Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, Novosibirsk 630090, Russia.
Department of Natural Sciences, Novosibirsk State University, Novosibirsk 630090, Russia.
Molecules. 2023 Feb 16;28(4):1904. doi: 10.3390/molecules28041904.
One of the key problems in the design of therapeutic and diagnostic oligonucleotides is the attachment of small-molecule ligands for targeted deliveries in such a manner that provides the controlled release of the oligonucleotide at a certain moment. Here, we propose a novel, convenient approach for attaching ligands to the 5'-end of the oligonucleotide via biodegradable, acid-labile phosphoramide linkage. The method includes the activation of the 5'-terminal phosphate of the fully protected, support-bound oligonucleotide, followed by interaction with a ligand bearing the primary amino group. This technique is simple to perform, allows for forcing the reaction to completion by adding excess soluble reactant, eliminates the problem of the limited solubility of reagents, and affords the possibility of using different solvents, including water/organic media. We demonstrated the advantages of this approach by synthesizing and characterizing a wide variety of oligonucleotide 5'-conjugates with different ligands, such as cholesterol, aliphatic oleylamine, and -anisic acid. The developed method suits different types of oligonucleotides (deoxyribo-, 2'-O-methylribo-, ribo-, and others).
在治疗性和诊断性寡核苷酸设计中，一个关键问题是通过小分子配体的连接来实现靶向递送，以便在特定时刻控制寡核苷酸的释放。在这里，我们提出了一种通过可生物降解的酸不稳定磷酰胺键将配体连接到寡核苷酸 5'-端的新方法。该方法包括对完全保护的、连接在载体上的寡核苷酸的 5'-末端磷酸进行活化，然后与带有伯氨基的配体相互作用。该技术操作简单，可以通过添加过量的可溶性反应物来迫使反应完全进行，消除了试剂溶解度有限的问题，并提供了使用不同溶剂的可能性，包括水/有机介质。我们通过合成和表征具有不同配体（胆固醇、脂肪族油胺和 - 茴香酸）的各种寡核苷酸 5'-缀合物，证明了该方法的优势。所开发的方法适用于不同类型的寡核苷酸（脱氧核糖、2'-O-甲基核糖、核糖等）。
