Liu Xun, Zhao Ziyin, Wu Fan, Chen Yongbing, Yin Lichen
Department of Thoracic Surgery, The Second Affiliated Hospital of Soochow University, Suzhou, 215004, China.
Institute of Functional Nano and Soft Materials (FUNSOM), Jiangsu Key Laboratory of Carbon-Based Functional Materials and Devices, Soochow University, Suzhou, 215123, China.
Adv Mater. 2022 Feb;34(8):e2108116. doi: 10.1002/adma.202108116. Epub 2022 Jan 17.
Cytosolic protein delivery is a prerequisite for protein-based biotechnologies and therapeutics on intracellular targets. Polymers that can complex with proteins to form nano-assemblies represent one of the most important categories of materials, because of the ease of nano-fabrication, high protein loading efficiency, no need for purification, and maintenance of protein bioactivity. Stable protein encapsulation and efficient intracellular liberation are two critical yet opposite processes toward cytosolic delivery, and polymers that can resolve these two conflicting challenges are still lacking. Herein, hyperbranched poly(β-amino ester) (HPAE) with backbone-embedded phenylboronic acid (PBA) is developed to synchronize these two processes, wherein PBA enhanced protein encapsulation via nitrogen-boronate (N-B) coordination while triggered polymer degradation and protein release upon oxidation by H O in cancer cells. Upon optimization of the branching degree, charge density, and PBA distribution, the best-performing A2-B3-C2-S -P is identified, which mediates robust delivery of various native proteins/peptides with distinct molecular weights (1.6-430 kDa) and isoelectric points (4.1-10.3) into cancer cells, including enzymes, toxins, antibodies, and CRISPR-Cas9 ribonucleoproteins (RNPs). Moreover, A2-B3-C2-S -P mediates effective cytosolic delivery of saporin both in vitro and in vivo to provoke remarkable anti-tumor efficacy. Such a potent and universal platform holds transformative potentials for protein pharmaceuticals.
胞质蛋白递送是基于蛋白质的生物技术和细胞内靶点治疗的先决条件。能够与蛋白质复合形成纳米组装体的聚合物是最重要的材料类别之一，因为其易于进行纳米制造、蛋白质负载效率高、无需纯化且能维持蛋白质生物活性。稳定的蛋白质封装和高效的细胞内释放是实现胞质递送的两个关键但相反的过程，而能够解决这两个相互矛盾挑战的聚合物仍然缺乏。在此，我们开发了一种主链嵌入苯硼酸（PBA）的超支化聚（β-氨基酯）（HPAE）来同步这两个过程，其中PBA通过氮-硼酸酯（N-B）配位增强蛋白质封装，同时在癌细胞中被H₂O₂氧化时引发聚合物降解和蛋白质释放。通过优化支化度、电荷密度和PBA分布，确定了性能最佳的A2-B3-C2-S₁-P，它能将各种具有不同分子量（1.6 - 430 kDa）和等电点（4.1 - 10.3）的天然蛋白质/肽，包括酶、毒素、抗体和CRISPR-Cas9核糖核蛋白（RNPs），强力递送至癌细胞。此外，A2-B3-C2-S₁-P在体外和体内均介导了皂草素的有效胞质递送，从而引发显著的抗肿瘤疗效。这样一个强大且通用的平台对蛋白质药物具有变革性潜力。
