Biomedical Polymers Laboratory, and Jiangsu Key Laboratory of Advanced Functional Polymer Design and Application, College of Chemistry, Chemical Engineering and Materials Science, Soochow University , Suzhou, 215123, People's Republic of China.
Laboratoire des Polymères, Institut des Matériaux and Institut des Sciences et Ingénierie Chimiques, École Polytechnique Fédérale de Lausanne (EPFL) , Bâtiment MXD, Station 12, CH-1015 Lausanne, Switzerland.
Biomacromolecules. 2018 Feb 12;19(2):555-562. doi: 10.1021/acs.biomac.7b01664. Epub 2018 Jan 12.
The clinical translation of protein drugs that act intracellularly is limited by the absence of safe and efficient intracellular protein delivery vehicles. Here, pH-sensitive coiled-coil peptide-cross-linked hyaluronic acid nanogels (HA-cNGs) were designed and investigated for targeted intracellular protein delivery to CD44 overexpressing MCF-7 breast cancer cells. HA-cNGs were obtained with a small size of 176 nm from an equivalent mixture of hyaluronic acid conjugates with GY(EIAALEK)GC (E3) and GY(KIAALKE)GC (K3) peptides, respectively, at pH 7.4 by nanoprecipitation. Circular dichroism (CD) proved the formation of coiled-coil structures between E3 and K3 peptides at pH 7.4 while fast uncoiling at pH 5.0. HA-cNGs showed facile loading of cytochrome C (CC) and greatly accelerated CC release under mild acidic conditions (18.4%, 76.8%, and 91.4% protein release in 24 h at pH 7.4, 6.0, and 5.0, respectively). Confocal microscopy and flow cytometry displayed efficient internalization of CC-loaded HA-cNGs and effective endosomal escape of CC in MCF-7 cancer cells. Remarkably, HA-cNGs loaded with saporin, a ribosome inactivating protein, exhibited significantly enhanced apoptotic activity to MCF-7 cells with a low IC of 12.2 nM. These coiled-coil peptide-cross-linked hyaluronic acid nanogels have appeared as a simple and multifunctional platform for efficient intracellular protein delivery.
细胞内作用的蛋白质药物的临床转化受到缺乏安全有效的细胞内蛋白质递送载体的限制。在这里，设计并研究了 pH 敏感的卷曲螺旋肽交联透明质酸纳米凝胶（HA-cNGs），用于靶向过表达 CD44 的 MCF-7 乳腺癌细胞的细胞内蛋白质递药。HA-cNGs 由透明质酸缀合物与 GY(EIAALEK)GC(E3)和 GY(KIAALKE)GC(K3)肽的等摩尔混合物在 pH 7.4 下通过纳米沉淀获得，粒径为 176nm。圆二色性 (CD) 证明了 E3 和 K3 肽在 pH 7.4 时形成卷曲螺旋结构，而在 pH 5.0 时快速解旋。HA-cNGs 易于负载细胞色素 C (CC)，并在温和酸性条件下（pH 7.4、6.0 和 5.0 时 24 小时分别释放 18.4%、76.8%和 91.4%的蛋白）大大加速 CC 的释放。共聚焦显微镜和流式细胞术显示负载 CC 的 HA-cNGs 的有效内化和 CC 在 MCF-7 癌细胞中的有效内体逃逸。值得注意的是，负载核糖体失活蛋白 saporin 的 HA-cNGs 对 MCF-7 细胞表现出显著增强的凋亡活性，IC50 低至 12.2 nM。这些卷曲螺旋肽交联透明质酸纳米凝胶已成为高效细胞内蛋白质递药的简单多功能平台。
