Post-SELEX chemical optimization of a trypanosome-specific RNA aptamer
Post-SELEX chemical optimization of a trypanosome-specific RNA aptamer
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DOI：
10.2174/138620708783398331
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发表时间：
2008-01-01
影响因子：
1.8
通讯作者：
Goeringer, H. Ulrich
中科院分区：
文献类型：
--
作者：
Adler, Annette;Forster, Nicole;Goeringer, H. Ulrich
非洲锥虫是昏睡病的病原体。用来控制和治疗这种疾病的治疗方法非常无效，因此，迫切需要开发改进的药物。最近，人们提出了设计新型杀锥虫药的新策略。其中包括依赖寄生虫特异性RNA适配子的技术。一种方法涉及适体引导的裂解化合物运输到寄生虫的溶酶体。该适配子被命名为2-16RNA，我们在此报道了该RNA在体内应用的优化。为了将适配子2-16转化为血清稳定试剂，产生了2脱氧-2‘F-和/或2’-脱氧-2‘-氨基-尿苷和胞苷取代的RNA。虽然2‘-NH2-DC/Du修饰的RNA是RNase抗性的，但它们在功能上是不活跃的。相反，2‘-F-DC/Du-取代的2-16RNA保留了与活体锥虫结合的能力(K-d=45 nM)，并与未修饰的RNA相同地被送到溶酶体。2‘-F-DC/Du-取代的2-16RNA具有热稳定性(T-m=75℃)，血清半衰期为3.4天。此外，适体2-16被聚乙二醇化以增加其血清保留时间。与聚乙二醇类聚合物的偶联
African trypanosomes are the causative agent of sleeping sickness. The therapeutics used to control and treat the disease are very ineffective and thus, the development of improved drugs is urgently needed. Recently, new strategies for the design of novel trypanocidals have been put forward. Among them are techniques that rely on parasite-specific RNA aptamers. One approach involves the aptamer-directed transport of lytic compounds to the lysosome of the parasite. The aptamer has been termed 2-16 RNA and here we report the optimization of the RNA for its applications in vivo. To convert aptamer 2-16 into a serum-stable reagent 2 deoxy-2'F-and/or 2'-deoxy-2'-NH2- uridine-and cytidine-substituted RNAs were generated. While 2'-NH2-dC/dU-modified RNAs were RNase-resistant, they were functionally inactive. By contrast, 2'-F-dC/dU-substituted 2-16 RNA retained its ability to bind to live trypanosomes (K-d=45 nM) and was routed to the lysosome identically to unmodified RNA. 2'-F-dC/dU-substituted 2-16 RNA is thermostable (T-m=75 degrees C) and has a serum half-life of 3.4 days. Furthermore, aptamer 2-16 was site-specifically PEGylated to increase its serum retention time. Conjugation with PEG polymers