文献：Herceptin-conjugated paclitaxel loaded PCL-PEG worm-like nanocrystal micelles for the combinatorial treatment of HER2-positive breast cancer

文献链接：https://xueshu.baidu.com/usercenter/paper/show?paperid=1p620e70pe0q0840kc4d0a50ux392701

作者：Jiahui Penga, Juan Chena, Fang Xie, Wei Baob, Hongyan Xuc, Hongxia Wangd, Yuhong Xua,

Zixiu Du

相关产品：

PCL5000-PEG2000-CHO 聚己内酯-聚乙二醇-甲醛

PCL2000-MPEG2000聚己内酯-甲氧基聚乙二醇

原文摘要：We have constructed Herceptin-conjugated, paclitaxel (PTX) loaded, PCL-PEG worm-like nanocrystal micelles (PTX@PCL-PEG-Herceptin) for the combinatorial therapy of HER2-positive breast cancer that exploit the specific targeting of Herceptin to HER2-positive breast cancer cells. Firstly, amphiphilic PCL2000-MPEG2000 and PCL5000- PEG2000-CHO were selected as the optimized matrix to wrap PTX that self-assembled into worm-like micelles with internal nanocrystal structures (PTX@PCL-PEG). Then the aldehydes of PCL5000-PEG2000-CHO exposed on the outside surface of PTX@PCL-PEG were utilized to react with the primary amines of Herceptin and formed stable, carbon-nitrogen single linkers (–C–N–) between the antibodies and nanoparticles. This study shows PTX@ PCL-PEG-Herceptin remained relatively stable in the circulation and in the tumor microenvironment, and rapidly targeted and entered into the HER2-overexpressing tumor cells while sparing normal tissues from the toxic effects. PTX@PCL-PEG-Herceptin shrank the tumors and prolonged survival time in a SKBR-3-tumor-xenograft, nude mice model more effectively than TAXOL®, PTX@PCL-PEG, Herceptin+TAXOL® and Herceptin+PTX@ PCL-PEG. Mechanistic studies showed that PTX@PCL-PEG-Herceptin entered into the HER2-positive tumor cells through the caveolin-mediated pathway. The conjugated Herceptin greatly enhanced the binding ability of the

nanoparticle to the targeted SKBR-3 cells. This novel strategy provides a rational and simple antibody-conjugated-nanoparticle platform for the clinical application of combinatorial anticancer treatment.

PCL-MPEG由PCL（聚己内酯）和MPEG（甲氧基聚乙二醇）组成。PCL具有良好的生物降解性和机械性能，而MPEG赋予了材料亲水性和生物相容性。PCL-PEG-CHO将PCL、PEG（聚乙二醇）与CHO（醛基）结合。CHO基团使其具备了可修饰性，方便与其他生物分子进行共价连接。PEG的存在增强了亲水性和生物相容性。选择两亲性的PCL2000-MPEG2000和PCL5000- PEG2000-CHO作为优化的基质，将PTX自组装成具有内部纳米晶体结构的（PTX@PCL-PEG）胶束。然后利用暴露在PTX@PCL-PEG外表面的PCL5000-PEG2000-CHO的醛与赫赛汀的伯胺反应，在抗体和纳米颗粒之间形成稳定的碳氮单连接体（-C-N-）制备PTX@PCL-PEG-赫赛汀。过程如下：

空白胶束的制备，PTX@PCL-PEG和PTX@PCL-槲皮素

PCL5000-PEG2000-CHO和PCL2000-MPEG2000，与PTX共溶解在氯仿中。然后在溶液中加入去离子水，超声形成白色、均匀的乳液。在真空下旋转蒸发去除氯仿，在水溶液中获得胶束。然后过滤，在生理盐水中透析去除未封装的化合物，用生理盐水代替水相，用于后续实验。

PTX@PCL-PEG-赫赛汀是由两个顺序反应制备

PTX@PCL-PEG（ PCL5000-PEG2000-CHO）和与醛的摩尔比5：1，然后4 μL氰基氢化钠（CH3BNNa，）。反应在桌面摇床上持续混合16小时。离心，上清中去除游离抗体。混合物在PBS中溶解，保存或冻干直到进一步使用。

结论：PCL2000-MPEG2000和PCL5000- PEG2000-CHO参与制备的PTX@PCL-PEG-赫赛汀作为一种ACNs，在体内循环和tumor微环境中与不可生物降解-c-n连接剂在纳米微环境中保持相对稳定。结果表明，该传递系统特异性靶向并进入her2阳性的tumor细胞，但在体内实验中不影响正常组织，显示赫赛汀具有靶向性，以及单抗细胞表面抗原直接与蠕虫样纳米晶胶束连接后的不变能力。赫赛汀与PTX@PCL-PEG-赫赛汀的蠕虫晶体形态的协同作用也比TAXOL®和PTX@PCL-PEG增加了注入tumor的化合物数量，并更有效地缩小了tumor。