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    InvivoChem Cat #: V1888
    CAS #: 152095-12-0Purity ≥98%

    Description: Dp44mT is a novel and potent iron chelator with DNA-damaging activity mediated by top2a inhibition and with selective antitumor activity. Dp44mT selectively inhibit topoisomerase IIα, with an GI50 value of ~100 nmol/L in the human breast cancer cell line MDA-MB-231. In control experiments, the Nalm-6 leukemic top2α+/- cells expressed ~57% as much levels of top2α enzyme as the wild type cells. Treated with Dp44mT at 100 nmol/L, compared with the top2α+/+ cells, top2α+/- cells showed partial resistance to the cytotoxic effects of the drug.After the exposure to Dp44mT at 100 nmol/L, the top2α+/+ cells showed 31.7%, while the top2α+/- cells only showed 9.4% sub-G1 containing cells. 

    References: Blood. 2004 Sep 1;104(5):1450-8; Cancer Res. 2009 Feb 1;69(3):948-57.

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    Molecular Weight (MW)285.37 
    CAS No.152095-12-0 
    Storage-20℃ for 3 years in powder form
    -80℃ for 2 years in solvent
    Solubility (In vitro)DMSO: 57 mg/mL (199.7 mM) 
    Water: <1 mg/mL
    Ethanol: 47 mg/mL (164.7 mM) 
    Solubility (In vivo)Propylene glycol: 1 mg/mL 
    SynonymsDp 44mt; 3-(Dipyridin-2-ylmethylideneamino)-1,1-dimethylthiourea

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    In Vitro

    In vitro activity: Dp44mT shows pronounced antiproliferative effects in SK-N-MC, SK-Mel-28, and MCF-7 cells with IC50 of 30 nM, 60 nM, and 60 nM, respectively, while no effects on normal MRC-5 fibroblasts. Dp44mT inhibits cellular Fe uptake from Fe-Tf in SK-N-MC neuroepithelioma and M109 cells, and induces cell apoptosis. In MDA-MB-231 cells, Dp44mT specifically targets topoisomerase topo2α, and thus causes DNA damage. Dp44mT, as a Pgp substrate, also overcomes multidrug resistance by the hijacking of lysosomal P-glycoprotein (Pgp).

    Kinase Assay: oligonucleotides are 5'-end labeled with [32P]ATP and T4 polynucleotide kinase. Labeling mixtures are subsequently centrifuged through Mini Quick Spin DNA columns (for pSK fragments) or Oligo columns (for oligonucleotides) to remove the unincorporated label. Annealing to the complementary strand of the oligonucleotides is done by heating the reaction mixture to 95°C and overnight cooling to room temperature in 10 mM Tris-HCl (pH 7.8), 100 mM NaCl, and 1 mM EDTA. DNA substrates (10 pmol/reaction) are incubated with 500 ng of top2a or top2h in the presence or absence of Dp44mT for the indicated times at 25°C in 10 μL of reaction buffer. Reactions are stopped by adding SDS (final concentration 0.5%). Samples are separated on 16% (for pSK DNA) or 20% (for the oligonucleotides) denaturing polyacrylamide gels (7 M urea). Imaging and quantitation are done using a PhosphorImager.

    Cell Assay: Cells (SK-N-MC, SK-Mel-28, MCF-7 and MRC-5 cells) are incubated in the presence and absence of DFO, 311, 3-aminopyridine-2-carboxaldehyde thiosemicarbazone, doxorubicin, and the DpT series of chelators (0-25 μM) for 72 hours at 37°C。 The effect of the chelators on proliferation is examined using the MTT assay.

    In VivoIn CD2F1 mice bearing M109 tumors, Dp44mT (0.4 mg/kg, i.v.) inhibits tumor growth in a dose-dependent manner. 
    Animal modelCD2F1 mice bearing M109 tumors 
    Formulation & DosageDissolved in Propylene glycol; 0.4 mg/kg, twice daily; i.v. injection 
    ReferencesBlood. 2004 Sep 1;104(5):1450-8; Cancer Res. 2009 Feb 1;69(3):948-57. 

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


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