Limiting the level of tertiary amines on polyamines leads to biocompatible nucleic acid vectors

Margarida Isabel Simão Carlos; Kai Zheng; Natalie Garrett; Natrah Arifin; David G Workman; Ilona Kubajewska; Abdulrahman A Halwani; Julian Moger; Qi Zhang; Andreas G Schätzlein; Ijeoma F Uchegbu

doi:10.1016/j.ijpharm.2017.04.059

Limiting the level of tertiary amines on polyamines leads to biocompatible nucleic acid vectors

Int J Pharm. 2017 Jun 30;526(1-2):106-124. doi: 10.1016/j.ijpharm.2017.04.059. Epub 2017 Apr 25.

Affiliations

¹ School of Pharmacy, University College London, 29-39 Brunswick Square, London, WC1N 1AX, UK.
² School of Pharmacy, University College London, 29-39 Brunswick Square, London, WC1N 1AX, UK; School of Chemical Engineering and Environment, Beijing Institute of Technology, 5 South, Zhongguancun Street, Beijing, 100081, PR China.
³ School of Physics, University of Exeter, Stocker Road, Exeter, EX4 4QL, UK.
⁴ School of Pharmacy, University College London, 29-39 Brunswick Square, London, WC1N 1AX, UK; School of Pharmacy, King Abdulaziz University, Abdullah Sulayman St., Jeddah, 80200, Saudi Arabia.
⁵ School of Chemical Engineering and Environment, Beijing Institute of Technology, 5 South, Zhongguancun Street, Beijing, 100081, PR China.
⁶ School of Pharmacy, University College London, 29-39 Brunswick Square, London, WC1N 1AX, UK; Nanomerics Ltd, Euro House, 1394 High Road, London, N20 9YZ, UK.
⁷ School of Pharmacy, University College London, 29-39 Brunswick Square, London, WC1N 1AX, UK; Nanomerics Ltd, Euro House, 1394 High Road, London, N20 9YZ, UK. Electronic address: ijeoma.uchegbu@ucl.ac.uk.

PMID: 28450169
DOI: 10.1016/j.ijpharm.2017.04.059

Abstract

We have designed an efficient, synthetic nucleic acid vector, which is relatively non-toxic. [N-(2-ethylamino)-6-O-glycolchitosan - EAGC] polymers were 10-50 fold less toxic than Lipofectamine 2000, able to complex DNA, mRNA and siRNA into positively charged (zeta potential=+40 - 50mV), 50-450nm nanoparticles. The level of tertiary amine N-2-ethylamino substitution (DS_tert) was inversely proportional to the IC50 of the EAGC polymers in the A431 cell line: IC50=6.18DS_tert^-0.9, r²=0.9991. EAGC polyplexes were stable against a heparin challenge, able to protect the nucleic acids from nuclease degradation and achieve levels of transfection comparable to Lipofectamine 2000 formulations. The relative biocompatibility of the vector allowed 10 fold higher doses of DNA (1μg compared to 0.1μg per well with Lipofectamine 2000) and siRNA (10.7μg per well vs 1.3μg with Lipofectamine 2000) to be applied to cells, when compared to Lipofectamine 2000. Finally intranasal application of EAGC - siRNA complexes resulted in siRNA transfer to the neurons of the olfactory bulb.

Keywords: Chitosan; DNA; Gene therapy; N-(2-ethylamino)-6-O-glycolchitosan; Nucleic acid delivery; Polymer; mRNA; siRNA.

MeSH terms

Animals
Biocompatible Materials / chemistry*
Cell Line
Cell Survival
DNA
Genetic Vectors / chemistry*
Humans
Lipids
Nucleic Acids / chemistry*
Polyamines / chemistry*
RNA, Messenger
RNA, Small Interfering
Rats, Sprague-Dawley
Transfection

Substances

Biocompatible Materials
Lipids
Lipofectamine
Nucleic Acids
Polyamines
RNA, Messenger
RNA, Small Interfering
DNA