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    ZCL278
    ZCL278

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    This product is for research use only, not for human use. We do not sell to patients.
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    InvivoChem Cat #: V1564
    CAS #: 587841-73-4Purity ≥98%

    Description: ZCL278 (ZCL-278; ZCL 278) is a potent, cell-permeable and selective Cdc42 GTPase inhibitor with potential anticancer activity. It inhibits Cdc42 GTPase with a Kd of 11.4 μM. ZCL278 inhibited Rac/Cdc42 phosphorylation in human metastatic prostate cancer PC-3 cells.

    References: Proc Natl Acad Sci U S A. 2013 Jan 22;110(4):1261-6.

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    Molecular Weight (MW)584.89
    FormulaC21H19BrClN5O4S2
    CAS No.587841-73-4
    Storage-20℃ for 3 years in powder form
    -80℃ for 2 years in solvent
    Solubility (In vitro)DMSO: 100 mg/mL (170.9 mM)
    Water: <1 mg/mL
    Ethanol: <1 mg/mL
    Other info

    Chemical Name: 2-(4-bromo-2-chlorophenoxy)-N-[[[4-[[(4,6-dimethyl-2-pyrimidinyl)amino]sulfonyl]phenyl]amino]thioxomethyl]-acetamide

    InChi Key: XKZDWYDHEBCGCG-UHFFFAOYSA-N

    InChi Code: InChI=1S/C21H19BrClN5O4S2/c1-12-9-13(2)25-20(24-12)28-34(30,31)16-6-4-15(5-7-16)26-21(33)27-19(29)11-32-18-8-3-14(22)10-17(18)23/h3-10H,11H2,1-2H3,(H,24,25,28)(H2,26,27,29,33)

    SMILES Code: O=C(NC(NC1=CC=C(S(=O)(NC2=NC(C)=CC(C)=N2)=O)C=C1)=S)COC3=CC=C(Br)C=C3Cl

    Synonyms

    ZCL-278; ZCL 278; ZCL278.


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    In Vitro

    In vitro activity: ZCL278 inhibits Cdc42 GTPase activity by the competition with GTP and inhibits Rac/Cdc42 phosphorylation in a time-dependent manner. ZCL278 (50 μM) inhibits Cdc42-mediated microspike formation and disrupts GM130-docked Golgi structures in serum-starved Swiss 3T3 fibroblasts. In addition, ZCL278 also suppresses Cdc42-mediated neuronal branching and growth cone dynamics as well as actin-based motility and migration in a metastatic prostate cancer PC-3 cell line without cytotoxicity.


    Kinase Assay: Inorganic phosphate produced as a result of GTPase activity was measured by using a p50RhoGAP or Cdc42GAP assay and absorbance-based detection method. Briefly, Cdc42 was preloaded with GTP or ZCL278 and incubated in the reaction buffer for 20 mins at 37 °C. GAP was then added for an additional 20 mins at 37 °C. Following a 10-min incubation in CytoPhos reagent, inorganic phosphate was detected at 650 nm.


    Kinase Assay: Lyophilized Cdc42 protein is reconstituted to 5 mg/mL in a buffer consisting of 50 mM Tris, 0.5 mM MgCl2, 50 mM NaCl, 3% (wt/vol) sucrose, and 0.6% dextran. The stock solution is then diluted to 1 μM in 5 mM phosphate buffer, pH 7.4. Into a quartz cuvettete containing Cdc42 solution, aliquots of ZCL278 are added and incubated for 5 min before each fluorescent measurement. The excitation wavelength is 275 nm, and the fluorescence of tryptophan at 350 nm is measured after each addition. The titration curve is fitted using the equimolar specific binding model in GraphPad, and the Kd is calculated.


    Cell Assay: In ZCL278-treated neurons, neuronal branching was significantly suppressed over the time course. In addition, ZCL278 markedly inhibited Cdc42-mediated growth cone motility.

    In VivoZCL278 reduces the JUNV RNA load in the spleen more than 33-fold, with JUNV RNA being undetectable in 5 out of 8 mice. These results are similar to those seen in Gabapentin-treated mice, demonstrating that ZCL278 can abrogate JUNV replication. Four-week-old C57BL/6 mice receive intravenous injections of Gabapentin or ZCL278 (100 μg/g, i.p.). At 1 h after treatment, the mice are inoculated intraperitoneally with JUNV Candid #1 (1×106 PFU) in no more than 1 mL with a 27 1/2-gauge needle. At the end of the experiment, the mice are sacrificed, their spleens are homogenized with a Dounce homogenizer and centrifuged to generate a cell pellet and supernatant, and RNA expression levels are determined.
    Animal modelMice
    Formulation & Dosage100 μg/g, i.p.
    References

    Proc Natl Acad Sci U S A. 2013 Jan 22;110(4):1261-6.


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

    ZCL278

    Characterization of ZCL278 functions. (A) ZCL278, but not ZCL197 or ZCL279, inhibits Cdc42-mediated microspike formation. Proc Natl Acad Sci U S A. 2013 Jan 22;110(4):1261-6.
     

    ZCL278

    ZCL278 inhibits neuronal branching and growth cone motility. Proc Natl Acad Sci U S A. 2013 Jan 22;110(4):1261-6.


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