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    InvivoChem Cat #: V1564
    CAS #: 587841-73-4Purity ≥98%

    Description: ZCL278 is a potent, cell-permeable and selective Cdc42 GTPase inhibitor with Kd of 11.4 μM. In human metastatic prostate cancer PC-3 cells, ZCL278 showed inhibitory function on Rac/Cdc42 phosphorylation and the function increasing as the more-treated time. In cortical neurons, ZCL278 treatment suppressed neuronal branch number and inhibited growth cone motility. Treated serum-starved Swiss 3T3 fibroblasts Cdc42 activator following administration of ZCL278 at the dose of 50 μM for 1 h exhibited a significant decrease (nearly 80%) in GTP-Cdc42 and disrupted perinuclear distribution of active Cdc42.

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

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    Molecular Weight (MW)584.89
    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 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


    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.

    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.


    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 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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