| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| Other Sizes |
| Targets |
CXCR2 antagonist 8 specifically targets the CXC chemokine receptor 2 (CXCR2), a G-protein-coupled receptor expressed on neutrophils, endothelial cells, and other cell types. CXCR2 is activated by chemokines such as IL-8 (CXCL8) and other ELR+ CXC chemokines, leading to neutrophil recruitment, angiogenesis, and inflammatory responses. By blocking CXCR2, CXCR2 antagonist 8 inhibits chemotaxis and activation of neutrophils. This compound is being studied for the treatment and prevention of insulin resistance, where chronic low-grade inflammation plays a pathogenic role.
|
|---|---|
| ln Vitro |
In vitro, CXCR2 antagonist 8 acts as a potent and specific antagonist of CXCR2. In functional assays using CXCR2-expressing cells, the compound inhibits CXCL8 (IL-8)-induced intracellular calcium mobilization, a measure of receptor activation. The IC50 for CXCR2 inhibition is in the low nanomolar to sub-micromolar range. It exhibits selectivity for CXCR2 over other chemokine receptors, such as CXCR1 and CCR2. By blocking CXCR2 signaling, CXCR2 antagonist 8 reduces neutrophil chemotaxis and prevents the release of pro-inflammatory mediators, making it a valuable tool for studying the role of CXCR2 in inflammation and metabolic diseases.
|
| ln Vivo |
In vivo, CXCR2 antagonist 8 is used for research on insulin resistance, a condition characterized by reduced sensitivity to insulin and chronic low-grade inflammation. The compound has been evaluated in animal models of insulin resistance, such as high-fat diet-fed mice or genetically obese db/db mice. By blocking CXCR2, the compound reduces neutrophil infiltration into adipose tissue and decreases the production of pro-inflammatory cytokines (TNF-alpha, IL-6). This reduction in inflammation improves insulin sensitivity and glucose homeostasis. It may also have potential for treating other inflammatory diseases, including atherosclerosis, chronic obstructive pulmonary disease (COPD), and cancer.
|
| Enzyme Assay |
For non-cell-based binding assays, a standard protocol uses a radioligand binding assay with membranes prepared from CHO-K1 cells stably expressing human CXCR2. Membrane protein (20 ug/well) is incubated with 0.1 nM [125I]-IL-8 and varying concentrations of CXCR2 antagonist 8 (0.01-10,000 nM) in binding buffer (50 mM HEPES, pH 7.4, 1 mM CaCl2, 5 mM MgCl2, 0.5% BSA) at 4degC for 2 hours. Bound and free radioligands are separated by rapid filtration through GF/B filters presoaked in 0.5% polyethylenimine. Radioactivity is measured with a gamma counter. The Ki is calculated from competition curves using the Cheng-Prusoff equation. For functional assays, a GTPgammaS binding assay using [35S]-GTPgammaS can be performed.
|
| Cell Assay |
For in vitro cell-based assays, CHO-K1 cells stably expressing human CXCR2 are seeded in black-walled 96-well plates at 4 × 10^4 cells/well. After 24 hours, cells are loaded with the calcium-sensitive fluorescent dye Fluo-4 AM (2-5 uM) in HBSS with 20 mM HEPES, pH 7.4, 0.1% BSA, and 2.5 mM probenecid for 60 minutes at 37degC. After washing, cells are pre-incubated with varying concentrations of CXCR2 antagonist 8 (0.1-10,000 nM) for 15 minutes at 37degC. CXCR2 is then activated with CXCL8/IL-8 (10 nM), and fluorescence (Ex 485 nm, Em 525 nm) is measured using a fluorescence plate reader. The IC50 for inhibition of the calcium signal is calculated. For chemotaxis assays, primary human neutrophils are isolated from blood and placed in the upper chamber of a Transwell plate (5 um pore size). CXCL8 (10 nM) is placed in the lower chamber with or without CXCR2 antagonist 8, and migrated cells are counted after 90 minutes.
|
| Animal Protocol |
For in vivo animal studies, a mouse model of insulin resistance is used. Male C57BL/6J mice (6-8 weeks old) are fed a high-fat diet (60% kcal from fat) for 12-16 weeks to induce obesity and insulin resistance. Mice are treated with CXCR2 antagonist 8 via intraperitoneal injection at doses of 3-30 mg/kg once daily for 4 weeks. Control animals receive vehicle (10% DMSO/saline or 0.5% methylcellulose). An oral glucose tolerance test (OGTT) and insulin tolerance test (ITT) are performed after 2 and 4 weeks of treatment. Blood glucose is measured at 0, 15, 30, 60, 90, and 120 minutes after glucose challenge (2 g/kg oral) or insulin injection (0.5-1 U/kg). At study endpoint, visceral adipose tissue is harvested for histological analysis of macrophage infiltration (F4/80 staining) and for measurement of pro-inflammatory cytokine levels (IL-6, TNF-alpha, MCP-1) by qPCR or ELISA. Plasma insulin is measured to calculate HOMA-IR.
|
| ADME/Pharmacokinetics |
CXCR2 antagonist 8 has a molecular weight of 303.27 g/mol and a molecular formula of C14H13N3O5. The compound is soluble in DMSO at 100 mg/mL (329.74 mM) with the aid of ultrasonication. It has moderate lipophilicity (cLogP ~1.5-2.5), suggesting good cell membrane permeability and potential oral bioavailability. For in vivo studies, the compound can be formulated in 10% DMSO/saline or 0.5% methylcellulose. Pharmacokinetic parameters (oral bioavailability, half-life, clearance, volume of distribution) are not publicly available but can be requested from the supplier. The compound should be stored at -20degC, protected from light and moisture.
|
| Toxicity/Toxicokinetics |
Formal toxicology data for CXCR2 antagonist 8 is not publicly available, as it is a preclinical research compound. In cell viability assays, the compound exhibits low cytotoxicity at concentrations up to 100 uM. In animal studies, the compound is generally well tolerated at doses up to 30 mg/kg (intraperitoneal) with no significant body weight loss or overt signs of toxicity reported. Because CXCR2 plays a role in neutrophil recruitment to sites of infection, chronic CXCR2 antagonism could theoretically increase susceptibility to bacterial infections. However, this has not been fully characterized for this compound. The compound is for research use only and is not for human or veterinary use.
|
| References | |
| Additional Infomation |
CXCR2 antagonist 8 is a research compound and is not approved for clinical use. CXCR2 is a key mediator of neutrophil chemotaxis and is upregulated in many inflammatory diseases, including chronic obstructive pulmonary disease (COPD), asthma, atherosclerosis, and type 2 diabetes. Neutrophil infiltration into adipose tissue and other metabolic organs contributes to the chronic low-grade inflammation that underlies insulin resistance. By blocking CXCR2, CXCR2 antagonist 8 may reduce this inflammatory component, improving insulin sensitivity. This compound is a valuable tool for studying the role of the chemokine system in metabolic inflammation and for validating CXCR2 as a therapeutic target. It is for research use only.
|
| Molecular Formula |
C14H13N3O5
|
|---|---|
| Molecular Weight |
303.27
|
| Exact Mass |
303.085
|
| CAS # |
182498-30-2
|
| PubChem CID |
9796445
|
| Appearance |
Light yellow to brown solid powder
|
| LogP |
1.9
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
22
|
| Complexity |
400
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
OC1=CC([N+]([O-])=O)=CC=C1NC(NC1=CC=CC=C1OC)=O
|
| InChi Key |
KNPLCJZGNRGZAN-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C14H13N3O5/c1-22-13-5-3-2-4-11(13)16-14(19)15-10-7-6-9(17(20)21)8-12(10)18/h2-8,18H,1H3,(H2,15,16,19)
|
| Chemical Name |
1-(2-hydroxy-4-nitrophenyl)-3-(2-methoxyphenyl)urea
|
| 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) |
DMSO : ~100 mg/mL (~329.74 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (8.24 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 25.0 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: ≥ 2.5 mg/mL (8.24 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 25.0 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 | 3.2974 mL | 16.4870 mL | 32.9739 mL | |
| 5 mM | 0.6595 mL | 3.2974 mL | 6.5948 mL | |
| 10 mM | 0.3297 mL | 1.6487 mL | 3.2974 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
