Supporting information

Multivalency: key feature in overcoming drug resistance with a cleavable cell-penetrating peptide-doxorubicin conjugate

Marco Lelle1,2,3, Christoph Freidel1, Stefka Kaloyanova1, Klaus Müllen1 and Kalina Peneva1,2*

1 Max Planck Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany

2 Institute of Organic and Macromolecular Chemistry, Jena Center of Soft Matter, Friedrich Schiller University Jena, Lessingstr. 8, 07743 Jena, Germany

3 Institute of Physiology II, University Hospital Jena, Kollegiengasse 9, 07743 Jena, Germany

*To whom correspondence should be addressed. Email:

Synthesized compounds:

1H NMR spectrum of compound 2 recorded in DMSO-d6 at 300 MHz

13C NMR spectrum of compound 2 recorded in DMSO-d6 at 75 MHz

1H NMR spectrum of compound 4 recorded in DMSO-d6 at 300 MHz

13C NMR spectrum of compound 4 recorded in DMSO-d6 at 75 MHz

1H NMR spectrum of compound 5 recorded in DMSO-d6 at 300 MHz

13C NMR spectrum of compound 5 recorded in DMSO-d6 at 75 MHz

1H NMR spectrum of compound 7 recorded in D2O at 700 MHz

1H NMR spectrum of compound 8 recorded in D2O at 700 MHz

1H NMR spectrum of compound 10 recorded in DMSO-d6 at 300 MHz

13C NMR spectrum of compound 10 recorded in DMSO-d6 at 75 MHz

1H NMR spectrum of compound 11 recorded in DMSO-d6 at 300 MHz

13C NMR spectrum of compound 11 recorded in DMSO-d6 at 75 MHz

1H NMR spectrum of compound 12 recorded in DMSO-d6 at 850 MHz

1H NMR spectrum of compound 13 recorded in D2O at 850 MHz

Intracellular trafficking of the peptide-drug conjugate 13 in Kelly-WT cells by fluorescence confocal laser scanning microscopy, 72 h after the incubation with 10 µM of the drug at 37°C. Fluorescence related to the fluorophore part of the anthracycline is illustrated by red color (scale bar= 15 µm).

Intracellular trafficking of the peptide-drug conjugate 13in Kelly-ADR cells by fluorescence confocal laser scanning microscopy, 72 h after the incubation with 10 µM of the drug at 37°C. Fluorescence related to the fluorophore part of the anthracycline is illustrated by red color (scale bar= 15 µm).

Investigation of the nuclear localizationof the peptide-drug conjugate 13 in Kelly-WT cells by fluorescence confocal laser scanning microscopy, 72 h after the incubation with 10 µM of the drug at 37°C.Drug-associated fluorescence is illustrated by red color, whereas the nuclear stain DRAQ5, which is applied 5 minutes prior to microscopy, is depicted in blue.Colocalizationof the anthracyclineand the nuclear stain is visualized by violet color (scale bar= 15 µm).

Investigation of the nuclear localizationof the peptide-drug conjugate 13in Kelly-ADR cells by fluorescence confocal laser scanning microscopy, 72 h after the incubation with 10 µM of the drug at 37°C.Drug-associated fluorescence is illustrated by red color, whereas the nuclear stain DRAQ5,which is applied 5 minutes prior to microscopy, is depicted in blue.Colocalizationof the anthracyclineand the nuclear stain is visualized by violet color (scale bar= 15 µm).

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