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    InvivoChem Cat #: V0761
    CAS #: 354812-17-2Purity ≥98%

    Description: SC-514 (SC 514; SC514) is a novel, potent, orally bioactive, selective and reversible, and ATP-competitive IKK-2 inhibitor with potential anti-inflammatory activity. It inhibits IKK2 with an IC50 of 3-12 μM, and does not inhibit other isoforms of  IKK. SC-514 has the potential for treating osteoclastogenesis. 

    References: J Biol Chem. 2003 Aug 29;278(35):32861-71.

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    Molecular Weight (MW)224.3
    CAS No.354812-17-2
    Storage-20℃ for 3 years in powder form
    -80℃ for 2 years in solvent
    Solubility (In vitro)DMSO: 45 mg/mL (200.6 mM)
    Water: <1 mg/mL
    Ethanol: <1 mg/mL
    Other info

    Chemical Name: 4-amino-[2,3'-bithiophene]-5-carboxamide

    SMILES Code: O=C(C1=C(N)C=C(C2=CSC=C2)S1)N           

    SynonymsSC 514; SC514; SC-514; 

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    In Vitro

    In vitro activity: SC-514 inhibits the native IKK complex or recombinant human IKK-1/IKK-2 heterodimer and IKK-2 homodimer similarly. SC-514 inhibits transcription of NF-κB-dependent IL-6, IL-8, and COX-2 genes in IL-1β-induced rheumatoid arthritis-derived synovial fibroblasts (RASFs) with IC50s of 20, 20 and 8 μM. 100 μM SC-514 blocks the phosphorylation and degradation of IκBα and also reduces the level of translocation of p65 into the nucleus in IL-1β-treated RASFs SC-514 does not inhibit the phosphorylation and activation of the IKK complex. SC-514 induces a delay but not a complete blockade in IκBα phosphorylation and degradation. SC-514 treatment cells shows a slightly slowed, decreased import of p65 into the nucleus and a faster export of p65 from the nucleus. SC-514 inhibits the phosphorylation of either IκBα or p65 similarly.

    Kinase Assay: IKK complexes are immunoprecipitated from IL-1β-treated RASF cell lysates (0.5-2 mg) using a NEMO antibody (3-10 μg) followed by the addition of protein A-agarose beads. Antibody complexes are pelleted by centrifugation and washed 3 times with 1 mL of cold whole-cell lysis buffer followed by 2 washes in kinase buffer (25 mM HEPES, pH 7.6, 2 mM MgCl2, 2 mM MnCl2, 10 mM NaF, 5 mM DTT, and 1 mM phenylmethylsulfonyl fluoride). 100-200 μg of immunoprecipitated IKK is analyzed for kinase activity in a reaction containing 10 μM biotinylated IκBα peptide as substrate and 1 μM [γ-33P]ATP (2500 Ci/mmol). After incubation at room temperature for 30 min, 25 μL of the reaction mixture is withdrawn and added to a SAM 96 biotin capture plate. After successive wash steps the plate was allowed to air-dry, and 25 μL of scintillation fluid is added to each well. Incorporation of [γ-33P]ATP is measured using a Top-Count NXT

    Cell Assay: SC-514 inhibits all forms of recombinant human IKK-2 with IC50 values in the 3–12μM range. It also inhibits the native IKK complex. SC-514 specifically binds at the ATP-binding site of IKK-2 and exerts a reversible and competitive inhibition with ATP. However, SC-514 shows non-competitive inhibition with the IκB site. As an inhibitor of IKK-2, SC-514 is found to block the phosphorylation and degradation of IκBα and reduce the translocation level of p65 into the nucleus in IL-1β-treated RASFs. Additionally, SC-514 shows dose-dependent inhibition in the transcription of NF-κB-induced genes, including IL-6, IL-8, and COX-2. Moreover, SC-514 shows efficacious in reduction of LPS-induced TNFα production in the acute model of inflammation. SC-514 is also reported to inhibit the osteoclastogenesis in BMM cells through attenuating RANKL-induced activation of NF-κB.

    In VivoSC-514 is efficacious in an acute model of inflammation, namely LPS-induced serum TNF-α production. SC-514 (50 mg/kg, i.p.) inhibits TNF-αproduction in vivo by ~70%.
    Animal modelRats
    Formulation & Dosage50 mg/kg; i.p. injection

    J Biol Chem. 2003 Aug 29;278(35):32861-71.

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

    Inhibition pattern of SC-514 at the ATP and IκB sites. J Biol Chem. 2003 Aug 29;278(35):32861-71.
    SC-514 dose-dependently inhibits IL-1β-induced NF-κB gene expression in RASF cells. J Biol Chem. 2003 Aug 29;278(35):32861-71.
    Inhibition of IKK-2 by SC-514 selectively blocks the NF-κB pathway. J Biol Chem. 2003 Aug 29;278(35):32861-71.
    SC-514 inhibits IκBα degradation in vitro in IL-1β-induced RASF cells although the immunoprecipitated IKK activity ex vivo demonstrates augmented kinase activity. J Biol Chem. 2003 Aug 29;278(35):32861-71.
    SC-514 inhibition of IKK-2 activity decreases the IκBα phosphorylation/degradation, as well as decreases the length of time NF-κB is in the nucleus. J Biol Chem. 2003 Aug 29;278(35):32861-71.
    SC-514 inhibits LPS-induced serum TNFα production in vivo in the rat. J Biol Chem. 2003 Aug 29;278(35):32861-71.


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