Roy & Diana Vagelos Laboratories, Department of Chemistry, University of Pennsylvania, Philadelphia, Pennsylvania 19104-6323, United States.
Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.
J Am Chem Soc. 2023 Aug 30;145(34):18760-18766. doi: 10.1021/jacs.3c07337. Epub 2023 Aug 22.
Delivery of nucleic acids with viral and synthetic vectors has pioneered genetic nanomedicine. Four-component lipid nanoparticles (LNPs) consisting of ionizable lipids, phospholipids, cholesterol, and PEG-conjugated lipids, assembled by microfluidic or T-tube, are the benchmark synthetic vector for delivery of mRNA. One-component multifunctional sequence-defined ionizable amphiphilic Janus dendrimer (IAJD) delivery systems for mRNA were developed by us to complement LNPs. IAJDs consist of multifunctional hydrophilic low-generation dendrons or minidendrons conjugated to hydrophobic dendrons. They were inspired by amphiphilic Janus dendrimers and glycodendrimers. IAJDs coassemble with mRNA into predictable-size vesicles, named dendrimersome nanoparticles (DNPs), by simple injection in acetate buffer, rather than by the complex technology required by LNPs. Assembly of DNPs by simple injection together with sequence design in the hydrophilic and hydrophobic modules of IAJDs endowed rapid screening to access discovery. Molecular design principles for targeted delivery were elaborated when the branching points of IAJDs were constructed from symmetrically and nonsymmetrically substituted plant phenolic acids interconnected by pentaerythritol (PE). Here, we report the first library containing simplified IAJDs constructed in only three steps from symmetrically trialkylated PE in the hydrophobic domain and four different piperazine-based ionizable amines in the hydrophilic part. Rapid coassembly with mRNA and screening led to the discovery of the two most active IAJDs targeting the spleen, liver, and lymph nodes, one predominantly to the spleen and liver and six delivering equally to the spleen, liver, lung, and lymph nodes. These IAJDs represent the simplest synthetic vectors and the first viral or synthetic system delivering equally to multiple organs.
病毒和合成载体递送核酸开创了遗传纳米医学。由可离子化脂质、磷脂、胆固醇和 PEG 连接脂质组成的四组分脂质纳米颗粒（LNPs），通过微流控或 T 型管组装，是递送 mRNA 的基准合成载体。我们开发了一种由多功能序列定义的可离子化两亲性 Janus 树枝状大分子（IAJD）组成的单一组分多功能信使 RNA 递药系统来补充 LNPs。IAJD 由多功能亲水性低代树枝状大分子或亚树枝状大分子与疏水性树枝状大分子偶联而成。它们的灵感来自于两亲性 Janus 树枝状大分子和糖树枝状大分子。IAJD 通过简单地在醋酸盐缓冲液中注射，与 mRNA 共组装成可预测大小的囊泡，称为树枝状大分子纳米颗粒（DNPs），而不是像 LNPs 那样需要复杂的技术。通过简单注射组装和 IAJD 亲水和疏水模块中的序列设计赋予了快速筛选以实现发现。当 IAJD 的分支点由通过季戊四醇（PE）互连的对称和非对称取代的植物酚酸构建时，详细阐述了用于靶向递送的分子设计原则。在这里，我们报告了第一个包含简化的 IAJD 的文库，这些 IAJD 仅由疏水部分中对称三烷基化的 PE 和亲水部分中四个不同的哌嗪基可离子化胺在三个步骤中构建而成。与 mRNA 的快速共组装和筛选导致发现了两种最活跃的靶向脾脏、肝脏和淋巴结的 IAJD，一种主要靶向脾脏和肝脏，六种则均匀地靶向脾脏、肝脏、肺和淋巴结。这些 IAJD 代表了最简单的合成载体和第一个能够均匀地靶向多个器官的病毒或合成系统。
