高黏液渗透和 ζ 电位变化的自乳化药物传递系统：一种用于囊性纤维化因果治疗的有效基因传递模型。
Highly mucus permeating and zeta potential changing self-emulsifying drug delivery systems: A potent gene delivery model for causal treatment of cystic fibrosis.
机构信息
Thiomatrix Forschungs- und Beratungs GmbH, 6020 Innsbruck, Austria.
Center for Chemistry and Biomedicine, Department of Pharmaceutical Technology, Institute of Pharmacy, University of Innsbruck, 6020 Innsbruck, Austria.
出版信息
Int J Pharm. 2019 Feb 25;557:124-134. doi: 10.1016/j.ijpharm.2018.12.048. Epub 2018 Dec 27.
AIM
It was the aim of the study to develop self-emulsifying drug delivery systems (SEDDS) with the ability to change their zeta potential towards higher values at the adsorption membrane and in this way facilitate the release of the DNA-cetrimonium complex and enhance transfection.
METHODS
Plasmid DNA was complexed via hydrophobic ion pairing utilizing various surfactants and the complex was incorporated into SEDDS achieving a payload of 1% (m/v). Log P of the complex was determined. SEDDS were characterized regarding droplet size, zeta potential, stability and toxicity. Alkaline phosphatase presented in the sputum of cystic fibrosis patients was quantified using 4-nitrophenyl phosphate disodium salt and 5-bromo-4-chloro-1H-indol-3-yl phosphate dipotassium salt as substrates. SEDDS containing 0.4% (m/v) 1,2-dipalmitoyl-sn-glycero-3-phosphate monosodium salt were characterized regarding their zeta potential changing properties utilizing isolated alkaline phosphatase and cystic fibrosis sputum. The mucus permeating properties of SEDDS were evaluated via Transwell method using cystic fibrosis sputum. Finally, the transfection efficiency of incorporated plasmid DNA was investigated.
RESULTS
Cetrimonium bromide showed the highest precipitation efficiency of 99.5 ± 2.72% for the complexation of pDNA. SEDDS containing propylene glycol, Capmul PG-8, Captex 300, Captex 355, Captex 8000, Cremophor EL, Cremophor RH-40 and Brij O10 showed stable emulsions with a droplet size between 20 and 100 nm and zeta potential <-3 mV over 4 h. SEDDS demonstrated highly protective effect against enzymatic degradation and moderate cell viability on freshly obtained pulmonary tissue. The pDNA-cetrimonium complex incorporated into SEDDS revealed a log P of about 2. A concentration of 0.879 ± 0.103 U/g alkaline phosphatase was found in the sputum of cystic fibrosis patients. SEDDS containing 1,2-dipalmitoyl-sn-glycero-3-phosphate monosodium salt showed a high potential of changing the zeta potential by applying isolated alkaline phosphatase as well as cystic fibrosis sputum along with high mucus permeating properties. Formulation C demonstrated the highest transfection efficiency with a 7.2-fold increased fluorescence intensity compared to naked pDNA.
CONCLUSION
The novel developed zeta potential changing SEDDS are opening versatile opportunities for the treatment of cystic fibrosis caused by gene mutation.
目的
本研究旨在开发具有在吸附膜处改变其表面zeta 电位至更高值的能力的自乳化药物传递系统（SEDDS），从而促进 DNA-十六烷基三甲基溴化铵复合物的释放并增强转染。
方法
利用各种表面活性剂通过疏水离子配对将质粒 DNA 进行复合，并将复合物掺入 SEDDS 中，实现载药量为 1%（m/v）。测定复合物的 log P。对 SEDDS 的粒径、zeta 电位、稳定性和毒性进行了表征。利用 4-硝基苯磷酸二钠盐和 5-溴-4-氯-1H-吲哚-3-基磷酸二钾盐作为底物，定量测定囊性纤维化患者痰液中的碱性磷酸酶。使用 1,2-二棕榈酰基-sn-甘油-3-磷酸单钠盐，对载药量为 0.4%（m/v）的 SEDDS 进行表面 zeta 电位变化特性的表征，方法是利用分离的碱性磷酸酶和囊性纤维化痰液。通过使用囊性纤维化痰液的 Transwell 方法，评估 SEDDS 的粘液渗透特性。最后，研究了掺入的质粒 DNA 的转染效率。
结果
十六烷基三甲基溴化铵对 pDNA 的复合物沉淀效率最高，达 99.5±2.72%。含有丙二醇、Capmul PG-8、Captex 300、Captex 355、Captex 8000、Cremophor EL、Cremophor RH-40 和 Brij O10 的 SEDDS 形成稳定的乳液，粒径在 20-100nm 之间，zeta 电位在 4 小时内均<-3mV。SEDDS 对新鲜获得的肺组织中的酶降解具有高度的保护作用，细胞活力适中。掺入 SEDDS 的 pDNA-十六烷基三甲基溴化铵复合物的 log P 约为 2。在囊性纤维化患者的痰液中发现碱性磷酸酶的浓度为 0.879±0.103U/g。含有 1,2-二棕榈酰基-sn-甘油-3-磷酸单钠盐的 SEDDS 具有通过应用分离的碱性磷酸酶以及囊性纤维化痰液改变表面 zeta 电位的高潜力，同时具有高粘液渗透性能。制剂 C 与裸 pDNA 相比，荧光强度增加了 7.2 倍，显示出最高的转染效率。
结论
新型的表面 zeta 电位变化 SEDDS 为治疗因基因突变引起的囊性纤维化提供了多种机会。
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