Lan Tao, Kandimalla Ekambar R
Idera Pharmaceuticals, Inc., Cambridge, MA, USA.
Methods Mol Biol. 2011;764:249-61. doi: 10.1007/978-1-61779-188-8_17.
Viral single-stranded (ss) RNA is the natural ligand for TLR7 and TLR8. Synthetic ssRNA has been shown to act as a ligand for TLR7 and TLR8. We have previously reported a novel RNA structure, referred to as stabilized immune modulatory RNA (SIMRA), in which two short phosphorothioate oligoribonucleotides were linked through their 3'-ends via a linker. SIMRA compounds had greater stability in serum than unmodified ssRNA and induced immune responses via TLR7 and/or TLR8. SIMRA compounds were synthesized using phosphoramidite chemistry on controlled-pore glass solid support derivatized with a linker. After cleavage from the solid support and removal of protecting groups, SIMRA compounds were purified on an anion-exchange HPLC followed by desalting/dialysis, and lyophilization. SIMRA compounds were characterized for their purity and sequence integrity by anion-exchange HPLC, capillary gel electrophoresis, polyacrylamide gel electrophoresis, and MALDI-TOF mass spectrophotometric analysis. As SIMRA compounds induce TLR7- and/or TLR8-mediated Th1-type immune responses, they have potential utility as therapeutic agents for a broad range of diseases, including cancer, infectious diseases, asthma, and allergies, and as adjuvants with vaccines.
病毒单链（ss）RNA是Toll样受体7（TLR7）和Toll样受体8（TLR8）的天然配体。合成的ssRNA已被证明可作为TLR7和TLR8的配体。我们之前报道了一种新型RNA结构，称为稳定免疫调节RNA（SIMRA），其中两个短的硫代磷酸寡核糖核苷酸通过连接子在其3'端相连。SIMRA化合物在血清中的稳定性高于未修饰的ssRNA，并通过TLR7和/或TLR8诱导免疫反应。SIMRA化合物是在连接子衍生化的可控孔径玻璃固相载体上采用亚磷酰胺化学合成的。从固相载体上切割并去除保护基团后，SIMRA化合物在阴离子交换高效液相色谱上进行纯化，随后进行脱盐/透析和冻干。通过阴离子交换高效液相色谱、毛细管凝胶电泳、聚丙烯酰胺凝胶电泳和基质辅助激光解吸电离飞行时间质谱分析对SIMRA化合物的纯度和序列完整性进行了表征。由于SIMRA化合物可诱导TLR7和/或TLR8介导的Th1型免疫反应，它们具有作为多种疾病治疗药物的潜在用途，包括癌症、传染病、哮喘和过敏，以及作为疫苗佐剂。
