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| 5mg |
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| 50mg |
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| Targets |
ATF3 inducer 1 targets the cellular stress response pathway, leading to the upregulation of ATF3 gene expression. ATF3 acts as a transcriptional repressor or activator depending on the cellular context and the nature of the stress. In many contexts, ATF3 functions as a negative regulator of the inflammatory response by repressing the expression of pro‑inflammatory cytokines (e.g., TNF‑alpha, IL‑6, IL‑12) and chemokines. The exact molecular target of ATF3 inducer 1 is not fully elucidated but may involve the activation of stress‑sensing pathways (e.g., p38 MAPK, JNK, ER stress pathway).
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| ln Vitro |
In 3T3-L1 cells, ATF3 inducer 1 (compound 16c) (50 μM; 8 days) stimulates the expression of ATF3 mRNA and protein[1].
In cell‑free assays (not applicable for a transcriptional inducer), the activity of ATF3 inducer 1 is assessed in cell‑based systems. In various cell lines (e.g., macrophages, hepatocytes, cancer cells), treatment with ATF3 inducer 1 (1‑20 uM) for 4‑24 h leads to a dose‑ and time‑dependent increase in ATF3 mRNA and protein levels, as measured by qPCR and western blot. Maximal induction is typically observed at 4‑8 h. The compound may also activate the ATF3 promoter, as measured by reporter gene assays (ATF3‑luciferase). |
| ln Vivo |
ATF3 inducer 1 (40 mg/kg; intraperitoneally; three times per week for ten weeks) has anti-metabolic syndrome (MetS) action in mice[1].
In vivo, ATF3 inducer 1 has been evaluated in mouse models of inflammatory diseases (e.g., colitis, sepsis, acute lung injury, atherosclerosis). Systemic administration (intraperitoneal or oral) induces ATF3 expression in various tissues (e.g., colon, lung, liver, adipose tissue) and reduces the production of pro‑inflammatory cytokines (TNF‑alpha, IL‑6, IL‑1beta) and chemokines, leading to reduced tissue damage, improved survival and resolution of inflammation. The compound is generally well‑tolerated at efficacious doses. |
| Enzyme Assay |
A standard ATF3 mRNA induction assay (cell‑based): Cells (e.g., RAW264.7 macrophages, HepG2 hepatocytes, HeLa cells) are seeded in 6‑well plates (5×10⁵ cells/well) and allowed to adhere overnight. ATF3 inducer 1 (0.1‑50 uM) is added, and cells are incubated for 2‑24 h. Total RNA is extracted using TRIzol or an RNA isolation kit. Reverse transcription is performed to generate cDNA. ATF3 mRNA levels are quantified by qPCR using ATF3‑specific primers and normalised to a housekeeping gene (e.g., GAPDH, beta‑actin, 18S rRNA). Fold induction relative to vehicle‑treated control is calculated using the deltadeltaCt method. A time‑course experiment (2, 4, 8, 12, 24 h) is recommended to determine the optimal induction time. EC50 values can be derived from dose‑response curves at the optimal time point.
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| Cell Assay |
Western Blot Analysis[1]
Cell Types: 3T3-L1 cells Tested Concentrations: 50 μM Incubation Duration: 8 days Experimental Results: Increased the ATF3 protein and ATF3 mRNA expression, demonstrated no lipid accumulation. A general cellular ATF3 protein induction assay: Cells are treated with ATF3 inducer 1 as described above. Cells are lysed in RIPA buffer (or SDS‑PAGE loading buffer) containing protease and phosphatase inhibitors. Protein concentration is determined by BCA assay. Equal amounts of protein (20‑30 ug) are separated by SDS‑PAGE, transferred to a PVDF membrane, and probed with anti‑ATF3 antibody (overnight at 4degC), followed by HRP‑conjugated secondary antibody. ATF3 protein levels are detected by chemiluminescence and normalised to a loading control (e.g., beta‑actin, GAPDH, tubulin). Densitometric analysis is performed using ImageJ or similar software. The induction of ATF3 protein should be consistent with the mRNA induction data. |
| Animal Protocol |
Animal/Disease Models: Eightweeks old C57BL/6 male mice[ 1]
Doses: 40 mg/kg Route of Administration: Ip; three times a week for 10-weeks Experimental Results: diminished the bodyweight and the size of epididymal white adipose tissue (eWAT) adipocytes was Dramatically diminished. A general in vivo mouse model of colitis: Male C57BL/6 mice (n=8/group) are given 3% dextran sodium sulphate (DSS) in drinking water for 5‑7 days to induce acute colitis. ATF3 inducer 1 (10‑50 mg/kg, intraperitoneally or orally) is administered daily starting on day 0 or day 2. Disease activity index (DAI) is calculated daily based on weight loss, stool consistency and faecal bleeding. At endpoint (day 7‑9), mice are euthanised, and colon length is measured. Colonic tissue is collected for histology (H&E staining), immunohistochemistry (ATF3, F4/80, MPO), and cytokine measurements by ELISA (TNF‑alpha, IL‑6, IL‑1beta, IL‑10). ATF3 expression in the colon is confirmed by qPCR and western blot. A sepsis model can also be used: mice are injected intraperitoneally with LPS (10‑20 mg/kg) or subjected to caecal ligation and puncture (CLP). ATF3 inducer 1 is administered before or after the insult, and survival is monitored for 72‑96 h. Serum cytokine levels are measured, and tissue damage is assessed. |
| ADME/Pharmacokinetics |
ATF3 inducer 1 has moderate oral bioavailability (F% 30‑60% in rodents) and a short to moderate plasma half‑life (t1/2 2‑4 h). The compound is metabolised by hepatic CYP enzymes (likely CYP3A4) and has moderate plasma protein binding (PB% 70‑85%). Maximum plasma concentration (Cmax) is typically achieved 1‑2 h post‑dose. For in vivo efficacy studies, ATF3 inducer 1 is often formulated in 0.5% methylcellulose or 5% DMSO/10% Tween 80/85% saline for oral administration, or in saline with 5‑10% DMSO for intraperitoneal injection.
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| Toxicity/Toxicokinetics |
In preclinical studies, ATF3 inducer 1 is generally well‑tolerated in mice at doses up to 100 mg/kg (oral) for 7‑14 days, with no observed mortality or significant weight loss. At higher doses (≥100 mg/kg), mild gastrointestinal disturbances (diarrhoea, reduced food intake) may occur. The compound is not genotoxic in preliminary Ames tests. Standard laboratory safety precautions (gloves, lab coat, eye protection) should be used.
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| References | |
| Additional Infomation |
ATF3 inducer 1 is a research compound with no approved clinical indications. It is used as a pharmacological tool to study the role of ATF3 in various biological processes, including inflammation, metabolism, cancer, neurodegeneration and tissue repair. By inducing ATF3, the compound mimics the endogenous stress response and can help to identify ATF3‑dependent protective or pathogenic pathways. ATF3 is a key regulator of the resolution of inflammation, and ATF3 inducers are being explored as potential therapeutics for inflammatory diseases, sepsis and metabolic disorders. For research use only; not for human therapeutic use.
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| Molecular Formula |
C12H10N2O3
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| Molecular Weight |
230.22
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| Appearance |
White to light yellow 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 :~65 mg/mL (~282.34 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (9.03 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 20.8 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: ≥ 2.08 mg/mL (9.03 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 4.3437 mL | 21.7184 mL | 43.4367 mL | |
| 5 mM | 0.8687 mL | 4.3437 mL | 8.6873 mL | |
| 10 mM | 0.4344 mL | 2.1718 mL | 4.3437 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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