4′‐ <i>C</i> ‐Aminoethoxy‐Modified DNAs Exhibit Increased Nuclease Resistance, Sustained RNase H Activity, and Inhibition of <i>KRAS</i> Gene Expression
4′‐ <i>C</i> ‐Aminoethoxy‐Modified DNAs Exhibit Increased Nuclease Resistance, Sustained RNase H Activity, and Inhibition of <i>KRAS</i> Gene Expression
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4′-<i>C</i>-氨基乙氧基修饰的 DNA 表现出增强的核酸酶抗性、持续的 RNase H 活性以及对 <i>KRAS</i> 基因表达的抑制
DOI：
10.1002/cbdv.202200125
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发表时间：
2022
期刊：
影响因子：
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通讯作者：
Ueno Yoshihito
中科院分区：
文献类型：
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作者：
Katsuzaki Yuki;Tsukimura Ryo;Chandela Akash;Chano Tokuhiro;Ueno Yoshihito
以脱氧胸苷为起始原料，完成了4 '-C-氨基乙氧基胸苷（AEoT）核苷亚磷酰胺的线性合成。将该类似物掺入几种寡核苷酸中，然后评价其作为反义寡核苷酸（ASO）的适用性。与未修饰的和4′-C-氨乙基胸苷（4′-AET）修饰的异源双链体相比，AEoT修饰的DNA/RNA双链体表现出改善的热稳定性。AEoT修饰的DNA的血清稳定性与未修饰的DNA相比显著增加了数倍。此外，发现修饰的DNA/RNA杂交体的RNA酶H依赖性切割持续存在。此外，修饰的反义寡核苷酸和未修饰的寡核苷酸对人肺癌细胞中的KRAS基因也显示出相对相当的抑制。这项研究加强了我们对4′-C-氨基乙氧基修饰的核苷酸作为阿索治疗剂的潜在应用的理解。
The linear synthesis of 4′‐C‐aminoethoxy thymidine (AEoT) nucleoside phosphoramidite was accomplished using deoxythymidine as the starting material. This analog was incorporated into several oligonucleotides, the applicability of which as antisense oligonucleotides (ASOs) was then evaluated. The AEoT‐modified DNA/RNA duplex exhibited improved thermal stability compared to unmodified and 4′‐C‐aminoethyl thymidine (4′‐AET) modified heteroduplexes. The serum stability of AEoT‐modified DNA was notably increased by several‐folds compared to that of unmodified DNA. Furthermore, RNase H‐dependent cleavage of the modified‐DNA/RNA hybrids was found to be sustained. In addition, the modified antisense and unmodified oligonucleotides also displayed relatively comparable inhibition of theKRASgene in human lung cancer cells. This study strengthens our understanding of the potential application of 4′‐C‐aminoethoxy‐modified nucleotides as ASO therapeutics.