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Pharmaceutical Research

, Volume 32, Issue 5, pp 1585–1603 | Cite as

Formulation and Pharmacokinetics of Thermosensitive Stealth® Liposomes Encapsulating 5-Fluorouracil

  • Chantal Al Sabbagh
  • Nicolas Tsapis
  • Anthony Novell
  • Patricia Calleja-Gonzalez
  • Jean-Michel Escoffre
  • Ayache Bouakaz
  • Hélène Chacun
  • Stéphanie Denis
  • Juliette Vergnaud
  • Claire Gueutin
  • Elias Fattal
Research Paper

ABSTRACT

Purpose

We optimize the encapsulation and investigate the pharmacokinetics of 5-Fluorouracil (5-FU) delivered by thermosensitive stealth® liposomes (TSLs) designed to trigger drug release upon hyperthermia using focused ultrasound (FUS).

Methods

5-FU was encapsulated into liposomes made of 1,2-Dipalmitoyl-sn-glycero-3-phosphocholine/cholesterol/1,2-Distearoyl-sn-glycero-3-phosphoethanolamine-N-PEG2000 either as a free molecule or complexed with copper-polyethylenimine. Heat-triggered drug release was evaluated using either a water bath or FUS. Formulation cytotoxicity was assessed on HT-29 cell line by MTS assay. Pharmacokinetics and biodistribution of 5-FU were evaluated in HT-29-tumor bearing mice.

Results

5-FU was easily encapsulated using the lipid hydration method (encapsulation efficacy of 13%) but poorly retained upon dilution. 5-FU complexation with copper-polyethylenimine improved 5-FU retention into liposomes and allowed to obtain an encapsulation efficacy of 37%. At 42°C, heat-triggered 5-FU release from TSLs was 63% using a water bath and 68% using FUS, within 10 min, whereas it remained below 20% for the non-thermosensitive formulation. The MTS assay revealed that formulation toxicity arose from 5-FU and not from the excipients. In addition, 5-FU complex encapsulation into TSLs induces a reduction of the IC50 from 115 down to 49 μM. Pharmacokinetics reveals a longer circulation of encapsulated 5-FU and a more important body exposure, although tumor passive targeting is not significantly higher than free 5-FU.

Conclusions

Complexation of 5-FU with copper-polyethylenimine appears an interesting strategy to improve 5-FU retention into TSLs in vitro and in vivo. TSLs allow heat-triggered release of the drug within 10 min at 42°C, a reasonable time for future in vivo experiments.

KEY WORDS

5-Fluorouracil copper-polyethylenimine complex focused ultrasound pharmacokinetics thermosensitive stealth® liposomes 

ABBREVIATIONS

[2-14C]-5-FU

14C-radiolabeled 5-Fluorouracil

5-FU

5-Fluorouracil

5-FU-Cu-PEI

5-Fluorouracil-copper-polyethylenimine complex

CHOL

Cholesterol

Cu

Copper (II) acetate monohydrate

Cu-PEI

Copper-polyethylenimine complex

dH

Hydrodynamic diameter

DLS

Dynamic light scattering

DPPC

1,2-Dipalmitoyl-sn-glycero-3-phosphocholine

DSPE-PEG2000

1,2-Distearoyl-sn-glycero-3-phosphoethanolamine-N-PEG2000

EDTA

Ethylenediaminetetraacetic acid

EPR

Enhanced permeation and retention

FBS

Fœtal bovine serum

FDA

Food and drug administration

FUS

Focused ultrasound

g

Gravitational acceleration

HPLC

High performance liquid chromatography

HT-29

Human colorectal adenocarcinoma cells

IC50

Half maximal inhibitory concentration

LTSLs

Low temperature sensitive liposomes

MTS

CellTiter 96® AQueous One Solution Cell Proliferation Assay

NTSLs

Non thermosensitive liposomes

PBS

Phosphate buffer saline

PDI

Polydispersity index

PEG2000

Polyethylen glycol 2 KDa

PEI

Polyethylenimine, ethylendiamine branched

psi

Pound per square inch

Tm

Phase transition temperature

TSLs

Thermosensitive liposomes

TTSLs

Traditional thermosensitive liposomes

UV/VIS

Ultraviolet/visible light domains

ζ-potential

Zeta potential

Notes

ACKNOWLEDGMENTS AND DISCLOSURE

This research work was funded by Agence Nationale de la Recherche (ANR-2010-TecSan-007). Institut Galien Paris-Sud is a member of the Laboratory of Excellence LERMIT supported by a grant from ANR (ANR-10-LABX-33). Authors would like to thank A. Maksimenko for help with the tumor model and H. Bénech for help with PK analysis.

Supplementary material

11095_2014_1559_MOESM1_ESM.docx (195 kb)
ESM 1 (DOCX 194 kb)

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Chantal Al Sabbagh
    • 1
    • 2
  • Nicolas Tsapis
    • 1
    • 2
  • Anthony Novell
    • 3
  • Patricia Calleja-Gonzalez
    • 1
    • 2
  • Jean-Michel Escoffre
    • 3
  • Ayache Bouakaz
    • 3
  • Hélène Chacun
    • 1
    • 2
  • Stéphanie Denis
    • 1
    • 2
  • Juliette Vergnaud
    • 1
    • 2
  • Claire Gueutin
    • 1
    • 2
  • Elias Fattal
    • 1
    • 2
  1. 1.Univ Paris-Sud, Institut Galien Paris-Sud, LabEx LERMIT, Faculté de PharmacieChâtenay-MalabryFrance
  2. 2.CNRS UMR 8612, Institut Galien Paris-Sud, LabEx LERMITChâtenay-MalabryFrance
  3. 3.Université François-Rabelais de Tours, Inserm, Imagerie et Cerveau UMR U 930ToursFrance

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