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A BODIPY-embedding miltefosine analog linked to cell-penetrating Tat(48-60) peptide favors intracellular delivery and visualization of the antiparasitic drug

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Abstract

Therapeutic application of many drugs is often hampered by poor or denied access to intracellular targets. A case in point is miltefosine (MT), an orally active antiparasitic drug, which becomes ineffective when parasites develop dysfunctional uptake systems. We report here the synthesis of a fluorescent BODIPY-embedding MT analogue with appropriate thiol functionalization allowing linkage to the cell-penetrating Tat(48-60) peptide through disulfide or thioether linkages. The resulting constructs are efficiently internalized into the otherwise MT-invulnerable R40 Leishmania strain, resulting in fast parasite killing, and hence successful avoidance of the resistance. In the disulfide-linked conjugate, an additional fluoro tag on the Tat moiety allows to monitor its reductive cleavage within the cytoplasm. Terminally differentiated cells such as peritoneal macrophages, impervious to MT unless infected by Leishmania, can uptake the drug in its Tat-conjugated form. The results afford proof-of-principle for using CPP vectors to avert drug resistance in parasites, and/or for tackling leishmaniasis by modulating macrophage uptake.

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Abbreviations

Boc:

Tert-Butyloxycarbonyl

BDP:

BODIPY: 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene

CPP:

Cell-penetrating peptide

DIEA:

Diisopropylethylamine

DIPCI:

Diisopropylcarbodiimide

Fmoc:

9-Fluorenylmethyloxycarbonyl

HBTU:

2-(1H-Benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate

Mmt:

4-Methoxytrityl

MT:

Miltefosine: hexadecylphosphocholine

Npys:

3-Nitro-2-pyridylsulfenyl

TFA:

Trifluoroacetic acid

TIS:

Triisopropylsilane

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Acknowledgments

Research supported by funds from the European Union (HEALTH-2007-223414 to L.R. and D.A.),VI PN de I+D+I 2008-2011, (PI09-01928, PI12-02706) and ISCIII-Subdirección General de Redes y Centros de Investigación Cooperativa (RICET RD 06/0021/0006, RD12/0018/0007 to L.R.) MICINN (CTQ2010-16457 to A.U.A, BIO2008-04487-CO3 to D.A.), MINECO (SAF2011-24899 to D.A.) and Generalitat de Catalunya (SGR2009-00492 to D.A.).

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Department of Experimental and Health Sciences, Pompeu Fabra University, Barcelona Biomedical Research Park, 08003, Barcelona, Spain

    Beatriz G. de la Torre & David Andreu

  2. Instituto de Química Orgánica General, CSIC, Juan de la Cierva 3, 28006, Madrid, Spain

    Valentín Hornillos & Francisco Amat-Guerri

  3. Instituto de Química Física “Rocasolano”, CSIC, Serrano 119, 28006, Madrid, Spain

    Valentín Hornillos & A. Ulises Acuña

  4. Centro de Investigaciones Biológicas, CSIC, Ramiro de Maeztu 9, 28040, Madrid, Spain

    Juan R. Luque-Ortega, M. A. Abengózar & Luis Rivas

Authors
  1. Beatriz G. de la Torre
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  2. Valentín Hornillos
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  3. Juan R. Luque-Ortega
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  4. M. A. Abengózar
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  5. Francisco Amat-Guerri
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  6. A. Ulises Acuña
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  7. Luis Rivas
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  8. David Andreu
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Corresponding authors

Correspondence to Luis Rivas or David Andreu.

Additional information

Dedicated to the memory of Prof. Francisco Amat-Guerri, who passed away during the preparation of this article.

B. G. de la Torre and V. Hornillos contributed equally to this work.

Electronic supplementary material

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726_2013_1661_MOESM1_ESM.docx

Supplementary material 1 (DOCX 449 kb) Synthesis of pyrroles 2 and 3, carbon numbering of MT analogue 1 and precursors; 1H- and 13C-NMR spectra of 1; HPLC and mass spectra of conjugates 7 and 8

Supplementary material 2 (DOCX 1746 kb)

Supplementary material 3 (DOCX 1273 kb)

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de la Torre, B.G., Hornillos, V., Luque-Ortega, J.R. et al. A BODIPY-embedding miltefosine analog linked to cell-penetrating Tat(48-60) peptide favors intracellular delivery and visualization of the antiparasitic drug. Amino Acids 46, 1047–1058 (2014). https://doi.org/10.1007/s00726-013-1661-3

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  • DOI: https://doi.org/10.1007/s00726-013-1661-3

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