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    BRD4770
    BRD4770

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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 #: V0394
    CAS #: 1374601-40-7Purity ≥98%

    Description: BRD4770 (BRD-4770) is a novel and selective inhibitor of G9a (a histone methyltransferase, also known as euchromatin histone methyltransferase 2 (EHMT2)) with anticancer activity. It inhibits G9a with an IC50 of 6.3 μM, and can induce cell senescence. It exhibits excellent antiproliferative activity against various cancer cell lines such as pancreatic cancer cells-PANC-1.  

    References: ACS Chem Biol. 2012 Jul 20;7(7):1152-7; Cell Death Dis. 2013 Jun 27;4:e690. 

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    Molecular Weight (MW)413.47
    FormulaC25H23N3O3
    CAS No.1374601-40-7
    Storage-20℃ for 3 years in powder form
    -80℃ for 2 years in solvent
    Solubility (In vitro)DMSO: 27 mg/mL (65.3 mM)
    Water:<1 mg/mL 
    Ethanol:<1 mg/mL 
    SMILESO=C(C1=CC=C2C(N=C(NC(C3=CC=CC=C3)=O)N2CCCC4=CC=CC=C4)=C1)OC
    Synonymsmethyl 2-benzamido-1-(3-phenylpropyl)-1H-benzo[d]imidazole-5-carboxylate; BRD-4770; BRD 4770; BRD4770; 


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    In Vitro

    In vitro activity: BRD4770 reduces cellular levels of di- and trimethylated H3K9 via inhibition of G9a, induces senescence, and inhibits both anchorage-dependent and -independent proliferation in the pancreatic cancer cell line PANC-1. The combination of gossypol and BRD4770 increases LC3-II levels and the autophagosome number, and thus acts in synergy to induce cell death in PANC-1 cells.


    Kinase Assay: Dissociation Enhanced Lanthanide Fluoro-ImmunoAssays (DELFIA) are performed in white, opaque 384-well plates coated with Neutravidin. Test compounds are diluted to 12 μg/mL in 50mM Tris-HCl pH 8.5 containing 4% DMSO and 10 μL is dispensed into the wells. Blank and control wells received only compound buffer. GST-G9a at 10 μg/mL and SAM at 40 μM are diluted in 50mM Tris HCl pH 8.5/10 mM DTT and added in a volume of 20 μL. Blank wells receives Tris/DTT buffer only. The reactions are initiated by the addition of 800 nM H3(1-20)-cysbiotin substrate in 50 mM Tris pH 8.5 in a volume of 10 μl, and incubated at room temperature for 60 minutes. The plates are washed 3 times with 100 μl of Wash Buffer (50mM Tris pH 7.4, 150 mM NaCl, 0.05% Tween 20, 0.2% BSA). Next, 50 μl of Fluoroimmunoassay (FI) Buffer (50 mM Tris HCl pH 7.8, 150 mM NaCl, 0.05% Tween 40, 25 μM DTPA, 0.2% BSA, 0.05% BGG) containing 5ng α-2X-di-meth H3-K9 and 5ng goat anti-rabbit Eu chelate is added to all wells of the plate, and the plate is incubated for an additional hour at room temperature. The plates are washed 3 times with 100 μL of Wash Buffer, and 50 μL of Enhancement Solution is added to each well. Time resolved fluorescence is measured after 45 minutes on a Viewlux Microplate Imager imaging for 15 seconds with a 354 μs window, 400 μs delay, excitation at 360 nm, and emission at 618 nm.


    Cell Assay: PANC-1 cells are seeded and treated with BRD4770 in 6-well plates for 72 h. Cells are trypsinized and tested for soft agar colony formation using CytoSelect 96-Well Cell Transformation Assay, using the CyQuant GR dye to measure total cellular nucleic acid levels. Fluorescence is detected with an Analyst HT plate reader using a 485/520 nm filter set.

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    References

    ACS Chem Biol. 2012 Jul 20;7(7):1152-7; Cell Death Dis. 2013 Jun 27;4:e690. 


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

    BRD4770

    Effects of BRD4770 inhibition on the ATM and ATR pathways.  2012 Jul 20;7(7):1152-7. 

    BRD4770

    Small-molecule inhibitors of G9a.

    BRD4770

    Comparison of genetic and small-molecule inhibition of G9a in PANC-1 cells.  2012 Jul 20;7(7):1152-7.


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