限制多胺上叔胺的含量可产生生物相容性核酸载体。

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

作者信息

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

机构信息

School of Pharmacy, University College London, 29-39 Brunswick Square, London, WC1N 1AX, UK.

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.

出版信息

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

DOI:10.1016/j.ijpharm.2017.04.059

PMID:28450169

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) was inversely proportional to the IC50 of the EAGC polymers in the A431 cell line: IC50=6.18DS, r=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.

摘要

我们设计了一种高效的合成核酸载体，其毒性相对较低。[N-(2-乙氨基)-6-O-乙二醇壳聚糖-EAGC]聚合物的毒性比Lipofectamine 2000低10至50倍，能够将DNA、mRNA和siRNA复合形成带正电荷（ζ电位=+40 - 50mV）、粒径为50 - 450nm的纳米颗粒。在A431细胞系中，叔胺N-2-乙氨基取代水平（DS）与EAGC聚合物的IC50成反比：IC50 = 6.18DS，r = 0.9991。EAGC多聚体对肝素攻击具有稳定性，能够保护核酸不被核酸酶降解，并且转染水平与Lipofectamine 2000制剂相当。与Lipofectamine 2000相比，该载体相对的生物相容性使得应用于细胞的DNA剂量（每孔1μg，而Lipofectamine 2000为每孔0.1μg）和siRNA剂量（每孔10.7μg，而Lipofectamine 2000为1.3μg）能够高出10倍。最后，鼻内应用EAGC - siRNA复合物导致siRNA转移至嗅球神经元。