生物响应可逆交联的多功能透明质酸-聚肽纳米载体用于乳腺癌的靶向治疗

In this project, novel reversibly crosslinked bio-responsive hyaluronic acid-polypeptide nanoparticles are designed and prepared for targeted chemotherapy of breast cancer. The nanoparticles can be readily formed by self-assembly of hyaluronic acid-g-poly(phenylalanine-co-histidine)-lipoic acid (HA-g-P(Phe-co-His)-LA) copolymers in water followed by crosslinking with a catalytic amount of 1,4-dithio-D,L-threitol. The reversibly crosslinked multifunctional nanoparticles have several unique features: (i) they have excellent biocompatibility and biodegradability; (ii) they are very stable under physiological conditions with minimal drug release; (iii) they can be actively targeted to CD44 receptor-overexpressing breast tumor; (iv) they can be efficiently internalized by CD44-positve breast cells via a receptor-mediated endocytosis mechanism; (v) they can efficiently escape from endosomes due to the proton sponge effect and fusogenic activity of histidine units; (vi) they can be rapidly de-crosslinked and destabilized under the cytoplasmic reductive conditions, resulting in fast drug release into the nuclei of CD44-positive breast cancer cells. These multiple-functional nanoparticles may achieve targeted delivery of potent hydrophobic anticancer drugs (doxorubicin, paclitaxel, camptothecin, etc) to breast cancer, resulting in safe and efficient chemotherapy of breast cancer.

本项目将研制生物响应可逆交联的多功能透明质酸-聚肽纳米载体用于乳腺癌的高效靶向化疗。该纳米载体由透明质酸-接枝-聚（苯丙氨酸-co-组氨酸）-硫辛酸（HA-g-P(Phe-co-His)-LA）共聚物在水溶液中自组装，然后用催化量的二硫苏糖醇交联制备得到。该纳米载体具有以下优异性能：（1）良好的生物相容性和生物降解性；（2）在血液循环中因交联很稳定，能有效抑制药物早释；（3）可主动靶向到CD44受体高表达的乳腺癌；（4）能通过CD44介导作用，高效选择性地内吞到乳腺癌细胞；（5）可通过组氨酸单元的质子海绵效应和融膜作用高效逃离内涵体；（6）能在细胞质和细胞核的还原环境下自动解交联，快速释放出药物。该多功能纳米载体可用于疏水化疗药物（阿霉素、紫杉醇、喜树碱等）的乳腺癌靶向释放，并有望最终实现安全高效的乳腺癌治疗。

项目针对当前纳米药物普遍存在血液循环时不稳定、肿瘤富集量不足、到达肿瘤后释放速度较慢或不完全等挑战，创新性地用透明质酸和聚肽构建了一系列生物响应性多功能纳米载体用于小分子抗肿瘤药物、蛋白质药物和核酸类药物的高效包载和靶向递送，实现了对多种皮下和原位肿瘤模型的高效低毒治疗。主要结果如下：（1）设计构建了生物响应可逆交联的透明质酸-聚多肽纳米粒，用于高效包载和靶向递送憎水抗肿瘤药物（阿霉素、紫杉醇、多西他赛等），实现了对人源MCF-7乳腺癌、A549非小细胞肺癌和4T1转移性乳腺癌的主动靶向治疗；（2）设计制备了一系列基于聚酯肽的胶束和囊泡纳米平台，实现了对小分子抗癌药物（多西他赛、阿霉素、单甲基奥瑞他汀E等）、蛋白质药物（细胞色素C、皂草毒素等）和siRNA的肿瘤靶向递送，在皮下B16F10黑色素瘤、HCT-116结直肠癌和原位A549肺癌等多种肿瘤模型中表现出了显著的抑瘤效果，具有较好的临床转化潜力；（3）用光点击化学法构建了多功能透明质酸纳米和微米凝胶，实现了对多种治疗性蛋白质药物（细胞色素C、果粒酶、皂草毒素、Herceptin等）的有效包裹和细胞内/外递送，为肿瘤的安全高效蛋白质疗法构建了新的递送平台；（4）开发了多种基于维他命E-寡聚多肽的具有良好生物相容性和功能性的表面活性剂，并用其构建了表面交联稳定、高效肿瘤靶向和生物响应性的多功能PLGA纳米载体，实现了对乳腺癌、肺癌和耐药肿瘤等模型的高效靶向化疗；（5）设计构建了基于聚酪氨酸的纳米粒子，实现了对蒽醌类疏水抗肿瘤药物的超高效率包载和生物响应性释放，在MD-MBA-231乳腺癌和HCT-116结直肠癌等模型中展现出较好的治疗效果。本项目构建的生物可降解聚多肽、透明质酸衍生物材料和多种纳米载药平台具有多功能、制备简单、创新性强、临床转化潜力大等优点，在生物医用材料和肿瘤治疗方面有重要的科学价值和应用前景。