Oncotarget

Research Papers:

Potentiating the cellular targeting and anti-tumor activity of Dp44mT via binding to human serum albumin: two saturable mechanisms of Dp44mT uptake by cells

Angelica M. Merlot, Sumit Sahni, Darius J.R. Lane, Ashleigh M. Fordham, Namfon Pantarat, David E. Hibbs, Vera Richardson, Munikumar R. Doddareddy, Jennifer A. Ong, Michael L.H. Huang, Des R. Richardson and Danuta S. Kalinowski _

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Oncotarget. 2015; 6:10374-10398. https://doi.org/10.18632/oncotarget.3606

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Abstract

Angelica M. Merlot1, Sumit Sahni1, Darius J.R. Lane1, Ashleigh M. Fordham1, Namfon Pantarat1, David E. Hibbs2, Vera Richardson1, Munikumar R. Doddareddy2, Jennifer A. Ong2, Michael L.H. Huang1, Des R. Richardson1,* and Danuta S. Kalinowski1,*

1 Molecular Pharmacology and Pathology Program, Department of Pathology and Bosch Institute, The University of Sydney, Sydney, NSW, Australia

2 Faculty of Pharmacy, The University of Sydney, Sydney, NSW, Australia

* These authors are contributed equally as senior authors

Correspondence to:

Des R. Richardson, email:

Danuta S. Kalinowski, email:

Keywords: Albumin, Dp44mT, Anti-tumor targeting, Human serum albumin

Received: January 26, 2015 Accepted: February 14, 2015 Published: March 15, 2015

Abstract

Di-2-pyridylketone 4,4-dimethyl-3-thiosemicarbazone (Dp44mT) demonstrates potent anti-cancer activity. We previously demonstrated that 14C-Dp44mT enters and targets cells through a carrier/receptor-mediated uptake process. Despite structural similarity, 2-benzoylpyridine 4-ethyl-3-thiosemicarbazone (Bp4eT) and pyridoxal isonicotinoyl hydrazone (PIH) enter cells via passive diffusion. Considering albumin alters the uptake of many drugs, we examined the effect of human serum albumin (HSA) on the cellular uptake of Dp44mT, Bp4eT and PIH. Chelator-HSA binding studies demonstrated the following order of relative affinity: Bp4eT≈PIH>Dp44mT. Interestingly, HSA decreased Bp4eT and PIH uptake, potentially due to its high affinity for the ligands. In contrast, HSA markedly stimulated Dp44mT uptake by cells, with two saturable uptake mechanisms identified. The first mechanism saturated at 5-10 µM (Bmax:1.20±0.04 × 107 molecules/cell; Kd:33±3 µM) and was consistent with a previously identified Dp44mT receptor/carrier. The second mechanism was of lower affinity, but higher capacity (Bmax:2.90±0.12 × 107 molecules/cell; Kd:65±6 µM), becoming saturated at 100 µM and was only evident in the presence of HSA. This second saturable Dp44mT uptake process was inhibited by excess HSA and had characteristics suggesting it was mediated by a specific binding site. Significantly, the HSA-mediated increase in the targeting of Dp44mT to cancer cells potentiated apoptosis and could be important for enhancing efficacy.


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