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Abstract

Cell-penetrating peptides (CPPs) are short transport peptides with a well-established ability for delivery of bioactive cargoes inside the cells both, in vitro and in vivo. CPPs enter unselectively in a wide variety of cell lines, this is a desirable property for most in vitro applications, however, in vivo e.g. in tumor models, specific targeted accumulation is required. In order to achieve tumor targeting, a known CPP, YTA4, was modified by prolonging it C-terminally with mainly negatively charged amino acids. Additionally, a matrix metalloproteinase-2 cleavage site was introduced between the CPP and the inactivating sequence. This new peptide, named NoPe, is an inactive pro-form of YTA4. It can be selectively cleaved and thereby activated by MMPs. We have conjugated an imaging agent, fluoresceinyl carboxylic acid, and a cytostatic agent methotrexate, to this activable pro-form. NoPe activation was demonstrated in vitro by recombinant MMP-2 cleavage and the cleavage of the attenuating sequence was abolished with MMP-2 specific inhibitor. Furthermore, the fluoresceinyl-NoPe is selectively accumulated in the tumor tissue in MDA-MB-231 tumor bearing mice after intravenous injection. Thus, this strategy proves to be successful for in vivo tumor imaging.

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Abbreviations

CPP:

Cell-penetrating peptide

LDH:

Lactate dehydrogenase

MMP:

Matrix metalloproteinase

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Acknowledgments

This work was supported by CePeP AB, Sweden, Cancer Foundation, Sweden, Swedish Research Council VR-NT, Stockholm Centre for Biomembrane Research.

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Correspondence to Ülo Langel.

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Mäe, M., Rautsi, O., Enbäck, J. et al. Tumour Targeting with Rationally Modified Cell-Penetrating Peptides. Int J Pept Res Ther 18, 361–371 (2012). https://doi.org/10.1007/s10989-012-9312-1

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  • DOI: https://doi.org/10.1007/s10989-012-9312-1

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