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In Vitro and In Vivo Efficacy of Self-Assembling RGD Peptide Amphiphiles for Targeted Delivery of Paclitaxel

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Abstract

Purpose

The objective of this work was to compare the efficacy of self-assembling cyclic and linear RGD peptide amphiphiles as carriers for delivering paclitaxel to αvβ3 integrin overexpressing tumors.

Methods

Linear (C18-ADA5-RGD) and cyclic (C18-ADA5-cRGDfK) peptide amphiphiles were synthesized and characterized for CMC, aggregation number and micelle stability using fluorescence spectroscopy methods. Size and morphology of micelles was studied using TEM. Fluorescence polarization and confocal microscopy assays were established to compare binding and internalization of micelles. The targeting efficacy was studied in A2058 cells using cytotoxicity assay as well as in vivo in melanoma xenograft mouse model.

Results

The linear and cyclic RGD amphiphiles exhibited CMC of 25 and 8 μM, respectively, formed nano-sized spherical micelles and showed competitive binding to αvβ3 integrin protein. FITC-loaded RGD micelles rapidly internalized into A2058 melanoma cells. Paclitaxel-loaded RGD micelles exhibited higher cytotoxicity compared with free drug in A2058 cells in vitro as well as in vivo.

Conclusion

Cyclic RGD micelles exhibited better targeting efficacy but were less effective compared to linear RGD micelles as drug delivery vehicle due to lower drug solubilization capacity and lesser kinetic stability. Results from the study proved the effectiveness of self-assembling low molecular weight RGD amphiphiles as carriers for targeted delivery of paclitaxel.

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Abbreviations

C16 (in amphiphile structures):

Palmitic acid

C18 (in amphiphile structures):

Stearic acid

ADA:

8-amino-3,6-dioxaoctanoic acid

HOBT:

Hydroxybenzotriazole

HATU:

2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium hexafluorophosphate

PyBOP:

Benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate

DIPEA:

N, N′-Diisopropylethylamine

DIC:

N, N′-Diisopropylcarbodiimide

DCM:

Dichloromethane

DMF:

N,N′-dimethyl formamide

TFA:

Trifluoroacetic acid

TIS:

Triisopropylsilane

MALDI-TOF:

Matrix assisted laser desorption/ ionization- time of flight

CMC:

Critical micellization concentration

Nagg :

Aggregation number

TEM:

Transmission electron microscopy

κ:

Molar solubilization capacity

FRET:

Forster resonance energy transfer

DII:

1,1′-dioctadecyl-3,3,3,3′, Tetramethylindocarbocyanine perchlorate

DIO:

3,3′-dioctadecyloxacarbocyanine perchlorate

FP:

Fluorescence polarization

mP:

Milli polarization units

FITC:

Fluorescein isothiocynate

IACUC:

Institutional animal care and use committee

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ACKNOWLEDGMENTS AND DISCLOSURES

Authors would like to thank Dr. Mamoun Alhamadsheh (Department of Pharmaceutics and Medicinal Chemistry) for his inputs on fluorescence polarization assay.

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Authors and Affiliations

  1. Department of Pharmaceutics and Medicinal Chemistry, Thomas J. Long School of Pharmacy and Health Sciences, University of the Pacific, Stockton, USA

    Poonam Saraf, Xiaoling Li & Bhaskara Jasti

  2. Department of Biology, College of the Pacific, University of the Pacific, Stockton, USA

    Lisa Wrischnik

Authors
  1. Poonam Saraf
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  2. Xiaoling Li
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  3. Lisa Wrischnik
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  4. Bhaskara Jasti
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Correspondence to Bhaskara Jasti.

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Saraf, P., Li, X., Wrischnik, L. et al. In Vitro and In Vivo Efficacy of Self-Assembling RGD Peptide Amphiphiles for Targeted Delivery of Paclitaxel. Pharm Res 32, 3087–3101 (2015). https://doi.org/10.1007/s11095-015-1689-z

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  • DOI: https://doi.org/10.1007/s11095-015-1689-z

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