WO2014109384A1 - Procédé de production de dérivé d'acide nucléique réticulé - Google Patents

Procédé de production de dérivé d'acide nucléique réticulé Download PDF

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WO2014109384A1
WO2014109384A1 PCT/JP2014/050325 JP2014050325W WO2014109384A1 WO 2014109384 A1 WO2014109384 A1 WO 2014109384A1 JP 2014050325 W JP2014050325 W JP 2014050325W WO 2014109384 A1 WO2014109384 A1 WO 2014109384A1
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group
compound
general formula
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Inventor
聡 小比賀
秀勲 三神山
庸 足立
直樹 三宅
昌浩 阪上
哲也 谷野
浩輔 阿南
関口　光明
典一 黒田
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Shionogi and Co Ltd
University of Osaka NUC
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Osaka University NUC
Shionogi and Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
    • C07H19/06—Pyrimidine radicals
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Definitions

  • the present invention relates to a method for producing an intermediate useful for the efficient production of amide-bridged nucleic acid derivatives.
  • the antisense method is known as one of the methods for treating diseases with nucleic acid drugs.
  • Antisense method introduces oligonucleotide (antisense strand) complementary to disease-related mRNA from the outside to form a double strand, thereby inhibiting the translation process of pathogenic RNA and treating or preventing disease. It is a technique to do.
  • Non-patent document 1 phosphorothioate in which an oxygen atom on a phosphorus atom is substituted with a sulfur atom
  • methylphosphonate non-patent document 2 in which a methyl group is substituted
  • BNA bridged nucleic acid
  • LNA locked nucleic acid
  • Obiga et al. are less susceptible to degradation by nucleases in vivo than conventional cross-linked types, have high binding affinity for target mRNA, and efficiently control the expression of specific genes.
  • Patent Document 4 a novel nucleic acid derivative in which an amide bond is introduced into a 2 ′, 4′-BNA cross-linked structure has been found (Patent Document 4).
  • An object of the present invention is to provide a method for efficiently producing an amide-bridged nucleic acid derivative useful as a molecule used in an antisense method.
  • the present inventors have found that an important intermediate for synthesizing an amide-bridged nucleic acid derivative can be efficiently synthesized by carrying out an aldol reaction under a specific condition, thereby completing the present invention. Further, the present invention was completed by finding a method for efficiently removing a ring by removing the protecting group of the intermediate.
  • the present invention (1) The following steps for subjecting a compound represented by the general formula (I) to an aldol reaction in the presence of a base:
  • R 1 and R 2 are the same or different and each represents a hydrogen atom, an amino group protecting group for nucleic acid synthesis, an alkyl group, an alkenyl group, a cycloalkyl group, an aryl group, an aralkyl group, an acyl group, a silyl group, Phosphate group, phosphate group protected with a protecting group for nucleic acid synthesis, -P (R 7 ) R 8 [wherein R 7 and R 8 are the same or different and protected with a hydroxyl group, a protecting group for nucleic acid synthesis A hydroxyl group, a mercapto group, a mercapto group protected with a protecting group for nucleic acid synthesis, an amino group, an alkoxy group having 1 to 5 carbon atoms, an alkylthio group
  • R 9 and R 10 are the same or different and are a hydroxyl group, a hydroxyl group protected with a protecting group for nucleic acid synthesis, a mercapto group, or a protecting group for nucleic acid synthesis.
  • Y 7 and Y 8 are hydroxyl-protecting groups for nucleic acid synthesis
  • R 5 ′ is an alkyl group, a hydrogen atom, an optionally substituted benzyl group, or an optionally substituted group.
  • An aromatic ring, R 6 ′ , R 7 ′ , R 8 ′ and R 9 ′ are a hydrogen atom or an alkyl group), and a method for producing a compound represented by the general formula (XIII), About.
  • numerator used for an antisense method can be manufactured efficiently. More specifically, an important intermediate can be produced with a minimum of raw material residues and by-products. Further, by using this intermediate, it has become possible to produce an amide-bridged nucleic acid derivative useful as an antisense molecule more efficiently than the conventional method. Furthermore, it has become possible to obtain an intermediate in a high yield by finding a method for removing the protecting group of the intermediate and efficiently cyclizing the intermediate.
  • nucleobase moiety of B is a structure in which, in addition to a hydrocarbon cyclic group, a carbon atom which is a constituent atom of the hydrocarbon ring is replaced with one or more hetero atoms such as a nitrogen atom, a sulfur atom or an oxygen atom Any heterocyclic group having a 5- to 20-membered ring and having aromaticity, including monocyclic and condensed rings.
  • the hydrocarbon ring include benzene, naphthalene, anthracene, phenanthrene, indane, indene, tetrahydronaphthylene, biphenylene and the like.
  • heterocyclic ring examples include pyrimidine nucleobase or purine nucleobase. Any ring may have one or more substituents selected from the following ⁇ group.
  • the pyrimidine nucleobase or purine nucleobase acts or substitutes for a base generally known as a constituent component of a nucleic acid (eg, guanine, adenine, cytosine, thymine, uracil) and other similar nucleic acid components. Includes every chemical structure you get.
  • a pyrimidine nucleobase or a purine nucleobase or a pyrimidine nucleobase or a purine nucleobase, which may have one or more substituents selected from the following ⁇ group.
  • a -yl group, a 2-oxo-pyrimidin-1-yl group, or a purin-9-yl group or a 2-oxo-pyrimidin-1-yl group having a substituent selected from the following group ⁇ is preferred.
  • ⁇ group hydroxyl group, hydroxyl group protected with a protecting group for nucleic acid synthesis, alkoxy group having 1 to 5 carbon atoms, mercapto group, mercapto group protected with a protecting group for nucleic acid synthesis, alkylthio group having 1 to 5 carbon atoms, amino An amino group protected with a protecting group for nucleic acid synthesis, an amino group substituted with an alkyl group having 1 to 5 carbon atoms, an alkyl group having 1 to 5 carbon atoms, and a halogen atom.
  • a group suitable as “optionally substituted purine nucleobase” is 6-aminopurin-9-yl (that is, adeninyl), 6 in which the amino group is protected with a protecting group for nucleic acid synthesis.
  • a group suitable as “an optionally substituted pyrimidine nucleobase” is 2-oxo-4-amino-1,2-dihydropyrimidin-1-yl (ie, cytosynyl), and the amino group is a nucleic acid.
  • purine nucleobases or pyrimidine nucleobases more preferably, 6-aminopurin-9-yl (ie, adeninyl), wherein the amino group is a protecting group for nucleic acid synthesis.
  • 6-aminopurin-9-yl, 2,6-diaminopurin-9-yl, 2-amino-6-chloropurin-9-yl, 2-amino protected with a protecting group for nucleic acid synthesis Amino-6-chloropurin-9-yl, 2-amino-6-fluoropurin-9-yl, 2-amino-6-fluoropurin-9-yl having an amino group protected with a protecting group for nucleic acid synthesis, 2 -Amino-6-bromopurin-9-yl, 2-amino-6-bromopurin-9-yl, 2-amino-6-hydroxypurin-9-yl in which the amino group is protected with a protecting group for nucleic acid synthesis ( That is, Guaninyl), Ami 2-amino-6-hydroxypurin-9-yl, 6-amino-2-methoxypurin-9-yl, 6-amino-2-chloropurin-9-yl, the group of which is,
  • Protecting group for amino group for nucleic acid synthesis and “protecting group for hydroxyl group for nucleic acid synthesis” and “hydroxyl group protected by protecting group for nucleic acid synthesis” are an amino group or a stable group during nucleic acid synthesis. Although it is not particularly limited as long as it can protect a hydroxyl group, specifically, it is stable under acidic or neutral conditions, and is obtained by a chemical method such as hydrogenolysis, hydrolysis, electrolysis and photolysis.
  • Examples of the protecting group that can be cleaved include formyl, acetyl, propionyl, butyryl, isobutyryl, pentanoyl, pivaloyl, valeryl, isovaleryl, octanoyl, nonanoyl, decanoyl, 3-methylnonanoyl, 8- Methylnonanoyl, 3-ethyloctanoyl, 3,7-dimethyloctanoyl, undecanoyl, dodecanoyl, tridecanoyl, te Radecanoyl, pentadecanoyl, hexadecanoyl, 1-methylpentadecanoyl, 14-methylpentadecanoyl, 13,13-dimethyltetradecanoyl, heptadecanoyl, 15-methylhexadecanoyl, octadecanoyl, 1-methylhepta De
  • Aryl group substituted by halogen atom, lower alkoxy group or nitro group includes “lower group substituted by halogen or tri-lower alkylsilyl group” such as 2,2,2-trichloroethoxycarbonyl, 2-trimethylsilylethoxycarbonyl “Alkoxycarbonyl group”; “alkenyloxycarbonyl group” such as vinyloxycarbonyl and aryloxycarbonyl; benzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 2 Examples include 1 to 2 “aralkyloxycarbonyl groups in which the aryl ring may be substituted with a lower alkoxy or nitro group”, such as nitrobenzyloxycarbonyl and 4-nitrobenzyloxycarbonyl.
  • the “protecting group for hydroxyl group” is preferably “aliphatic acyl group”, “aromatic acyl group”, “methyl group substituted with 1 to 3 aryl groups”, “lower alkyl, lower alkoxy, halogen”.
  • the protective group of “hydroxy group protected with a protective group for nucleic acid synthesis preferably an “aliphatic acyl group”, “aromatic acyl group”, 4-oxopentanoyl (levulinoyl
  • Alkyl group means a linear or branched alkyl group having 1 to 20 carbon atoms, and includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, s-butyl, tert-butyl, n -Pentyl, isopentyl, 2-methylbutyl, neopentyl, 1-ethylpropyl, n-hexyl, isohexyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3,3-dimethylbutyl, 2 Linear or branched having 1 to 6 carbon atoms such as 1,2-dimethylbutyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2-ethylbutyl In addition to chain alkyl groups (in the alky
  • alkenyl group refers to a straight or branched alkenyl group having 2 to 20 carbon atoms, and includes ethenyl, 1-propenyl, 2-propenyl, 1-methyl-2-propenyl, 1-methyl-1-propenyl.
  • cycloalkyl group refers to a cycloalkyl group having 3 to 10 carbon atoms, and examples thereof include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, norbornyl, adamantyl, and the like, preferably cyclopropyl, A cycloalkyl group having 3 to 8 carbon atoms such as cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and the like.
  • cycloalkyl group includes a heterocyclic group in which one or more methylenes on the ring of the cycloalkyl group are substituted with an oxygen atom, a sulfur atom, or a nitrogen atom substituted with an alkyl group, Examples thereof include a tetrahydropyranyl group.
  • aryl group means a monovalent substituent having 6 to 14 carbon atoms in which one hydrogen atom is removed from an aromatic hydrocarbon group, and examples thereof include phenyl, indenyl, naphthyl, phenanthrenyl, anthracenyl and the like. .
  • the aryl ring may be substituted with one or more groups such as a halogen atom, a lower alkyl group, a hydroxyl group, an alkoxy group, an aryloxy group, an amino group, a nitro group, trifluoromethyl, a phenyl group,
  • groups such as a halogen atom, a lower alkyl group, a hydroxyl group, an alkoxy group, an aryloxy group, an amino group, a nitro group, trifluoromethyl, a phenyl group
  • the optionally substituted aryl group include 2-methylphenyl, 2,6-dimethylphenyl, 2-chlorophenyl, 4-chlorophenyl, 2,4-dichlorophenyl, 2,5-dichlorophenyl, and 2-bromo.
  • Examples include phenyl, 4-methoxyphenyl, 4-chloro-2-nitrophenyl, 4-nitrophenyl, 2,4-dinitrophenyl, biphenyl and the like.
  • Preferable examples include a halogen atom, a phenyl group substituted with a lower alkoxy group nitro group, and a phenyl group.
  • the “aralkyl group” means an alkyl group having 1 to 6 carbon atoms substituted with an aryl group, and includes benzyl, ⁇ -naphthylmethyl, ⁇ -naphthylmethyl, indenylmethyl, phenanthrenylmethyl, anthracenyl “Methyl group substituted with 1 to 3 aryl groups” such as methyl, diphenylmethyl, triphenylmethyl, ⁇ -naphthyldiphenylmethyl, 9-anthrylmethyl, 4-methylbenzyl, 2,4,6 -Trimethylbenzyl, 3,4,5-trimethylbenzyl, 4-methoxybenzyl, 4-methoxyphenyldiphenylmethyl, 4,4'-dimethoxytriphenylmethyl, 2-nitrobenzyl, 4-nitrobenzyl, 4-chlorobenzyl, 4-bromobenzyl, “lower alkyl, lower alkoxy, halogen, such as 4-cyanobenzyl, In
  • methyl group substituted with 1 to 3 aryl groups “methyl substituted with 1 to 3 aryl groups with aryl ring substituted with lower alkyl, lower alkoxy, halogen, cyano group” Group, more preferably 4-methoxyphenyldiphenylmethyl, 4,4′-dimethoxytriphenylmethyl.
  • acyl group examples include formyl, acetyl, propionyl, butyryl, isobutyryl, pentanoyl, pivaloyl, valeryl, isovaleryl, octanoyl, nonanoyl, decanoyl, 3-methylnonanoyl, 8-methylnonanoyl, 3-ethyloctanoyl, 3,7-dimethyl Octanoyl, undecanoyl, dodecanoyl, tridecanoyl, tetradecanoyl, pentadecanoyl, hexadecanoyl, 1-methylpentadecanoyl, 14-methylpentadecanoyl, 13,13-dimethyltetradecanoyl, heptadecanoyl, 15-methylhexayl Alkylcarbonyl groups such as decanoyl, octadecanoyl, 1-methylheptadecanoy
  • Halogenoarylcarbonyl group 2,4,6-trimethylbenzoyl, lower alkylated arylcarbonyl group such as 4-toluoyl, lower alkoxylated arylcarbonyl group such as 4-anisoyl, 2-carboxybenzoyl, 3-carb Carboxylated arylcarbonyl groups such as xylbenzoyl, 4-carboxybenzoyl, nitrated arylcarbonyl groups such as 4-nitrobenzoyl, 2-nitrobenzoyl; lower alkoxycarbonylated arylcarbonyl groups such as 2- (methoxycarbonyl) benzoyl
  • an “aromatic acyl group” such as an arylated arylcarbonyl group such as 4-phenylbenzoyl, preferably a formyl, acetyl, propionyl, butyryl, isobutyryl, pentanoyl, pivaloyl, benzoyl group.
  • sil group examples include “tri-lower alkylsilyl group” such as trimethylsilyl, triethylsilyl, isopropyldimethylsilyl, t-butyldimethylsilyl, methyldiisopropylsilyl, methyldi-t-butylsilyl, triisopropylsilyl, diphenylmethylsilyl, “Tri-lower alkylsilyl group substituted with 1 to 2 aryl groups” such as butyldiphenylbutylsilyl, diphenylisopropylsilyl, phenyldiisopropylsilyl, and the like, preferably trimethylsilyl, triethylsilyl, triisopropylsilyl , T-butyldimethylsilyl, t-butyldiphenylsilyl, and more preferably trimethylsilyl.
  • tri-lower alkylsilyl group
  • the “protecting group” of the “phosphate group protected with a protecting group for nucleic acid synthesis” is not particularly limited as long as it can stably protect the phosphate group during nucleic acid synthesis, but is not specifically limited. Refers to a protecting group that is stable under acidic or neutral conditions and can be cleaved by chemical methods such as hydrogenolysis, hydrolysis, electrolysis and photolysis.
  • the protecting group of the “mercapto group protected with a protecting group for nucleic acid synthesis” is not particularly limited as long as it can stably protect the mercapto group during nucleic acid synthesis. Or a protecting group that is stable under neutral conditions and can be cleaved by a chemical method such as hydrogenolysis, hydrolysis, electrolysis and photolysis. , An alkylthio group such as ethylthio and tert-butylthio, an arylthio group such as benzylthio, and the like, and a “aliphatic acyl group” or “aromatic acyl group” are preferred. And more preferably a benzoyl group.
  • C 1-5 alkoxy group examples include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, s-butoxy, tert-butoxy and n-pentoxy. Is a methoxy or ethoxy group.
  • alkylthio group having 1 to 5 carbon atoms examples include methylthio, ethylthio, propylthio, isopropylthio, butylthio, isobutylthio, s-butylthio, tert-butylthio, and n-pentylthio.
  • C 1-6 cyanoalkoxy group refers to a group substituted by the cyano group of the above “C 1-5 alkoxy group”. Examples of such a group include cyanomethoxy, 2-cyano Examples thereof include ethoxy, 3-cyanopropoxy, 4-cyanobutoxy, 3-cyano-2-methylpropoxy, and 1-cyanomethyl-1,1-dimethylmethoxy, and a 2-cyanoethoxy group is preferable.
  • amino group substituted with an alkyl group having 1 to 5 carbon atoms examples include, for example, methylamino, ethylamino, propylamino, isopropylamino, butylamino, isobutylamino, s-butylamino, tert-butylamino, dimethyl Amino, diethylamino, dipropylamino, diisopropylamino, dibutylamino, diisobutylamino, di (s-butyl) amino, di (tert-butyl) amino can be mentioned, preferably methylamino, ethylamino, dimethylamino , Diethylamino or diisopropylamino group.
  • C 1-5 alkyl group examples include methyl, ethyl, propyl, isopropyl, isopropyl, butyl, isobutyl, s-butyl, tert-butyl, n-pentyl, etc. , Methyl or ethyl group.
  • halogen atom examples include a fluorine atom, a chlorine atom, a bromine atom or an iodine atom, and preferably a fluorine atom or a chlorine atom.
  • the protecting group of the “amino group protected with a protecting group for nucleic acid synthesis” is not particularly limited as long as it can stably protect an amino group during nucleic acid synthesis.
  • a protecting group that is stable under neutral conditions and can be cleaved by chemical methods such as hydrogenolysis, hydrolysis, electrolysis and photolysis, such as formyl, acetyl, propionyl, butyryl, isobutyryl, pentanoyl, Pivaloyl, valeryl, isovaleryl, octanoyl, nonanoyl, decanoyl, 3-methylnonanoyl, 8-methylnonanoyl, 3-ethyloctanoyl, 3,7-dimethyloctanoyl, undecanoyl, dodecanoyl, tridecanoyl, tetradecanoyl, pentadecanoyl, hexadecana Noyl, 1-methylpenta
  • arylcarbonyl group such as lower alkoxycarbonylated arylcarbonyl group such as 2- (methoxycarbonyl) benzoyl, arylated arylcarbonyl group such as 4-phenylbenzoyl; “Lower alkoxycarbonyl groups” such as xoxycarbonyl, ethoxycarbonyl, t-butoxycarbonyl, isobutoxycarbonyl; “halogen or tri-lower alkylsilyl groups such as 2,2,2-trichloroethoxycarbonyl, 2-trimethylsilylethoxycarbonyl” A lower alkoxycarbonyl group substituted with a “alkenyloxycarbonyl group” such as vinyloxycarbonyl or aryloxycarbonyl; benzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, 4-nitrobenzyloxy Examples thereof include 1 to 2 “aralkyloxycarbon
  • Nucleoside analog means a non-natural type of “nucleoside” in which a purine base or a pyrimidine base and a sugar are bonded, and a purine base or an aromatic heterocycle other than purine and pyrimidine and an aromatic hydrocarbon ring. A substance that can be substituted with a pyrimidine base is combined with a sugar.
  • the “salt” refers to a salt of the compound (III) of the present invention, and is preferably a salt such as sodium salt, potassium salt or lithium salt.
  • Alkali earth metal salts such as alkali metal salts, calcium salts and magnesium salts, metal salts such as aluminum salts, iron salts, zinc salts, copper salts, nickel salts and cobalt salts; inorganic salts such as ammonium salts, t- Octylamine salt, dibenzylamine salt, morpholine salt, glucosamine salt, phenylglycine alkyl ester salt, ethylenediamine salt, N-methylglucamine salt, guanidine salt, diethylamine salt, triethylamine salt, dicyclohexylamine salt, N, N'-di Benzylethylenediamine salt, chloroprocaine salt, procaine salt, diethanolamine salt, N-benzyl -Amine salts such as organic salts such as phene
  • the “reagent used for protecting a hydroxyl group for nucleic acid synthesis” refers to a reagent used for introducing the “protecting group for a hydroxyl group for nucleic acid synthesis” into a sugar hydroxyl group constituting a nucleic acid.
  • tert-butyldiphenylsilyl group In the case of tert-butyldiphenylsilyl group; tert-butyldimethylsilyl chloride or tert-butyldiphenylsilyl trifluoromethanesulfonate 5) in the presence of a base such as imidazole, N-methylimidazole, 4,4-dimethylaminopyridine, triethylamine, etc.
  • a base such as imidazole, N-methylimidazole, 4,4-dimethylaminopyridine, triethylamine, etc.
  • a benzoyl group in the presence of a base such as pyridine, triethylamine or potassium carbonate, benzoyl chloride, benzoic anhydride ii) a base such as triethylamine or N-methylmorpholine and EDC (1-ethyl-3- (3 In the presence of a condensing agent such as -dimethylaminopropyl) carbodiimide) hydrochloride, DCC (N, N'-dicyclohexylcarbodiimide), in the case of benzoic acid 6) levonoyl group; In the presence of a condensing agent such as DCC, levulinic acid 7) In the case of 2-ethoxyethyl group: ethoxy vinyl ether is applicable in the presence of an acid catalyst such as pyridium paratoluenesulfonic acid or toluenesulfonic acid.
  • an acid catalyst such as pyridium paratoluenesulf
  • aldol reaction represented by the following formula:
  • Aldol reaction is also referred to as aldol condensation or aldol addition. This is a reaction in which a carbonyl compound having a hydrogen atom at the ⁇ -position is reacted with another carbonyl compound to form a ⁇ -hydroxycarbonyl compound. An equilibrium reaction that occurs with either an acid or base catalyst.
  • Examples of the carbonyl compound to be reacted with the starting material in the present invention include ketone compounds such as acetone and aldehyde compounds such as formaldehyde, preferably formalin or paraformaldehyde.
  • a base usually, it is carried out in the presence of a base.
  • the base used at this time include inorganic bases such as sodium hydroxide and sodium carbonate, and organic bases such as N-methylmorpholine, DBU, triethylamine and pyridine.
  • organic bases such as N-methylmorpholine, DBU, triethylamine and pyridine.
  • An organic base having a pKa of a conjugate acid of 6 to 10 is preferable, and examples thereof include triethylamine and N-methylmorpholine.
  • a water-soluble solvent such as alcohol, acetonitrile or N, N′-dimethylformamide (DMF) is preferable.
  • a suitable combination is 1 to 3 volumes of an aqueous formalin solution, 1 to 3 volumes of N-methylmorpholine as a base, and 2 to 6 volumes of acetonitrile as a solvent. This is because as a result of carrying out the aldol reaction on the compound of the general formula (I) in this combination, it was possible to minimize the residual amount of the raw material (I) and the amount of by-products after the reaction.
  • reaction temperature and reaction time are not particularly limited, but preferably the reaction temperature is 50 ° C. to 100 ° C., and the reaction time is 1 to 24 hours.
  • Further embodiments of the present invention also include a condensation reaction represented by the following formula: More specifically, the starting material compound and the desilylating agent are reacted, and the condensing agent is reacted with the reaction solution.
  • the “desilylating agent” is a reaction aid added to cause the elimination reaction of the silyl group.
  • various acids such as hydrochloric acid, acetic acid, paratoluenesulfonic acid, fluorination Fluoride ions such as tetrabutylammonium, hydrofluoric acid and cesium fluoride can be mentioned.
  • a “condensation agent” is a reaction aid added to cause an amino group and a carboxyl group to undergo dehydration condensation.
  • EDC hydrochloride (1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride)
  • DCC dicyclohexylcarbodiimide
  • DIC diisopropylcarbodiimide
  • CDI carbonyldiimidazole
  • PyBop [(benzotriazole- 1-yloxy) tripyrrolidinophosphonium hexafluorophosphate]
  • HATU [O- (7-azabenzotriazol-1-yl) -N, N, N ′, N′-tetramethyluronium hexafluorophosphate]
  • DMT- Reagents used for peptide synthesis such as MM [4- (4,6-dimethoxy-1,3,5-triazin-2-yl) -4-methyl
  • a further aspect of the invention also includes a transglycosylation reaction represented by the following formula:
  • transglycosylation reaction refers to a reaction for substituting the base moiety of a nucleic acid derivative.
  • the general formula in the present invention includes both a cis form and a trans form.
  • a trans form is preferable.
  • the solid in the reaction process in this invention, the solid can be maintained and it is an industrially very useful method.
  • the compound represented by the above general formula (I) (corresponding to A-5 below) can be obtained, for example, by the following method.
  • R 1 , R 2 , R 3 , R 4 , R 5 , R 6 and B in the formula are as defined above.
  • P 1 has the same meaning as the above-mentioned “protecting group for hydroxyl group in nucleic acid synthesis”.
  • a trityl group and a benzyl group which may be substituted with a methoxy group are preferable.
  • the compound represented by the general formula (A-1) in the formula can be synthesized by the method described in Organic Letters, 7, 1569-1572 (2005).
  • the compound represented by the general formula A-1 is converted into 1.0 equivalent to 10.0 equivalents, preferably 3.0 equivalents to 6.0 equivalents of R 3 -Hal (in a solvent such as DMF, acetonitrile, dichloromethane, etc.).
  • Hal is halogen
  • an organic base eg, N-methylimidazole
  • an inorganic base eg, sodium
  • Step 2 the compound represented by formula (A-2) is reacted with a strong acid such as hydrochloric acid to obtain a compound A-3 in which the hydroxyl protecting group P 1 bonded to the 4′-position is deprotected. It is a process.
  • This step can be performed according to the method described in Protective Groups in Organic Synthesis, Theodora W Green (John Wiley & Sons) and the like.
  • the compound represented by the general formula A-2 is converted into a strong acid (1.0 equivalent to 10.0 equivalents, preferably 2.0 equivalents to 4.0 equivalents) in a solvent such as methanol, ethanol, ethyl acetate, or dioxane.
  • An aqueous solution such as hydrochloric acid and sulfuric acid, an organic acid such as paratoluenesulfonic acid, etc.) at 0 ° C. to 100 ° C., preferably 20 ° C. to 30 ° C., for 0.5 to 24 hours to react with the general formula (A-3) Can be obtained.
  • the compound represented by the general formula A-3 is 1.0 equivalent to 5.0 equivalents in a solvent such as tetrahydrofuran, dichloromethane, acetonitrile, DMF, or a mixed solvent such as water-tetrahydrofuran and water-toluene, preferably Is 2.0 equivalents to 4.0 equivalents of (R 2 ) 2 O and 1.0 equivalents to 5.0 equivalents, preferably 2.0 equivalents to 4.0 equivalents of an organic base (such as pyridine) or inorganic Reaction with a base (for example, sodium hydride, sodium hydroxide, sodium carbonate, potassium carbonate, cesium carbonate, sodium bicarbonate, etc.) at -20 ° C to 50 ° C, preferably 0 ° C to 30 ° C for 0.5 to 24 hours To obtain a compound represented by the general formula (A-4).
  • a solvent such as tetrahydrofuran, dichloromethane, acetonitrile, DMF, or a mixed solvent such as
  • This step is a step of obtaining a compound (A-5) having a carbonyl group by oxidizing the hydroxyl group bonded to the 4′-position of the compound represented by the general formula (A-4).
  • This step can be performed according to the method described in Experimental Chemistry Course 5th edition Volume 17 (Yamazen) and the like.
  • the compound represented by the general formula A-4 is added in an amount of 1.0 equivalent to 5.0 equivalent, preferably 2.0 equivalent to 4.0 equivalent of EDC and 1.0 equivalent to 3. equivalent in a solvent such as DMSO.
  • an organic acid eg, pyridinium paratoluenesulfonate
  • This step is a step of obtaining the compound represented by the general formula (A-6) by subjecting the compound represented by the general formula (A-5) to the aldol reaction shown in claim 1 of the present invention.
  • the compound represented by the general formula (A-5) is 1.0 to 20.0 times volume or 1.0 equivalent to 20.
  • a carbonyl compound represented by 0 equivalent of R 5 C ( ⁇ O) R 6 preferably a 1.0- to 3.0-fold volume formalin aqueous solution, and the like, and a 1.0- to 20.0-fold volume or 1.0 Equivalent to 20.0 equivalents of an organic base, preferably 1.0 to 3.0 volumes of an organic base with a pKa of 6 to 10 (for example, N-methylmorpholine, etc.) and 20 ° C. to 100 ° C.
  • the compound represented by the general formula (A-6) can be obtained by reacting preferably at 70 ° C. to 80 ° C. for 0.5 to 24 hours.
  • R 4 of the compound represented by the general formula (A-6) obtained by the above method is a hydrogen atom, the following oxidation reaction is performed.
  • This step can be performed according to the method described in Experimental Chemistry Course, 5th edition, volume 17 (Yamazen).
  • a compound represented by the general formula (A-6) is mixed with a buffer solution such as an aqueous solution of sodium dihydrogen phosphate in a solvent such as ethyl acetate, acetonitrile, or tert-butanol, and 1.0 equivalent to 20.0 equivalents of 2 -1.0 equivalent to 10.0 equivalents, preferably 2.0 equivalents to 5.0 equivalents of sodium chlorite in the presence of methyl-2-butene, squalene, etc., and 0 ° C to 60 ° C, preferably 10
  • a compound represented by the general formula (A-7) can be obtained by reacting at a temperature of from 40 ° C. to 40 ° C. for 0.5 to 24 hours.
  • R 4 of the compound represented by the general formula (A-6) is an alkoxy group
  • a solvent such as methanol, ethanol, 1,4-dioxane, tetrahydrofuran, or a mixed solvent thereof
  • a 0.1 N to 10 N aqueous alkali solution sodium hydroxide aqueous solution, potassium hydroxide aqueous solution, etc.
  • a compound represented by A-7 can be obtained.
  • R 1 , R 2 , R 3 , R 4 , R 5 , R 6 and B in the formula are as defined above.
  • P 2 has the same meaning as the above-mentioned “protecting group for hydroxyl group in nucleic acid synthesis”.
  • a 4,4′-dimethoxytrityl group, a trityl group, a TBDPS group or a TBS group is preferable.
  • P 3 represents a “phosphoramidite group”.
  • “Phosphoramidite group” means a group represented by the formula —P (OR 11 ) (NR 12 ) (wherein R 11 is an alkyl group having 1 to 6 carbon atoms or a cyanoalkyl group having 1 to 7 carbon atoms).
  • R 12 represents an alkyl group having 1 to 6 carbon atoms.), Preferably a group represented by the formula —P (OC 2 H 4 CN) (N (iPr) 2 ) It is a group represented by —P (OCH 3 ) (N (iPr) 2 ).
  • Step 7 is a step wherein compound A-8 is obtained by deprotecting the protecting group R 2 substituted on the amino group of the compound represented by formula (A-7). This step can be performed according to the method described in Protective Groups in Organic Synthesis, Theodora W Green (John Wiley & Sons) and the like.
  • Step 8 is a step for obtaining a compound represented by the general formula (A-9) in which the 2′-position and the 4′-position are cyclized by an amide bond from the compound represented by the general formula (A-8). .
  • other condensing agents or HOBt, HOSu, etc. are added to these condensing agents) and reacted at ⁇ 20 to 120 ° C., preferably 0 to 80 ° C. for 0.5 to 24 hours.
  • a compound represented by the formula (A-9) can be obtained.
  • This step is a general formula (A-10) wherein the 5′-position hydroxyl group of the compound represented by the general formula (A-9) is protected with a protecting group P 2 (for example, 4,4′-dimethoxytrityl group).
  • P 2 for example, 4,4′-dimethoxytrityl group.
  • the compound represented by the general formula (A-9) is dissolved in a solvent such as dichloromethane or toluene in the presence of a base such as pyridine, triethylamine, diisopropylethylamine, DABCO (1,4-diazabicyclo [2.2.2] octane).
  • a base such as pyridine, triethylamine, diisopropylethylamine, DABCO (1,4-diazabicyclo [2.2.2] octane).
  • a pyridine solvent 1.0 equivalent to 5 equivalents, preferably 1.0 equivalent to 1.5 equivalents of 4,4′-dimethoxytrityl chloride is added, and 0 ° C. to 80 ° C., preferably 20 ° C. to 30 ° C.
  • a compound represented by the general formula (A-10) in which P 2 is a 4,4′-dimethoxytrityl group can be obtained by reacting at 0.5 ° C
  • Step 10 This step is a step of obtaining a compound represented by the general formula (A-11) in which the hydroxyl protecting group R 3 bonded to the 3′-position of the compound represented by the general formula (A-10) is deprotected. It is. This step can be performed according to the method described in Protective Groups in Organic Synthesis, Theodora W Green (John Wiley & Sons) and the like.
  • R 3 of the compound represented by the general formula (A-10) is, for example, a silyl protecting group (TBS group, TBDPS group, TES group, etc.), in a solvent such as tetrahydrofuran, methanol, ethanol, dioxane, 1.0 Equivalent to 10.0 equivalents, preferably 1.1 equivalents to 3.0 equivalents of a desilylating agent (fluorine-containing reagents such as tetrabutylammonium fluoride, ammonium fluoride, cesium fluoride, trifluoroacetic acid, hydrochloric acid, etc.
  • the compound represented by the general formula (A-11) can be obtained by reacting with an acid at 0 to 100 ° C., preferably 20 to 80 ° C. for 0.1 to 24 hours.
  • R 3 of the compound represented by the general formula (A-10) is, for example, a benzyl protecting group (Bn group, p-methoxybenzyl group, etc.), it is dissolved in a solvent such as methanol, tetrahydrofuran, ethanol, dioxane, toluene and the like. Or palladium-carbon powder or palladium hydroxide-carbon powder in a mixed solvent thereof, and the mixture is reacted at 0 ° C. to 40 ° C. for 1 hour to 24 hours in a hydrogen stream, thereby obtaining a compound represented by the general formula (A-11) Can be obtained.
  • a solvent such as methanol, tetrahydrofuran, ethanol, dioxane, toluene and the like.
  • palladium-carbon powder or palladium hydroxide-carbon powder in a mixed solvent thereof, and the mixture is reacted at 0 ° C. to 40 ° C. for 1 hour to 24 hours in
  • This step is represented by the general formula (A-12) in which the 3′-position hydroxyl group is amidated by reacting the compound represented by the general formula (A-11) with an amidite forming reagent.
  • This is a step of obtaining a compound.
  • the compound represented by the general formula (A-11) is mixed with 2-cyanoethyl-N, N-diisopropyl in a solvent such as acetonitrile, dichloromethane, tetrahydrofuran or the like in the presence of a base such as diisopropylethylamine or triethylamine.
  • Step 1 This step comprises the step of preparing a compound represented by the general formula (A-10-2) in which the hydroxyl protecting group R 3 bonded to the 3′-position of the compound represented by the general formula (A-9) is deprotected. It is a process to obtain.
  • the compound represented by the general formula (A-10-2) can be obtained by the same method as the third step (steps A-10 to A-11) of the general production method (1) of the four amidites.
  • nucleobase is adenine (A), guanine (G), or cytosine (C) and the amino group is unprotected
  • silylation with a silylating agent such as TMS chloride or TBS chloride in the presence of the base
  • An acylating agent such as benzoyl chloride, phenoxyacetyl chloride, various acid anhydrides, or an iminating agent such as 1,1-dimethoxy-N, N′-dimethylmethanamine is ⁇ 20 ° C. to 120 ° C., preferably 0 ° C. to 50 ° C.
  • reacting with an acid treatment or a desilylating agent to obtain a compound represented by the general formula (A-10-2) in which the amino group of the nucleobase is protected. be able to.
  • Second Step the 5′-position hydroxyl group of the compound represented by the general formula (A-10-2) is protected with a protecting group R 7 (usually a 4,4′-dimethoxytrityl group).
  • R 7 usually a 4,4′-dimethoxytrityl group.
  • This is a step of obtaining a compound represented by 11).
  • the compound represented by the general formula (A-11) can be obtained by the same method as the second step (steps A-9 to A-10) of the general production method (1) of the four amidites.
  • R 1 , R 2 , R 3 , R 5 , R 6 and B are as defined above.
  • R 7 is a hydroxyl-protecting group used for nucleic acid synthesis such as 4,4′-dimethoxytrityl group (or trityl group, TBDPS group, TBS group).
  • Step 1 This step is a step for obtaining a compound represented by the general formula (A-13) by removing the protecting groups at the 2′-position and the 3′-position from the compound represented by the general formula (A-7).
  • the protecting group at the 2 ′ position is a trifluoroacetyl group, it is 1.0 equivalent to 5 equivalents, preferably 1.0 equivalents to 3. equivalents, in a solvent such as tetrahydrofuran, methanol, acetonitrile, pyridine, or a mixed solvent thereof.
  • the compound represented by the general formula (A-13) can be obtained by reacting for a period of time.
  • the characteristic of this reaction is that the trifluoroacetyl group, which is not normally removed by the desilylating agent, is removed, but the nucleobase protecting group such as benzoyl group is not removed. It is a method that makes it possible to improve.
  • R 2 is an acyl protecting group containing a trifluoroacetyl group or a carbamate protecting group
  • a solvent such as methanol, ethanol, 1,4-dioxane, tetrahydrofuran, or a mixed solvent thereof.
  • 1N-10N alkaline solution sodium hydroxide aqueous solution, potassium hydroxide-methanol solution, etc.
  • the compound represented by the general formula (A-13) can be obtained by removing the protecting group at the 'position and then desilylating with TBAF or the like.
  • R 2 is a tertiary butyloxycarbonyl group (BOC group)
  • an acid such as trifluoroacetic acid or hydrogen chloride and ⁇ 20 ° C.
  • a solvent such as methanol, ethanol, 1,4-dioxane, tetrahydrofuran, or without solvent.
  • R 2 is a benzyl-based protecting group (CBz group, Bn group, MPM group, etc.), in a solvent such as methanol, tetrahydrofuran, ethanol, dioxane, toluene, or a mixed solvent thereof, palladium-carbon powder or water
  • a solvent such as methanol, tetrahydrofuran, ethanol, dioxane, toluene, or a mixed solvent thereof
  • a solvent such as methanol, tetrahydrofuran, ethanol, dioxane, toluene, or a mixed solvent thereof
  • palladium-carbon powder or water By adding palladium oxide-carbon powder and reacting at 0 ° C. to 40 ° C. for 1 to 24 hours under a hydrogen stream, the protecting group at the 2′-position is removed, followed by desilylation with TBAF or the like.
  • a compound represented by the general formula (A-13) can be obtained.
  • This step is a step of obtaining a compound represented by the general formula (A-14) in which the 2′-position and the 4′-position are cyclized by amide bonds from the compound represented by the general formula (A-13). .
  • 1.0 equivalent to 10 equivalents preferably 1.0 equivalent to 3 equivalents of DCC, EDC, CDI (carbonyldiimidazole) in a solvent such as N, N′-dimethylformamide, tetrahydrofuran, acetonitrile, or a mixed solvent thereof.
  • a solvent such as N, N′-dimethylformamide, tetrahydrofuran, acetonitrile, or a mixed solvent thereof.
  • other condensing agents or HOBt, HOSu, etc. are added to these condensing agents) and reacted at ⁇ 20 ° C. to 120 ° C., preferably 25 ° C.
  • a compound represented by the formula (A-14) can be obtained.
  • silylation with silylating agents such as TMS chloride, TBS chloride, etc. in the presence of the base results in benzoyl
  • An acylating agent such as chloride, phenoxyacetyl chloride, various acid anhydrides, or an iminating agent such as 1,1-dimethoxy-N, N′-dimethylmethanamine is used at ⁇ 20 ° C. to 120 ° C., preferably 0 ° C. to 50 ° C.
  • the compound represented by the general formula (A-14) in which the amino group of the nucleobase is protected can be obtained by reacting with an acid treatment or a desilylating agent. .
  • This step is a general formula (A-11) wherein the 5′-position hydroxyl group of the compound represented by the general formula (A-14) is protected with a protecting group R 7 (for example, 4,4′-dimethoxytrityl group).
  • R 7 for example, 4,4′-dimethoxytrityl group.
  • B becomes adenine (A), guanine (G ), Cytosine (C), or a compound represented by the general formula (A-7) of these derivatives.
  • Q 1 , Q 2 , and Q 3 in the formula have the same meaning as the aforementioned “alkyl group having 1 to 5 carbon atoms”.
  • This step is a step represented in the compound represented by the above general formula (A-10) from a thymidine-type compound in which the nucleobase part (B) is thymine or a thymine derivative (A-10t). ) Is a step of obtaining a cytidine-type compound which is cytosine or a cytosine derivative (A-10c).
  • a base such as triethylamine, imidazole, N, N-dimethylaminopyridine, and a sulfonyl chloride such as 2,4,6-triisopropylbenzenesulfonyl chloride in a solvent such as acetonitrile, dichloromethane, pyridine, or a mixed solvent thereof;
  • aqueous ammonia is added, and 0 ° C. to 80 ° C., preferably 0 ° C. to 30 ° C., 0.5 to By reacting for 8 hours, a compound represented by the general formula (A-10c) can be obtained.
  • Step 2 This step is a step of obtaining a compound represented by the general formula (A-11c) by further modifying the amino group of cytosine or cytosine derivative from the compound represented by the general formula (A-10c).
  • Bases such as triethylamine, imidazole and N, N-dimethylaminopyridine, and acyl chlorides such as benzoyl chloride and acetyl chloride in a solvent such as N, N′-dimethylformamide, pyridine, acetonitrile, dichloromethane or a mixture thereof
  • it is represented by the general formula (A-11c) by reacting with an acid anhydride such as benzoic anhydride or acetic anhydride at ⁇ 20 ° C. to 80 ° C., preferably 0 ° C. to 30 ° C. for 0.1 to 48 hours. Can be obtained.
  • Step 1 Synthesis of Compound 2
  • a suspension of Compound 1 (4.40 g, 8.05 mmol) in tetrahydrofuran (26 mL) under a nitrogen atmosphere was synthesized according to the method described in Organic Letters, 7, 1569-1572 (2005).
  • saturated aqueous sodium hydrogen carbonate (13 mL) and Cbz chloride (1.26 mL, 8.86 mmol) were added under ice-cooling, and the mixture was stirred at room temperature for 1 hr.
  • Water (13 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (75 mL). The organic layer was washed with saturated brine (25 mL) and dried over sodium sulfate.
  • Step 3 Synthesis of Compound 4 To a chloroform (40 mL) solution of the crude product of Compound 3 obtained in Step 2, 4 mol / L hydrogen chloride / ethyl acetate solution (2.00 mL, 8.00 mmol) was added under ice cooling, The mixture was stirred for 4.5 hours while warming to room temperature. Further, a 4 mol / L hydrogen chloride / ethyl acetate solution (1.34 mL, 5.36 mmol) was added under ice cooling, and the mixture was stirred for 30 minutes.
  • Step 5 Synthesis of Compound 6
  • triethylamine (2.00 mL, 14.4 mmol) and 37% formalin aqueous solution (2.00 mL, 26.9 mmol) were added. It added at room temperature and stirred at 80 degreeC for 4 hours.
  • 1 mol / L hydrochloric acid (20 mL) was added to the reaction solution under ice-cooling, and the mixture was extracted with ethyl acetate (30 mL and 20 mL). The organic layer was washed with water (20 mL) and saturated brine (20 mL), and dried over sodium sulfate. The solvent was distilled off under reduced pressure to obtain a crude product of compound 6.
  • Step 6 Synthesis of Compound 7 Crude product of Compound 6 obtained in Step 5, sodium dihydrogen phosphate dihydrate (551 mg, 3.53 mmol) and 2-methyl-2-butene (1.87 mL, 17.7 mmol) )
  • sodium dihydrogen phosphate dihydrate 551 mg, 3.53 mmol
  • 2-methyl-2-butene (1.87 mL, 17.7 mmol)
  • tert-butanol 10 mL
  • water 3 mL
  • sodium chlorite 800 mg, 8.85 mmol
  • the mixture was stirred for 2.5 hours.
  • To the reaction solution was added 0.2 mol / L aqueous sodium hydroxide solution (20 mL), and the mixture was washed with toluene (20 mL).
  • Step 1 Synthesis of Compound 9 DMF (3.9 L) suspension of Compound 1 (1300 g, 2.53 mol) according to the method described in Organic Letters, 7, 1569-1572 (2005) under nitrogen atmosphere A solution of TBS chloride (1717 g, 11.39 mol) in DMF (2.6 L) was added dropwise at 5 to 15 ° C. Subsequently, sodium iodide (1138 g, 7.59 mol) was added at 10 to 20 ° C., and N-methylimidazole (999 mL, 12.7 mol) was added dropwise at the same temperature. After stirring at 20-30 ° C. for 6 hours, the mixture was allowed to stand overnight.
  • the reaction mixture was poured into a mixture of ethyl acetate (3.9 L) and ice water (10.4 L), and ethyl acetate (2.6 L) and water (2.6 L) were further added.
  • the organic layer was washed successively with 1 mol / L hydrochloric acid (6.5 L), 10% aqueous sodium carbonate solution (6.5 L), and 10% brine (6.5 L).
  • the solvent was distilled off under reduced pressure, and methanol (2.6 L) was added to the obtained residue (2.68 kg).
  • the solvent was distilled off again under reduced pressure to obtain a crude product of compound 9 (2.62 kg).
  • Step 2 Synthesis of Compound 10
  • 35% hydrochloric acid (633 mL, 7.59 mol) was added dropwise at 20-30 ° C. Stir for 4 hours.
  • the reaction mixture was poured into a mixture of diisopropyl ether (6.5 L) and ice water (3.9 L), and diisopropyl ether (6.5 L) and water (2.6 L) were further added.
  • the aqueous layer was washed with diisopropyl ether (3.25 L), then a solution of sodium carbonate (1073 g, 10.1 mol) in water (6.5 L) was added and extracted with ethyl acetate (19.5 L and 6.5 L), The organic layer was dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and tetrahydrofuran (2.6 L) was added to the obtained residue. The solvent was distilled off again under reduced pressure to obtain a crude product of compound 10 (1.08 kg).
  • the physical property data of the obtained compound 10 were as follows.
  • Step 3 Synthesis of Compound 11 To a solution of the crude product of Compound 10 obtained in Step 2 (17.7 g) and pyridine (10.1 mL, 125 mmol) in tetrahydrofuran (80 mL) under a nitrogen atmosphere at 0 to 10 ° C. Acetic anhydride (17.8 mL, 125 mmol) was added dropwise and stirred for 1 hour. The reaction mixture was poured into a 20% aqueous sodium carbonate solution (112 mL) under ice-cooling, and tetrahydrofuran (24 mL) and water (48 mL) were added.
  • Step 4 Synthesis of Compound 12 Under a nitrogen atmosphere, a solution of compound 11 (450 g, 935 mmol) in DMSO (1.35 L) was charged with EDC (538 g, 2800 mmol), DMSO (675 mL), pyridinium paratoluenesulfonate (235 g, 935 mmol) and DMSO (225 mL) was sequentially added, and the mixture was stirred at 20-30 ° C. for 2.5 hours. Subsequently, ethyl acetate (4.5 L), water (2.25 L) and ethyl acetate (2.25 L) were sequentially added at 10 to 20 ° C.
  • Step 5 Synthesis of Compound 13 First, conditions for the aldol reaction performed in Step 5 were examined. Specifically, the combination of the type / amount of the base used, the type / amount of the reagent, the type / amount of the solvent, the reaction temperature, and the reaction time were examined, and the target product, by-product formation rate, and raw material residual rate were determined by HPLC. Analyzed in The results are shown in the following table.
  • Step 6 (2R, 3S, 4R, 5R) -3- (tert-Butyldimethylalkyloxy) -2-hydroxymethyl-5- (5-methyl-2,4-dioxo-3,4-dihydropyrimidine-1 (2H) -yl) -4- (2,2,2-trifluoro-N-methylacetamide) tetrahydrofuran-2-carboxylate (compound 14) in ethyl acetate solution containing compound 13 obtained in synthesis step 5 To the mixture, sodium dihydrogen phosphate dihydrate (292 g, 1870 mmol) in water (1.35 L), squalene (898 mL, 1870 mmol), and acetonitrile (1.35 L) were sequentially added.
  • aqueous layers were combined and washed with ethyl acetate (900 mL), and then ethyl acetate (5.4 L) and a 2 mol / L aqueous hydrochloric acid solution (2.25 L) were added to the aqueous layer.
  • the organic layer was washed with 10% brine (2.25 L) and dried over anhydrous sodium sulfate.
  • the mixture was filtered, and the drying agent was washed with ethyl acetate (1.35 L).
  • potassium acetate (82.5 g, 841 mmol) and ethanol (338 mL) were added. The mixture was stirred at 20-30 ° C. for 1 hour.
  • Step 1 Synthesis of Compound 15 1 mol / L TBAF-tetrahydrofuran solution (0.888 mL, 0.888 mmol) was added dropwise to a suspension of compound 14 (455 mg, 0.807 mmol) in pyridine (4 mL) under a nitrogen atmosphere. After stirring at ⁇ 70 ° C. for 9 hours, the mixture was allowed to stand at room temperature overnight. The next day, the mixture was further stirred at 60 to 70 ° C. for 9 hours, and then allowed to stand overnight at room temperature. After adding EDC hydrochloride (310 mg, 1.62 mmol) and stirring at 60-70 ° C. for 9 hours, EDC hydrochloride (310 mg, 1.62 mmol) was added and allowed to stand overnight at room temperature.
  • EDC hydrochloride 310 mg, 1.62 mmol
  • DMTr (4,4′-dimethoxytrityl) chloride (547 mg, 1.61 mmol) was added and stirred at room temperature for 5 hours, and then DMTr chloride (547 mg, 1.61 mmol) was further added and stirred for 2 hours.
  • 5% Aqueous sodium hydrogen carbonate solution (10 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (10 mL), and washed 3 times with water (5 mL). Each water wash was extracted with ethyl acetate (5 mL).
  • Step 2 Synthesis of Compound 16 (T Amidite) Compound 16 was synthesized according to a method according to the method described in WO2011 / 052436A1.
  • the route from compound 14 to compound 15 can also be synthesized by a method that does not go through a deTFA reaction as follows.
  • Step 1 Synthesis of Compound 30
  • Step 2 Synthesis of Compound 31
  • the crude product of Compound 30 (196 mg) obtained in the previous step was suspended in N, N′-dimethylformamide (1.4 ml), EDC hydrochloride (76 mg) was added, and room temperature For 4 hours.
  • the reaction mixture was filtered to remove inorganic substances (washed with N, N′-dimethylformamide (1.5 ml)), and then water (3.3 ml) was added to precipitate a slurry solid. .
  • Step 3 Synthesis of Compound 32
  • Compound 32 was obtained by treating Compound 31 with 4,4′-dimethoxytrityl chloride in pyridine.
  • Step 4 Conversion of Compound 32 to Compound 15
  • Compound 15 was obtained by treating Compound 32 with a tetrabutylammonium fluoride-tetrahydrofuran solution.
  • Step 1 The conditions of the transglycosylation reaction performed in the synthesis step 1 of Compound 17 were examined. Specifically, the combination amount of the reagent and the type of the solvent was examined, and the target product, by-product formation rate, and the raw material residual rate were analyzed by HPLC. The results are shown in the following table.
  • N O-bis (trimethylsilyl) was added to a suspension of compound 14 (170 g, 271 mmol) and 6-N-benzoyladenine (64.9 g, 271 mmol) in cyclopentyl methyl ether (1700 mL) under a nitrogen atmosphere.
  • Acetamide (466 mL, 1900 mmol) and TMSOTf (98 mL, 543 mmol) were added and stirred at 70-73 ° C. for 4 hours.
  • Tetrahydrofuran (2920 mL), 2 mol / L hydrochloric acid (1700 mL) and water (1020 mL) were added to the reaction solution under ice cooling, and then the organic layer and aqueous layer were partitioned.
  • the organic layer was 0.1 mol / L hydrochloric acid (1700 mL).
  • Chloroform (340 mL) was added to the residue obtained by distilling off the solvent under reduced pressure, and the resulting slurry obtained by stirring for 20 minutes was filtered.
  • the solid collected by filtration was washed with chloroform (510 mL) and then air-dried to obtain Compound 17 (165 g, yield 80.9%).
  • Step 2 Synthesis of Compound 18 Under a nitrogen atmosphere, 1 mol / L TBAF-tetrahydrofuran solution (14.9 mL, 14.9 mmol) was added dropwise to a solution of Compound 17 (4.75 g, 7.44 mmol) in tetrahydrofuran (25 mL). Stir at ⁇ 65 ° C for 3.5 hours. EDC hydrochloride (1.71 g, 8.92 mmol) was added and stirred at 60-65 ° C. for 1 hour. The solvent was distilled off under reduced pressure, acetonitrile (50 mL) was added to the resulting residue, and the solvent was distilled off again under reduced pressure three times to obtain a crude product of compound 18 (9.12 g).
  • Step 3 Synthesis of Compound 19
  • DABCO 2.09 g, 18.6 mmol
  • DMTr chloride 5. 04 g, 14.9 mmol
  • Step 4 Synthesis of Compound 20 (A Amidite) Under a nitrogen atmosphere, a suspension of compound 19 (107 g, 150 mmol) and DIEA (79 mL, 450 mmol, 3.0 eq.) In a pyridine (428 mL, 4 V) suspension under ice cooling under 2-cyanoethyl-N, N-diisopropylchlorophosphoro Amidite (53.6 mL, 240 mmol, 1.6 eq.) was added dropwise and stirred at room temperature for 2 hours. Under ice-cooling, 5% aqueous sodium hydrogen carbonate solution (1.1 L) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (1.1 L).
  • Step 1 Synthesis of Compound 33 Under a nitrogen atmosphere, a suspension of compound 14 (5.0 g, 8.07 mmol) and adenine (1.091 g, 8.07 mmol) in cyclopentyl methyl ether (50 mL) was added to N, O-bis (trimethylsilyl) acetamide (13. 85 mL, 56.5 mmol) and TMSOTf (2.92 mL, 16.15 mmol) were added and stirred at 70 ° C. for 4 hours.
  • Step 2 Synthesis of Compound 34
  • a solution of compound 33 (309 mg, 0.578 mmol) in ethanol (3 ml) was added 1 mol / L lithium borohydride tetrahydrofuran solution (1.156 ml) at 0 ° C., and the mixture was stirred at room temperature for 3 hours.
  • a 4 mol / L hydrogen chloride-dioxane solution (0.578 ml) was added to the reaction mixture, and the mixture was stirred at room temperature for 1 hr, EDC hydrochloride (222 mg, 1.156 mmol.) was added, and the mixture was stirred at 65 ° C. for 2 hr. After allowing to cool, the obtained solid was collected by filtration, washed with methanol, and then air-dried to obtain Compound 34 (187 mg, yield 77%).
  • Step 3 Synthesis of Compound 35 Under a nitrogen stream, compound 34 (149 mg, 0.354 mmol) and methanol (4.5 mL) were added to a 20 ml Falcon tube, followed by addition of ammonium fluoride (52.5 mg, 1.417 mmol) at room temperature, and 60 ° C. And heated at reflux for 8 hours. After allowing to cool, methanol (1.5 ml) and tetrahydrofuran (2 ml) were added, and the resulting slurry was collected by filtration to give compound 35 (89.5 mg, yield 89.5%) as a white solid. .
  • Step 4 Synthesis of Compound 36 An azeotropic dehydration operation of compound 35 (8.00 g, 19.9 mmol) with pyridine (80 ml) was repeated three times with 500 ml of 1 L eggplant colben, and then suspended in pyridine (80 ml). N, N′-dimethylformamide-dimethylacetal (8.0 mL) was added and stirred at room temperature for 1 hour. The resulting orange solution was concentrated as it was, and the residue was subjected to a single azeotropic dehydration operation with pyridine (80 ml) and then dissolved in pyridine (80 ml).
  • Methanol 5.0 ml was added dropwise to the reaction solution, stirred for 40 minutes, and diluted with ethyl acetate.
  • the organic layer was washed twice with saturated aqueous sodium hydrogen carbonate solution and saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated. Again, it was dissolved in pyridine (100 ml), 25% aqueous ammonia (30 ml) was added, and the mixture was stirred at room temperature for 20 hours.
  • Step 5 Conversion of Compound 36 to A (Bz) -DMTr Form (Compound 19) Under a nitrogen stream, HOBt (1.38 g, 10.19 mmol), dichloromethane (20 mL), N-methylmorpholine (2 .17 mL, 19.72 mmol) was added. Thereafter, BzCl (1.15 mL, 9.86 mmol) was added dropwise at room temperature to prepare Bz-OBt. Under a nitrogen stream, Compound 36 (2.00 g, 3.29 mmol), pyridine (20 mL), and N-methylmorpholine (2.17 mL, 19.72 mmol) were added to a 100 mL eggplant flask B.
  • Step 1 Synthesis of Compound 37
  • Compound 37 was obtained by treating Compound 14 with trimethylsilyldiazomethane-n-hexane solution / methanol.
  • Step 2 Synthesis of Compound 38
  • Compound 38 was obtained by subjecting Compound 37 to the same conditions as in Step 2 of Example 2.
  • Step 3 Synthesis of Compound 39 Under a nitrogen stream, Compound 37 (78.0 mg, 0.10 mmol), N-benzoyladenine (47.8 mg, 0.20 mmol), 1,2-dichloroethane (1 mL), BSA were placed in a 10 mL eggplant flask. (0.15 mL, 0.60 mmol) was added. The reaction solution was heated to 60 ° C. and stirred for 40 minutes, and then allowed to cool to room temperature. Trimethylsilyl trifluoromethanesulfonate (0.020 mL, 0.11 mmol) was added at room temperature, and the mixture was heated to reflux for 4 hours.
  • Step 4 Synthesis of Compound 40
  • Compound 38 was obtained by treating Compound 39 with NaOH / methanol.
  • Step 5 Conversion of Compound 40 to Compound 35
  • Compound 35 was obtained by carrying out under the same conditions as in Step 3 of Example 4.
  • Step 6 Conversion from Compound 35 to A (Bz) amidite (Compound 20) The step was carried out under the same conditions as in Step 5 of the method not passing through the deTFA reaction of Example 4 and Step 4 of Example 4. [Example 5]
  • Step 1 Synthesis of Compound 33 The reaction was performed under the same conditions as in Step 1 of the method of Example 4 without passing through the deTFA reaction.
  • Step 2 Conversion from Compound 33 to Compound 35 Synthesized by the same route as in Step 2 of Example 4.
  • Step 3 Conversion from Compound 35 to Compound 36
  • the reaction was carried out under the same conditions as in Step 4 of the method not passing through the deTFAization reaction of Example 4.
  • PacCl (3.85 mL) was added dropwise at room temperature over 3 minutes and then stirred for 20 minutes to prepare Pac-OBt.
  • the above-mentioned trimethylsilylation reaction solution was added to this reaction solution and stirred overnight, and the reaction was incomplete, so the same amount of Pac-Bt solution was added, and the mixture was further stirred for 23 hours.
  • reaction solution was poured into a saturated aqueous sodium hydrogen carbonate solution, the organic layer was separated, and washed successively with a saturated aqueous sodium hydrogen carbonate solution and saturated brine. Each aqueous layer was re-extracted with dichloromethane, and the organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was azeotroped with toluene to remove pyridine, dried, dissolved in tetrahydrofuran (50 ml), added with hydrogen fluoride / triethylamine complex (4 ml), and stirred at room temperature for 40 minutes.
  • reaction mixture was diluted with ethyl acetate, washed twice with saturated aqueous sodium hydrogen carbonate solution and then with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated.
  • the obtained residue was purified by silica gel column chromatography (ethyl acetate / acetone, 100/0 ⁇ 80/20) to give compound 42 (9.91 g, purity 96%, yield 84%) as a beige powder.
  • reaction mixture was diluted with ethyl acetate, washed successively with saturated aqueous sodium hydrogen carbonate solution and saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated.
  • a (amidine) amidite can be synthesized from compound 40 by the following route.
  • Step 1 Synthesis of Compound 44
  • Compound 40 (1.82 g, 2.99 mmol) and pyridine (20 mL) were added to a 100 mL eggplant flask under a nitrogen stream. Thereafter, 1,1-dimethoxy-N, N-dimethylmethanamine (1.192 ml, 8.97 mmol) was added and stirred at room temperature for 30 minutes. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (dichloromethane / methanol, 10/0 ⁇ 9/1) to obtain compound 44 (1.74 g, 87.7%) as white. Obtained as a foamy solid.
  • Step 2 Synthesis of Compound 45 Under a nitrogen stream, Compound 44 (1.70 g, 2.56 mmol), N, N-diisopropylethylamine (2.68 mL, 15.37 mmol), and chloroform (17 mL) were added to a 100 mL eggplant flask. Thereafter, 2-cyanoethyl-N, N-diisopropylchlorophosphoramidite (1.714 ml, 7.68 mmol) was added at 0 ° C., and the mixture was warmed to room temperature and stirred for 4 hours. Saturated multistory water was added to the reaction solution to stop the reaction.
  • Step 1 Synthesis of Compound 21 N, O-bis (trimethylsilyl) was added to a suspension of compound 14 (15.0 g, 26.6 mmol) and guanine (4.02 g, 26.6 mmol) in cyclopentyl methyl ether (135 mL) under a nitrogen atmosphere.
  • Acetamide (65.1 mL, 266 mmol) and cyclopentyl methyl ether (7.5 mL) were added and stirred at 60-70 ° C. for 10 minutes before TMSOTf (11.83 g, 53.20 mmol) and cyclopentyl methyl ether (7.5 mL). ) was added and stirred for 2 hours.
  • Step 2 Synthesis of Compound 22 Under a nitrogen atmosphere, a solution of compound 21 (14.0 g, 23.9 mmol) and N-methylimidazole (7.60 mL, 95.4 mmol) in DMA (70 mL) was added with a 1 mol / L TBAF-tetrahydrofuran solution. (28.6 mL, 28.6 mmol) and DMA (14 mL) were added and stirred at 70-80 ° C. for 8 hours. EDC hydrochloride (13.72 g, 71.55 mmol) and DMA (7 mL) were added at 20-30 ° C. and stirred for 4 hours.
  • N-methylimidazole (7.60 mL, 95.4 mmol) was added to the reaction solution, TBS chloride (14.38 g, 95.40 mmol) was added at 20 ° C. or lower, and the mixture was stirred at room temperature for 2 hours.
  • Ethyl acetate (280 mL) and 10% brine (140 mL) were added to the reaction solution, and the organic layer was washed twice with water (140 mL). Each washing solution was extracted with ethyl acetate (140 mL).
  • Step 3 Synthesis of Compound 23 Trimethylsilyl chloride (9.33 g, 85.9 mmol) was suspended in a pyridine (37.5 mL, 464 mmol) suspension of Compound 22 (7.50 g, 17.2 mmol) at 10 to 20 ° C. under a nitrogen atmosphere. ) was added dropwise, followed by stirring at room temperature for 30 minutes. Next, phenoxyacetyl chloride (3.52 g, 20.6 mmol) was added dropwise at 0 to 5 ° C., followed by stirring at room temperature for 1 hour.
  • the reaction mixture was diluted with ethyl acetate (150 mL), and a mixed solution of 10% brine (37.5 mL) and concentrated hydrochloric acid (30.0 mL) was added dropwise at 0 to 10 ° C.
  • the organic layer was washed twice with 1 mol / L hydrochloric acid (75 mL) and water (75 mL), and then ethyl acetate (37.5 mL) and 2 mol / L hydrochloric acid methanol solution (38.7 mL, 77.4 mmol) were sequentially added. Stir at room temperature for 1 hour.
  • Step 4 Synthesis of compound 24 (G amidite) Under a nitrogen atmosphere, a solution of compound 23 (500 mg, 1.10 mmol) in DMA (3.5 mL) was added DMTr chloride (1.11 g, 3.29 mmol) and DABCO (369 mg, 3 .21 mmol) was added and stirred at room temperature for 2.5 hours. DABCO (369 mg, 3.21 mmol) and 2-cyanoethyldiisopropylchlorophosphoramidite (0.587 mL, 2.63 mmol) were added to the reaction solution at 10 ° C. or lower, and the mixture was stirred at room temperature for 1 hour.
  • the route from compound 14 to compound 21a can also be synthesized by a method that does not go through a deTFA reaction as follows.
  • Step 1 Synthesis of Compound 46 Under a nitrogen stream, cyclopentyl methyl ether of Compound 14 (5.56 g, 8.88 mmol) and 2-acetamido-9H-purin-6-yl-diphenylcarbamate (3.79 g, 9.77 mmol) (50 mL, 10 V)
  • N, O-bis (trimethylsilyl) acetamide BSA, 21.75 mL, 89 mmol
  • trimethylsilyl trifluoromethanesulfonate (3.95 mL, 17 .76 mmol) was added and heated at the same temperature for 4 hours.
  • Step 2 Synthesis of Compound 47
  • a solution of crude compound 46 (9.52 g, corresponding to 8.88 mmol) in methanol (25 ml) was added 28% aqueous ammonia (25 ml), and the mixture was stirred at room temperature for 4 hours.
  • ethyl acetate 250 ml
  • 10% brine 60 ml
  • the precipitated slurry-like solid was collected by filtration.
  • Step 4 Conversion of Compound 48 to Compound 21a Methanol (1.5 mL) and ammonium fluoride (17 mg, 0.458 mmol) were added to compound 48 (100 mg, 0.229 mmol) at room temperature, and 4.5 hours at 70 ° C. Heated to reflux. After cooling to room temperature, the precipitate was collected by filtration, washed with methanol (0.5 ml), and air-dried to obtain Compound 21a (51 mg, yield 68.9%).
  • Process 1 It implemented on the same conditions as the process 1 of the comparative example 1.
  • Process 2 It carried out on the same conditions as the process 2 of the comparative example 1.
  • Process 3 Compound 38 (12 mg, 15 umol) and ON, N-diphenylcarbamoyl-N-acetylguanine (12 mg, 31 umol) were azeotroped with toluene in an eggplant flask (10 mL) (1 mL, once). After drying under a vacuum line, 1,2-dichloroethane (1 mL) was added under a nitrogen atmosphere. N, O-bis (trimethylsilyl) acetamide (BSA, 38 ⁇ L, 154 ⁇ mol) was added, heated to 60 ° C. and stirred for 10 minutes.
  • N-diphenylcarbamoyl-N-acetylguanine (12 mg, 31 umol) were azeotroped with toluene in an eggplant flask (10 mL) (1 mL, once). After drying under a vacuum line, 1,2-dichloroethane (1 mL) was added under a nitrogen atmosphere. N, O-bis (trimethyl
  • TMSOTf trimethylsilyl trifluoromethanesulfonate
  • Process 5 The synthesis was carried out by the same route as in Step 3 of the route not passing through the deTFAation reaction of Example 6.
  • Step 6 The compound was synthesized by the same route as in Step 3 of Example 6.
  • Step 7 It implemented on the same conditions as the process 4 of Example 6.
  • Step 1 Synthesis of Compound 21
  • the compound 21 was prepared under the same conditions as in Step 1 of Example 6.
  • Step 2 Synthesis of Compound 21a The reaction was performed under the same conditions as in Step 2 of Example 6.
  • Step 3 Synthesis of Compound 23 Under a nitrogen stream, Compound 21a (19.5 g, 60.5 mmol) was made into a slurry solution with pyridine (200 ml) and dichloromethane (20 ml), and chlorotrimethylsilane (65.7 g, 605 mmol) was cooled with ice. Dropping was performed. After the dropwise addition, the internal temperature was raised to room temperature, the mixture was stirred for 1.5 hours, the reaction solution was cooled again to ice cooling, and PacCl (11.5 g, 67.4 mmol) was added dropwise.
  • Step 4 Synthesis of Compound 23a Pyridine (200 g) and 4,4′-dimethoxytrityl chloride (21.5 g, 63.5 mmol) were added to the residue obtained by subjecting the crude product of Compound 23 to pyridine azeotropy (60 ml ⁇ 3 times). The mixture was further stirred at room temperature for 2.5 hours. The reaction mixture was added to ethyl acetate (400 ml) and saturated aqueous sodium hydrogen carbonate solution (500 ml), and the layers were separated, the aqueous layer was re-extracted with ethyl acetate (100 ml), and the resulting organic layers were combined and saturated.
  • Step 5 Synthesis of compound 24 To compound 23a (28.3 g, 3 73 mmol), acetonitrile (220 ml) and tetrahydrofuran (30 ml) were added under a nitrogen stream, diisopropylammonium tetrazolide (7.34 g, 42.9 mmol), and then 2-cyanoethyl N, N, N ′, N′-tetraisopropyl phosphoramidite (16.9 g, 55.9 mmol) was added. After stirring at 30 ° C. for 4 hours, the reaction solution was added to ethyl acetate (200 ml) and saturated aqueous sodium hydrogen carbonate (20 ml) for liquid separation.
  • Step 1 to Step 5 The steps can be performed under the same conditions as in Example 6. [Example 9]
  • Step 1 Synthesis of Compound 58
  • Compound 58 can be obtained by adding levulinic acid, EDC hydrochloride and 4-dimethylaminopyridine to a dichloromethane solution of compound 23 (synthesized by the method described in Example 6) and stirring overnight at room temperature.
  • Step 1 Synthesis of Compound 60 To a N, N-dimethylformamide solution of compound 21a (synthesized by the method described in Example 6), tert-butyl-diphenylsilyl chloride and imidazole are added and reacted at room temperature to form a TBDPS at the 5′-position hydroxyl group. After the product is obtained, compound 60 can be obtained by adding trimethylsilyl chloride and phenoxyacetyl chloride to the pyridine solution and reacting at room temperature.
  • Step 2 Synthesis of Compound 61
  • Compound 61 can be synthesized in the same manner as in Step 4 of Example 6. [Example 11]
  • Step 1 Synthesis of Compound 25 108.1 g of Compound 15 (Purity: 92.53%, Content: 100 g, 0.167 mol), DMF 300 ml, Imidazole 47.8 g (4.2 eq.), TBSCl 51.6 g (2.05 eq) .) was added and stirred at around 25 ° C. for 23 hours.
  • the solution was separated by adding 600 ml of AcOEt and 600 ml of water, and the aqueous layer was re-extracted with 300 ml of AcOEt.
  • the organic layers were mixed, washed with 600 ml of 5% NaHCO 3 and 600 ml of 5% NaCl, and the solvent was recovered.
  • Step 2 Synthesis of Compound 26 Compound 25 132.6 g (corresponding to 0.167 mol), MeCN 800 ml, TEA 50.7 g (3 eq.), DMAP 2.0 g (0.1 eq.), TPBSO2Cl 65.8 g (1.3 eq. ) And stirred at around 25 ° C. for 22 hours. 25% NH 3 aq. 800 ml was added and stirred at around 25 ° C. for 2 hours. MeCN was roughly recovered, and 400 ml of AcOEt and 400 ml of water were added for liquid separation, and the aqueous layer was re-extracted with 300 ml of AcOEt.
  • Step 3 192.8 g (corresponding to 0.167 mol) of Compound 26 of Compound 27, 500 ml of DMF, and 75.6 g ( 2 eq.) Of Bz 2 O were added, and the mixture was stirred at around 25 ° C. for 22 hours.
  • AcOEt 1000 ml, 5% NaHCO 3 1000 ml was added for liquid separation, and the aqueous layer was re-extracted with AcOEt 500 ml.
  • the organic layers were mixed, washed with 500 ml of 5% NaHCO 3 and 500 ml of 5% NaCl, and the solvent was recovered to obtain crude compound 5.
  • Step 4 Synthetic compound 27 124.1 g (0.152 mol), THF 869 ml, 1M-TBAF / THF 182 ml (1.2 eq.) Were added, and the mixture was stirred for 2 hours. The solvent was recovered, and liquid separation was performed by adding 620 ml of AcOEt and 620 ml of 5% NaHCO 3 . The solvent was recovered by washing with 620 ml of 5% NaCl. After removing the solvent by toluene 620 ml ⁇ 3 for the purpose of removing TBSOH, the solidified amorphous was loosened to obtain the intended compound 28 (119.2 g, yield 119% from compound 27).
  • Step 5 Synthesis of Compound 29 (MeC (Bz) amidite) 108.2 g (equivalent to 0.138 mol) of Compound 28, 564 ml of dichloromethane and 35.7 g (2 eq.) Of DIPEA were charged and 2-cyanoethyldiisopropylchlorophosphoramidite at 5 ° C. or lower. 49.0 g (1.5 eq.) was added dropwise. Then, it stirred at 25 degreeC vicinity for 2 hours. 564 ml of 5% NaHCO 3 was added for liquid separation, and the solvent was recovered to obtain crude MeC (Bz).
  • the reaction mixture was partitioned by adding tetrahydrofuran (20 mL) and 1 mol / L hydrochloric acid (20 mL) under ice-cooling, and the organic layer was washed with 0.1 mol / L hydrochloric acid (110 mL) and water (110 mL), and then added to anhydrous magnesium sulfate. After drying, the solvent was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform / methanol) to obtain Compound 62 (84 mg, yield 75%). 1 H-NMR was observed as a 71:29 rotamer mixture.
  • Step 1 Synthesis of Compound 25 Synthesis was performed under the same conditions as in Step 1 of Example 11.
  • Step 5 Synthesis of Compound 66
  • the reaction mixture was partitioned by adding tetrahydrofuran (20 mL) and 1 mol / L hydrochloric acid (20 mL) under ice-cooling, and the organic layer was washed with 0.1 mol / L hydrochloric acid (110 mL) and water (110 mL), and then added to anhydrous magnesium sulfate. After drying, the solvent was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform / methanol) to obtain Compound 67 (80 mg, yield 75%).
  • Step 1 Synthesis of Compound 102 N, N′-dimethylformamide solution (10.5 L, 10.5 L) of Compound 101 (3.5 kg, 4.782 mol) derived from D-glucose by the method described in WO 2011/052436 3V) was added potassium carbonate (1652 g, 11.95 mol, 2.5 eq) and paramethoxybenzyl chloride (823.8 g, 5.260 mol, 1.1 eq), and the mixture was stirred at room temperature for 19 hours. Ethyl acetate 35.00L (10v / w) and city water 35.00L (10v / w) were added to this reaction liquid, and extraction and liquid separation were performed to obtain an organic layer I and an aqueous layer I.
  • the aqueous layer I was extracted with 10.50 L (3 v / w) of ethyl acetate to obtain an organic layer II.
  • the obtained organic layers I and II were combined, washed with 24.50 L (7 v / w) of water and 24.50 L (7 v / w) of 10% brine, concentrated and dried under reduced pressure to give compound 102 (4470 g).
  • the crude product was obtained as a brown oil.
  • the obtained compound 102 was not purified, and the yield was quantitative, and it was advanced to the next step.
  • Step 2 Synthesis of Compound 103
  • Compound 102 (crude, 4470 g) was dissolved in tetrahydrofuran (16.30 L, 4 v / w), and city water (407.5 g, 0.1 w / w) was added.
  • city water 407.5 g, 0.1 w / w
  • n-tributylphosphine (1258 g, 6.218 mol, 1.3 eq) was added dropwise at a liquid temperature of 30 ° C. or lower, and the mixture was stirred at the same temperature for 5 hours.
  • Step 3 Synthesis of Compound 104
  • Compound 103 (crude, 5557 g) was dissolved in dichloromethane (7.900 L (2 v / w)), and pyridine (945.6 g, 11.95 mol, 2.5 eq) was added.
  • TFAA (1507 g, 7.175 mol, 1.5 eq) was added dropwise at a liquid temperature of 35 ° C. or lower, and the mixture was stirred at the same temperature for 2 hours.
  • the reaction was diluted with CH 2 Cl 2 (19.75 L, 5 v / w), cold 10% Na 2 CO 3 aq. (11.85 L, 3 v / w), extraction and liquid separation were performed, and the obtained organic layer was added with 5% NaHCO 3 aq.
  • Step 4 Synthesis of Compound 105
  • Compound 104 + n-Bu 3 PO (4392 g) was dissolved in 10.60 L (3 v / w) of DMF at a liquid temperature of 50 ° C., and potassium carbonate (1062 g, 7.684 mol, 2 eq) was added.
  • potassium carbonate (1062 g, 7.684 mol, 2 eq) was added.
  • iodomethane (818.3 g, 5.765 mol, 1.5 eq) was added dropwise at a liquid temperature of 55 ° C. or lower, and the mixture was stirred at the same temperature for 7 hours and at room temperature for 15 hours.
  • Step 5 Synthesis of Compound 106
  • Crude product (4386 g) of Compound 105 was dissolved in acetonitrile (25.20 L, 7 v / w), and the solution temperature was 30 ° C. or less, and 7586 g (13.84 mol, 3.6 eq) + city of CAN A water (9.0000 L, 2.5 v / w) solution was added dropwise over 30 minutes, and the mixture was stirred at the same temperature for 2 hours.
  • Ethyl acetate (36.00 L, 10 v / w) and city water (25.20 L, 7 v / w) were added to this reaction solution for extraction and liquid separation, and the resulting organic layer was washed with city water (25.20 L, 7 v / w).
  • a 5% sodium hydrogen oxyacid aqueous solution (25.20 L, 7 v / w) was added thereto, and the resulting insoluble matter was filtered (AcOEt 8.000 L (2.2 v / w) was used for washing).
  • Step 1 Synthesis of Compound 107
  • Compound 100 (2.38 mmol) derived from D-glucose by the method described in WO 2011/052436 was dissolved in methylamine (20 mL, 2.0 mol / L THF solution, 40 mmol). And heated to 60 ° C. using a microwave and stirred for 8 hours. The reaction solution was concentrated to dryness to obtain Compound 107. The obtained compound was not purified and proceeded to the next reaction as it was.
  • Step 2 Conversion of Compound 107 to Compound 106
  • Compound 107 obtained in the previous step was dissolved in dichloromethane (10 mL), pyridine (481 ⁇ l, 5.96 mmol) was added, and TFAA (504 ⁇ L, 3.58 mmol) was added dropwise at room temperature. Stir for 2 hours. Saturated aqueous ammonium chloride solution was added and extracted. The obtained organic layer was washed with a saturated aqueous sodium hydrogen carbonate solution and saturated brine, and dried over magnesium sulfate.
  • Step 1 Synthesis of Compound 107 Under a nitrogen stream, Compound 106 (599.60 g, 734.85 mmol), Bz-adenine (351.82 g, 2 eq.), 1,2-DCE (3000 ml, 5 v / w) were added to a 20 L flask and dissolved by stirring. did. BSA (899.50 g, 6 eq.) Was added to this solution, the temperature was raised, and the mixture was stirred for 1 hour while maintaining at 49 ° C. After cooling to 24 ° C., TMSOTf (49.55 g, 0.3 eq.) Was added, the temperature was raised again, and stirring was continued for 5.5 hours while maintaining the temperature at 60 ° C.
  • the reaction solution was cooled to 5 ° C., and 8% aqueous sodium hydrogen carbonate solution (900 g) was added dropwise. After completion of the dropwise addition, this was stirred at room temperature for 15 hours (beige slurry).
  • DCM (7800 g) was added thereto, and the mixture was stirred at room temperature for 1 hour, and the solid was separated by filtration through celite.
  • the filtered residue was slurried again with DCM (4800 g) and the solid was filtered off through celite filtration. Further, the residue was slurry washed with DCM (3000 g), and the solid was separated by suction filtration (this operation was repeated twice).
  • Step 2 Synthesis of Compound 108 Under a nitrogen stream, 1.0 mol / L-TBAF / THF solution (766.44 g, 1.2 eq.) was added to a THF (1290 ml, 2 v / w) solution of Compound 107 (645 g, 694.24 mmol) at room temperature. The mixture was stirred at 24 ° C. for 21 hours. The reaction solution was concentrated under reduced pressure, dissolved in ethyl acetate (1743 g), transferred to a 10 L water washing kettle, and washed with city water (2142 g). The organic layer was washed again with 5% brine (1290 g) and the brine was extracted again with ethyl acetate (322 g).
  • Step 3 Synthesis of Compound 109
  • Compound 108 (402 g, 582.04 mmol), acetonitrile (3216 ml, 8 v / w), 1 mol / l aqueous acetic acid solution (543 ml, 1.35 v / w) in a 10 L four-necked flask under a nitrogen stream 1 mol / l sodium acetate aqueous solution (1085 ml, 2.7 v / w), TEMPO (9.09 g, 0.1 eq.), 80% sodium chlorite (138.13 g, 2.1 eq.), 9% hypochlorous acid Sodium chlorate (6.0 ml, 0.015 v / w) was sequentially added, the internal temperature was raised to 36 ° C., and the mixture was stirred while keeping warm.
  • Step 4 Synthesis of Compound 110 Under a nitrogen stream, 25% aqueous ammonia (468 ml, 1 v / w) was added to a solution of compound 109 (468 g, content 410 g, 582.04 mmol) in methanol (2800 ml, 6 v / w) at room temperature. After that, the temperature was raised to 39 ° C., and the mixture was stirred while keeping warm. After stirring for 6 hours, 25% aqueous ammonia (140 ml, 0.3 v / w) was added. After further stirring for 16 hours, 25% aqueous ammonia (94 ml, 0.2 v / w) was added.
  • Step 5 Synthesis of Compound 111
  • Compound 110 (470 g, contents 294 g, 582.04 mmol) and DMF (1800 ml, 6.1 v / w) were added to a 10 L four-necked flask under a nitrogen stream, and the mixture was stirred while keeping at 24 ° C. .
  • EDC hydrochloride (169 g, 1.5 eq.) was added and stirred while keeping warm.
  • EDC hydrochloride 23 g, 0.2 eq.
  • city water (5124 g) was added dropwise over 15 minutes.
  • Step 6 Synthesis of Compound 112 Under a nitrogen stream, Compound 111 (225 g, 462.47 mmol) and acetic acid (1125 ml, 5 v / w) were added to a 3 L flask, and the mixture was heated to 50 ° C. and dissolved by stirring. 20% Pd (OH) 2 -C (44.98 g, 0.2 w / w) was added thereto, and the inside of the system was replaced with hydrogen, followed by stirring while keeping hydrogen blown into the reaction solution. After stirring for 6 hours, 20% Pd (OH) 2 —C (22.49 g, 0.1 w / w) was added.
  • the route from compound 106 to compound 110 can also be synthesized by the following method.
  • Step 1 Synthesis of Compound 113
  • the compound 113 can be synthesized under the same conditions as in Step 2 of the synthesis of A (Bz) amidite in Reference Example 1 described above.
  • Step 2 Synthesis of Compound 114
  • the compound 114 can be synthesized under the same conditions as in Step 3 of the synthesis of A (Bz) amidite in Reference Example 1 described above.
  • Step 3 Derivation of Compound 114 to Compound 110
  • Compound 114 (14 mg, 0.023 mmol) was dissolved in 1 mL of dichloroethane, and 16 mg (0.069 mmol) of N-benzoylguanine and 42 mg of (E) -trimethylsilyl N-trimethylsilylacetamide (0. 21 mmol) was added and heated to 60 ° C. and stirred for 20 minutes. After the reaction solution was lowered to room temperature, TMSOTf (4 ⁇ l, 0.023 mmol) was added, and the mixture was heated again to 60 ° C. and stirred for 6 hours.
  • Step 1 Synthesis of Compound 115 Under a nitrogen stream, ON, N-diphenylcarbamoyl-N-isobutylguanine (93.0 g, 223 mmol), 1,2-dichloroethane (760 ml) was charged, and BSA (136. 2 g, 670 mmol) was added dropwise. The internal temperature was heated to 60 ° C. and stirred for 30 minutes. Thereafter, the internal temperature was lowered to ice-cooling, compound 106 (152 g, 186 mmol) was added, and after washing with 1,2-dichloroethane (150 ml), TMSOTf (l2.4 g, 55.8 mmol) was added dropwise. It was.
  • Step 2 Synthesis of Compound 116 After adding ammonium fluoride (77.3 g, 2.09 mol) to a methanol (500 g) solution of Compound 115 (192.5 g, 1.74 mmol) at room temperature, the bath temperature was raised to 68 ° C. For 3.5 hours. After cooling to room temperature, the reaction solution was added to water (1 L) and ethyl acetate (1.5 L). Liquid separation was performed, and the aqueous layer was extracted with ethyl acetate (500 ml ⁇ 2). The organic layers were combined, washed with saturated brine (500 ml), and concentrated.
  • Step 3 Synthesis of Compound 117
  • Compound 116 (87 g) was dissolved in acetonitrile (440 g) at room temperature, and 1 mol / L acetic acid (87 ml) and 1 mol / L sodium acetate (174 ml) were added. Further TEMPO (1.8 g, 11.6 mmol) was added followed by 80% sodium chlorite (NaClO2, 27.5 g, 243 mmol) and 12% sodium hypochlorite (NaClO, 1.5 ml). The mixture was stirred at around 28 ° C. for 2.5 hours, and 12% sodium hypochlorite (NaClO, 1.5 ml) was added (the reaction solution changed from amber to brown).
  • Step 4 Synthesis of Compound 118
  • Crude compound 117 113 g was dissolved in methanol (400 g), 25% aqueous ammonia (200 ml) was added, and the mixture was stirred at 40 ° C. overnight.
  • the reaction mixture was concentrated and azeotroped with toluene (200 ml ⁇ 3 times) to obtain a crude product of Compound 118 (112.8 g).
  • Step 5 Synthesis of Compound 119
  • Compound 118 (112.8 g, corresponding to 115.7 mmol) was dissolved in N, N′-dimethylformamide (440 g) at room temperature under a nitrogen stream, and EDC hydrochloride (33.3 g, 173) was dissolved. .6 mmol) was added.
  • EDC hydrochloride (17.8 g, 92.8 mmol) was added while stirring as it was and confirming the progress of the reaction. After confirming the completion of the reaction, the reaction solution was dispersed under stirring with water (1760 g) and ethyl acetate (25 g).
  • Step 6 Conversion from Compound 119 to Compound 21a Under a nitrogen stream, compound 119 (40 g, 79.6 mmol) was mixed with acetic acid (400 ml) at room temperature, palladium hydroxide (12.5 g) was added, and then hydrogen atmosphere was added. Stir for 24 hours at 30 ° C. Since crystals precipitated in the reaction system, water (80 g) was added, and the mixture was further stirred for 2 days. Then, the catalyst was filtered off, and the filtrate was concentrated.
  • the route from compound 106 to compound 118 can also be synthesized by the method described in the following scheme.
  • Example of synthesis of T amidite Conversion of compound 106 to compound 14b can be carried out by the method described in the following scheme.
  • the compound 14b can be synthesized by the method described in Example 3 or International Publication No. 2011/052436.
  • Step 1 Synthesis of Compound 124 Chemische Berichte, 101 (7), 2289-93 (1968). To obtain Compound 124 from D-glucosamine hydrochloride.
  • Step 2 Synthesis of Compound 125 Under a nitrogen stream, a suspension of sodium hydride (0.89 g, 22.2 mmol) in N, N′-dimethylformamide (30 mL) was charged with Compound 124 (5.2 g, 17.1 mmol) in N, N′-dimethylformamide (30 mL) was added dropwise and stirred for 30 minutes. Subsequently, benzyl bromide (2.4 mL, 20.5 mmol) was added dropwise to the reaction solution at 7-8 ° C., and the mixture was stirred at room temperature for 90 minutes. A saturated aqueous ammonium chloride solution was added, and the mixture was extracted with tert-butyl methyl ether.
  • Step 3 Synthesis of Compound 126
  • N, O-bistrimethylsilylacetamide (4.7 mL, 18.9 mmol) was added to a suspension of thymine (1.43 g, 11.4 mmol) in acetonitrile (15 mL) at 80 ° C. Stirred for 60 minutes under.
  • a solution of compound 125 (3.0 g, 7.6 mmol) in acetonitrile (15 mL) and tin tetrachloride (0.89 mL, 7.6 mmol) were added, and the mixture was stirred at 80 ° C. for 6 hours. After cooling to room temperature, saturated multistory water was added, and the precipitated solid was removed by Celite filtration.
  • the reaction mixture was concentrated under reduced pressure, extracted from the residue with ethyl acetate, and washed with water and saturated brine.
  • the crude product 125a (4.1g) was obtained by drying with sodium sulfate and concentrating under reduced pressure.
  • the crude product 126 (4.1 g) was dissolved in methanol (30 mL) and water (10 mL), camphorsulfonic acid (0.73 g, 3.1 mmol) was added at room temperature, and the mixture was stirred for 2 hours.
  • the mixture was neutralized by adding a 1 mol / L aqueous sodium hydroxide solution and then concentrated under reduced pressure.
  • Step 4 Synthesis of Compound 127
  • sodium periodate (2.3 g, 10.6 mmol) was added to a tetrahydron-water mixed solution of compound 126 (3.4 g, 7.1 mmol) (1: 1, 30 mL). And stirred for 1 hour.
  • Ethylene glycol (0.4 mL, 7.1 mmol) was added and stirred for a while, and the resulting white solid was removed by filtration.
  • the organic layer was extracted with ethyl acetate, washed with water and saturated brine, and dried over sodium sulfate. The organic layer was distilled off under reduced pressure to obtain a crude product 127 (3.6 g).
  • Step 6 Synthesis of Compound 129
  • 2-methyl-2-butene (7.8 mL, 7.8 mL) was added to a mixed solution of crude product 128 (3.6 g) in tert-butanol-water-tetrahydrofuran (5: 5: 1, 33 mL).
  • 75 mmol sodium dihydrogen phosphate (2.7 g, 22.5 mmol) and sodium chlorite (1.4 g, 15 mmol) were added and stirred for 2 hours 30 minutes.
  • a 1 mol / L hydrochloric acid aqueous solution was added to adjust the reaction solution to pH 2 to 3, followed by extraction with chloroform.
  • the organic layer was washed with water and saturated brine, and dried over sodium sulfate.
  • the organic layer was distilled off under reduced pressure to obtain a crude product 129 (3.7 g).
  • Step 7 Synthesis of Compound 130 A 2 mol / L trimethylsilyldiazomethane solution (4.9 mL, 9.7 mmol) was added dropwise to a toluene-methanol solution (2: 1, 30 mL) of the crude product 129 (3.7 g) at room temperature. And stirred for 40 minutes. Acetic acid (0.2 mL) was added, and the reaction mixture was concentrated under reduced pressure. The obtained solid was recrystallized from ethyl acetate / methanol to obtain compound 130 (1.54 g, 3.1 mmol) as a white solid.
  • step 3 The conditions for the glycosylation reaction performed in step 3 were examined. Specifically, the combination of acid catalyst and solvent was examined, and the product and the remaining amount of raw material were analyzed by HPLC or the like. As a result, it has been found that the combination shown in Condition 7 as shown in the following table is the optimum condition that gives the highest product yield and the lowest raw material residual rate and byproduct production rate.
  • Step 6 Compound 153 (480 mg, 0.97 mmol) obtained in the fifth step was dissolved in pyridine (1.5 ml). Under ice-cooling, mesyl chloride (0.18 ml, 2.28 mmol) was added, and the mixture was stirred at 0 ° C. for 30 minutes. The reaction solution was diluted with ethyl acetate, extracted with water. The obtained organic layer was washed 3 times with 2 mol / L aqueous hydrochloric acid, then washed with a saturated aqueous sodium hydrogen carbonate solution and saturated brine, and dried over magnesium sulfate. The solvent was distilled off under reduced pressure to obtain Compound 154 (520 mg, 93% yield) as amorphous.
  • Step 7 Compound 154 (515 mg, 0.90 mmol) obtained in step 6 was dissolved in acetonitrile (5 ml). Diazabicycloundecene (0.18 ml, 1.20 mmol) was added at room temperature, and the mixture was stirred at room temperature for 7 hours. The reaction mixture was diluted with ethyl acetate under ice-cooling, and extracted with 5% aqueous citric acid solution. The obtained organic layer was washed with 5% aqueous citric acid solution, saturated aqueous sodium hydrogen carbonate solution and saturated brine, and dried over magnesium sulfate. The solvent was distilled off under reduced pressure to obtain a pale yellow oil.
  • Step 9 Compound 122 (17 mg, 0.90 mmol) obtained in the eighth step was dissolved in a mixed solvent of tetrahydrofuran (1 ml) and methanol (0.2 ml). 20% Parazimed carbon (8 mg) was added and stirred for 2 hours under hydrogen atmosphere. Nitrogen substitution was performed, the catalyst was removed by filtration, and the solvent was distilled off from the filtrate under reduced pressure. Recrystallization of the obtained colorless oil (methanol / chloroform) gave Compound 14b (10 mg, 93% yield) as a white solid.
  • T amidite (compound 16) and MeC (Bz) amidite (compound 29) can be obtained from compound 14b by the method described in Example 3, Example 11 or International Publication No. 2011/052436.
  • Compound 122 was converted to 1) NaOH aq.
  • Compound 156 was obtained in two steps by treatment with / THF 2) (CF 3 CO) 2 O / pyridine.
  • Compound 156 can be derived into A (Bz) amidite (Compound 20) and G (Pac) amidite (Compound 24) by the method of Reference Example 1.

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Abstract

La présente invention concerne un procédé pour la production d'un dérivé d'acide nucléique réticulé par amide, qui est utile comme molécule en vue d'une utilisation comme anti-sens, avec une efficacité élevée. Un intermédiaire qui est important pour la synthèse d'un dérivé d'acide nucléique réticulé par amide peut être synthétisé avec une efficacité élevée par mise en œuvre d'une réaction aldolique dans des conditions spécifiées. L'intermédiaire peut être fermé en un cycle avec une efficacité élevée par élimination d'un groupe protecteur à partir de celui-ci.
PCT/JP2014/050325 2013-01-10 2014-01-10 Procédé de production de dérivé d'acide nucléique réticulé Ceased WO2014109384A1 (fr)

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WO2017047816A1 (fr) * 2015-09-18 2017-03-23 田辺三菱製薬株式会社 Acide nucléique réticulé guna, procédé de production de ce dernier, et composé intermédiaire
WO2018123925A1 (fr) 2016-12-28 2018-07-05 第一三共株式会社 Médicament thérapeutique pour le syndrome d'alport
WO2019009299A1 (fr) 2017-07-05 2019-01-10 国立大学法人大阪大学 OLIGONUCLÉOTIDE ANTISENS ENA POUR L'INHIBITION DE L'EXPRESSION DE L'α-SYNUCLÉINE
WO2019172286A1 (fr) 2018-03-09 2019-09-12 第一三共株式会社 Agent thérapeutique pour la maladie du stockage du glycogène de type ia
WO2019240164A1 (fr) 2018-06-13 2019-12-19 第一三共株式会社 Agent thérapeutique contre le dysfonctionnement myocardique
WO2021010301A1 (fr) 2019-07-12 2021-01-21 第一三共株式会社 OLIGONUCLÉOTIDE ANTISENS PERMETTANT DE MODIFIER L'ÉPISSAGE pré-ARNm DE DUX4
WO2021049504A1 (fr) 2019-09-10 2021-03-18 第一三共株式会社 Conjugué galnac-oligonucléotide pour une utilisation d'administration ciblée sur le foie, et son procédé de production
WO2023176862A1 (fr) 2022-03-16 2023-09-21 第一三共株式会社 ARNsi POUR SUPPRIMER L'EXPRESSION DU RÉCEPTEUR 2 DE LA TRANSFERRINE
WO2023176863A1 (fr) 2022-03-16 2023-09-21 第一三共株式会社 Oligonucléotide chimiquement modifié ayant une activité d'arni
US11963974B2 (en) 2017-03-10 2024-04-23 National Center For Child Health And Development Antisense oligonucleotide and composition for prevention or treatment of glycogen storage disease type Ia
WO2024185775A1 (fr) 2023-03-06 2024-09-12 スペラファーマ株式会社 Dérivé de benzoyle substitué par alcényloxy à longue chaîne et procédé de synthèse d'oligonucléotides l'utilisant
WO2026034588A1 (fr) * 2024-08-07 2026-02-12 株式会社大阪合成有機化学研究所 Cristal de composé réticulé par un amide et son procédé de production

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Cited By (15)

* Cited by examiner, † Cited by third party
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WO2017047816A1 (fr) * 2015-09-18 2017-03-23 田辺三菱製薬株式会社 Acide nucléique réticulé guna, procédé de production de ce dernier, et composé intermédiaire
US10961269B2 (en) 2015-09-18 2021-03-30 Mitsubishi Tanabe Pharma Corporation Bridged nucleic acid GuNA, method for producing same, and intermediate compound
WO2018123925A1 (fr) 2016-12-28 2018-07-05 第一三共株式会社 Médicament thérapeutique pour le syndrome d'alport
US11963974B2 (en) 2017-03-10 2024-04-23 National Center For Child Health And Development Antisense oligonucleotide and composition for prevention or treatment of glycogen storage disease type Ia
WO2019009299A1 (fr) 2017-07-05 2019-01-10 国立大学法人大阪大学 OLIGONUCLÉOTIDE ANTISENS ENA POUR L'INHIBITION DE L'EXPRESSION DE L'α-SYNUCLÉINE
US11958878B2 (en) 2018-03-09 2024-04-16 Daiichi Sankyo Company, Limited Therapeutic agent for glycogen storage disease type IA
WO2019172286A1 (fr) 2018-03-09 2019-09-12 第一三共株式会社 Agent thérapeutique pour la maladie du stockage du glycogène de type ia
WO2019240164A1 (fr) 2018-06-13 2019-12-19 第一三共株式会社 Agent thérapeutique contre le dysfonctionnement myocardique
WO2021010301A1 (fr) 2019-07-12 2021-01-21 第一三共株式会社 OLIGONUCLÉOTIDE ANTISENS PERMETTANT DE MODIFIER L'ÉPISSAGE pré-ARNm DE DUX4
WO2021049504A1 (fr) 2019-09-10 2021-03-18 第一三共株式会社 Conjugué galnac-oligonucléotide pour une utilisation d'administration ciblée sur le foie, et son procédé de production
WO2023176863A1 (fr) 2022-03-16 2023-09-21 第一三共株式会社 Oligonucléotide chimiquement modifié ayant une activité d'arni
WO2023176862A1 (fr) 2022-03-16 2023-09-21 第一三共株式会社 ARNsi POUR SUPPRIMER L'EXPRESSION DU RÉCEPTEUR 2 DE LA TRANSFERRINE
WO2024185775A1 (fr) 2023-03-06 2024-09-12 スペラファーマ株式会社 Dérivé de benzoyle substitué par alcényloxy à longue chaîne et procédé de synthèse d'oligonucléotides l'utilisant
EP4678626A1 (fr) 2023-03-06 2026-01-14 Spera Pharma, Inc. Dérivé de benzoyle substitué par alcényloxy à longue chaîne et procédé de synthèse d'oligonucléotides l'utilisant
WO2026034588A1 (fr) * 2024-08-07 2026-02-12 株式会社大阪合成有機化学研究所 Cristal de composé réticulé par un amide et son procédé de production

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