Synthesis of 2'-O-[2-(N-Methylcarbamoyl)ethyl]ribonucl eosides Using Oxa-Michael reaction and chemical and biological properties of oligonucleotide derivatives incorporating these modified ribonucleosides.
Synthesis of 2'-O-[2-(N-Methylcarbamoyl)ethyl]ribonucl eosides Using Oxa-Michael reaction and chemical and biological properties of oligonucleotide derivatives incorporating these modified ribonucleosides.
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使用 Oxa-Michael 反应合成 2-O-[2-(N-甲基氨基甲酰基)乙基]核糖核苷以及掺入这些修饰核糖核苷的寡核苷酸衍生物的化学和生物学特性。
DOI：
10.1021/jo101963z
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Sekine M
中科院分区：
文献类型：
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作者：
Yamada T;Okaniwa N;Saneyoshi H;Ohkubo A;Seio K;Nagata T;Aoki Y;Takeda S;Sekine M
为了开发新的2′-O-修饰的核苷类寡核苷酸作为核酸药物，我们合成了三种2′-羟基修饰的核苷衍生物，（甲氧羰基）乙基（MOCE），2-（N-甲基氨基甲酰基）乙基（MCE），和2-（N，N-二甲基氨基甲酰基）乙基（DMCE）基团作为关键中间体，通过适当保护的核糖核苷的oxa-Michael反应（U，C，A，G）衍生物。其中，2 '-O-MCE核糖核苷在碱性条件下最稳定。为了研究2′-O-修饰对含2′-O-修饰的核糖核苷的寡核苷酸的核酸酶抗性及其与互补RNA和DNA链的杂交亲和力的影响，成功合成了2′-O-MCE-核糖核苷亚磷酰胺衍生物，并进行了2′-O-MCE-寡核苷酸和2′-O-甲基-含2′-O-MCE-核糖核苷的寡核苷酸的合成。由此获得的2′-O-MCE-寡核苷酸和具有2′-O-MCE和2′-O-甲基基团的嵌合寡核苷酸显示出互补的RNA链和比相应的2′-O-甲基化种类高得多的核酸酶抗性。最后，我们将2′-O-MCE-核糖核苷掺入反义2′-O-甲基-寡核糖核苷酸中，以检测其在与小鼠dystrophin前体mRNA相关的剪接反应中的外显子跳跃活性。外显子跳跃试验表明，这些2′-O-甲基寡核苷酸的2′-O-甲基化修饰比相应的2′-O-甲基化寡核苷酸硫代磷酸酯衍生物具有更好的效果。
To develop oligonucleotides containing new 2′-O-modified ribonucleosides as nucleic acid drugs, we synthesized three types of ribonucleoside derivatives modified at the 2′-hydroxyl group with 2-(methoxycarbonyl)ethyl (MOCE), 2-(N-methylcarbamoyl)ethyl (MCE), and 2-(N,N-dimethylcarbamoyl)ethyl (DMCE) groups, as key intermediates, via the oxa-Michael reaction of the appropriately protected ribonucleoside (U, C, A, and G) derivatives. Among them, the 2′-O-MCE ribonucleosides were found to be the most stable under basic conditions. To study the effects of the 2′-O-modification on the nuclease resistance of oligonucleotides incorporating the 2′-O-modified ribonucleosides and their hybridization affinities for the complementary RNA and DNA strands, 2′-O-MCE-ribonucleoside phosphoramidite derivatives were successfully synthesized and subjected to the synthesis of 2′-O-MCE-oligonucleotides and 2′-O-methyl-oligonucleotides incorporating 2′-O-MCE-ribonucleosides. The 2′-O-MCE-oligonucleotides and chimeric oligomers with 2′-O-MCE and 2′-O-methyl groups thus obtained demonstrated complementary RNA strands and much higher nuclease resistances than the corresponding 2′-O-methylated species. Finally, we incorporated the 2′-O-MCE-ribonucleosides into antisense 2′-O-methyl-oligoribonucleotides to examine their exon-skipping activities in splicing reactions related to pre-mRNA of mouse dystrophin. The exon-skipping assay of these 2′-O-methyl-oligonucleotide incorporating 2′-O-MCE-uridines showed better efficacies than the corresponding 2′-O-methylated oligoribonucleotide phosphorothioate derivatives.