Nie Hui-Jun, Hu Hao, Qi Xinming, Zhou Yin-Jue, Liu Lu, Chen Xiao-Hua
State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
Center for Drug Safety Evaluation and Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
JACS Au. 2025 Feb 24;5(3):1402-1412. doi: 10.1021/jacsau.5c00012. eCollection 2025 Mar 24.
Oligonucleotide-based therapies, especially ligand-conjugated siRNAs, offer significant therapeutic potential for a wide array of diseases. However, conventional solid-phase synthesis and current postsynthetic in-solution conjugation methods face notable challenges related to efficiency, accessibility, and the scalability of diverse ligand-oligonucleotide conjugates. Herein, we introduce a novel strategy for highly efficient, rapid, and modular assembly of GalNAc-siRNA conjugates based on light-induced primary amine and -nitrobenzyl alcohol cyclization (PANAC) chemistry. Leveraging the advantages of PANAC photoclick chemistry and modular conjugation linkers, our method enables the direct assembly of trivalent GalNAc (tGalNAc) with commercially available primary-amine-modified siRNAs. This approach demonstrates the efficient and rapid assembly of therapeutically relevant oligonucleotides with ligands of interest, offering operational simplicity and practicality; thus, it effectively overcomes the limitations of existing methods. More importantly, the developed siRNA-tGalNAc conjugates showed a robust gene silencing effect superior to the parent siRNA conjugate, highlighting the effectiveness of our method in generating and screening siRNA conjugates to enhance in vivo potency. Overall, our method enables modular and rapid assembly of therapeutically relevant oligonucleotide-tGalNAc conjugates using readily accessible oligonucleotides and commercially available tGalNAc-amine ligands. This approach expands the toolkit for generating ligand-oligonucleotide conjugates, providing a general and efficient platform with broad applicability, thereby advancing the optimization and development of oligonucleotide-based therapeutics.
基于寡核苷酸的疗法，尤其是配体偶联的小干扰RNA（siRNA），对多种疾病具有显著的治疗潜力。然而，传统的固相合成方法以及当前合成后在溶液中的偶联方法，在效率、可及性以及各种配体 - 寡核苷酸偶联物的可扩展性方面面临着显著挑战。在此，我们介绍一种基于光诱导伯胺和对硝基苄醇环化（PANAC）化学的新型策略，用于高效、快速且模块化地组装N - 乙酰半乳糖胺（GalNAc） - siRNA偶联物。利用PANAC光点击化学和模块化偶联接头的优势，我们的方法能够将三价GalNAc（tGalNAc）与市售的伯胺修饰的siRNA直接组装。这种方法展示了将治疗相关的寡核苷酸与感兴趣的配体进行高效且快速的组装，具有操作简单性和实用性；因此，它有效地克服了现有方法的局限性。更重要的是，所开发的siRNA - tGalNAc偶联物显示出比亲本siRNA偶联物更强的基因沉默效果，突出了我们的方法在生成和筛选siRNA偶联物以增强体内效力方面的有效性。总体而言，我们的方法能够使用易于获取的寡核苷酸和市售的tGalNAc - 胺配体，模块化且快速地组装治疗相关的寡核苷酸 - tGalNAc偶联物。这种方法扩展了生成配体 - 寡核苷酸偶联物的工具包，提供了一个具有广泛适用性的通用且高效的平台，从而推动了基于寡核苷酸疗法的优化和发展。
