文献：Which one performs better for targeted lung cancer combination therapy: pre- or post bombesin-decorated nanostructured lipid carriers?

文献链接：https://pubmed.ncbi.nlm.nih.gov/26455787/

作者：Jiahui Du and Ling Li

相关产品：DOTAP

原文摘要：

Purpose: The co-delivery of gene and drugs has the potential to treat cancer. The aim of this study was to compare post-bombesin decorated nanostructured lipid carriers (NLC) carrying both doxorubicin (DOX) and DNA with pre-bombesin decorated NLC for lung cancer therapy. Methods: Post-bombesin decorated NLC were prepared by two steps. First, DOX and DNA-loaded NLC (DOX-DNA-NLC) was prepared. Second, Bombesin-NH2 (BN-NH2) was added into DOX-DNA-NLC to react with stearic acid-polyethylene glycol-COOH (SA-PEG-COOH) loaded in NLC. Pre-bombesin decorated NLC were prepared by two steps. First, Bombesin (BN)-conjugated ligands were synthesized. Second, DOX and DNA were loaded into BN decorated NLC. Their average size, zeta potential, drug and gene loading were evaluated. NCl H460 human non-small lung cancer cells (NCl-H460 cells) were used for the testing of in vitro transfection efficiency and in vitro cytotoxicity. In vivo transfection efficiency and anti-tumor effect of NLC were evaluated on mice bearing NCl-H460 cells model.

Results: Post-bombesin decorated NLC has a particle size of 128 nm, DOX encapsulation efficiency (EE) of 85% and DNA EE of 91%. Pre-bombesin decorated NLC has a particle size of 101 nm, DOX EE of 86% and DNA EE of 92%. Post-bombesin decorated NLC displayed more stable and remarkably higher transfection efficiency and better anti-tumor ability than pre-bombesin decorated NLC both in vitro and in vivo.

Conclusion: Post-bombesin decorated NLC could function as better carriers to improve the cell targeting and nuclear targeting ability. The resulting nanomedicine could be a promising active targeting drug/gene therapeutic system for lung cancer therapy.

DOTAP被用于体内外核酸和蛋白质的输送。DOTAP是一种季铵盐，具有带正电荷的两亲性类脂化合物的特性。它是一种阳离子脂质，可用于瞬时或稳定转染DNA（如质粒、bacmids）和修饰核酸（如反义寡核苷酸），并且可以不使用辅助脂质。DOTAP在阳离子脂质体研究中相当常见，是制备阳离子脂质体常用的正电荷脂质之一。

图为：Pre-BN-DOX-DNA-NLC的制备

DOTAP在DOX的阳离子NLC（DOX-NLC）的制备中的用途：

DOX是通过与两种等量的TEA在二甲亚砜（DMSO）中过夜搅拌获得的。DOX-NLC采用溶剂扩散法制备。简单地说，通过将GMS、油酸、大豆卵磷脂和PEG-SA混合制备脂质分散体。将DOX溶解于乙醇中，加入到脂质分散体中，得到有机相。同时，将含有Tween 80和DOTAP的水溶液加热，然后立即将有机相加入到水溶液中搅拌。将分散体在室温下冷却并搅拌，直到有机溶剂完全蒸发形成NLC。球团涡旋并在蒸馏水中重新悬浮，洗涤几次，通过膜过滤，用氢氧化钠调节pH值。

图为：Free DOX 的体外DOX释放谱

结论：基于脂质的方案由不同的阳离子脂质和辅助脂质组成，如本研究中使用的DOTAP。阳离子纳米颗粒可以克服这种物理障碍，因为它们会穿透tumor组织。此外，阳离子纳米颗粒通过纳米颗粒与质膜之间的静电相互作用来诱导抗cancer作用。阳离子纳米颗粒利用物理静电应力刺激细胞膜，而不管生物阻力机制如何。所有测试的配方都带有正电荷，允许载体通过静电引力吸附在带负电荷的细胞膜上。