文献：Functionalized docetaxel-loaded lipid-basednanosuspensions to enhance antitumor efficacy in vivo.

文献链接：https://www.semanticscholar.org/paper/Functionalized-docetaxel-loaded-to-enhance-efficacy-Pang-Wang/10b9a1a8fcfd45c3e0b04fc201d4f3afe4dd033e

作者：Xiuping Pang ,Tianqi Wang ,Dandan Jiang ,Weiwei Mu ,Bo Zhang ,Na Zhang

相关产品：DSPE-PEG2000-NHS  磷脂-聚乙二醇2000-活性酯

原文摘要：Purpose: To further enhance the antitumor efficacy through targeted delivery, DTX loaded lipid-based-nanosuspensions (DTX-LNS) were prepared and functionalized by PEGylation or NGR modification to develop DSPE-PEG2000 modified DTX-LNS (P-DTX-LNS) or DSPE-PEG2000-NGR modified DTX-LNS (N-DTX-LNS), respectively.

Methods: Based on our previous work, functionalized DTX-LNS including P-DTX-LNS and N-DTX-LNS were prepared using thin-film hydration, and then characterized. Release behavior, stability in vitro, cytotoxicity and cellular uptake of functionalized LNS were observed. To demonstrate tumor targeting efficiency of functionalized DTX-LNS, in vivo real-time and ex vivo imaging study were conducted. Furthermore, therapeutic efficacy in vivo was evaluated in an H22-bearing mice model.

Results: Functionalized DTX-LNS 100–110 nm in diameter were successfully prepared and exhibited good stability under various conditions. In vitro release studies demonstrated that DTX was released from functionalized DTX-LNS steadily and reached approximately 95% at 48 hrs. Functionalized DTX-LNS showed dose-dependent cytotoxicity and time-dependent internalization in human hepatocellular liver carcinoma cells (HepG2) cells. In vivo real-time and ex vivo imaging results indicated that tumor targeting efficiencies of P-DiR-LNS and NDiR-LNS were 29.9% and 34.3%, respectively. Moreover, evaluations of in vivo antitumor efficacy indicated that functionalized DTX-LNS effectively inhibited tumor growth with low toxicity.

Conclusion: The functionalized LNS exhibited suitable particle size, nearly spherical structure, enough drug loading and great potentials for large-scale production. The results in vitro and in vivo demonstrated that functionalized LNS could realize tumor targeting and antitumor efficacy. Consequently, functionalized DTX-LNS could be expected to be used for tumor targeting therapy.

DSPE-PEG2000-NHS可作为有用的自组装试剂，用于制备聚乙二醇化脂质体或胶束。在脂质双分子层靶向化合物递送中，该试剂能够提升化合物的溶解度，并借助PEG提供的隐形性能，延长循环半衰期，同时减少非特异性蛋白质结合或细胞粘附。其NHS基团能够与含胺分子发生共轭反应，实现生物分子的修饰或连接。DSPE-PEG2000-NGR结合了DSPE磷脂、PEG链以及NGR靶向肽。NGR是一种能够与tumor新生Blood vessels内皮细胞上的CD13受体结合的靶向肽，因此该分子具有tumor靶向性。作为化合物递送载体，DSPE-PEG2000-NGR能够实现化合物的靶向递送，提高化合物在tumor部位的浓度，同时减少全身有害性。DSPE-PEG2000-NHS在许多方面有应用，例如在DSPE-PEG2000-NGR合成过程中，其具体制备过程如下：

图为：DSPE-PEG2000-NGR的合成

DSPE-PEG2000-NHS在DSPE-PEG2000-NGR合成中的应用：

通过DSPEPEG2000-NHS和NGR进行酰胺化反应，合成了活性靶向材料DSPE-PEG2000-NGR。将溶解在蒸馏水中的过量NGR加入到二甲基甲酰胺（DMF）中的DSPE-PEG2000-NHS中。冗余的NGR通过透析去除。产物经自由冷冻后得到，通过核磁共振（1 H-NMR）确定其结构。

DSPEPEG2000-NHS在DSPE-PEG2000- NGR合成及表征中的应用：

为了制备N-DTX-LNS，通过酰胺化反应合成了活性靶向材料DSPE-PEG2000-NGR。DSPEPEG2000-NHS、NGR和DSPE-PEG2000-NGR的1 H NMR谱如图。在DSPE-PEG2000-NGR中出现了信号，这归因于NGR中靠近硫原子的亚甲基。结果表明，NGR与DSPE-PEG2000-NHS成功连接，活性靶向材料DSPE-PEG2000- NGR被用于实验。

图为：DSPE-PEG2000-NGR的表征

结论：DSPE-PEG2000-NHS 中的 NHS（N - 羟基琥珀酰亚胺）基团是一种高活性的酯类基团，它能够与含有伯胺基（-NH₂）的分子发生酰胺化反应。而在合成 DSPE-PEG2000-NGR 的过程中，NGR（Asn-Gly-Arg，天冬酰胺 - 甘氨酸 - 精氨酸短肽序列）中存在的伯胺基可以与 DSPE-PEG2000-NHS 的 NHS 基团发生反应。所以，DSPE-PEG2000-NHS 为 NGR 与 DSPE-PEG2000 的连接提供了必要的活性反应位点，使得两者能够顺利结合，这是合成 DSPE-PEG2000-NGR 的基础。DSPE-PEG2000-NHS 充当了连接 DSPE-PEG2000 和 NGR 的桥梁。DSPE-PEG2000 本身具有良好的生物相容性和两亲性，可以在生物体系中稳定存在并发挥作用。NGR 是一种具有靶向功能的短肽，能够特异性地识别并结合某些tumor细胞表面过度表达的受体。通过 DSPE-PEG2000-NHS 的连接作用，将 DSPE-PEG2000 和 NGR 结合在一起，形成具有特定功能的 DSPE-PEG2000-NGR 分子。这样的分子既具备了 DSPE-PEG2000 的稳定性和生物相容性，又拥有了 NGR 的靶向功能，为后续的生物应用提供了基础。