Multiple triphenylphosphonium cations shuttle a hydrophilic peptide into mitochondria

Mol Pharm. 2009 Jul-Aug;6(4):1138-44. doi: 10.1021/mp900032r.

Abstract

A variety of diseases are related to mitochondrial dysfunction. Hence, the ability to transport drugs to mitochondria that are otherwise cell impermeable would be of great therapeutic potential. Triphenylphosphonium (TPP) cations have been shown to accumulate in mitochondria when attached to small molecules. Here we report on the consequence of increasing the number of TPP moieties that are covalently linked to a model hydrophilic peptide Hemagglutinin A (HA). By extending the HA peptide with l-lysine amino acids to which the TPP's are covalently linked through the epsilon-amine, we have systematically synthesized the HA peptide with 0-3 TPP's. All peptides were subsequently labeled with FITC at the N-terminus. Cellular uptake and mitochondrial localization of the HA-TPP conjugates in HeLa cells were profoundly augmented with increasing number of TPPs, suggesting that this approach is applicable for the delivery of peptides. Furthermore, confocal microscopy demonstrated that the peptides localize to mitochondria. Importantly, all peptide conjugates did not show apparent toxicity at concentrations that are several orders of magnitude higher than those used for HA peptide delivery.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cations / chemical synthesis
  • Cations / chemistry*
  • Cations / pharmacology
  • Flow Cytometry
  • Fluorescein-5-isothiocyanate
  • HeLa Cells
  • Hemagglutinins / metabolism*
  • Humans
  • Mitochondria / metabolism*
  • Organophosphorus Compounds / chemical synthesis
  • Organophosphorus Compounds / chemistry*
  • Organophosphorus Compounds / pharmacology
  • Peptide Fragments / metabolism*
  • Trityl Compounds / chemical synthesis
  • Trityl Compounds / chemistry*
  • Trityl Compounds / pharmacology

Substances

  • Cations
  • Hemagglutinins
  • Organophosphorus Compounds
  • Peptide Fragments
  • Triphenylmethyl Compounds
  • Fluorescein-5-isothiocyanate