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| 5mg |
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| Other Sizes |
| Targets |
NOX2-IN-2 diTFA targets the p47phox-p22phox protein-protein interaction (PPI). NOX2 (NADPH oxidase 2) is an enzyme complex that generates reactive oxygen species (ROS) in phagocytes. The p47phox and p22phox subunits are critical for the activation of the NOX2 complex. By inhibiting the interaction between these two proteins with a Ki of 0.24 microM, NOX2-IN-2 diTFA prevents the assembly of the active NOX2 complex and blocks NOX2-dependent ROS production.
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| ln Vitro |
NOX2-IN-2 diTFA is a potent inhibitor of NOX2 with a Ki of 0.24 microM for targeting the p47phox-p22phox protein-protein interaction. This indicates high affinity and potency for disrupting the PPI. In cellular assays, the compound effectively inhibits ROS production by NOX2, as demonstrated by a reduction in ROS levels in NOX2-expressing cells. It is a selective inhibitor that targets the protein-protein interaction rather than the enzyme‘s active site.
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| ln Vivo |
Specific in vivo activity data for NOX2-IN-2 diTFA is not provided. As a NOX2 inhibitor, it is expected to be effective in animal models of diseases driven by oxidative stress and inflammation, such as inflammatory bowel disease (IBD), ischemia-reperfusion injury, and neuroinflammation. By blocking NOX2-derived ROS production, the compound may reduce tissue damage and inflammation in these models.
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| Enzyme Assay |
The binding of NOX2-IN-2 diTFA to the p47phox-p22phox interaction site can be assessed using a fluorescence polarization (FP) or AlphaScreen assay. Procedure: Biotinylated p22phox peptide (10 nM) is incubated with p47phox protein (20 nM) in assay buffer (25 mM HEPES, pH 7.4, 100 mM NaCl, 0.1% BSA, 0.01% Tween-20). Varying concentrations of NOX2-IN-2 diTFA (0.001-1000 nM) are added. After 1-2 hours, streptavidin-coated donor beads and acceptor beads (for AlphaScreen) or fluorescein-labeled streptavidin (for FP) are added. The signal is measured, and the IC50 is calculated. The reported Ki is 0.24 microM.
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| Cell Assay |
The inhibition of NOX2-derived ROS production can be assessed in a cell-based dihydroethidium (DHE) or DCFH-DA assay. Procedure: Human leukocytes (e.g., neutrophils or macrophages) or NOX2-expressing cell lines (e.g., THP-1 differentiated macrophages) are seeded in 96-well plates (1x10⁵ cells/well). Cells are pre-treated with varying concentrations of NOX2-IN-2 diTFA (0.01-100 uM) for 30 minutes. ROS production is stimulated by adding PMA (phorbol 12-myristate 13-acetate, 100 nM) or opsonized zymosan. The probe DCFH-DA (10 uM) is added, and fluorescence (excitation 485 nm, emission 530 nm) is measured kinetically for 30-60 minutes. The IC50 for inhibition of ROS production is calculated. Cell viability is assessed by MTT assay.
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| Animal Protocol |
The in vivo efficacy of NOX2-IN-2 diTFA can be evaluated in a mouse model of acute lung injury (ALI) or inflammatory bowel disease (IBD). Procedure: For the ALI model, male C57BL/6J mice (8 weeks) are administered NOX2-IN-2 diTFA intranasally at doses of 0.5, 1, and 5 mg/kg, 30 minutes before intratracheal instillation of LPS (10 ug/mouse) to induce ALI. After 6 hours, mice are euthanized, and bronchoalveolar lavage fluid (BALF) is collected. Neutrophil infiltration in BALF (cell count and differential) and protein content are measured. Lung tissue is harvested for measurement of ROS levels (DHE staining) and MPO activity. A reduction in these parameters indicates efficacy.
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| ADME/Pharmacokinetics |
Specific PK data for NOX2-IN-2 diTFA is not provided. As a small molecule with a molecular weight of 699.56 g/mol, it is relatively large and may have moderate oral bioavailability. The compound is soluble in DMSO at 125 mg/mL. For in vivo studies, it is likely formulated in vehicles such as DMSO:PEG300:Saline for intraperitoneal or intranasal administration. The diTFA salt form is used to enhance solubility and stability. It is stored as a powder at 4degC.
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| Toxicity/Toxicokinetics |
Specific toxicology data for NOX2-IN-2 diTFA is not provided. As a research chemical, it is not intended for in vivo use in humans. Since NOX2 is involved in host defense, inhibiting it could potentially increase susceptibility to infections. Standard safety precautions for handling research chemicals should be followed. The compound is for research use only.
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| References | |
| Additional Infomation |
NOX2-IN-2 diTFA (compound 33) is a potent NOX2 inhibitor targeting the p47phox-p22phox protein-protein interaction with a Ki of 0.24 microM. It effectively suppresses the production of reactive oxygen species (ROS) generated by NOX2 in cells. It is used in research on inflammatory diseases and oxidative stress. This product is for research use only and is not an approved drug.
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| Molecular Formula |
C29H27F6N7O7
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| Molecular Weight |
699.56
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| Exact Mass |
699.188
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| CAS # |
2648709-80-0
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| PubChem CID |
156407711
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
4
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| Rotatable Bond Count |
10
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| Heavy Atom Count |
49
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| Complexity |
730
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1=CC2=C(C=CC(=N2)N)C=C1OCCOCCN3C=C(N=N3)COC4=CC5=C(C=C4)N=C(C=C5)N.C(=O)(C(F)(F)F)O.C(=O)(C(F)(F)F)O
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| InChi Key |
JGVQFWXGKWGPBE-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C25H25N7O3.2C2HF3O2/c26-24-7-1-17-13-20(3-5-22(17)28-24)34-12-11-33-10-9-32-15-19(30-31-32)16-35-21-4-6-23-18(14-21)2-8-25(27)29-23;2*3-2(4,5)1(6)7/h1-8,13-15H,9-12,16H2,(H2,26,28)(H2,27,29);2*(H,6,7)
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| Chemical Name |
6-[2-[2-[4-[(2-aminoquinolin-6-yl)oxymethyl]triazol-1-yl]ethoxy]ethoxy]quinolin-2-amine;bis(2,2,2-trifluoroacetic acid)
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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 Note: (1). Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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 : ~125 mg/mL (~178.68 mM; with ultrasonication)
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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 | 1.4295 mL | 7.1473 mL | 14.2947 mL | |
| 5 mM | 0.2859 mL | 1.4295 mL | 2.8589 mL | |
| 10 mM | 0.1429 mL | 0.7147 mL | 1.4295 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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