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4'-C-Methoxy-2'-deoxy-2'-fluoro 修改后的核酸改善了代谢稳定性,并产生了有效的RNAi介导的基因沉默

Elise Malek-Adamian¹, Dale C Guenther², Shigeo Matsuda²

¹Department of Chemistry, McGill University , 801 Sherbrooke Street West, Montreal, Quebec H3A 0B8, Canada.

Journal of the American Chemical Society

|September 23, 2017

概括

新型4'- C- methoxy修饰的尿素类似物增强了siRNA核酶的抵抗力和基因沉默功效. 在治疗RNA干扰应用中,C4'α表具有前景.

科学领域:

医学化学
核酸化学
有关RNA干扰

背景情况:

治疗小干扰RNA (siRNAs) 面临核酶降解和低于最佳功效的挑战.
为了提高siRNA的稳定性和有效性,正在研究尿素核酸的糖分.

研究的目的:

设计和合成新的4阶段改性2阶段-脱氧-2阶段-uridine (2阶段-FU) 类似物.
评估4个阶段C- methoxy (4阶段OMe) epimers (α和β) 对siRNAs核酶抵抗,热稳定性和RNA干扰 (RNAi) 活性的影响.

主要方法:

4个阶段OMe表体的立体选择和非立体选择合成.
H 核磁共振分析和计算建模,以研究糖和形状效应.
热稳定性测试和核酶抗性研究.
修改RNA结构的X射线晶体学
在含有改性单体的siRNA中评估RNAi介导的基因沉默.

主要成果:

合成的4 étaire-C4 étaireα表是立体选择性的;这两种表均可通过非立体选择性的途径获得.
H 核磁共振和计算研究表明,4 个阶段的OMe 组会诱导α- epimer 中的东北糖偏差.
该α-epimer表现出与未经修改的核酸相比的热稳定性,而β-epimer则导致显著的不稳定性.
由于替代剂与酸盐组的接近,这两种4个阶段的OMe表体都表现出核酶耐药性的增加.
包含C4 étaireα- epimer单体的siRNAs有效调解RNAi,其效率与2 étaire- FU修改的siRNAs相似.

结论:

4 étaire-C-methoxy修饰,特别是C4 étaireα表,增强了尿素类似物中的核酶耐药性.
它具有良好的热稳定性,并具有强大的RNAi活性,使其成为治疗性siRNA开发的有希望的候选者.
结构和构造洞察力解释了核酶抵抗的观察到的改善.

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