AbstractThe integration of fluorine atoms into biologically active organic compounds has proved to be a vital technique in small molecule drugs. This technique can substantially enhance crucial properties, including metabolic stability, lipophilicity, and bioavailability, often with a mere addition of a single fluorine atom or a trifluoromethyl group. Over the past few decades, this concept has also been applied in nucleic acid chemistry. A commonly employed 2′‐OH substitution is the introduction of a 2′‐deoxy‐2′‐fluoro (2′‐F) group. The strong electronegativity of fluorine prompts the modified siRNA to readily adopt a C3′‐endo conformation, resulting in significant advantages in terms of binding affinity. To enrich the toolbox of chemical modification of oligonucleotides, the replacement of the 2′‐OH with the 2′‐O‐trifluoromethyl group has been developed in RNA analog synthesis. Oligodeoxynucleotides containing the 2′‐O‐trifluoromethyl group can greatly increase the thermal stability of DNA/RNA duplexes depending on the position and amount of the modification. Moreover, 2′‐O‐trifluoromethylated oligodeoxynucleotide also exhibited a slightly higher resistance to snake venom phosphodiesterase than the unmodified oligodeoxynucleotide. The 2′‐O‐trifluoromethylated oligonucleotides can emerge as a label to study RNA structure and function as well, or to develop DNA/RNA‐based diagnostics. Hence, it is necessary to report an effective method for the synthesis, deprotection, purification, and characterization of oligonucleotides bearing a 2′‐O‐trifluoromethyl group. © 2024 Wiley Periodicals LLC.Basic Protocol 1: Preparation of 6‐N‐benzoyl‐5′‐O‐dimethoxytrityl‐2′‐O‐trifluoromethyl adenosine 3′‐(2‐cyanoethyl N,N‐diisopropyl)phosphoramiditeBasic Protocol 2: Preparation of 4‐N‐acetyl‐5′‐O‐dimethoxytrityl‐2′‐O‐trifluoromethyl cytidine 3′‐(2‐cyanoethyl N,N‐diisopropyl)phosphoramiditeBasic Protocol 3: Preparation of 2‐N‐isobutyryl‐5′‐O‐dimethoxytrityl‐2′‐O‐trifluoromethyl guanine 3′‐(2‐cyanoethyl N,N‐diisopropyl)phosphoramiditeBasic Protocol 4: Preparation of 5′‐O‐dimethoxytrityl‐2′‐O‐2‐trifluoromethyl uridine 3′‐(2‐cyanoethyl N,N‐diisopropyl) phosphoramiditeBasic Protocol 5: Solid‐phase synthesis of 2′‐O‐trifluoromethylated RNA analogsBasic Protocol 6: Deprotection and purification of 2′‐O‐trifluoromethyl‐RNAs

中文翻译：

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2′-O-三氟甲基核苷酸的合成及其在RNA类似物制备中的应用


摘要将氟原子整合到生物活性有机化合物中已被证明是小分子药物的重要技术。该技术通常只需添加一个氟原子或一个三氟甲基即可显着增强关键特性，包括代谢稳定性、亲脂性和生物利用度。在过去的几十年里，这个概念也被应用在核酸化学中。常用的 2'-OH 取代是引入 2'-脱氧-2'-氟 (2'-F) 基团。氟的强电负性促使修饰后的 siRNA 很容易采用 C3′-内切构象，从而在结合亲和力方面具有显着的优势。为了丰富寡核苷酸化学修饰的工具箱，将2′-OH替换为2′-氧‐三氟甲基已在 RNA 类似物合成中得到发展。含有 2'- 的寡脱氧核苷酸氧‐三氟甲基可以极大地提高 DNA/RNA 双链体的热稳定性，具体取决于修饰的位置和数量。此外，2′-氧‐三氟甲基化寡脱氧核苷酸还表现出比未修饰的寡脱氧核苷酸稍高的对蛇毒磷酸二酯酶的抵抗力。 2′-氧‐三氟甲基化寡核苷酸也可以作为标签来研究 RNA 结构和功能，或开发基于 DNA/RNA 的诊断。因此，有必要报道一种有效的方法来合成、去保护、纯化和表征带有2'的寡核苷酸。 ‐O ‐三氟甲基。 © 2024 Wiley 期刊有限责任公司。基本协议1 ： 6‐的制备氮‐苯甲酰基‐5′‐氧‐二甲氧基三苯甲基‐2′‐氧-三氟甲基腺苷 3′-(2-氰乙基氮,氮‐二异丙基)亚磷酰胺基本协议2 : 4-的制备氮‐乙酰基‐5′‐氧‐二甲氧基三苯甲基‐2′‐氧-三氟甲基胞苷 3′-(2-氰乙基氮,氮‐二异丙基)亚磷酰胺基本协议3 : 2-的制备氮‐异丁酰基‐5′‐氧‐二甲氧基三苯甲基‐2′‐氧-三氟甲基鸟嘌呤 3′-(2-氰乙基氮,氮‐二异丙基)亚磷酰胺基本协议4 : 5′-的制备氧‐二甲氧基三苯甲基‐2′‐氧‐2-三氟甲基尿苷 3′-(2-氰乙基氮,氮‐二异丙基）亚磷酰胺基本协议5 : 2′-的固相合成氧‐三氟甲基化RNA类似物基本协议6 ：2′-的脱保护和纯化氧‐三氟甲基‐RNA