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Characterization of Polyion Complex Micelles Designed to Address the Challenges of Oligonucleotide Delivery

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Abstract

Purpose

To optimize oligonucleotide (ODN)-based polyion complex micelles (PICMs) by studying the effects of polymer composition and length on their properties.

Methods

Atom transfer radical polymerization was used to synthesize copolymers with increasing hydrophilic nonionic and cationic block lengths. PICMs were prepared by mixing the copolymers and ODN at various nitrogen-to-phosphate (N/P) ratios and characterized by gel electrophoresis and dynamic light scattering. The stability of the complexes towards dissociation was tested using a competitive assay with heparin. Finally, protection of the incorporated ODN against DNAse I degradation was evaluated.

Results

A library of copolymers composed of poly(ethylene glycol) (PEG) and poly(aminoethyl methacrylate) (PAEMA) and/or poly((dimethylamino)ethylmethacrylate) (PDMAEMA) was synthesized. All polymers efficiently interacted with the ODN at N/P ratios approaching 1.5. Narrowly distributed but easily dissociable PICMs were obtained using PEG 5000 and short DMAEMA chains. Shortening the PEG block to 2000, increasing the number of cationic units and using AEMA produced more stable complexes but at the cost of colloidal properties. All polymers were able to protect the ODN from nuclease degradation.

Conclusions

PEG 3000-based PICMs possess good colloidal properties, intermediate stability towards dissociation and adjustable buffering capacity, making them potentially useful for the delivery of nucleic acid drugs.

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Abbreviations

AEMA:

2-aminoethyl methacrylate

AEMABoc:

2-(N-(tert-butoxycarbonyl)amino)ethyl methacrylate

ATRP:

atom transfer radical polymerization

β :

buffering capacity

DLS:

dynamic light scattering

DMAEMA:

2-(N,N-dimethylamino)ethyl methacrylate

EDTA:

ethylenediaminetetraacetic acid

EO:

ethylene oxide

EtBr:

ethidium bromide

GPC:

gel permeation chromatography

1H NMR:

proton nuclear magnetic resonance

M n :

number-average molecular weight

M w :

weight-average molecular weight

MW:

molecular weight

N/P:

nitrogen-to-phosphate

ODN:

oligodeoxyribonucleotide

PEG:

poly(ethylene glycol)

PEI:

poly(ethyleneimine)

PI:

polydispersity index

PICMs:

polyion complex micelles

siRNA:

small interfering RNA

SD:

standard deviation

THF:

tetrahydrofuran

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Acknowledgments

This work was financially supported by the Natural Sciences and Engineering Research Council of Canada (NSERC) and the Canada Research Chair Program. M.H.D. further acknowledges graduate research scholarships from NSERC and Fonds Québécois de la Recherche sur la Nature et les Technologies.

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  1. Faculty of Pharmacy, University of Montreal, P.O. Box 6128, Downtown Station, Montreal, Quebec, H3C 3J7, Canada

    Marie-Hélène Dufresne, Mahmoud Elsabahy & Jean-Christophe Leroux

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  1. Marie-Hélène Dufresne
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  2. Mahmoud Elsabahy
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  3. Jean-Christophe Leroux
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Correspondence to Jean-Christophe Leroux.

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Dufresne, MH., Elsabahy, M. & Leroux, JC. Characterization of Polyion Complex Micelles Designed to Address the Challenges of Oligonucleotide Delivery. Pharm Res 25, 2083–2093 (2008). https://doi.org/10.1007/s11095-008-9591-6

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