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“Click” Conjugation of Peptide on the Surface of Polymeric Nanoparticles for Targeting Tumor Angiogenesis

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ABSTRACT

Purpose

Angiogenesis plays a critical role in tumor growth. This phenomena is regulated by numerous mediators such as vascular endothelial growth factor (VEGF). CBO-P11, a cyclo-peptide, has proven to specifically bind to receptors of VEGF and may be used as targeting ligand for tumor angiogenesis. We herein report the design of novel nanoparticles conjugated to CBO-P11 in order to specifically target tumor site.

Methods

The conjugation of CBO-P11 on the surface of poly(vinylidene fluoride) (PVDF) nanoparticles was investigated using the copper(I)-catalyzed Huisgen 1,3-dipolar cycloaddition known as “click” reaction. CBO-P11 was modified with a near-infrared cyanine dye bearing an alkyne function, allowing both “click” coupling on azido-modified nanoparticles and fluorescence labelling. Each step of this nanodevice construction was judiciously performed in aqueous solution and successfully characterized. The cytotoxicity of nanoparticles was evaluated in human brain endothelial cell line and their affinity for VEGF receptors was determined via fluorescence-based uptake assays on porcine aortic endothelial cell line.

Results

Nanoparticles were found to be spherical, dense, monodisperse and stable. No cytotoxicity was observed after four days of incubation demonstrating the biocompatibility of nanoparticles. Fluorescence highlighted the specific interaction of these functionalized nanoparticles for VEGF receptors, suggesting that the targeting peptide bioactivity was retained.

Conclusions

These results demonstrate the potential of these functionalized nanoparticles for targeting tumor angiogenesis and their possible use as multifunctional plateform for cancer treament if coupled with therapeutic agents.

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Abbreviations

AA:

acrylic acid

ACN:

acetonitrile

DCM:

dichloromethane

DIEA:

diisopropylethylamine

DLS:

dynamic light scattering

DMEM:

Dulbecco’s modified eagle medium

EDC:

ethyl-3(3dimethylaminopropyl)carbodiimide

EDTA:

ethylenediaminetetraacetic acid

EGM-2:

endothelial growth medium

ESI:

electrospray ionization

FBS:

fetal bovine serum

FESEM:

field emission scanning electron microscope

FTIR:

fourier transform infrared

HCMEC:

human brain endothelial cells

HOBt:

N-hydroxybenzotriazole

HPLC:

high-performance liquid chromatography

HUVEC:

human umbilical vein endothelial cells

IC50 :

half-maximal inhibitory concentration

Ip:

polydispersity index

MALDI-TOF:

matrix-assisted laser desorption/ionization-time of flight

mTEG:

modified tetraethyleneglycol

NIR:

near-infrared

NMP:

N-methylpyrrolidone

NMR:

nuclear magnetic resonance

PAA:

poly(acrylic acid)

PAE:

porcine aortic endothelial cells

PBS:

phosphate-buffered saline

POA:

perfluorooctanoic acid

PVDF:

poly(vinylidene fluoride)

Rg:

radius of gyration

Rh:

hydrodynamic radius

SANS:

small-angle neutron scattering

SLS:

static light scattering

t-BuOH:

tertio-butanol

TFA:

trifluoroactic acid

THF:

tetrahydrofurane

TIS:

triisopropylsilane

UV:

ultra-violet

VEGF:

vascular endothelial growth factor

VEGFR:

receptor of VEGF

VF2:

vinylidene fluoride

WST-1:

4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate

XPS:

X-ray photoelectron spectroscopy

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ACKNOWLEDGMENTS

We are grateful to François Bauer (Piezotech SA) for the PVDF nanoparticles synthesis. We also thank Dr. Christophe Schatz (Laboratoire de Chimie des Polymères Organiques, UMR CNRS 5629, University of Bordeaux, France) for dynamic light scattering experiments and La Ligue Contre le Cancer for its financial support.

Author information

Authors and Affiliations

  1. CNAB, Chimie Bio-Organique, UMR CNRS 5084, Université de Bordeaux 2, 33076, Bordeaux, France

    Stéphanie Deshayes, Victor Maurizot & Gérard Déléris

  2. Laboratoire des Solides Irradiés, UMR CNRS-CEA 7642 Ecole Polytechnique, 28 route de Saclay, 91128, Palaiseau, France

    Stéphanie Deshayes, Marie-Claude Clochard, Cécile Baudin & Thomas Berthelot

  3. Laboratoire de Radiolyse, Service de Chimie Moléculaire, UMR CNRS-CEA 3299, 91191, Gif-Sur-Yvette, France

    Stéphane Esnouf

  4. Laboratoire Léon Brillouin, UMR CNRS-CEA 12, 91191, Gif-Sur-Yvette, France

    Didier Lairez

  5. Laboratoire des Mécanismes Moléculaires de l’Angiogenèse, INSERM U920 Université de Bordeaux 1, 33405, Talence, France

    Michel Moenner

Authors
  1. Stéphanie Deshayes
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  2. Victor Maurizot
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Corresponding author

Correspondence to Marie-Claude Clochard.

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Deshayes, S., Maurizot, V., Clochard, MC. et al. “Click” Conjugation of Peptide on the Surface of Polymeric Nanoparticles for Targeting Tumor Angiogenesis. Pharm Res 28, 1631–1642 (2011). https://doi.org/10.1007/s11095-011-0398-5

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  • DOI: https://doi.org/10.1007/s11095-011-0398-5

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