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.
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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