsense-mediated decay. The compound has been used in studies of mRNA processing and gene regulation. It serves as an essential chemical probe for investigating the complex functions of eIF4A3 and the EJC. The compound is not approved for clinical use and is intended for research applications only.
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| Enzyme Assay |
In vitro enzyme assays for eIF4A3-IN-8 typically involve measuring the inhibition of eIF4A3 ATPase or helicase activity. The compound is dissolved in DMSO to prepare stock solutions, which are then diluted in assay buffer to the desired concentrations (typically 0-100 μM). For ATPase assays, recombinant eIF4A3 protein is incubated with ATP and varying concentrations of the compound. The amount of inorganic phosphate released is measured using a malachite green assay or a coupled enzyme system. For helicase assays, RNA or DNA substrates with a fluorescent label are used, and unwinding is measured by fluorescence polarization or gel electrophoresis. IC50 values are calculated by plotting the percentage of inhibition against the compound concentration. The compound's solubility in DMSO is 100 mg/mL (570.84 mM).
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| Cell Assay |
In vitro cell-based assays using eIF4A3-IN-8 are conducted in cell culture models to study the effects of eIF4A3 inhibition on mRNA processing and gene regulation. Cells are treated with the compound at concentrations ranging from 1-100 μM for 24-72 hours. After treatment, cells are harvested for RNA extraction and analysis of mRNA splicing, export, and stability. The expression of EJC target genes is measured by qPCR or RNA sequencing. Protein levels of EJC components and downstream effectors are analyzed by Western blot. Cell viability is assessed using MTT or CCK-8 assays. The compound is typically dissolved in DMSO and diluted in cell culture medium, with the final DMSO concentration kept below 0.1%. The compound's effects on cell proliferation and gene expression are concentration-dependent.
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| Animal Protocol |
In vivo animal experiments with eIF4A3-IN-8 are not well-documented as the compound is primarily used as a chemical probe in cell-based studies. The compound's role as a research tool for studying eIF4A3 and EJC function suggests that it may be used in animal models to investigate the in vivo functions of these proteins. However, specific in vivo protocols for this compound are not widely reported in the available literature. The compound is intended for research use only and is not approved for clinical applications. When used in animal studies, the compound would likely be formulated in appropriate vehicles such as DMSO, PEG, or saline for administration. Dosing would depend on the specific experimental design and the desired level of eIF4A3 inhibition.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of eIF4A3-IN-8 have not been extensively characterized as the compound is primarily used as a research tool rather than a therapeutic agent. The compound is soluble in DMSO at 100 mg/mL (570.84 mM) and can be prepared as stock solutions for in vitro studies. For cell culture applications, the compound is diluted in medium to working concentrations. The compound is stable as a powder at -20°C for up to 3 years and in solution at -80°C for up to 1 year. The compound has a molecular weight of 175.18 g/mol and a purity of ≥99%. For long-term storage, the powder should be kept sealed in a dry place at 2-8°C. Solution stability is maintained at -20°C for 3-6 months and at -80°C for up to 12 months.
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| Toxicity/Toxicokinetics |
The toxicity of eIF4A3-IN-8 has not been extensively characterized as the compound is primarily used as a research tool for in vitro studies. The compound is intended for research use only and is not for human use. Standard laboratory safety precautions should be followed when handling the compound, including the use of gloves and eye protection. The compound is a small-molecule inhibitor of eIF4A3, and its toxicity in cell culture is likely concentration-dependent. At higher concentrations, the compound may affect cell viability due to inhibition of essential cellular processes mediated by eIF4A3 and the EJC. The compound is not classified as a highly toxic substance but should be handled with appropriate care.
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| References | |
| Additional Infomation |
eIF4A3-IN-8 (CAS 18474-59-4) is a selective ATP-competitive inhibitor of eukaryotic initiation factor 4A3 (eIF4A3). The compound is chemically defined as 6-methyl-1H-indole-2-carboxylic acid and has the molecular formula C10H9NO2 and a molecular weight of 175.18 g/mol. eIF4A3 is a DEAD-box RNA helicase and a core component of the exon junction complex (EJC) that plays critical roles in mRNA splicing, export, translation, and nonsense-mediated decay. eIF4A3-IN-8 serves as a valuable chemical probe to elucidate the specific functions of eIF4A3 and the EJC in gene regulation. The compound has a purity of ≥99%. It is soluble in DMSO at 100 mg/mL (570.84 mM). The compound is stored as a powder at -20°C for up to 3 years and in solution at -80°C for up to 1 year. It is intended for research use only.
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| Molecular Formula |
C10H9NO2
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| Molecular Weight |
175.1840
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| Exact Mass |
175.063
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| CAS # |
18474-59-4
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| PubChem CID |
3163396
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| Appearance |
Off-white to light brown solid powder
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| Density |
1.3±0.1 g/cm3
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| Boiling Point |
411.5±25.0 °C at 760 mmHg
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| Melting Point |
220°C
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| Flash Point |
202.7±23.2 °C
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| Vapour Pressure |
0.0±1.0 mmHg at 25°C
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| Index of Refraction |
1.696
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| LogP |
2.77
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
2
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| Rotatable Bond Count |
1
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| Heavy Atom Count |
13
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| Complexity |
217
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| Defined Atom Stereocenter Count |
0
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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 : ~100 mg/mL (~570.84 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (14.27 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 (14.27 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 (14.27 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 | 5.7084 mL | 28.5421 mL | 57.0841 mL | |
| 5 mM | 1.1417 mL | 5.7084 mL | 11.4168 mL | |
| 10 mM | 0.5708 mL | 2.8542 mL | 5.7084 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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