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    InvivoChem Cat #: V0831
    CAS #: 108409-83-2Purity ≥98%

    Description: FH535 (FH-535; FH 535) is a novel and potent dual inhibitor of the Wnt/β-catenin and the PPAR (γ/δ) pathways with potential antitumor activity. It shows potent anti-proliferative activity in vitro against various cancer cells such as colon, lung, and hepatocellular carcinomas but not normal fibroblasts. FH535 also exhibits excellent in vivo antitumor efficacy in pancreatic cancer xenografts.

    References: Mol Cancer Ther. 2008;7(3):521-9; Int J Toxicol. 2012;31(4):380-9; PLoS One. 2014;9(6):e99272. 

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    Molecular Weight (MW)361.20
    CAS No.108409-83-2
    Storage-20℃ for 3 years in powder form
    -80℃ for 2 years in solvent
    Solubility (In vitro)DMSO: 72 mg/mL (199.3 mM)
    Water: <1 mg/mL
    Ethanol: <1 mg/mL
    SMILES Code O=S(C1=CC(Cl)=CC=C1Cl)(NC2=CC=C([N+]([O-])=O)C=C2C)=O
    SynonymsFH-535; FH535; FH 535;

    Chemical Name: 2,5-dichloro-N-(2-methyl-4-nitrophenyl)-benzenesulfonamide


    InChi Code: InChI=1S/C13H10Cl2N2O4S/c1-8-6-10(17(18)19)3-5-12(8)16-22(20,21)13-7-9(14)2-4-11(13)15/h2-7,16H,1H3

    SMILES Code: O=S(C1=CC(Cl)=CC=C1Cl)(NC2=CC=C([N+]([O-])=O)C=C2C)=O

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    In Vitro

    In vitro activity: FH535 is a potent inhibitor of the Wnt/β-catenin signaling pathway and also a dual PPARγ and PPARδ antagonist. In vitro, FH535 can inhibit the growth of colon, lung, and hepatocellular carcinoma but not normal fibroblasts.  FH535 selectively inhibits cell proliferation and potentiates imatinib-induced apoptosis in myeloid leukemia cell lines. It is reported that FH535 has ability to inhibit growth, migration, and invasion of TN breast cancer cell lines (MDA-MB-231 and HCC38) without affecting adhesive abilities of cells to type I collagen. FH535 antagonizes β-Catenin/Tcf–mediated transcription, and inhibits recruitment of the coactivators GRIP1 and β-catenin to PPARδ and PPARγ. FH535 shows selective anti-proliferation effect on some cancer cells expressing high or active Wnt/β-catenin pathway. FH535 increases cigarette smoke condensate cytotoxicity, and causes changes in β-catenin and EGR-1 signaling. FH535 has potential therapeutic value in treatment of liver cancer by targeting liver cancer stem cells and hepatocellular carcinoma cell lines.

    Kinase Assay: Three copies of the optimized or mutated Tcf-binding element from TOPFLASH or FOPFLASH driving a secreted alkaline phosphatase reporter gene are cloned into pCEP4 plasmid, replacing the cytomegalovirus promoter. The plasmids are transfected into HepG2 cells, and hygromycin-resistant clones are pooled. Library screening is done at 20 μmol/L concentration in HepG2 serum-free media. Hits are tested in the HCT116 cell line for inhibition of TOPFLASH luciferase activity but not for inhibition of a reporter activity controlled from β-actin promoter. 

    Cell Assay:  Cell viability is determined by the modified 3H-thymidine incorporation assay. Briefly, cells (HCT116, SW48, RKO, LoVo, COLO205, IEC6, A427, HCC15, NCI-H1703, A549, HepG2, Hep3b, Huh7, Fibroblasts) are plated in 96-well microplates for 24 h and treated in triplicate with various concentrations of the test compound. After 48 h of compound exposure, the cells are incubated for an additional 48 h in compound-free medium. The cells are then incubated in medium containing 3H-thymidine for 24 h, washed and mixed with the scintillant in the 96-well plate. Individual wells are counted with a 96-well scintillation counter and the LC50 is calculated.

    In VivoFH535 (at 25 mg/kg, i.p.) demonstrates a potent anti-tumor effect on pancreatic cancer xenografts in mice. FH535 also represses angiogenesis in pancreatic cancer xenografts.
    Animal model Female BALB/c athymic nude mice (Four weeks old)
    Formulation & Dosage Prepared in 100 μL DMSO/DMEM (1:1); 25 mg/kg; i.p.
    ReferencesMol Cancer Ther. 2008 Mar;7(3):521-9. doi: 10.1158/1535-7163.MCT-07-2063; Int J Toxicol. 2012 Jul-Aug;31(4):380-9; PLoS One. 2014 Jun 18;9(6):e99272;  2015 Jul 6;8:1651-70. 

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


    FH535 inhibits β-catenin dependent transcriptional activation in HCC cell lines. PLoS One. 2014 Jun 18;9(6):e99272.


    FH535 reduces cyclin D1 and Survivin protein levels in Huh7 cells. PLoS One. 2014 Jun 18;9(6):e99272.


    FH535 reduces cyclin D1 and survivin mRNA levels in LCSC and in HCC cell lines. PLoS One. 2014 Jun 18;9(6):e99272.


    FH535 inhibited pancreatic cancer cell migration.  2015 Jul 6;8:1651-70.


    FH535 inhibited pancreatic cancer cell invasion.  2015 Jul 6;8:1651-70.



    Inhibitory effect of FH535 on pancreatic cancer cell growth.  2015 Jul 6;8:1651-70.


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