Hepatocyte-Specific Delivery of siRNAs Conjugated to Novel Non-nucleosidic Trivalent N-Acetylgalactosamine Elicits Robust Gene Silencing in Vivo
Hepatocyte-Specific Delivery of siRNAs Conjugated to Novel Non-nucleosidic Trivalent N-Acetylgalactosamine Elicits Robust Gene Silencing in Vivo
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DOI：
10.1002/cbic.201500023
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发表时间：
2015-04-13
期刊：
影响因子：
3.2
通讯作者：
Manoharan, Muthiah
中科院分区：
文献类型：
--
作者：
Rajeev, Kallanthottathil G.;Nair, Jayaprakash K.;Manoharan, Muthiah
我们最近证明，由于脱唾液酸糖蛋白受体（ASGPR）介导的摄取，与三触角N-乙酰半乳糖胺（GalNAc）缀合的siRNA可在肝脏中诱导强大的RNAi介导的基因沉默。开发了基于非核苷接头的新型单价 GalNAc 单元，可在固相合成条件下产生简化的三价 GalNAc 缀合寡核苷酸。与之前优化的三触角 GalNAc 构建体相比，使用单价 GalNAc 构建模块合成寡核苷酸缀合物所需的合成步骤更少。重新设计的三价 GalNAc 配体保持了最佳价态、空间方向和糖部分之间的距离，以便 ASGPR 正确识别。通过将三价 GalNAc 连续共价连接到有义链的 3 端来合成 siRNA 缀合物，并产生具有与亲本三价 GalNAc 缀合物设计相似的体外和体内效力的缀合物。
We recently demonstrated that siRNAs conjugated to triantennary N-acetylgalactosamine (GalNAc) induce robust RNAi-mediated gene silencing in the liver, owing to uptake mediated by the asialoglycoprotein receptor (ASGPR). Novel monovalent GalNAc units, based on a non-nucleosidic linker, were developed to yield simplified trivalent GalNAc-conjugated oligonucleotides under solid-phase synthesis conditions. Synthesis of oligonucleotide conjugates using monovalent GalNAc building blocks required fewer synthetic steps compared to the previously optimized triantennary GalNAc construct. The redesigned trivalent GalNAc ligand maintained optimal valency, spatial orientation, and distance between the sugar moieties for proper recognition by ASGPR. siRNA conjugates were synthesized by sequential covalent attachment of the trivalent GalNAc to the 3-end of the sense strand and resulted in a conjugate with in vitro and in vivo potency similar to that of the parent trivalent GalNAc conjugate design.