Pharmaceutical Research

, Volume 30, Issue 11, pp 2832–2842 | Cite as

Mitochondrial Delivery of Doxorubicin by Triphenylphosphonium-Functionalized Hyperbranched Nanocarriers Results in Rapid and Severe Cytotoxicity

  • Theodossis A. Theodossiou
  • Zili Sideratou
  • Maria E. Katsarou
  • Dimitris Tsiourvas
Research Paper

ABSTRACT

Purpose

To develop a novel hyperbranched polymer-based nanocarrier for efficient drug delivery to cell mitochondria. Also to study for the first time the cytotoxic effect of doxorubicin via mitochondria-specific delivery system.

Methods

We introduced alkyltriphenylphosphonium groups (TPP) to a poly(ethylene imine) hyperbranched polymer (PEI). We harnessed the hydrophobic assembly of these alkylTPP functionalized PEI molecules into ~100 nm diameter nanoparticles (PEI-TPP) and further encapsulated the chemotherapy agent doxorubicin (DOX), to produce the mitotropic nanoparticles PEI-TPP-DOX.

Results

By administering PEI-TPP-DOX to human prostate carcinoma cells DU145, we found that: (i) PEI-TPP-DOX specifically localized at cell mitochondria as revealed by the inherent DOX fluorescence; (ii) in contrast to the slow apoptotic cell death incurred by DOX over the period of days at micromolar concentrations, PEI-TPP-DOX triggered rapid and severe cytotoxicity within few hours of incubation and at submicromolar incubation concentrations. This cytotoxicity was mainly found to be of a necrotic nature, not precluding autophagy related death pathways to a smaller extent.

Conclusions

We have elaborated a versatile mitotropic nanocarrier; furthermore, using this platform, we have developed a mitochondrial-doxorubicin formulation with exceptional cytocidal properties, even in nanomolar concentrations.

KEY WORDS

cytotoxicity doxorubicin hyperbranched poly(ethylene imine) mitochondrial drug delivery triphenylphosphonium 

ABBREVIATIONS

ANT

Adenine nucleotide translocator

ANTI-A

Antimycin A

ATR

Atractyloside potassium salt

CASP8

Z-IETD-FMK

CASP9

Z-LEHD-FMK

CCCP

Carbonyl cyanide 3-chlorophenylhydrazone

CSA

Cyclosporine A

DIPEA

N,N-diisopropylethylamine

DLS

Dynamic light scattering

DOX

Doxorubicin

EB

Ethidium bromide

EPR

Enhanced permeability and retention

ETC

Electron transport chain

FBS

Fetal bovine serum

HBTU

2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium

HOBt

N-hydroxybenzotriazole

LDH

Lactate dehydrogenase

L-NAME

-nitro-L-arginine methyl ester hydrochloride

MnTmPyP

Mn(III)tetrakis(1-methyl-4-pyridyl)porphyrin

MPTP

Mitochondrial permeability transition pore

MTT

Thiazolyl blue tetrazolium bromide

MYXO

Myxothiazol

NADH

β-Nicotinamide adenine dinucleotide reduced disodium salt

OLIGO

Oligomycin

PBS

Phosphate buffer saline

PEI

Hyperbranched poly(ethylene imine)

PNNAG

p-nitrophenyl-N-acetyl-β-d-glucosaminide

ROS

Reactive oxygen species

ROT

Rotenone

RUR

Ruthenium red

STS

Staurosporine

TB

Trypan blue

TPP

Triphenylphosphonium cation

TTFA

Thenoyltrifluoroacetone

UA

Uric acid

WORT

Wortmannin

Supplementary material

11095_2013_1111_MOESM1_ESM.doc (1.9 mb)
ESM 1(DOC 1914 kb)

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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Theodossis A. Theodossiou
    • 1
  • Zili Sideratou
    • 1
  • Maria E. Katsarou
    • 1
  • Dimitris Tsiourvas
    • 1
  1. 1.Department of Physical Chemistry, IAMPPNMNCSR “Demokritos”Ag. ParaskeviGreece

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