2′-O-[2-[(N,N-dimethylamino)oxy]ethyl]-modified oligonucleotides inhibit expression of mRNA in vitro and in vivo
2′-O-[2-[(N,N-dimethylamino)oxy]ethyl]-modified oligonucleotides inhibit expression of mRNA in vitro and in vivo
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DOI：
10.1093/nar/gkh220
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发表时间：
2004-01-01
影响因子：
14.9
通讯作者：
Manoharan, M
中科院分区：
文献类型：
--
作者：
Prakash, TP;Johnston, JF;Manoharan, M
本文报道了2 ′-O-[2-[（N，N-二甲氨基）氧基]乙基]（2 ′-O-DMAOE）修饰的寡核苷酸的合成及其反义活性。2 '-O-DMAOE修饰的寡核苷酸在小鼠中显示出上级的代谢稳定性。在末端具有2 '-O-DMAOE-修饰的核苷残基和在中心区域具有2'-脱氧核苷残基的硫代磷酸寡核苷酸“gapmer”在细胞培养物中对两个靶点的mRNA表达显示出剂量依赖性抑制。“Gapmer”寡核苷酸具有一个或两个2 '-O-修饰区和允许RNA酶H消化靶mRNA的2'-脱氧寡核苷酸硫代磷酸酯区。为了确定体内效力和功效，用2 '-O-DMAOE缺口聚物处理BalbC小鼠，并观察到靶向C-raf mRNA表达的剂量依赖性降低。在整个序列中具有2 '-O-DMAOE修饰的寡核苷酸在HUVEC细胞中非常有效地降低了细胞间粘附分子-1（ICAM-1）蛋白的表达，IC 50为1.8 nM。这些均匀修饰的2 ′-O-DMAOE寡核苷酸对ICAM-1蛋白表达的抑制可能是由于选择性干扰翻译起始复合物的形成。这些结果表明，当采用RNA酶H依赖性或RNA酶H非依赖性靶减少机制时，2 ′-O-DMAOE修饰的寡核苷酸可用于基于反义的治疗。
Synthesis and antisense activity of oligonucleotides modified with 2'-O-[2-[(N,N-dimethylamino)oxy] ethyl] (2'-O-DMAOE) are described. The 2'-O-DMAOE-modified oligonucleotides showed superior metabolic stability in mice. The phosphorothioate oligonucleotide 'gapmers', with 2'-O-DMAOE- modified nucleoside residues at the ends and 2'-deoxy nucleosides residues in the central region, showed dose-dependent inhibition of mRNA expression in cell culture for two targets. 'Gapmer' oligonucleotides have one or two 2'-O-modified regions and a 2'-deoxyoligonucleotide phosphorothioate region that allows RNase H digestion of target mRNA. To determine the in vivo potency and efficacy, BalbC mice were treated with 2'-O-DMAOE gapmers and a dose-dependent reduction in the targeted C-raf mRNA expression was observed. Oligonucleotides with 2'-O-DMAOE modifications throughout the sequences reduced the intercellular adhesion molecule-1 (ICAM-1) protein expression very efficiently in HUVEC cells with an IC50 of 1.8 nM. The inhibition of ICAM-1 protein expression by these uniformly modified 2'-O-DMAOE oligonucleotides may be due to selective interference with the formation of the translational initiation complex. These results demonstrate that 2'-O-DMAOE- modified oligonucleotides are useful for antisense-based therapeutics when either RNase H-dependent or RNase H-independent target reduction mechanisms are employed.