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    InvivoChem Cat #: V1490
    CAS #: 182498-32-4Purity ≥98%

    Description: SB225002 (SB 225002; SB-225002) is a novel, potent, and selective non-peptide antagonist of chemokine receptor CXCR2 with potential anti-inflammatory activity. It inhibits CXCR2 with an IC50 of 22 nM for inhibiting the binding of interleukin 125I-IL-8 to CXCR2. SB 225002 showed >150-fold selectivity over CXCR1 and four other 7-TMRs tested. In vitro, SB 225002 potently inhibited human and rabbit neutrophil chemotaxis induced by both IL-8 and GROalpha. In vivo, SB 225002 selectively blocked IL-8-induced neutrophil margination in rabbits. The present findings suggest that CXCR2 is responsible for neutrophil chemotaxis and margination induced by IL-8. SB225002 is also reported to be a mitotic inhibitor that promotes mitotic catastrophe in chemo-sensitive and -resistant ovarian cancer cells independent of p53 status in vitro. SB225002 induces apoptosis in both wild-type and p53-deficient ovarian cancer (OVCA) cells through alternative mechanisms. This selective antagonist will be a useful tool compound to define the role of CXCR2 in inflammatory diseases where neutrophils play a major role. 

    References: J Biol Chem. 1998 Apr 24;273(17):10095-8; Cancer Res. 2006 Mar 15;66(6):3071-7; Surgery. 2014 Apr;155(4):640-9.

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    Molecular Weight (MW)352.14
    CAS No.182498-32-4
    Storage-20℃ for 3 years in powder form
    -80℃ for 2 years in solvent
    Solubility (In vitro)DMSO: 70 mg/mL (198.7 mM)
    Water:<1 mg/mL
    Ethanol: 3 mg/mL warmed (8.5 mM)
    Solubility (In vivo)2% DMSO+Castor oil: 10mg/mL  

    SB225002; SB 225002; SB225002;

    Chemical Name: 1-(2-bromophenyl)-3-(2-hydroxy-4-nitrophenyl)urea

    SMILES Code: O=C(NC1=CC=C([N+]([O-])=O)C=C1O)NC2=CC=CC=C2Br

    Exact Mass: 350.98547 

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    In Vitro

    In vitro activity: In vitro, SB225002 inhibits GROα-stimulated calcium mobilization, and potently inhibits human and rabbit neutrophil chemotaxis induced by both IL-8 and GROalpha. SB 225002 substantially reduces the levels of phosphorylated ERK1/2, and decreases cell proliferation in WHCO1 cells. SB225002 also shows the antitumor activity as a microtubule inhibitor.

    Kinase Assay: Assays are performed in 96-well microtiter plates where the reaction mixture contains 1.0 μg/ml membrane protein in 20 mM Bis-Tris-propane, pH 8.0, with 1.2 mM MgSO4, 0.1 mM EDTA, 25 mM NaCl, and 0.03% CHAPS and SB 225002 (10 mM stock in Me2SO) added at the indicated concentrations, the final Me2SO concentration is<1% under standard binding conditions. Binding is initiated by addition of 0.25 nM 125I-IL-8 (2,200 Ci/mmol). After 1-h incubation at room temperature the plate is harvested using a Tomtec 96-well harvester onto a glass fiber filtermat blocked with 1% polyethyleneimine, 0.5% BSA and washed three times with 25 mM NaCl, 10 mM Tris·HCl, 1 mM MgSO4, 0.5 mM EDTA, 0.03% CHAPS, pH 7.4. The filter is dried, sealed in a sample bag containing 10 ml of Wallac 205 Betaplate liquid scintillation fluid, and counted with a Wallac 1205 Betaplate liquid scintillation counter. 

    Cell Assay: Three esophageal squamous cell carcinoma cell lines WHCO1, WHCO5, and WHCO6 originally established from surgical biopsies of primary esophageal squamous cell carcinomas are cultured in DMEM containing 10% FCS at 37°C in a humidified atmosphere of 5% CO2. MTT assays are carried out using the Cell Proliferation kit I. Briefly, 1.5 × 103 cells are plated in 96-well plates in a final volume of 180 μL DMEM per well. SB 225002 (antagonist of CXCR2, 400 nM) is added to cells and 0.001% DMSO (solvent) is added as a control. After the indicated incubation period, 18 μL of the MTT labeling reagent (final concentration 0.5 mg/mL) is added to each well and incubated for 4 hours in a humidified atmosphere. One hundred eighty microliters of the solubilization solution are added to each well and the plates were left overnight at 37°C. The spectrophotometric absorbance of samples is measured at 595 nm using a microtiter plate reader.

    In VivoIn rabbits, SB225002 selectively blocks IL-8-induced neutrophil margination. In mouse intrahepatic cholangiocellular carcinoma model, SB225002 (1 mg/kg i.p.) suppresses the growth of transplanted subcutaneous tumors. In addition, SB225002 also displays long-lasting antinociceptive effects, and reduces TNBS-induced colitis in mouse models. 
    Animal modelRabbits
    Formulation & DosageDissolved in DMSO; 5.5 μg/kg/min; Cannula in the external jugular vein

    J Biol Chem. 1998 Apr 24;273(17):10095-8; Cancer Res. 2006 Mar 15;66(6):3071-7; Surgery. 2014 Apr;155(4):640-9.

    These protocols are for reference only. InvivoChem does not independently validate these methods.



    Competition binding of 125I-IL-8, [3H]FMLP, [3H]LTB4, [3H]LTD4, or 125I-C5a by SB 225002 to appropriate membranes expressing either cloned or primary receptors.  1998 Apr 24;273(17):10095-8.



    Effect of SB 225002 on IL-8-, GROα-, or C5a-induced human neutrophil chemotaxis.  1998 Apr 24;273(17):10095-8. 


    Effect of SB 225002 on IL-8- and fMLP-induced neutrophil margination in rabbits.  1998 Apr 24;273(17):10095-8.


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