Title

Diblock Copolymers with Tunable PH Transitions for Gene Delivery

Abstract

A series of diblock copolymers containing an endosomal-releasing segment composed of diethylaminoethyl methacrylate (DEAEMA) and butyl methacrylate (BMA) were synthesized via reversible addition-fragmentation chain transfer (RAFT) polymerization. The materials were designed to condense plasmid DNA (pDNA) through electrostatic interactions with a cationic poly(N,N-dimethylaminoethyl methacrylate) (DMAEMA) first block. The pDMAEMA was employed as a macro chain transfer agent (macroCTA) for the synthesis of a series in which the relative feed ratios of DEAEMA and BMA were systematically varied from 20% to 70% BMA. The resultant diblock copolymers exhibited low polydispersity (PDI ≤ 1.06) with similar molecular weights (M = 19.3-23.1kDa). Dynamic light scattering (DLS) measurements in combination with 1H NMR D 2O studies demonstrated that the free copolymers assemble into core-shell micelles at physiological pH. Reduction of the solution pH to values representative of endosomal/lysosomal compartments induced an increase in the net cationic charge of the core through protonation of the DEAEMA residues. This protonation promotes micelle destabilization and exposure of the hydrophobic BMA residues that destabilize biological membranes. The pH value at which this micelle-to-unimer transition occurred was dependent on the hydrophobic content of the copolymer, with higher BMA-containing copolymer compositions exhibiting pH-induced transitions to the membrane-destabilizing state at successively lower pH values. The ability of the diblock copolymers to deliver pDNA was subsequently investigated using a GFP expression vector in two monocyte cell lines. High levels of DNA transfection were observed for the copolymer compositions exhibiting the sharpest pH transitions and membrane destabilizing activities, demonstrating the importance of tuning the endosomal-releasing segment composition.

Department(s)

Materials Science and Engineering

Comments

The authors gratefully acknowledge the National Institutes of Health ( NIBIB EB2991 , NIAID AI074661 ) and Center for the Intracellular Delivery of Biologics which is supported by the Washington State Life Science Discovery Fund (grant number 2496490 ) for funding.

Keywords and Phrases

Cell culture; Copolymer; DNA; Drug delivery; Gene therapy; Micelle

International Standard Serial Number (ISSN)

0142-9612

Document Type

Article - Journal

Document Version

Citation

File Type

text

Language(s)

English

Rights

© 2012 Elsevier, All rights reserved.

PubMed ID

22169826

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