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    Combretastatin A4 (CA-4; CRC 87-09)
    Combretastatin A4 (CA-4; CRC 87-09)

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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 #: V1610
    CAS #: 117048-59-6Purity ≥98%

    Description: Combretastatin A4 (CA-4; CRC-87-09)  is a highly potent tubulin inhibitor with potential antitumor activity. It belongs to the so called microtubule-targeting agent (MTA) or microtubule disrupting agent and acts by binding to β-tubulin with a Kd of 0.4 μM. 

    References: J Med Chem. 2014 Apr 24;57(8):3369-81; Magn Reson Med. 2016 Feb;75(2):866-72.


    Publications Citing Use of InvivoChem Combretastatin A4 (CA-4; CRC 87-09)

    1) Eur J Med Chem. 2019 Dec 15;184:111732. doi: 10.1016/j.ejmech.2019.111732;

    2) Eur J Med Chem. 2021 Mar 5;213:113058. doi: 10.1016/j.ejmech.2020.113058

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    Molecular Weight (MW)316.35
    FormulaC18H20O5
    CAS No.117048-59-6
    Storage-20℃ for 3 years in powder form
    -80℃ for 2 years in solvent
    Solubility (In vitro)DMSO: 63 mg/mL (199.1 mM)
    Water:<1 mg/mL
    Ethanol: 34 mg/mL (107.5 mM)
    Solubility (In vivo)5% DMSO+50% PEG 300+ddH2O: 30mg/mL 
    SynonymsCombretastatin A-4; Combretastatin A 4; Combretastatin A4; CRC 87-09; CRC 87-09; CRC 87-09; CA-4; CA4; CA 4; 


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    In Vitro

    In vitro activity: Combretastatin A4 inhibits the growth of MDA-MB-231, A549, Hela, HL-60, SF295, HCT-8, MDA-MB435, PC3M, OVCAR-8, NCI-H358M, and lymphocyte cells with IC50 of 2.8, 3.8, 0.9, 2.1, 6.2, 5.3, 7.9, 4.7, 0.37, 8, and 3.2 nM, respectively. 1 μM Combretastatin A4 inhibits tubulin polymerization by 35%, and 10 μM nearly completely blocks tubulin polymerization. Combretastatin A4 demonstrates great relative binding capacity, reaching 78% of colchicine binding


    Kinase Assay: Colchicine (1.2 μ M) is incubated with tubulin (1.3 mg/mL) in the incubation buffer (80 mM PIPES, 2.0 mM MgCl2, 0.5 mM EGTA, pH 6.9) at 37℃ for 1 h. Varying concentrations (0.1 − 125 μ M) of Combretastatin A4 are used to compete with colchicine originally bound to tubulin. After incubation, the filtrate is obtained. The ability of the analogue to inhibit the binding of colchicine is expressed as a percentage of control binding in the absence of any competitor. 


    Cell Assay: MDA-MB-231, A549, and HeLa cells are grown in DMEM medium (115 units/mL of penicillin G, 115 μg/mL of streptomycin, and 10% fetal bovine serum). Cells are seeded in 96-well plates (5000 cells/well) containing 50 μL of growth medium for 24 h. After medium removal, 100 μL of fresh medium containing individual analogue compounds at different concentrations is added to each well and incubated at 37 ℃ for 72 h. After 24 h of culture, the cells are supplemented with 50 μL of analogue compounds dissolved in DMSO (less than 0.25% in each preparation). After 72 h of incubation, 20 μL of resazurin is added for 2 h before recording fluorescence at 560 nm (excitation) and 590 nm (emission) using a Victor microtiter plate fluorimeter. The IC50 is defined as the compound concentration required to inhibit cell proliferation by 50% in comparison with cells treated with the maximum amount of DMSO (0.25%) and considered as 100% viability.

    In VivoIn NT2 and MDA-MB-231 mammary tumor model, administration of Combretastatin A4 (100 mg/kg, i.p.) induces a significant decrease in lipids R1 and a reduction in tumor pO2 measured by electron paramagnetic resonance (EPR) oximetry. Combretastatin A4 (100 mg/kg, i.p.) significant decreases the Ktrans in male NMRI mice.
    Animal modelFVB/N or nude NMRI female mice bearing NT2 and MDA-MB-231 tumors  
    Formulation & DosageDissolved in DMSO; 100 mg/kg; i.p. injection
    ReferencesJ Med Chem. 2014 Apr 24;57(8):3369-81; Magn Reson Med. 2016 Feb;75(2):866-72.


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

    Combretastatin A4

    Effects of CA-4 and 4h on interphase microtubules. J Med Chem. 2014 Apr 24;57(8):3369-81.
     
    Combretastatin A4
    Flow cytometric analysis of cell cycle distributions of HeLa cells treated with pyridine-bridged CA-4 analogues 4h and 4s. J Med Chem. 2014 Apr 24;57(8):3369-81.


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