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

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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.

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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

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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.

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Authors and Affiliations

  1. Univ Paris-Sud, Institut Galien Paris-Sud, LabEx LERMIT, Faculté de Pharmacie, 5 rue J.B. Clément, 92296, Châtenay-Malabry, France

    Chantal Al Sabbagh, Nicolas Tsapis, Patricia Calleja-Gonzalez, Hélène Chacun, Stéphanie Denis, Juliette Vergnaud, Claire Gueutin & Elias Fattal

  2. CNRS UMR 8612, Institut Galien Paris-Sud, LabEx LERMIT, 5 rue J.B. Clément, 92296, Châtenay-Malabry, France

    Chantal Al Sabbagh, Nicolas Tsapis, Patricia Calleja-Gonzalez, Hélène Chacun, Stéphanie Denis, Juliette Vergnaud, Claire Gueutin & Elias Fattal

  3. Université François-Rabelais de Tours, Inserm, Imagerie et Cerveau UMR U 930, Tours, France

    Anthony Novell, Jean-Michel Escoffre & Ayache Bouakaz

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  1. Chantal Al Sabbagh
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Correspondence to Elias Fattal.

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Al Sabbagh, C., Tsapis, N., Novell, A. et al. Formulation and Pharmacokinetics of Thermosensitive Stealth® Liposomes Encapsulating 5-Fluorouracil. Pharm Res 32, 1585–1603 (2015). https://doi.org/10.1007/s11095-014-1559-0

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