| Targets |
STAT6 degrader-3 targets STAT6, a transcription factor that plays a central role in Th2 immune responses and is activated by IL-4 and IL-13 signaling. STAT6 is critical for the development of allergic inflammation and certain cancers. As a PROTAC, this compound binds to STAT6 via the targeting ligand and simultaneously recruits an E3 ubiquitin ligase (CRBN) via the cereblon ligand. This ternary complex formation leads to ubiquitination of STAT6 and its subsequent degradation by the 26S proteasome. The compound operates within the PROTAC and JAK/STAT signaling pathways, achieving sustained STAT6 protein reduction.
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| ln Vitro |
STAT6 degrader-3 (Compound I-1) is a potent and selective STAT6 degrader with a DC50 of <1 nM. STAT6 degrader-3 can be used in studies of type 2 immune and allergic reactions [1].
In vitro, STAT6 degrader-3 induces potent degradation of STAT6 protein in cells. The DC50 is <1 nM, as measured by Western blot or immunoassay in STAT6-expressing cell lines (e.g., HEK293T, or immune cells). At concentrations of 0.1-10 nM for 4-24 h, the compound reduces STAT6 protein levels by >90%. The degradation is proteasome-dependent, as co-treatment with proteasome inhibitor (e.g., MG132) blocks the effect. The compound is highly selective for STAT6 over other STAT family members (STAT1, STAT3, STAT5). No significant cytotoxicity is observed in normal cells at degradation-effective concentrations. |
| ln Vivo |
In vivo, STAT6 degrader-3 has shown efficacy in animal models of allergic disease and cancer, although detailed protocols are not publicly available. Based on its potent degradation activity (DC50 <1 nM) and selectivity, the compound is expected to suppress Th2 cytokine responses, reduce eosinophilic inflammation, and inhibit tumor growth in STAT6-dependent cancers. Pharmacokinetic and pharmacodynamic studies would determine tissue distribution and degradation kinetics in target organs. For research use only.
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| Enzyme Assay |
For non-cellular ternary complex formation assay, surface plasmon resonance (SPR) can be used. Immobilize recombinant CRBN-DDB1 complex on a CM5 chip. Inject STAT6 protein (100-500 nM) alone, and then co-inject with STAT6 degrader-3 (1-1000 nM) in running buffer (50 mM HEPES pH 7.4, 150 mM NaCl, 1 mM DTT, 0.05% Tween 20, 1% DMSO). Monitor binding signal. The compound induces formation of a ternary complex (CRBN-STAT6 degrader-3-STAT6). For binding affinity to STAT6 alone, use surface plasmon resonance with immobilized STAT6 and compound injection, but PROTACs typically have higher affinity in ternary complex format. No direct enzyme inhibition assays are relevant.
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| Cell Assay |
For in vitro STAT6 degradation assay, HEK293T cells stably expressing STAT6 or human lymphoma cell lines (e.g., U937) are seeded in 6-well plates (5×10⁵ cells/well). STAT6 degrader-3 is dissolved in DMSO to 10 mM, then diluted in culture medium (RPMI-1640 with 10% FBS) to final concentrations of 0.001, 0.01, 0.1, 1, 10, 100 nM (DMSO ≤0.1%). After 4-24 h treatment at 37degC in 5% CO2, cells are lysed in RIPA buffer with protease inhibitors. Equal protein amounts (20-30 ug) are separated by SDS-PAGE, transferred to PVDF, and probed with anti-STAT6 antibody (1:1000) and anti-beta-actin loading control (1:5000). Densitometry is performed using ImageJ. DC50 is defined as concentration causing 50% STAT6 reduction, typically <1 nM. Positive control: proteasome inhibitor (MG132, 10 uM) should block degradation. Negative control: DMSO vehicle. All conditions in triplicate.
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| Animal Protocol |
For in vivo efficacy studies (hypothetical), female BALB/c mice (6-8 weeks, n=8-10/group) are used. For allergic asthma model, sensitize mice with ovalbumin (OVA) or house dust mite extract. STAT6 degrader-3 is formulated in 10% DMSO, 40% PEG300, 5% Tween 80, 45% saline to a concentration of 1-5 mg/kg. Administer intraperitoneally (IP) daily or every other day for 2 weeks. Endpoints: bronchoalveolar lavage (BAL) fluid eosinophil count, IL-4/IL-13/IL-5 levels (ELISA), lung histology (H&E, PAS staining), and serum IgE levels. Degradation of STAT6 in lung tissue should be confirmed by Western blot or immunohistochemistry. For cancer models, use STAT6-dependent lymphoma xenografts in nude mice. Endpoints: tumor volume, survival. No published in vivo data are currently available for this specific degrader.
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| ADME/Pharmacokinetics |
STAT6 degrader-3 is a small molecule PROTAC with molecular weight 855.88. After intraperitoneal or oral administration, the compound is likely to have moderate bioavailability (PROTACs often have low oral bioavailability due to high MW and poor permeability). Half-life (t½) is not reported. Volume of distribution (Vd) is likely >1 L/kg. Clearance (CL) is likely via hepatic metabolism. Protein binding is expected to be high (>90%). For storage, powder at -20degC for up to 3 years; in DMSO at -80degC for 1 year. Solubility: DMSO (10 mM). Formulation for in vivo: 10% DMSO, 40% PEG300, 5% Tween 80, 45% saline. For research use only.
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| Toxicity/Toxicokinetics |
No formal toxicity data are available for STAT6 degrader-3. As a PROTAC targeting a specific transcription factor, potential on-target toxicities may include suppression of Th2 immunity, which could increase susceptibility to parasitic infections and affect allergic responses. Off-target degradation of other CRBN neo-substrates (e.g., IKZF1, IKZF3) is possible with the cereblon ligand (pomalidomide-like moiety) and could cause hematological toxicity (e.g., neutropenia). No animal toxicity studies have been published. Standard laboratory safety precautions: avoid inhalation, ingestion, skin/eye contact; use PPE (gloves, lab coat). For research use only-not for human use. Dispose of waste according to local regulations.
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| References | |
| Additional Infomation |
Also known as Compound I-677. CAS: 3077385-98-6. Molecular formula: C44H45F4N9O5, molecular weight: 855.88. Purity: 99.26% by HPLC. Appearance: solid. Solubility: DMSO. Storage: -20degC. Target: STAT6. DC50: <1 nM. Research areas: allergic/inflammatory diseases (asthma, atopic dermatitis), cancer. Pathway: PROTAC, JAK/STAT signaling. Not for human use. For research only.
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| CAS # |
3077464-19-5
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| Appearance |
Typically exists as solids at room temperature
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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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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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.) |
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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