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METHOD FOR PRODUCING CAPPED RNA

Foreign code F210010583
File No. (J1036-03)
Posted date 2021年11月2日
Country 世界知的所有権機関(WIPO)
International application number 2021JP006360
International publication number WO 2021172204
Date of international filing 令和3年2月19日(2021.2.19)
Date of international publication 令和3年9月2日(2021.9.2)
Priority data
  • 特願2020-032889 (2020.2.28) JP
Title METHOD FOR PRODUCING CAPPED RNA
Abstract A method for producing a capped RNA that is a 5' end cap-modified RNA, said method being characterized by comprising reacting an activated cap compound represented by formula (1) with a monophosphate RNA that is monophosphorylated at the 5' end. (In the formula, L represents a leaving group.) Preferably, the activated cap compound is a compound represented by formula (2).
Outline of related art and contending technology BACKGROUND ART
In eukaryotic mRNA and the like, a 5 'cap structure in which 7-methylguanylate is 5'-5 'bonded to the 5' terminal via a triphosphate bond is known. A cap structure is known to facilitate translation of mRNA, and there is a demand for efficient introduction of a cap structure into mRNA in order to efficiently synthesize a protein of interest in a protein expression system or the like.
Synthesis of mRNA includes a chemical synthesis method based on an enzymatic transcription method and an amidite method, but the latter has a great advantage in that it can freely introduce) contributing to the improvement of the stability and translatability of the chemically modified (mRNA essential in mRNA medicine. On the other hand, there is no technique by which a cap structure can be easily introduced to the chemically synthesized mRNA. Only a technique is known in chemical synthesis in which the 5 'end is diphosphorylated, followed by enzymatically introducing a cap (e.g., NpL 1). Fig. 16 is a conceptual diagram illustrating this conventional cap introduction method. In this method, RNA diphosphorylated by transcriptional synthesis or chemical synthesis using an CAP enzyme is prepared, and a cap structure is further introduced to the 5 'end using the CAP enzyme.
In addition, another known method of CAP oxidation is a method in which guanine is methylated using an enzyme by solid-phase synthesis and monophosphate activation (for example, see Non-Patent Document 2). According to the method of this document, an imidazole group is introduced into the monophosphate group at the 5 'terminal of RNA, and a diphosphate group such as GDP (guanidinediphosphate) is reacted with the imidazole group to introduce a CAP structure at the 5' terminal of RNA.
Furthermore, another method is also known (see, for example, Non-Patent Document 3). This method also introduces a CAP structure to the 5 'terminal of RNA by introducing an imidazole group to the triphosphate group at the 5' terminal of RNA by solid phase synthesis, and reacting the imidazole group with a monophosphate group such as GMP (guanine phosphate).
  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
  • Inventor
  • ABE Hiroshi
  • KIMURA Yasuaki
  • ABE Naoko
IPC(International Patent Classification)
Specified countries National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DJ DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS IT JO JP KE KG KH KN KP KR KW KZ LA LC LK LR LS LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SA SC SD SE SG SK SL ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN WS ZA ZM ZW
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW KM ML MR NE SN ST TD TG
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