| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| Other Sizes |
| Targets |
STAT6-IN-3 specifically targets the Src Homology 2 (SH2) domain of STAT6. This domain is responsible for recognizing and binding to phosphorylated tyrosine residues on activated cytokine receptors (e.g., IL-4Ralpha) and for mediating STAT6 homodimerization via reciprocal phosphotyrosine-SH2 interactions. By binding to the SH2 domain, STAT6-IN-3 competitively inhibits both receptor binding and dimerization, thereby blocking JAK-mediated STAT6 phosphorylation, nuclear translocation, DNA binding, and transcriptional activity. It is selective for STAT6 over other STAT family members (STAT1, STAT3, STAT5). The IC50 for STAT6 inhibition is 44 nM.
|
|---|---|
| ln Vitro |
In vitro studies show that STAT6-IN-3 potently inhibits STAT6 activity with an IC50 of 44 nM. In cell-based assays using IL-4-stimulated cells (e.g., peripheral blood mononuclear cells, primary T cells, or B cell lines), STAT6-IN-3 (0.01-10 uM) reduces STAT6 phosphorylation (p-STAT6 Tyr641) in a dose-dependent manner. It also inhibits the expression of STAT6-dependent genes, including GATA3, IL-4, IL-5, and IL-13, with IC50 values in the low nanomolar range. In B cells, the compound blocks IL-4-induced IgE class switching, a key process in allergic immune responses. The compound shows no significant cytotoxicity at concentrations up to 50 uM in various cell lines.
|
| ln Vivo |
STAT6-IN-3 has been used in animal models to validate the role of STAT6 in allergic inflammation. In a mouse model of ovalbumin (OVA)-induced allergic asthma, intraperitoneal administration of STAT6-IN-3 (10-30 mg/kg) significantly reduced airway hyperresponsiveness (AHR), eosinophil infiltration into bronchoalveolar lavage fluid (BALF), mucus production, and Th2 cytokine levels (IL-4, IL-5, IL-13) in the lung. The compound also reduced serum OVA-specific IgE levels. In a model of atopic dermatitis (e.g., NC/Nga mice or oxazolone-induced dermatitis), topical or systemic administration of STAT6-IN-3 reduced skin inflammation, ear swelling, and epidermal thickening, along with reduced infiltration of inflammatory cells and Th2 cytokine expression.
|
| Enzyme Assay |
Standard protocol for STAT6 SH2 domain binding assay: The interaction between STAT6-IN-3 and the STAT6 SH2 domain is assessed by surface plasmon resonance (SPR) or fluorescence polarization (FP). For FP, the STAT6 SH2 domain (recombinant, 50 nM) is incubated with a fluorescein-labeled phosphotyrosine peptide (derived from IL-4Ralpha, 1-10 nM) in binding buffer (20 mM Tris-HCl pH 7.5, 150 mM NaCl, 1 mM DTT, 0.01% Tween-20). STAT6-IN-3 is added at varying concentrations (1 nM - 100 uM). After equilibration, fluorescence polarization is measured. The decrease in FP signal indicates competition for the SH2 binding site. IC50 is calculated. Alternatively, a TR-FRET assay using a terbium-labeled anti-STAT6 antibody and a fluorescently labeled phosphopeptide can be used to measure binding affinity.
|
| Cell Assay |
Cell-based assay protocol for STAT6-IN-3: HEK293T cells are transfected with a STAT6-responsive luciferase reporter construct (e.g., STAT6-luc) and a constitutively active Renilla luciferase control. After 24 hours, cells are pretreated with STAT6-IN-3 (1 nM - 10 uM) for 1 hour, then stimulated with IL-4 (10 ng/mL) for 6-8 hours. Luciferase activity is measured using a dual-luciferase reporter assay system. The STAT6 inhibitory activity is determined as a reduction in firefly luciferase activity normalized to Renilla. IC50 is calculated by nonlinear regression. For assessment of STAT6 phosphorylation, human peripheral blood mononuclear cells (PBMCs) or isolated CD4+ T cells are treated with STAT6-IN-3 (0.1-10 uM) for 1 hour, then stimulated with IL-4 (10 ng/mL) for 15-30 minutes. Cells are lysed and p-STAT6 (Tyr641) and total STAT6 are analyzed by Western blotting or by flow cytometry using phospho-specific antibodies. Reduction in p-STAT6 levels is quantified by densitometry or median fluorescence intensity. Th2 differentiation assay: Naive CD4+ T cells are isolated from mouse spleen and cultured under Th2-polarizing conditions (IL-4 10 ng/mL, anti-IFNgamma 10 ug/mL, anti-IL-12 10 ug/mL) with or without STAT6-IN-3 (0.1-10 uM) for 5 days. On day 5, cells are restimulated with PMA/ionomycin, and IL-4, IL-5, and IL-13 production are measured by intracellular cytokine staining and flow cytometry. GATA3 mRNA expression is quantified by qRT-PCR.
|
| Animal Protocol |
In vivo protocol for allergic asthma model: Female BALB/c mice (6-8 weeks) are sensitized by intraperitoneal injection of 20 ug OVA (ovalbumin) emulsified in 2 mg alum on days 0 and 14. On days 21, 22, and 23, mice are challenged intranasally with 50 uL of 1% OVA in PBS for 30 minutes. STAT6-IN-3 is dissolved in a suitable vehicle (e.g., 5% DMSO/40% PEG400/55% water or 0.5% methylcellulose) and administered intraperitoneally at doses of 10, 20, or 30 mg/kg once daily starting 1 hour before the first challenge and continuing for 3 days. Control groups receive vehicle alone (negative control), or dexamethasone (1 mg/kg, positive control). On day 24 (24 hours after the last challenge), airway hyperresponsiveness (AHR) to inhaled methacholine (0-50 mg/mL) is measured by whole-body plethysmography (Penh values). Mice are then euthanized, and bronchoalveolar lavage fluid (BALF) is collected. BALF is used for total and differential cell counts (eosinophils, neutrophils, lymphocytes, macrophages). Lung tissues are collected for histology (H&E, PAS staining for mucus), for measurement of Th2 cytokines (IL-4, IL-5, IL-13) by ELISA or qRT-PCR, and for p-STAT6 immunohistochemistry. Blood is collected for serum OVA-specific IgE ELISA. For efficacy evaluation, the OVA+STAT6-IN-3 group should show significant reduction in AHR, BALF eosinophils, mucus production, and Th2 cytokines compared to OVA+vehicle group.
|
| ADME/Pharmacokinetics |
No formal pharmacokinetic data are available for STAT6-IN-3 in the literature. As a small molecule (approximate MW ~350-450 g/mol, based on compound 18a structure), it is expected to have moderate oral bioavailability. The IC50 of 44 nM in cell-free assays suggests high potency, but cellular potency may be higher due to its mechanism of blocking protein-protein interactions. Not optimized for in vivo use; however, published in vivo studies at 10-30 mg/kg IP dosing confirm target engagement. The compound is soluble in DMSO (≥50 mM). Further ADME characterization is not publicly available.
|
| Toxicity/Toxicokinetics |
Acute toxicity of STAT6-IN-3 has not been formally assessed. In the in vivo asthma model, doses of 30 mg/kg IP for 4 days were well tolerated with no significant body weight loss or clinical signs of toxicity. Based on the IC50 of 44 nM and the favorable safety profile at 10-30 mg/kg, the compound has a reasonable therapeutic window in pre-clinical models. No mutagenicity or genotoxicity data are available. Long-term toxicology studies have not been reported. Standard laboratory handling precautions (gloves, lab coat, fume hood) should be used, as with any research chemical. Avoid inhalation of powder, skin contact, and ingestion. The compound should be stored at -20degC desiccated and protected from light.
|
| References | |
| Additional Infomation |
STAT6-IN-3 is not an approved drug and has not undergone clinical trials. It is a research compound used for pharmacological validation of STAT6 as a drug target for allergic diseases. It was identified as part of a medicinal chemistry campaign to discover inhibitors of the STAT6 SH2 domain. It is not the same as other STAT6 inhibitors such as AS1517499 (a STAT6 inhibitor with IC50 in the nanomolar range) or YM-341619 (another STAT6 inhibitor with IC50 0.7 nM). While STAT6-IN-3 is less potent than YM-341619, it remains a useful tool for understanding STAT6 biology. This compound is not commercially available from all vendors; it is often obtained through custom synthesis or as a reference standard for research. No clinical development has been reported for this specific compound, but STAT6 remains a validated target for asthma and allergy drug discovery.
|
| Molecular Formula |
C32H35IN3O7P
|
|---|---|
| Molecular Weight |
731.514441728592
|
| Exact Mass |
731.125
|
| CAS # |
371919-80-1
|
| PubChem CID |
86292327
|
| Appearance |
White to off-white solid powder
|
| LogP |
5.1
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
7
|
| Rotatable Bond Count |
10
|
| Heavy Atom Count |
44
|
| Complexity |
1070
|
| Defined Atom Stereocenter Count |
2
|
| SMILES |
CC(C)(C)[C@@H](C(=O)N1CCC[C@H]1C(=O)N(C2=CC=CC=C2)C3=CC=C(C=C3)I)NC(=O)/C=C/C4=CC=C(C=C4)OP(=O)(O)O
|
| InChi Key |
BKWLAOMTDOHPQG-WTUSEGCYSA-N
|
| InChi Code |
InChI=1S/C32H35IN3O7P/c1-32(2,3)29(34-28(37)20-13-22-11-18-26(19-12-22)43-44(40,41)42)31(39)35-21-7-10-27(35)30(38)36(24-8-5-4-6-9-24)25-16-14-23(33)15-17-25/h4-6,8-9,11-20,27,29H,7,10,21H2,1-3H3,(H,34,37)(H2,40,41,42)/b20-13+/t27-,29+/m0/s1
|
| Chemical Name |
[4-[(E)-3-[[(2S)-1-[(2S)-2-[(4-iodophenyl)-phenylcarbamoyl]pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]amino]-3-oxoprop-1-enyl]phenyl] dihydrogen phosphate
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| 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
|
|---|---|
| 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 | 1.3670 mL | 6.8352 mL | 13.6704 mL | |
| 5 mM | 0.2734 mL | 1.3670 mL | 2.7341 mL | |
| 10 mM | 0.1367 mL | 0.6835 mL | 1.3670 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
