公开(公告)号：US5837852A

公开(公告)日：1998-11-17

申请号：US136214

申请日：1993-10-14

申请人： Thomas D. Y. Chung ,  Christopher W. Cianci ,  Moira Hagen ,  Mark Krystal ,  Richard J. Colonno

发明人： Thomas D. Y. Chung ,  Christopher W. Cianci ,  Moira Hagen ,  Mark Krystal ,  Richard J. Colonno

IPC分类号： A61K38/00 ,  C07H21/00 ,  C12N15/113 ,  C12N5/00 ,  A61K48/00 ,  C12N15/00 ,  C07N14/00

CPC分类号： C12N15/1131 ,  C07H21/00 ,  A61K38/00 ,  C12N2310/13 ,  C12N2310/317

摘要： Novel capped oligonucleotides useful in treatment of influenza infection. A synthetically derived 67-nucleotide RNA substrate, containing a �.sup.32 P! labeled cap-1 structure was used to analyze parameters of influenza virus endonuclease activity. This substrate was specifically cleaved by the influenza virus polymerase to yield a single capped 11-nucleotide fragment capable of directly priming transcription. An analysis of systematic truncations of this RNA substrate in cleavage, elongation, and binding reactions demonstrated that the minimum chain length required for cleavage was one nucleotide past the cleavage site. In contrast, the minimum chain length required for priming activity was found to be 9 nucleotides, while a chain length of at least 4 nucleotides was required for efficient binding. Based on these chain length requirements, the present inventors show that a pool of capped oligonucleotides--too short to prime transcription but long enough to bind with high affinity to the viral polymerase--are potent inhibitors of cap-dependent in vitro transcription.

摘要翻译： 用于治疗流感感染的新型封端寡核苷酸。 使用含有[32P]标记的cap-1结构的合成衍生的67-核苷酸RNA底物来分析流感病毒核酸内切酶活性的参数。 该底物被流感病毒聚合酶特异地切割，得到能够直接引发转录的单一封端的11-核苷酸片段。 在裂解，延伸和结合反应中对该RNA底物的系统截短的分析表明，切割所需的最短链长度是经过切割位点的一个核苷酸。 相比之下，发现引发活性所需的最短链长度为9个核苷酸，而需要至少4个核苷酸的链长才能有效结合。 基于这些链长度要求，本发明人表明，封闭的寡核苷酸库 - 太短而不能提供转录但足够长以高度结合到病毒聚合酶 - 是帽依赖性体外转录的有效抑制剂。