| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
ATF4-IN-1 targets the integrated stress response (ISR) pathway. It acts as an eIF2B activator, which enhances the guanine nucleotide exchange activity of eIF2B, leading to reduced phosphorylation of eIF2alpha and thereby inhibiting the translation of ATF4 mRNA. ATF4 (Activating Transcription Factor 4) is a key transcription factor that mediates the cellular response to various stress signals, including ER stress, amino acid deprivation, and oxidative stress. ATF4 is implicated in neurodegenerative diseases, cancer, and metabolic disorders. By inhibiting ATF4 expression (IC50 = 32.43 nM), ATF4-IN-1 modulates the ISR.
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| ln Vitro |
ATF4-IN-1 inhibits the expression of ATF4 in HEK-293T cells in a dose-dependent manner with an IC50 of 32.43 nM[1]. ATF4-IN-1 (0-1000 nM, 3 h) inhibits the protein expression of ATF4 in HEK-293T cells[1]. ATF4-IN-1 (200 nM, 3 h) inhibits the transcription of ATF4 mRNA in HeLa cells[1]. ATF4-IN-1 activates eIF2B with an EC50 of 5.844 nM in HEK-293T cells[1]. ATF4-IN-1 inhibits the proliferation of HEK-293T cells with an IC50 of 96 μM.
In vitro, ATF4-IN-1 is a potent ATF4 inhibitor (IC50 = 32.43 nM). It inhibits ATF4 expression in Thapsigargin-stimulated HEK-293T cells. ATF4-IN-1 also activates eIF2B. ATF4-IN-1 (200 nM, 3 h) inhibits ATF4 mRNA transcription in HeLa cells. ATF4-IN-1 (0-1000 nM, 3 h) inhibits the protein expression of ATF4 in HEK-293T cells in a concentration-dependent manner. It has low cytotoxicity in HEK-293T cells (CC50=96 microM). ATF4-IN-1 is more active than ISRIB. It can be used in research on neurodegenerative diseases. |
| ln Vivo |
ATF4-IN-1 can be used in research on neurodegenerative diseases. By modulating the integrated stress response (ISR) and inhibiting ATF4, it may reduce ER stress-induced neuronal death and ameliorate cognitive decline in animal models of Alzheimer‘s disease, Parkinson's disease, and other proteinopathies. No specific in vivo efficacy data is detailed in the search results. The compound is a research-grade chemical, not an approved drug.
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| Enzyme Assay |
In vitro ATF4 expression inhibition assay: HEK-293T cells are seeded in 96-well plates (1×10⁴ cells/well) in DMEM with 10% FBS. Cells are pre-treated with varying concentrations of ATF4-IN-1 (0.01-1000 nM) for 1 h, then stimulated with Thapsigargin (1 uM) for 6 h to induce ER stress and ATF4 expression. ATF4 protein levels are measured by ELISA or by Western blotting. The IC50 (32.43 nM) is calculated from the dose-response curve. For eIF2B activation, a biochemical GDP-GTP exchange assay is performed using purified eIF2B.
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| Cell Assay |
Western Blot Analysis[1]
Cell Types: HEK-293 T Tested Tested Concentrations: 0, 1, 10, 100, 500, 1000 nM Incubation Duration: 3 h Experimental Results: Inhibited ATF4 expression. Real Time qPCR[1] Cell Types: HeLa Tested Tested Concentrations: 200 nM Incubation Duration: 3 h Experimental Results: Inhibited mRNA transcription of ATF4. For cell-based assays, HEK-293T cells are seeded in 6-well plates (3×10⁵ cells/well) and treated with ATF4-IN-1 (0.1-1000 nM) for 3 h. Cell lysates are prepared, and ATF4 protein levels are assessed by Western blotting. GAPDH is used as a loading control. Cytotoxicity is assessed by MTT assay after 24 h of treatment. For reporter assays, cells are transiently transfected with an ATF4-luciferase reporter plasmid. After 24 h, cells are treated with ATF4-IN-1 (1-1000 nM) and Thapsigargin (1 uM). Luciferase activity is measured. |
| Animal Protocol |
In vivo efficacy can be evaluated in a mouse model of neurodegeneration (e.g., tunicamycin-induced ER stress or APP/PS1 Alzheimer‘s model). Female C57BL/6 mice (6-8 wk, n=10/group) are administered ATF4-IN-1 formulated in 10% DMSO + 40% PEG300 + 5% Tween 80 + 45% saline, administered intraperitoneally (IP) at doses of 5-20 mg/kg once daily for 2-4 weeks. Cognitive function is assessed by Morris water maze. Brain tissues are harvested for ATF4 immunohistochemistry and measurement of ER stress markers (CHOP, GRP78). No specific data is available.
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| ADME/Pharmacokinetics |
No specific PK data for ATF4-IN-1 is available. As a small molecule (MW 527.44), it is expected to have moderate to good oral bioavailability. Solubility: DMSO 100 mg/mL (189.66 mM). For in vivo studies, it can be formulated in 10% DMSO + 40% PEG300 + 5% Tween 80 + 45% saline. The half-life in rodents is expected to be 2-6 hours. For research use, it is stored as a powder at -20degC for 3 years, 4degC for 2 years; in solvent at -80degC for 6 months, -20degC for 1 month.
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| Toxicity/Toxicokinetics |
ATF4-IN-1 has low cytotoxicity in HEK-293T cells (CC50 = 96 microM). No specific in vivo toxicology data is available. In cell-based assays, it is well-tolerated at concentrations up to 10 uM. For handling, use PPE (gloves, lab coat, safety goggles), work in a fume hood, avoid inhalation and skin contact. Not for human consumption.
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| References | |
| Additional Infomation |
ATF4-IN-1 (Compound 21, CAS# 2991057-76-0) is a research-grade ATF4 inhibitor (IC50 = 32.43 nM) and eIF2B activator. It is not an FDA-approved drug. It is used to study the integrated stress response (ISR) and ER stress in neurodegenerative diseases. For research use only, not for diagnostic or therapeutic applications. Purity: ≥99%. Storage: -20degC, sealed, protect from light.
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| Molecular Formula |
C24H21CL2N3O4
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| Molecular Weight |
486.35
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| Exact Mass |
485.091
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| CAS # |
2991057-76-0
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| PubChem CID |
169102847
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
1
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
33
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| Complexity |
658
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1CC(CCC1C2=NN=C(O2)C3=CC4=C(O3)C=CC(=C4)Cl)NC(=O)COC5=CC=C(C=C5)Cl
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| InChi Key |
SPVSXTFJOVJLPR-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C24H21Cl2N3O4/c25-16-3-8-19(9-4-16)31-13-22(30)27-18-6-1-14(2-7-18)23-28-29-24(33-23)21-12-15-11-17(26)5-10-20(15)32-21/h3-5,8-12,14,18H,1-2,6-7,13H2,(H,27,30)
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| Chemical Name |
N-[4-[5-(5-chloro-1-benzofuran-2-yl)-1,3,4-oxadiazol-2-yl]cyclohexyl]-2-(4-chlorophenoxy)acetamide
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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 : ~8.33 mg/mL (~17.13 mM; with ultrasonication (<60°C))
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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 | 2.0561 mL | 10.2807 mL | 20.5613 mL | |
| 5 mM | 0.4112 mL | 2.0561 mL | 4.1123 mL | |
| 10 mM | 0.2056 mL | 1.0281 mL | 2.0561 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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