公开(公告)号：US20050267300A1

公开(公告)日：2005-12-01

申请号：US11099430

申请日：2005-04-05

申请人： Muthiah Manoharan ,  Michael Jung ,  Kallanthottathil Rajeev ,  Rajendra Pandey ,  Gang Wang

发明人： Muthiah Manoharan ,  Michael Jung ,  Kallanthottathil Rajeev ,  Rajendra Pandey ,  Gang Wang

IPC分类号： C07D285/00 ,  C07D285/01 ,  C07H21/00 ,  C07H21/04

CPC分类号： C07H21/00 ,  C07H21/04 ,  Y02P20/55

摘要： The present invention relates to processes and reagents for oligonucleotide synthesis and purification. One aspect of the present invention relates to compounds useful for activating phosphoramidites in oligonucleotide synthesis. Another aspect of the present invention relates to a method of preparing oligonucleotides via the phosphoramidite method using an activator of the invention. Another aspect of the present invention relates to sulfur-transfer agents. In a preferred embodiment, the sulfur-transfer agent is a 3-amino-1,2,4-dithiazolidine-5-one. Another aspect of the present invention relates to a method of preparing a phosphorothioate by treating a phosphite with a sulfur-transfer reagent of the invention. In a preferred embodiment, the sulfur-transfer agent is a 3-amino-1,2,4-dithiazolidine-5-one. Another aspect of the present invention relates to compounds that scavenge acrylonitrile produced during the deprotection of phosphate groups bearing ethylnitrile protecting groups. In a preferred embodiment, the acrylonitrile scavenger is a polymer-bound thiol. Another aspect of the present invention relates to agents used to oxidize a phosphite to a phosphate. In a preferred embodiment, the oxidizing agent is sodium chlorite, chloroamine, or pyridine-N-oxide. Another aspect of the present invention relates to methods of purifying an oligonucleotide by annealing a first single-stranded oligonucleotide and second single-stranded oligonucleotide to form a double-stranded oligonucleotide; and subjecting the double-stranded oligonucleotide to chromatographic purification. In a preferred embodiment, the chromatographic purification is high-performance liquid chromatography.

摘要翻译： 本发明涉及寡核苷酸合成和纯化的方法和试剂。 本发明的一个方面涉及可用于在寡核苷酸合成中活化亚磷酰胺的化合物。 本发明的另一方面涉及使用本发明的活化剂通过亚磷酰胺方法制备寡核苷酸的方法。 本发明的另一方面涉及硫转移剂。 在优选的实施方案中，硫转移剂是3-氨基-1,2,4-二噻唑烷-5-酮。 本发明的另一方面涉及通过用本发明的硫转移试剂处理亚磷酸酯制备硫代磷酸酯的方法。 在优选的实施方案中，硫转移剂是3-氨基-1,2,4-二噻唑烷-5-酮。 本发明的另一方面涉及清除在带有乙腈基保护基团的磷酸酯基团去保护过程中产生的丙烯腈的化合物。 在优选的实施方案中，丙烯腈清除剂是聚合物结合的硫醇。 本发明的另一方面涉及用于将亚磷酸盐氧化成磷酸盐的试剂。 在优选的实施方案中，氧化剂是亚氯酸钠，氯胺或吡啶-N-氧化物。 本发明的另一方面涉及通过使第一单链寡核苷酸和第二单链寡核苷酸退火以形成双链寡核苷酸来纯化寡核苷酸的方法; 并对双链寡核苷酸进行色谱纯化。 在优选的实施方案中，色谱纯化是高效液相色谱法。

公开(公告)号：US20050026861A1

公开(公告)日：2005-02-03

申请号：US10865245

申请日：2004-06-10

申请人： Ekambar Kandimalla ,  Lakshmi Bhagat ,  Rajendra Pandey ,  Dong Yu ,  Sudhir Agrawal

发明人： Ekambar Kandimalla ,  Lakshmi Bhagat ,  Rajendra Pandey ,  Dong Yu ,  Sudhir Agrawal

IPC分类号： A01N43/04 ,  A61K31/70 ,  A61K48/00 ,  C07H21/02 ,  C07H21/04 ,  C12N20060101

CPC分类号： C12N15/117 ,  C12N2310/17 ,  C12N2310/315 ,  C12N2310/3183 ,  C12N2310/321 ,  C12N2310/336 ,  C12N2310/53 ,  C12N2320/51 ,  C12N2310/3521

摘要： The invention provides immunostimulatory oligonucleotides having at least one CpG dinucleotide and a secondary structure at the 5′- or 3′-end. These oligonucleotides have either reduced or improved immunostimulatory properties. The invention establishes that 5′-terminal secondary structures affect immunostimulatory activity significantly more than those at the 3′-end. The invention also provides methods for increasing or decreasing the immunostimulatory activity of a CpG-containing nucleic acid.

摘要翻译： 本发明提供了在5'-或3'-末端具有至少一个CpG二核苷酸和二级结构的免疫刺激性寡核苷酸。 这些寡核苷酸具有降低的或改善的免疫刺激性质。 本发明确定5'-末端二级结构比3'-末端显着地影响免疫刺激活性。 本发明还提供了增加或降低含CpG的核酸的免疫刺激活性的方法。