WO2001000641A1 - Nouveaux dérivés de bicyclonucléoside - Google Patents

Nouveaux dérivés de bicyclonucléoside Download PDF

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Publication number
WO2001000641A1
WO2001000641A1 PCT/JP2000/004092 JP0004092W WO0100641A1 WO 2001000641 A1 WO2001000641 A1 WO 2001000641A1 JP 0004092 W JP0004092 W JP 0004092W WO 0100641 A1 WO0100641 A1 WO 0100641A1
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group
amino
nucleic acid
acid synthesis
protected
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PCT/JP2000/004092
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English (en)
Japanese (ja)
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Takeshi Imanishi
Satoshi Kohiga
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Sankyo Company, Limited
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Priority to AU54290/00A priority Critical patent/AU5429000A/en
Publication of WO2001000641A1 publication Critical patent/WO2001000641A1/fr

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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
    • 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
    • C07H19/10—Pyrimidine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids
    • 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
    • C07H19/16—Purine radicals
    • 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
    • C07H19/16—Purine radicals
    • C07H19/20—Purine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids
    • 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 an intermediate for producing a non-naturally occurring oligonucleotide analog having excellent antisense or antigene activity and being stable in vivo.
  • the present invention relates to a novel oligonucleotide analogue containing one or more of the bicyclonucleoside structures.
  • BACKGROUND ART Oligonucleotides having excellent antisense or antigene activity and being stable in vivo are expected to be useful drugs.
  • IS1S has developed ISISSIS222, a chicken-type oligonucleotide, for the treatment of human cytomegalovirus retinitis. It is sold in the United States as Vitravene TM .
  • non-natural type oligonucleotide analogs that is, the ability to form a stable complementary strand with m-RNA or DNA, the stability to various nucleases, Considering the occurrence of side effects due to non-specific binding to various proteins, etc., non-natural type that has more excellent antisense or antigene activity, is stable in vivo, and has few side effects Origonucleotide analogues are desired.
  • a compound which is an intermediate for the production of a non-natural type oligonucleotide analog and is related to the intermediate of the present invention is described in JP-A-10-195998.
  • B is a pyrimidine or purine nucleobase or an analog thereof, and Y 1 and Y 2 are the same or different and represent a hydrogen atom or a hydroxyl-protecting group.
  • 2-5A which is known as an in vivo substance having antiviral activity, has three or more hydroxyl groups of 2 'and 5' of adenosine linked by a phosphodiester group, and a tri-terminal group at the 5 'end. Oligonucleotide to which phosphoric acid is bound.
  • INF interferon
  • dsRNA double-stranded form of RNA that the virus takes up during the growth process.
  • the enzyme synthesizes 2-5A in the presence of ATP.
  • 2-5A converts inactive RNaseL (RNA degrading enzyme) to active RNaseL, and this active RNaseL will By degrading the m-RNA of the virus, the growth of the virus in the cell is stopped.
  • 2-5A plays an important role in the expression of the antiviral effect of INF, but is degraded by 2 ', 5'-phosphodiesterase to ATP or AMP in which the effect is not expressed .
  • An object of the present invention is to provide an intermediate for producing a non-naturally occurring oligonucleotide derivative which has excellent antisense or antigene activity and is stable in vivo, and also has an antiviral activity.
  • An object of the present invention is to provide a novel bicyclonucleoside derivative, which is an intermediate for producing a 2-5A analog known as an in vivo substance.
  • the inventor of the present invention has conducted intensive studies to solve the above-mentioned problem, and as a result, a novel bicyclonucleoside derivative having 3′-amino-3′-N, 4′-C-methylene has been developed. That the derivative is useful as an intermediate for producing a nucleonucleotide analog; that the derivative is useful as an intermediate for producing a 2-5A analog; The present inventors have found that a novel oligonucleotide analog having one or more reoside structures has excellent nuclease resistance and can be an excellent antisense or antigene drug, and thus completed the present invention. . DISCLOSURE OF THE INVENTION The novel bicyclonucleoside derivative of the present invention
  • R 4 a R 4b
  • R 4 and R 4b are the same or different and each is a hydroxyl group, a hydroxyl group protected with a protecting group for nucleic acid synthesis, a mercapto group, a mercapto group protected with a protecting group for nucleic acid synthesis, Amino group, amino group protected with a protecting group for nucleic acid synthesis, alkoxy group having 1 to 6 carbon atoms, alkylthio group having 1 to 6 carbon atoms, cyanoalkoxy group having 1 to 7 carbon atoms or 1 carbon atom Represents an amino group substituted with 6 to 6 alkyl groups).
  • R 2 represents a hydrogen atom, an alkyl group having 1 to 6 carbon atoms or a protecting group for nucleic acid synthesis of an amino group
  • B represents a purine 91-yl or 2-oxo-1,2-dihydropyrimidin-11-yl group which may have a substituent selected from the following group: And a pharmacologically acceptable salt thereof.
  • a mercapto group protected with a protecting group for nucleic acid synthesis An alkylthio group having 1 to 6 carbon atoms,
  • bicyclonucleoside derivatives of the present invention preferred are:
  • R 1 is a hydrogen atom, an aliphatic acyl group, an aromatic acyl group, a methyl group substituted with 1 to 3 aryl groups, lower alkyl, lower alkoxy, halogen or cyano group.
  • R 1 is a hydrogen atom, an acetyl group, a benzoyl group, a benzyl group, a p-methoxybenzyl group, a dimethoxytrityl group, a monomethoxytrityl group or a tert-butyldiphenylsilyl group.
  • R 2 is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, an aliphatic acyl group, an aromatic acyl group, or an aryl ring having 1 to 2 lower alkoxy or nitro groups. Is a substituted or unsubstituted arylalkyloxycarbonyl group,
  • R 2 may be substituted with a hydrogen atom, an alkyl group having 1 or 2 carbon atoms, an aliphatic acyl group, or 1 or 2 lower alkoxy or 2-port groups in the aryl ring.
  • R 2 is a hydrogen atom, a methyl group, a trifluoroacetyl group or a benzyloxycarbonyl group.
  • R 3 is a hydrogen atom, an aliphatic acyl group, an aromatic acyl group, a methyl group substituted with 1 to 3 aryl groups, a lower alkyl, a lower alkoxy, a halogen or a cyano group, and the aryl ring is Methyl group, silyl group, phosphoramidite group, phosphonyl group substituted with 1 to 3 substituted aryl groups 8)
  • R 3 is a hydrogen atom, an acetyl group, a benzoyl group, a benzyl group, or a p-methoxybenzene.
  • B is a 6-amino-purine-9-yl group (that is, an adenylyl group), a 6-amino-purine-9-yl group in which an amino group is protected by a protecting group for nucleic acid synthesis, 2, 6-diaminopurine 9-yl group, 2-amino-6-chloropurine-9-yl group, 2-amino-6-amino-amino group protected with a protecting group for nucleic acid synthesis, 2-amino-6-chloro-purine 9-yl group, 2— Amino-6-fluorinyl-1-yl group and 2-amino-6-fluoropurine-9-yl group, 2-amino-6-bromopurine-9-yl group in which the amino group is protected with a protecting group for nucleic acid synthesis.
  • 2-amino-6-bromopurin-9-yl group and 2-amino-6-hydroxypurine-9-yl group in which the amino group is protected by a protecting group for nucleic acid synthesis ie, guaninyl group
  • the amino group being protected with a protecting group for nucleic acid synthesis is protected by a protecting group for nucleic acid synthesis.
  • Droxypurine—9-yl, amino and hydroxyl groups protected with a protecting group for nucleic acid synthesis 2—amino-6—hydroxypurine-9-yl, 6-amino-2-methoxypurine-19-yl, 6-Amino-2—chloropurine 9-yl group, 6—Amino-2_fluoroprin-19-yl group, 2, 6—Dimethoxypurine-19-yl group, 2,6-Dichloropurine-19-1 6-mercaptopurine 9-yl group, 2-oxo-1 4-amino-1,2, -dihydropyrimidine-1 1-yl group (ie, cytosyl group), and amino group are protecting groups for nucleic acid synthesis.
  • B-force 6 Benzoylaminopurine-1-9-yl group, adenylyl group, 2-isobutyryllamino 6-Hydroxypurine-19-yl group, Gua2-norre group, 2— Oxo-1 4-benzoylamino_1,2-dihydropyrimidine-1—yl group, cytosyl group, 2-oxo-5-methyl-4-benzoylamino-1,2,2-dihydropyrimidine-1 1-yl group , A 5-methylcytosyl group, a perashell group or a thyminyl group.
  • R 1 is selected from 2) or 3) above
  • R 2 is selected from 4) to 6) above
  • R 3 is selected from 7) to 8) above
  • B is selected from 9) to 10) above. Any combination obtained in this way is also suitable, particularly preferably a combination of 2), 4), 7) and 9) and a combination of 3), 6), 8) and 10).
  • novel oligonucleotides of the present invention are:
  • R 2 represents a hydrogen atom, an alkyl group having 1 to 6 carbon atoms or a protecting group for nucleic acid synthesis of an amino group
  • B represents a purine 9-yl group or a 2-oxo-11,2-dihydroxypyrimidine-11-yl group which may have a substituent selected from the following ⁇ group: And / or pharmacologically acceptable salts thereof containing one or more of the following structures: However, when two or more of the above structures are contained, ⁇ is the same or different between the structures.
  • a mercapto group protected with a protecting group for nucleic acid synthesis A mercapto group protected with a protecting group for nucleic acid synthesis
  • oligonucleotide analog means a non-natural type in which a nucleoside unit in a natural type oligonucleotide is substituted with one or two or more of the above-mentioned structure (la) or (lb),
  • a sugar derivative in which the sugar moiety is modified for example, a phosphate derivative in which the phosphate diester binding moiety is thiolated, an ester in which the terminal phosphate moiety is esterified, a purine base
  • the amide body in which the above amino group has been amidated may also be contained as another nucleoside or nucleotide unit.
  • novel oligonucleotide analogs of the present invention preferred are
  • R 2 is a hydrogen atom, C 1 to 6 alkyl group having a carbon aliphatic Ashiru group, an aromatic Ashiru group, or, ⁇ re Ichiru ring by 1 to 2 lower alkoxy or di Toro group substituted Oligonucleotide analogs and pharmaceutically acceptable salts thereof, which are optionally substituted aralkyloxycarbonyl groups,
  • R 2 is an aralkyl whose aryl group may be substituted with a hydrogen atom, an alkyl group having 1 or 2 carbon atoms, an aliphatic acyl group, or 1 or 2 lower alkoxy or nitro groups.
  • R 2 is a hydrogen atom, a methyl group, a trifluoroacetyl group or a benzyloxycarbonyl group, an oligonucleotide analogue and a pharmacologically acceptable salt thereof,
  • B is a 6-aminopurin-9-yl group (that is, an adenyl group), a 6-aminopurin-9-yl group in which the amino group is protected by a protecting group for nucleic acid synthesis, 2, 6-diaminopurine-1-yl, 2-amino-6-chloropurine-9-yl and amino groups protected with a protecting group for nucleic acid synthesis 2-amino-6-chloropurine-9— 2-ylamino-6-fluoropurine 9 _ 2-amino-6-fluoropurine-19-yl, 2-amino-6-bromopurine-9-yl, and amino groups whose nucleic acid and amino groups are protected with protecting groups for nucleic acid synthesis Protected 2-protected 2-amino-6-bromopurine 9-yl, 2-amino-6-hydroxypurine-19-yl (that is, guaninyl) and amino groups protect nucleic acid synthesis.
  • 2-amino-6-hydroxypurin-1-yl, amino- and hydroxyl-protected 2-amino-6-hydroxypurin-1-yl-6,6-protected with a protecting group for nucleic acid synthesis Amino 1 2 —Methoxyprin 1 9—yl group, 6—Amino 1—2-Chloropurine 9-yl group, 6 —Amino 2 —Fluoropurine 9-yl group, 2, 6—Dimethoxypurine 1 9 —Y 2,6—dichrolic pudding 6-mercaptopurine 9-yl group, 2-oxo-4-amino-1,2,2-dihydropyrimidine-11-yl group (ie, cytosinyl group), and the amino group was protected with a protecting group for nucleic acid synthesis.
  • any combination obtained by selecting R 2 from the above 2) to 4) and selecting B from the above 5) to 6) is also suitable. Particularly preferred are a combination of 2) and 5), a combination of 3) and 5) and a combination of 4) and 6).
  • the “protecting group for hydroxyl group synthesis of nucleic acid” in the definition of R 1 and R 3 is not particularly limited as long as it can stably protect the hydroxyl group during nucleic acid synthesis. Examples thereof include formyl, acetyl, and propionyl.
  • Benzyl, ⁇ -naphthoyl, and arylcanolebonyl groups such as
  • benzoinole Arylcarbonyl groups, lower alkylated arylcarbonyl groups such as 2,4,6-trimethylbenzoinole, 4-tonoleyl, and lower alkoxylated aryl groups such as 4-anisole Carbonyl group, 2-carboxybenzoyl,
  • 3-carboxybenzoyl 4-carbonyl carbonyl groups such as carboxybenzoyl, 4-nitrobenzoyl, 2-nitrobenzyl carbonyl groups such as nitrobenzoyl; 2 — Lower alkoxycarbonylated arylcarbonyl groups such as (methoxycarbonyl) benzoyl,
  • aromatic acyl such as aryloxycarbonyl such as phenylbenzoyl
  • Tri-lower alkylsilyl groups such as trimethylsilyl, triethylsilyl, isopropyldimethylsilyl, t-butyldimethylsilyl, methyldiisopropylpropionyl ⁇ ⁇ ⁇ , methyldi-t-butylsilyl, triisopropylsilyl, diphenylmethylsilinole, t — "Silyl groups” such as tri-lower alkylsilyl groups substituted with one or two aryl groups, such as butyldiphenylsilyl, diphenylisoprovirsilyl, and phenyldiisopropylylsilyl;
  • “Lower alkoxymethyl groups” such as methoxymethyl, 1,1-dimethyl-1-methoxymethyl, ethoxymethyl, propoxymethyl, isopropoxymethyl, butoxymethyl, t-butoxymethyl;
  • Halogenated tinole group such as trichloroethyl; benzyl, ⁇ -naphthylmethyl, —naphthylmethyl, diphenylenomethyl, triphenylmethyl, 1-naphthyldiphenylmethyl, 9-anthrylmethyl "A methyl group substituted with 1 to 3 aryl groups";
  • a "lower alkoxycarbonyl group substituted by a halogen or tri-lower alkylsilyl group such as 2,2,2-trichloromouth ethoxycarbonyl, 2-trimethylsilylethoxycarbonyl;
  • R 1 is preferably an aliphatic acyl group, an aromatic acyl group, a methyl group substituted with 1 to 3 aryl groups, a lower alkyl, or a lower alkoxy.
  • R 3 preferably, aliphatic Ashiru group, an aromatic Ashiru group, 1 to 3 of a methyl group substituted by Ariru group, a lower alkyl, Ariru ring by lower alkoxy, halogen Moshiku the Shiano group substituted A methyl group or a silyl group substituted with one to three aryl groups, more preferably an acetyl group, a benzoyl group, a benzinole group, a p-methoxybenzinole group, or a tert-butyl group. It is a diphenylsilyl group.
  • phosphate group protected by a protecting group for nucleic acid synthesis refers to a group capable of stably protecting a phosphate group during nucleic acid synthesis.
  • Cylindrometed lower alkyl groups such as 2-cyanoethyl, 2-cyano-1,1,1-dimethylethyl;
  • Ethyl groups substituted by silyl groups such as 2-methyldiphenylsilylethyl, 2-trimethylsilylethyl, 2-triphenylsilylethyl; 2,2,2_trichloroethynole , 2,2,2-Tribromoethylenol, 2,2,2-Trifluoroethylenol, 2,2,2-Trichloro-1,1dimethinole "Hexogenated lower alkyl groups” such as ethyl;
  • Cycloalkyl groups such as cyclopentinole, cyclopentinole, cyclopentinole, cyclohexinole, cyclopentyl, nonolebourinole, adamantinole
  • Aryl groups such as feninole, indeninole, naphtinole, fenance reninole, anthraceninole;
  • alkyl group having 1 to 6 carbon atoms in the definition of R 2 includes, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutynole, s-butynole, tert-butynole, Examples thereof include a linear or branched alkyl group having 1 to 6 carbon atoms such as pentynole and hexyl, preferably an alkyl group having 1 to 4 carbon atoms, and more preferably a carbon atom having 1 to 6 carbon atoms. Number 1 or 2 Alkyl groups, most preferably a methyl group.
  • protecting group for nucleic acid synthesis of an amino group in the definition of R 2 is not particularly limited as long as it can stably protect the amino group during nucleic acid synthesis.
  • examples thereof include formyl and acetyl. , Propionyl, butyryl, isobutyryl, pentanoyl, pinocyl, ylon, linole, octanol, decanol, 8-methylnonanoyl, 3-ethylnonanol, 3,7-dimethyl Octanoyl, Pendecanoyl, Tridecanoyl, Hexadecanyl, 14-Methylpentadecanoyl, 13-, 13-Dimethyltetradecanol, 1-Methylheptadecanoyl, Nonadecanol, Eikosanol and Henikosanol Alkylcarbonyl groups, carboxylated alkyls such as succinoyl, gluta
  • Benzoyl, ⁇ -naphthyl, mono-naphthyl, and other aryl groups such as 2-naphthoyl, 2-bromobenzoinole, 4-chlorobenzoyl, and phenol-like groups such as benzoyl, 2,4, 6-lower alkylated arylarylcarbonyl groups such as trimethylbenzoyl, 4- tonoreoyl, lower alkoxylated arylylcarbonyl groups such as 4-anisolenoyl, 2-carboxybenzoyl.
  • a “lower alkoxycarboyl group substituted by a halogen or tri-lower alkylsilyl group such as 2,2,2-trichloromouth ethoxycarbonyl, 2-trimethylsilinoleethoxycarboel;
  • Group '' or ⁇ an aralkyloxycarbonyl group in which the aryl ring may be substituted with one or two lower alkoxy or two-port groups '', and particularly preferably trifluorocarbonyl. Cetyl group or base Nji Ruo alkoxycarbonyl group.
  • halogen atom in the definition of the above group include a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom. It is a chlorine atom.
  • a straight-chain or branched-chain alkyl group having 1 to 6 carbon atoms such as sobutynole, s-butynole, tert-butynole, pentynole, and hexyl can be mentioned, and is preferably a methyl group or an ethyl group. .
  • the protecting group of nucleic acid synthesis "hydroxyl groups are protected with a protective group of nucleic acid synthesis" in the non-group defined above R 4 a and R 4 b sequence, the definitive definition of R 1 and R 3 'hydroxyl nucleic acid
  • the same groups as those exemplified in "Synthetic protecting group” can be mentioned, preferably “aliphatic acyl” or “aromatic acyl", and more preferably benzoyl.
  • Is a protective group for nucleic acid synthesis "mercapto group which is protected by a protecting group of nucleic acid synthesis" in the shed group defined above R 4 a and R 4 b sequence, for example, in the definition of R 1 and R 3
  • protecting groups for nucleic acid synthesis of hydroxyl group "groups that form disulfides” such as alkynolethio groups such as methylthio, ethylthio, and tert-butylthio, and arylthio groups such as benzylthio It is preferably an "aliphatic acyl group” or an "aromatic acyl group", and more preferably a benzoyl group.
  • Is a protective group for nucleic acid synthesis "Amino groups protected by protecting groups of the nucleic acid synthesis" in the non-group defined above R 4 a and R 4 b sequence, that put on the above definitions of R 2 "Amino group
  • Examples of the group include the same groups as those exemplified in the above “Protecting group for nucleic acid synthesis”, preferably an “aliphatic acyl group” or an “aromatic acyl group”, and more preferably a benzoyl group. .
  • R 4 a and R 4 b and ⁇ group definition for example, main butoxy, et butoxy, n- propoxy, isopropoxy, n - butoxy, Examples thereof include straight-chain or branched alkoxy groups having 1 to 6 carbon atoms such as isobutoxy, s-butoxy, tert-butoxy, pentyloxy, and hexyloxy, and preferably methoxy or ethoxy. Group.
  • R 4 b is set to "having 1 to 6 ⁇ alkylthio group having a carbon" in the ⁇ group definitions, for example, methylthio, Echiruchio, Puropiruchi O, Lee Sopuropiruchio, butylthio, isobutylthio, s - butylthio, tert- Examples thereof include a butylthio, pentylthio and hexylthio group, and a methylthio or ethylthio group is preferable.
  • R 4 is set to "having 1 to 6 ⁇ alkylthio group having a carbon in the ⁇ group definitions, for example, methylthio, Echiruchio, Puropiruchi O, Lee Sopuropiruchio, butylthio, isobutylthio, s - butylthio, tert- Examples thereof include a butylthio, pentylthio and hexylthi
  • R 4 b "alkylamino amino group which is substituted by the number 1 to 6 ⁇ alkyl group having a carbon" in the ⁇ group definitions, for example, Mechiruami Roh, Echiruamino, Puropiruamino, isopropyl ⁇ Mino, Buchiruamino, Isobuchiruamino S-butylamino, tert-butylamino, pentylamino, hexylamino, dimethylamino, getylamino, dipropylamino, diisopropylamino, dibutylamino, diisobutylamino, di (s-butyl) amino, di (tert-butyl) amino And dipentylamino and dihexylamino groups, preferably a methylamino, ethylamino, dimethylamino or getylamino group.
  • said R 3 may be, for example, Shianome butoxy, Shianoe butoxy, Xia Roh propyl O carboxymethyl, Shianobuchiru Okishi, cited Kishiruokishi group Xia Roh pliers Ruo carboxymethyl, to Shiano And is preferably a 2-cyanoethoxy group.
  • the “pharmacologically acceptable salt thereof” means that the bicyclonucleoside derivative (1) of the present invention and the oligonucleotide analog containing the above structure (la) or (lb) can be made into a salt.
  • a salt preferably such an alkali metal salt such as a sodium salt, a potassium salt, or a lithium salt; an alkaline earth metal salt such as a calcium salt or a magnesium salt; or aluminum.
  • Metal salts such as salts, iron salts, zinc salts, copper salts, nickel salts, cobalt salts; inorganic salts such as ammonium salts, t-octylamine salts, dibenzylamine salts, morpholine salts, dalcosamine salts, and phenyl.
  • Glycine alkyl ester salt ethylene diamine salt, N-methyl dalcamine salt, guanidine salt, getylamine salt, triethylamine , Dicyclohexylaminate, N, N'-dibenzylethylenediamine, chlorinated pro-diamine-pro-diyne, diethanolamine, N-benzyl-phenethylamine, Amines, such as organic salts such as piperazine, tetramethylammonium, and tris (hydroxymethyl) aminomethane; hydrofluorides, hydrochlorides, hydrobromides, hydroiodides Inorganic salts such as hydrohalides, nitrates, perchlorates, sulfates, and phosphates; and lower salts such as methanesulfonate, trifluoromethanesulfonate, and ethanesulfonate.
  • a sodium salt, a potassium salt and a triethylamine salt are preferable, and a bicyclonucleoside derivative (1) is preferred. In this case, a free body is preferable.
  • the bicyclonucleoside derivative (1) of the present invention and the oligonucleotide analog containing the above structure (1a) or (lb) are allowed to stand in the air. By doing so, water may be absorbed, adsorbed water may be attached, or a hydrate may be formed. Such salts are also included in the present invention. Furthermore, the bicyclonucleoside derivative (1) of the present invention and the oligonucleotide analog containing the above structure (1a) or (lb) may absorb some other solvent and become a solvate in some cases. Such salts are also included in the present invention. Specific examples of the bicyclonucleoside derivative (1) of the present invention include, for example, compounds shown in Tables 1 and 2 below. In Tables 1 and 2,
  • B n is a benzyl group
  • B z is a benzoyl group
  • C bz is a benzyloxycarbonyl group
  • Me is a methyl group
  • PMB n is p-methoxybenzyl Group
  • ⁇ iPr '' is isopropyl group
  • ⁇ MMT r '' is 4-methoxytriphenylmethyl group
  • ⁇ DMTr '' is 4,4'-dimethoxytriphenylmethyl group
  • T fa is a trifluoroacetyl group
  • TMTr is 4, 4 ', 4 "— trimethoxytriphenylmethyl group
  • TMS is a trimethylsilyl group
  • T DMS indicates a tert-butyldimethylsilyl group
  • TDPS indicates a tert-
  • preferred compounds include compound numbers 1-3, 1-4, 1-5, 1-8, 1-10, 1-11, 1-17, 1-18, 1-20, 1-21, 1-22, 1-23, 1-24, 1-25, 1-28, 1-30,
  • Further preferred compounds include Compound Nos. 1-4, 1-23, 2-5, 2-6, 2-8, 2-16, 2-17, 2-42, 2-56, 2-59 , 2-86, 2-89, 2-108, 2-140, 2-142 and 2-143.
  • the most preferred compounds include
  • R 8 represents a protecting group for nucleic acid synthesis of a hydroxyl group, and is preferably t-butyldimethylsilyl, trimethylsilyl, triethylsilyl, or triisopropylsilyl.
  • Silyl groups such as ril, dimethylisopropylsilyl, getylisopropylsilyl, t-butinoresiphenylsiligure, diphenylmethylinoresilisle, triphenylinoresilinole, 4-methoxy rifiphenylmethyl, 4 A trityl group such as 4,4 ', 4 ", 4"-trimethoxytriphenylmethyl; an aralkyl group such as benzyl or p-methoxybenzyl; more preferably, t-butyl. It is a diphenylsilyl group.
  • R 9 represents a leaving group (provided that it functions as a protecting group on nitrogen), and is preferably a lower alkylsulfonyl group such as methanesulfonyl or ethanesulfonyl, or a trifluoromethanesulfonyl group.
  • a arylsulfonyl group such as a halogen-substituted lower alkynolenorfonyl group and p-toluenesulfonyl, and more preferably a methanesulfonyl or p-toluenesulfonyl group.
  • R 10 represents a protecting group for nucleic acid synthesis, preferably an aralkyloxycarbonyl group such as a benzyloxycarbonyl group; a dichloroacetyl group, a trichloroacetyl group, or a trifluoroacetyl group. And more preferably a benzyloxycarbonyl group or a trifluoroacetyl group.
  • R 11 represents a protecting group for nucleic acid synthesis of a hydroxyl group, and is preferably formyl, acetyl, propionyl, butyryl, isobutyryl, pentanoyl, pivaloyl, nokrelyl, isonokrillyl, octanoyl, decanol, 8-methylnonanoyl, 3-ethylooctanoyl, 3,7-dimethyloctanoyl, pendecanoyl, tridecanoyl, hexadecanoyl, 14-methylpentadecanoyl, 13,3-13-dimethyltetradecanoyl, 1 Alkylcarbonyl groups such as monomethylheptadecanoyl, nonadecanoyl, eicosanoyl and henicosanoyl-for succinoinole, glutaroyl, and ajiboyl Such as carboxyalkylcarbonyl groups
  • Benzoyl, ⁇ -naphthyl,] 3-Naphthyl-like arylcanoleponyl group 2—bromobenzoinole, 4-chlorobenzoyl group such as benzoyl, 2,4,6—triyl
  • Lower alkylated arylcarbonyl groups such as methinobenzoyl, 4- tonoreoyl, lower alkoxylated arylcarbonyl groups such as 4-anisole, 2-carboxybenzoyl.
  • aromatic acyl group more preferably an “aliphatic acyl group”, and particularly preferably an acetyl group.
  • Beta 1 is the following alpha 1 substituent a good purine have one selected from the group 9 - I le or 2 - Okiso one 1, 2-dihydrazide mud pyrimidine one 1 - shows the I le radical .
  • Method A is a method for obtaining the target compound (1) using the compound (2) as a starting material. Hereinafter, each step of the method A will be described in detail.
  • the compound (3) is produced by reducing the azido group of the compound (2), which can be produced by the method B described below, to an amino group in an inert solvent in the presence of hydrogen and a catalyst. It is a process.
  • the solvent to be used is not particularly limited as long as it does not participate in the reaction.
  • alcohols such as methanol, ethanol, and isopropanol, getyl ether, tetrahydrofuran, and dioxane are used.
  • Such as ethers aromatic hydrocarbons such as toluene, benzene, and xylene; aliphatic hydrocarbons such as hexane and cyclohexane; esters such as ethyl acetate and propyl acetate; formamide and dimethylform Amides, such as amide, dimethylacetamide, N-methyl-2-pyrrolidone, hexamethylphosphotriamide, fatty acids such as formic acid and acetic acid, water, or a mixed solvent thereof is used.
  • aromatic hydrocarbons such as toluene, benzene, and xylene
  • aliphatic hydrocarbons such as hexane and cyclohexane
  • esters such as ethyl acetate and propyl acetate
  • formamide and dimethylform Amides such as amide, dimethylacetamide, N-methyl-2-pyrrolidone, hexamethylphosphotriamide, fatty acids such as formic acid
  • the catalyst to be used is not particularly limited as long as it is usually used in a catalytic reduction reaction, but is preferably palladium carbon, palladium black, Raney nickel, platinum oxide, platinum black, or rhodium.
  • the pressure at which nilphosphine rhodium monochloride or palladium monosulfate is used is not particularly limited, but is usually 1 to 10 atm.
  • reaction temperature and reaction time vary depending on the type of the starting material, solvent and catalyst, but are usually 0 ° C to 100 ° C (preferably 20 ° to 40 ° ⁇ ), 5 minutes. It is 48 hours (preferably 1 hour to 10 hours).
  • the target compound (3) of this reaction is collected from the reaction mixture according to a conventional method. For example, it can be obtained by removing the catalyst by filtration and distilling off the solvent.
  • This step is a step of producing a compound (4) by reacting the compound (3) produced in the step A-1 with a leaving group-introducing reagent in an inert solvent in the presence of a base catalyst.
  • Solvents used include, for example, aliphatic hydrocarbons such as hexane, heptane, rigoline, petroleum ether; aromatic hydrocarbons such as benzene, toluene and xylene; methylene chloride Norogenated hydrocarbons such as carbon, tetrachloroform, carbon tetrachloride, dichloroethane, cyclobenzene, and dichlorobenzene; ethyl formate, ethyl acetate, propyl citrate, butyl acetate, and getyl carbonate.
  • —Ethers such as ter, tetrahydrofuran, dioxane, dimethoxyethane, and ethylene glycol dimethyl ether; such as acetone, methylethylketone, methylisobutylketone, isophorone, cyclohexanone; Ketones; Nitro compounds such as Nitroethane and Nitrobenzene; Nitrils such as Acetonitrile and Isobutyronitrile; Formamide, N, N-dimethylformamide, N Amides, such as N, N-dimethylacetamide, N-methyl-2-pyrrolidone, N-methylpyrrolidinone, and hexamethylphosphotriamide; sulfoxides, such as sulfolane; But preferably with methylene chloride. is there.
  • the base catalyst used is preferably a base such as triethylamine, pyridine or dimethylaminopyridine.
  • Examples of the leaving group-introducing reagent used include alkynolesulfoninolehalides such as methanesulfonyl chloride and ethanesnolefonyl bromide; and p-toluenesulfonyl chloride. And arylsulfonyl halides, preferably methanesulfonyl chloride and p-toluenesulfonyl chloride.
  • the reaction temperature varies depending on the starting compound used, the solvent, the leaving group-introducing reagent, and the base catalyst.
  • the reaction temperature is usually from 0 ° C to 50 ° C, and preferably from 10 ° C to 40 ° C. C.
  • the reaction time varies depending on the starting compound used, the solvent, the leaving group-introducing reagent, the base catalyst and the reaction temperature, but is usually from 10 minutes to 24 hours, preferably from 1 to 15 hours. .
  • the target compound (4) of this reaction is collected from the reaction mixture according to a conventional method.
  • the reaction mixture is neutralized, the reaction mixture is concentrated, an immiscible organic solvent such as water and ethyl acetate is added, and after washing with water, the organic layer containing the target compound is separated and dried over anhydrous sodium sulfate or the like. Thereafter, it is obtained by distilling off the solvent.
  • the obtained compound can be further purified, if necessary, by a conventional method, for example, recrystallization, silica gel column chromatography or the like.
  • This step is a step of cyclizing compound (4) produced in step A-2 in an inert solvent in the presence of a base catalyst to produce compound (5).
  • the solvent used is not particularly limited as long as it does not hinder the reaction and dissolves the starting material to some extent.
  • Ethers such as dimethylene glycol dimethyl ether can be mentioned, and more preferably, tetrahydrofuran.
  • Examples of the base catalyst used include, for example, lithium metal hydride, sodium hydride, and alkali metal hydrides such as lithium hydride, and preferably, sodium hydride. Um.
  • the reaction temperature is generally 0 ° C. to 50 ° C., preferably 0 ° to 30 ° C.
  • the reaction time is generally 1 hour to 3 days, preferably 3 hours. Or two days.
  • the target compound (5) of this reaction is collected from the reaction mixture according to a conventional method. For example, concentrate the reaction mixture, add an immiscible organic solvent such as water and ethyl acetate, wash with water, separate the organic layer containing the target compound, dry over anhydrous sodium sulfate, etc., and evaporate the solvent Obtained by:
  • the obtained compound can be further purified, if necessary, by a conventional method, for example, recrystallization, silica gel column chromatography or the like.
  • step A-3 the amino group of compound (5) produced in step A-3 is deprotected in an inert solvent in the presence of a base catalyst to produce compound (6).
  • the solvent used is not particularly limited as long as it does not participate in the reaction, but is preferably water; alcohols such as methanol, ethanol and n-propanol; and tetrahydrofuran and dioxane.
  • Organic solvents such as ethers or mixed solvents of water and the above-mentioned organic solvents, more preferably alcohols.
  • the base used is not particularly limited as long as it does not affect the other parts of the compound, but is preferably a metal alkoxide such as sodium methoxide, sodium carbonate, or carbonate.
  • Alkali metal carbonates such as potassium and lithium carbonate, sodium hydroxide, hydroxide water, lithium hydroxide
  • Alkali metal hydroxides such as ammonia, ammonia water, and ammonia such as concentrated ammonium hydroxide are used.
  • the reaction temperature varies depending on the starting compound, solvent and the like used, but is usually from 0 ° C to the reflux temperature of the solvent used, and preferably from 10 ° C to the reflux temperature of the solvent used.
  • the reaction time varies depending on the starting compound used, the solvent, the reaction temperature and the like, but is usually 10 minutes to 24 hours, preferably 30 minutes to 10 hours.
  • the target compound (6) for the reaction is collected from the reaction mixture according to a conventional method. For example, under cooling, dilute the reaction solution with ether, add an immiscible organic solvent such as water and ethyl acetate, wash with water, separate the organic layer containing the target compound, and dry with anhydrous sodium sulfate or the like. It is obtained by distilling off the solvent.
  • the obtained compound can be further purified, if necessary, by a conventional method, for example, recrystallization, silica gel column chromatography or the like.
  • This step is a step of producing a compound (7) by reacting the compound (6) produced in the step A-4 with a protecting group-introducing reagent in an inert solvent in the presence of a base catalyst.
  • solvent used examples include aliphatic hydrocarbons such as hexane, heptane, lignin, and petroleum ether; aromatic hydrocarbons such as benzene, toluene, and xylene; methylene chloride Norogenated hydrocarbons such as chloroform, carbon tetrachloride, dichloroethane, chlorobenzene, and dichlorobenzene; esters such as ethyl formate, ethyl acetate, propyl acetate, butyl acetate, and getyl carbonate; Ethers such as monoter, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxetane, and ethylene glycol dimethyl ether; acetone, methylinoethyl ketone, methyl isobutyl ketone, isophorone, cyclohexanone; Ketones such as; Nitroethane, Nitro compounds such as
  • the base catalyst used is preferably a base such as triethylamine, pyridine or dimethylaminopyridine.
  • protecting group-introducing reagent used include, for example, aralkyloxycarbonyl halides such as benzyloxycarbonyl chloride; dichloroacetic anhydride, trichloroacetic anhydride, trifluoroacetic anhydride.
  • aralkyloxycarbonyl halides such as benzyloxycarbonyl chloride
  • dichloroacetic anhydride trichloroacetic anhydride
  • trifluoroacetic anhydride examples include halogeno lower alkyl anhydrides, such as benzyloxycarbonyl anhydride and trifluoroacetic anhydride.
  • the reaction temperature varies depending on the starting compound used, the solvent, the protecting group-introducing reagent, and the base catalyst, but is usually from 0 ° C to 50 ° C, and preferably from 0 ° C to 30 ° C. .
  • the reaction time varies depending on the starting compound used, the solvent, the protecting group-introducing reagent, the base catalyst, and the reaction temperature, but is usually from 10 minutes to 24 hours, preferably from 30 minutes to 15 hours. is there.
  • the target compound (7) of this reaction is collected from the reaction mixture according to a conventional method.
  • the reaction solution is neutralized, the reaction mixture is concentrated, an immiscible organic solvent such as water and ethyl acetate is added, and after washing with water, the organic layer containing the target compound is separated and dried with anhydrous sodium sulfate or the like. After drying, it is obtained by distilling off the solvent.
  • the obtained compound can be further purified, if necessary, by a conventional method, for example, recrystallization, silica gel column chromatography or the like. (A-6 processes)
  • This step is a step of producing a compound (8) by reacting the compound (7) produced in the step A-5 with an acid anhydride in a solvent in the presence of an acid catalyst.
  • Solvents used include, for example, ethers such as getyl ether, dioxane, and tetrahydrofuran; ditriles such as acetonitrile and isobuchi nitro; formamide, Amides such as N, N-dimethylformamide, N, N-dimethylacetamide, N-methyl-2-pyrrolidone, N-methylpyrrolidinone, hexamethylphosphorotriamide; acetic acid Such an organic acid may be mentioned, and acetic acid is preferred.
  • the acid catalyst to be used for example, inorganic acids such as hydrochloric acid, sulfuric acid, and nitric acid can be used, and sulfuric acid (particularly, concentrated sulfuric acid) is preferable.
  • Examples of the acid anhydride to be used include anhydrides of lower aliphatic carboxylic acids such as acetic anhydride and propionic anhydride, and preferred is anhydrous acetic acid.
  • the reaction temperature varies depending on the starting compound, solvent, acid catalyst and acid anhydride used, but is usually from 0 ° C to 50 ° C, preferably from 10 ° to 40 ° C.
  • the reaction time varies depending on the used starting compound, solvent, acid catalyst, acid anhydride and reaction temperature, but is usually from 10 minutes to 12 hours, and preferably from 30 minutes to 3 hours. .
  • the target compound (8) of this reaction is collected from the reaction mixture according to a conventional method. For example, neutralize the reaction solution, add an immiscible organic solvent such as water and ethyl acetate, wash with water, separate the organic layer containing the target compound, dry with sodium sulfate anhydride and dry the solvent. Is obtained by distillation.
  • an immiscible organic solvent such as water and ethyl acetate
  • the obtained compound can be further purified by a conventional method, for example, recrystallization, silica gel chromatography, or the like.
  • This process is carried out in an inert solvent in the presence of an acid catalyst, Compound (8) having a desired substituent prepared according to the literature (H. Vorbrueggen, K. Kro liki ewicz and B, Bennua, Chem. Ber., 1 14, 1234-1255 (1981)).
  • This is a step of producing a compound (9) by reacting a trimethylsilylated compound corresponding to a purine or pyrimidine which may be present.
  • Solvents used include aromatic hydrocarbons such as benzene, toluene, and xylene; methylene chloride, chloroform, carbon tetrachloride, 1,2-dichloroethane, cyclobenzene, and dichlorobenzene.
  • G hydrocarbons; nitritols such as acetonitrile and isobutyronitrile; formamide, N, N-dimethylformamide, N, N-dimethylacetamide, N-methyl-12-pyro Amides such as ridone, N-methylpyrrolidinone, and hexamethylphosphorotriamide; the ability to include carbon sulfide and the like; 1,2-dichloroethane is preferred.
  • the acid catalyst used for example, can be mentioned A1 C1 3, SnCl 4, T iCl 4, ZnCl 2, BF 3, a Lewis acid catalyst such as triflate Ruo b methanesulfonic Santo Re methylsilyl, suitably Is trimethylsilyl trifluoromethanesulfonate.
  • the reaction temperature varies depending on the starting compound, solvent and acid catalyst used, but is usually 0 to 100 ° C, preferably 30 to 80 ° C.
  • the reaction time varies depending on the used starting compound, solvent, acid catalyst and reaction temperature, but is usually 1 hour to 3 days, preferably 1 hour to 2 days.
  • the target compound (9) of the reaction is collected from the reaction mixture according to a conventional method. For example, neutralize the reaction solution, add an immiscible organic solvent such as water and ethyl acetate, wash with water, separate the organic layer containing the target compound, dry with sodium sulfate anhydride and dry the solvent. Is obtained by distillation.
  • an immiscible organic solvent such as water and ethyl acetate
  • the obtained compound can be further purified, if necessary, by a conventional method, for example, recrystallization, silica gel column chromatography or the like. (A—8 steps)
  • step A-7 the compound (9) produced in step A-7 is reacted with a deprotecting reagent in an inert solvent, and if necessary, alkylation is carried out to produce compound (1). It is.
  • the method of deprotection varies depending on the type of protecting group. However, even if it is a method that does not cause other side reactions, it is not particularly limited, but it is possible to use “Protective Groups in Organic Synthesis” ( Theodora W. Greene, 1981, published by A Wiley-Interscience Publication).
  • deprotection can be performed sequentially by appropriately combining these methods.
  • the protecting group is (1) an "aliphatic or aromatic acyl group", (2) a "methyl group substituted by 1 to 3 aryl groups” or a “lower alkyl, lower alkoxy, halogen, cyano,
  • a methyl group substituted with one to three aryl groups in which an aryl group is substituted with a group, or (3) a "silyl group” the following methods can be used.
  • reaction is usually carried out by reacting a base in an inert solvent.
  • the solvent to be used is not particularly limited as long as it is one used in a usual hydrolysis reaction.
  • water alcohols such as methanol, ethanol, n-propanol, tetrahydrofuran
  • An organic solvent such as ethers such as dioxane or a mixed solvent of water and the above organic solvent is used, and preferably, alcohols.
  • the base used is not particularly limited as long as it does not affect the other parts of the compound, but is preferably a metal alkoxide such as sodium methoxide; sodium carbonate, carbonate Alkali metal carbonates, such as lithium and lithium carbonate; alkaline metal hydroxides or aqueous ammonia, such as sodium hydroxide, lithium hydroxide, lithium hydroxide, and sodium hydroxide Ann Ammonia, such as methanol, is used, and is preferably alkali metal carbonate.
  • a metal alkoxide such as sodium methoxide
  • sodium carbonate, carbonate Alkali metal carbonates such as lithium and lithium carbonate
  • alkaline metal hydroxides or aqueous ammonia such as sodium hydroxide, lithium hydroxide, lithium hydroxide, and sodium hydroxide
  • Ann Ammonia such as methanol, is used, and is preferably alkali metal carbonate.
  • reaction temperature and reaction time vary depending on the starting material, solvent and base used, and are not particularly limited.However, in order to suppress a side reaction, the reaction is usually performed at 0 ° C to 150 ° C and 1 to 10 ° C. Implemented for hours.
  • the target compound (1) of this reaction is collected from the reaction mixture according to a conventional method.
  • the reaction mixture is concentrated, an immiscible organic solvent such as water and ethyl acetate is added, washed with water, an organic layer containing the target compound is separated, dried with anhydrous sodium sulfate, and the solvent is removed. Obtained by distillation.
  • the obtained compound can be further purified by a conventional method, for example, recrystallization or silica gel column chromatography.
  • the protecting group is ⁇ a methyl group substituted by 1 to 3 aryl groups '' or ⁇ 1 to 3 aryl groups wherein the aryl ring is substituted by lower alkyl, lower alkoxy, halogen, or cyano groups.
  • the reaction is performed using a reducing agent in an inert solvent.
  • the solvent used is methanol, ethanol, or ethanol.
  • Alcohols such as ethanol; ethers such as getyl ether, tetrahydrofuran, and dioxane; aromatic hydrocarbons such as toluene, benzene, and xylene; hexane, and hexane.
  • aromatic hydrocarbons such as toluene, benzene, and xylene
  • hexane, and hexane is methanol, ethanol, or ethanol.
  • ethers such as getyl ether, tetrahydrofuran, and dioxane
  • aromatic hydrocarbons such as toluene, benzene, and xylene
  • hexane hexane
  • hexane such as aliphatic hydrocarbons
  • esters such as ethyl acetate and propyl acetate
  • organic acids such as acetic acid
  • the reducing agent to be used is not particularly limited as long as it is generally used in a catalytic reduction reaction.
  • the reducing agent is palladium on carbon, Raney nickel, platinum oxide, platinum black, rhodium oxide.
  • Examples include aluminum oxide, triphenylphosphine rhodium monochloride, and palladium monosulfate.
  • the pressure is not particularly limited, but it is usually 1 to 10 atm.
  • the reaction temperature is 0 ° C. to 60 ° C., and preferably 20 to 40 ° C. ( :
  • the reaction time is 10 minutes to 24 hours, preferably 1 to 3 hours.
  • the target compound (1) of this reaction is collected from the reaction mixture according to a conventional method. For example, remove the reducing agent from the reaction mixture, add an immiscible organic solvent such as water and ethyl acetate, wash with water, separate the organic layer containing the target compound, and dry with anhydrous sodium sulfate or the like. It is obtained by distilling off the solvent.
  • an immiscible organic solvent such as water and ethyl acetate
  • the obtained compound can be further purified, if necessary, by a conventional method, for example, recrystallization, silica gel chromatography, or the like.
  • the reaction can be performed using an acid.
  • examples of the solvent used include aromatic hydrocarbons such as benzene, toluene, and xylene; methylene chloride, chloroform, carbon tetrachloride, 1,2-dichloroethane, and cyclobenzene. And halogenated hydrocarbons such as dichlorobenzene; alcohols such as methanol, ethanol, isopropanol, and ⁇ ert-butanol; and alcohols such as acetonitrile and isobutyronitrile.
  • aromatic hydrocarbons such as benzene, toluene, and xylene
  • halogenated hydrocarbons such as dichlorobenzene
  • alcohols such as methanol, ethanol, isopropanol, and ⁇ ert-butanol
  • alcohols such as acetonitrile and
  • Such trilinoles formamide, N, N-dimethylformamide, N, N-dimethylacetamide, N-methyl-1-pyrrolidone, N-methylpyrrolidinone, and hexamethylphosphorotriamide.
  • amides include organic acids such as acetic acid, preferably organic acids (particularly, acetic acid) or alcohols (particularly, acetic acid). It is a tert- butanol Ichiru).
  • the acid used is preferably acetic acid or trifluoroacetic acid.
  • the reaction temperature is 0 ° C to 6 0 ° C, suitably 2 0 to 4 0 ° c reaction time is C is 1 0 minutes to 2 4 hours, preferably from 1 to 3 It is time.
  • the target compound (1) of this reaction is collected from the reaction mixture according to a conventional method.
  • the reaction mixture is neutralized, an immiscible organic solvent such as water and ethyl acetate is added, and after washing with water, the organic layer containing the target compound is separated. It is obtained by drying with sodium hydrogen sulfate and distilling off the solvent.
  • the obtained compound can be further purified, if necessary, by a conventional method, for example, recrystallization, silica gel column chromatography or the like.
  • the protecting group is a “silyl group”
  • a compound that generates a fluorine cation such as tetrabutylammonium fluoride, hydrofluoric acid, hydropyridine monopyridine, and potassium fluoride.
  • acetic acid, methanesulfonic acid It can be removed by treatment with an organic acid such as para-toluenesulfonic acid, trifluoroacetic acid, trifrenoleolomethanesnolefonic acid or an inorganic acid such as hydrochloric acid.
  • reaction when removing with a fluorine anion, the reaction may be accelerated by adding an organic acid such as formic acid, acetic acid or propionic acid.
  • organic acid such as formic acid, acetic acid or propionic acid.
  • the solvent used is not particularly limited as long as it does not hinder the reaction and dissolves the starting materials to some extent.
  • it is dimethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, or the like.
  • Ethers such as dimethoxetane and methylenglycol dimethyl ether; nitriles such as acetonitrile and isobutyronitrile; water; organic acids such as acetic acid; and mixed solvents thereof. .
  • the reaction temperature is from 0 ° C. to 100 ° C., preferably from 20 ° to 70 ° C.
  • the reaction time is 5 minutes to 48 hours, preferably 1 to 24 hours.
  • the target compound (1) of this reaction is collected from the reaction mixture according to a conventional method. For example, it can be obtained by distilling off the solvent and purifying by silica gel column chromatography.
  • the compound in which R 2 is an alkyl group having 1 to 6 carbon atoms can be produced by deprotecting R 1 ° by the above-mentioned method and then using an alkylating agent and a base.
  • the solvent to be used is not particularly limited as long as it does not inhibit the reaction and dissolves the starting material to some extent.
  • ethers such as getyl ether, dioxane, and tetrahydrofuran are used.
  • chlorinated hydrocarbons such as methylene chloride, chloroform, carbon tetrachloride, dichloroethane, cyclobenzene, and dichlorobenzene, and mixed solvents thereof.
  • alkylating agent alkyl iodide is preferred.
  • organic bases such as triethylamine, pyridine, dimethylaminopyridine, and 1,8-diazavicic [5,4,0] indene-7-ene are preferable.
  • the reaction temperature is — 30. C. to 40 ° C., and preferably ⁇ 10 to 20 ° C.
  • the reaction time is from 1 to 100 hours, preferably from 12 to 48 hours.
  • the target compound (1) of this reaction is collected from the reaction mixture according to a conventional method. For example, it can be obtained by distilling off the solvent and purifying by silica gel column chromatography.
  • 13 ⁇ 4 1 Ya 13 ⁇ 4 3 is, in the case of the phosphoric acid group or a protected phosphoric acid group, further, additionally performs the following method.
  • a compound (1) in which R 1 or R 3 is hydrogen is reacted with a desired phosphorylating reagent (for example, 2-chlorophenylphosphorobistriazolide) in an inert solvent.
  • a desired phosphorylating reagent for example, 2-chlorophenylphosphorobistriazolide
  • the solvent to be used is not particularly limited as long as it does not inhibit the reaction. Usually, an aromatic amine such as pyridine is used.
  • the reaction temperature is not particularly limited up to 120 to 100 ° C., but is usually carried out at room temperature.
  • the reaction time varies depending on the solvent and the reaction temperature, but is 1 hour when the reaction is carried out at room temperature using pyridine as a reaction solvent.
  • the target compound is, for example, appropriately neutralized the reaction mixture, If a solute is present, remove it by filtration, add an immiscible organic solvent such as water and ethyl acetate, wash with water, separate the organic layer containing the target compound, dry over anhydrous magnesium sulfate, etc. (The obtained target compound can be further purified, if necessary, by a conventional method, for example, recrystallization, reprecipitation or chromatography).
  • the protecting group on the phosphate group can be removed by a method usually used in nucleic acid synthesis.
  • R 1 or R 3 is a group represented by the above formula (P (R 4 a ) R 4 b ), the following method is additionally performed.
  • a compound in which R 1 or R 3 is hydrogen in an inert solvent in the presence of a deoxidizing agent is treated with a compound represented by the formula X—P (R ′′ a 2 ) R 4 b 2 (wherein R 4 a 2 and R 4 b 2 are the same or different and each is a hydroxyl group protected with a protecting group for nucleic acid synthesis, a mercapto group protected with a protecting group for nucleic acid synthesis, an amino group protected with a protecting group for nucleic acid synthesis, An alkoxy group having 1 to 6 carbon atoms, an alkylthio group having 1 to 6 carbon atoms, an amino group substituted with a cyano alkoxy group having 1 to 7 carbon atoms or an alkyl group having 1 to 6 carbon atoms, X represents a halogen atom).
  • Suitable phosphating agents to be used are morpholine methoxyphosphine, morpholine cyanoethoxy phosphine, chloromethylamino methoxyphosphine, chlorodimethinoleaminosinoethoxy phosphine, and It is a phosphine such as lorodiisopropylaminomethoxyphosphine and chlorodiisopropylaminophosphinoethoxyphosphine, and more preferably, is chloromorphinoxymethoxyphosphine, chloromorpholinoacetoxyphosphine, and chlorodimethoxyphosphine.
  • the solvent used is not particularly limited as long as it does not affect the reaction, but is preferably an ether such as tetrahydrofuran, getyl ether, or dioxane.
  • deoxidizing agent used examples include heterocyclic amines such as pyridine and dimethylaminopyridine, and aliphatic amines such as trimethylamine, triethylamine and diisopropylethylamine. Preferably, they are aliphatic amines (particularly diisopropylethylamine).
  • the reaction temperature is not particularly limited, but is usually from 150 to 50 ° C., and preferably room temperature.
  • the reaction time varies depending on the starting materials, reagents, temperature and the like to be used, but is usually 5 minutes to 30 hours, and preferably 30 minutes when reacted at room temperature.
  • the target compound is neutralized, for example, by appropriately neutralizing the reaction mixture. If there is an insoluble substance, it is removed by filtration, and then an immiscible organic solvent such as water and ethyl acetate is added. after washing with water, dried over separating the organic layer containing the desired compound. anhydrous magnesium sulfate and the like, the target compound obtained c obtained by distilling off the solvent, if necessary, a conventional method, for example, recrystallization, It can be further purified by precipitation or close chromatography.
  • an immiscible organic solvent such as water and ethyl acetate
  • protecting groups on each functional group can be removed by methods commonly used during nucleic acid synthesis.
  • the above compound (2) can be produced by the method B described below.
  • R 12 represents a protecting group for nucleic acid synthesis of a hydroxyl group, and is preferably an arylcanolebonyl group such as benzoyl, ⁇ -naphthyl, or) 3-naphthyl; , 6-trimethylbenzoyl, 4 — lower alkylated arylcarbonyl groups such as toluoyl, 4 — arylated arylylcarbonyl groups such as phenylbenzoyl, etc. And more preferably a benzoyl group.
  • the starting compound (10) used in the above method can be produced by the following method. That is, using commercially available diacetone-D-glucose as a starting material, literature (0. T. Schmidt, Methods in Carbohydr. Chem., 4, 318 (1964); JS Brimacombe and 0. A. Ching, Carbhyd. Res. , 8, 82 (1968); TF Tarn and B. Fraser-Reid, Can. J. Chem., 57, 2818 (1979); SA Suzhkov, Nucleosides & Nucleotides, 13, 2283 (1994)).
  • the compound (10) can be produced according to the method.
  • each step of the method B will be described in detail. ⁇ Law B>
  • This step is a step of producing a compound (11) by deprotecting the protecting group of the primary hydroxyl group of the starting compound (10) in an inert solvent in the presence of a base.
  • the solvent to be used is not particularly limited as long as it is one used in a usual hydrolysis reaction.
  • water alcohols such as methanol, ethanol, n-propanol, tetrahydrofuran
  • An organic solvent such as an ether such as dioxane or a mixed solvent of water and the above organic solvent is used, and preferably, alcohols.
  • the base used is not particularly limited as long as it does not affect the other parts of the compound, but is preferably a metal alkoxide such as sodium methoxide; sodium carbonate, carbonate Alkali metal carbonates such as potassium and lithium carbonate; alkaline metal hydroxides or aqueous ammonia such as sodium hydroxide, potassium hydroxide, lithium hydroxide, and potassium hydroxide; concentrated ammonia Ammonia such as mono-methanol is used, more preferably alkaline metal carbonate.
  • a metal alkoxide such as sodium methoxide
  • sodium carbonate, carbonate Alkali metal carbonates such as potassium and lithium carbonate
  • alkaline metal hydroxides or aqueous ammonia such as sodium hydroxide, potassium hydroxide, lithium hydroxide, and potassium hydroxide
  • concentrated ammonia Ammonia such as mono-methanol is used, more preferably alkaline metal carbonate.
  • the reaction temperature and reaction time vary depending on the starting material, the solvent, the base used, and the like, and are not particularly limited. However, in order to suppress a side reaction, the reaction is usually performed at 0 ° C to 150 ° C and 1 Implemented for hours.
  • the target compound (11) of this reaction is collected from the reaction mixture according to a conventional method.
  • the reaction solution is neutralized, the reaction mixture is concentrated, an immiscible organic solvent such as water and ethyl acetate is added, and after washing with water, the organic layer containing the target compound is separated, and anhydrous sodium sulfate, etc. After drying with, the solvent is distilled off.
  • the obtained compound can be further purified, if necessary, by a conventional method, for example, recrystallization, silica gel column chromatography or the like.
  • the compound (2) is produced by reacting the compound (11) produced in the step B-1 with a hydroxyl-protecting agent in an inert solvent in the presence of a base. .
  • the solvent used is not particularly limited as long as it does not hinder the reaction and dissolves the starting material to some extent.
  • examples thereof include aliphatic hydrocarbons such as hexane and heptane; benzene and toluene.
  • Aromatic hydrocarbons such as xylene and xylene: Halogenated hydrocarbons such as methylene chloride, chloroform, carbon tetrachloride, dichloroethane, methanol, benzene and dichloromethane; ethyl formate, ethyl acetate, acetic acid Esters such as propyl, butyl acetate, and getyl carbonate; ethers such as getyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxetane, and diethylene glycol dimethyl ether; Nitrils such as Ril, Isopyronitrile; Formamide, N, Amides such
  • the base to be used is not particularly limited as long as it is used as a base in a usual reaction.
  • Organic bases such as gin, quinoline, N, N-dimethylaniline and N, N-getylaniline can be mentioned, and preferably, triethylamine.
  • Protecting agents used include, for example, t-butyldimethylsilyl chloride, trimethylsilyl chloride, triethylsilyl chloride, triethylsilyl bromide, triisopropylsilyl chloride, Dimethyl isoprovirsilyl chloride, Jethyluisoprovirsilyl chloride, t-butyl Silyl halides, such as diphenylsilyl chloride, diphenylmethylsilyl chloride, triphenylsilinolechloride, 4—methoxytriphenylmethyl chloride, 4,4'-dimethoxide Triphenyl halides such as trifluoromethyl chloride, 4,4 ', 4 "-trimethoxytriphenylmethyl chloride, benzyl chloride, benzyl bromide, p-methoxybenzyl bromide And aralkylhalides such as t-butyldiphenylsilyl chloride.
  • the reaction is usually carried out at a temperature of from 120 ° C to the reflux temperature of the solvent used, preferably from 0 ° C to the reflux temperature of the solvent used.
  • the reaction time varies depending mainly on the reaction temperature, the starting compound, the base used or the type of the solvent used, but is usually from 10 minutes to 3 days, preferably from 1 hour to 24 hours. .
  • the target compound (2) of this reaction is collected from the reaction mixture according to a conventional method. For example, neutralize the reaction solution, add an immiscible organic solvent such as water and ethyl acetate, wash with water, separate the organic layer containing the target compound, dry with sodium sulfate anhydride and dry the solvent. Is obtained by distillation.
  • an immiscible organic solvent such as water and ethyl acetate
  • an oligonucleotide analog having the above structure (1a) or (1b) can be produced by the method C or the method D described below.
  • R 13 represents a protecting group for nucleic acid synthesis of a hydroxyl group (particularly, a trityl group which may be substituted by a methoxy group)
  • R 14 represents a phosphonyl group
  • the compound (1c) produced by the method A in an inert solvent (the compound corresponds to the compound (1) obtained by deprotecting R 1 and R 3 in step A-8) ), And reacting the protecting reagent to produce compound (12).
  • the solvent used preferably, aromatic hydrocarbons such as benzene, toluene, and xylene; methylene chloride, chloride, carbon tetrachloride.
  • the protecting reagent used can be selectively protected at only the 5'-position and is acidic. There is no particular limitation as long as it can be removed under neutral conditions. Triarylmethyl halides such as LID, monomethoxytrityl chloride and dimethoxytrityl chloride.
  • the base to be used may be a heterocyclic amide such as pyridine, dimethylaminopyridine, pyrrolidinopyridine, or an aliphatic tertiary such as trimethylamine or triethylamine.
  • Amines are preferred, with pyridin, dimethylamino pyridine, and pyrrolidino pyridine being preferred.
  • the base When a liquid base is used as the solvent, the base itself acts as a deoxidizing agent, so there is no need to add a base again.
  • the reaction temperature is usually from 0 to 150 ° C, preferably from 20 to 100 ° C, depending on the starting materials, reagents, solvents and the like used.
  • the reaction time varies depending on the raw materials, solvent, reaction temperature, etc. used, but is usually 1 to 100 hours. Preferably, it is 2 to 24 hours.
  • the target compound (12) of this reaction is collected from the reaction mixture according to a conventional method. For example, concentrate the reaction mixture, add an immiscible organic solvent such as water and ethyl acetate, wash with water, separate the organic layer containing the target compound, dry over anhydrous sodium sulfate, etc., and evaporate the solvent Obtained by:
  • the obtained compound can be further purified by a conventional method, for example, recrystallization, silica gel chromatography, or the like.
  • step C-11 the compound (12) produced in step C-11 is reacted with a monosubstituted monochloro (alkoxy) phosphine or a disubstituted monoalkoxyphosphine usually used for amidation in an inert solvent.
  • step C-11 a step of producing a compound (Id) which is one of the bicyclonucleoside derivatives of the present invention.
  • the solvent to be used is not particularly limited as long as it does not affect the reaction, but is preferably a ether such as tetrahydrofuran, getylether, or dioxan. .
  • Examples of the mono-substituted mono- (alkoxy) phosphines used include, for example, chloro (morpholino) methoxyphosphine, chloro (morpholino) cyanoethoxyphosphine, and chloro (dimethylamino) ) Phosphines such as methoxy phosphine, chloro mouth (dimethylamino) cyano ethoxy phosphine, clo mouth (diisopropyl amino) methoxy phosphine, and cro mouth (diisopropyl amino) cyano ethoxy phosphine.
  • chloro (morpholino) methoxyphosphine chloro (morpholino) cyanoethoxyphosphine
  • chloro (dimethylamino) ) Phosphines such as methoxy phosphine, chloro mouth (dimethylamino) cyano ethoxy phosphine, clo mouth
  • the cro-mouth (morpholino) methoxy phosphine, the cro-mouth (morpholino) cyanoethoxy phosphine, the clo mouth (disopropylamino) methoxy phosphine, the cro mouth (diisopropylamino) cyano-ethoxy phosphine are preferred. It is.
  • a deoxidizing agent is used.
  • the deoxidizing agent used is a heterocyclic ring such as pyridine or dimethylaminopyridine.
  • Amines, trimethylamine, trie Aliphatic amines such as tilamine and diisopropylethylamine are preferable, and aliphatic amines (particularly, diisopropylethylamine) are preferred.
  • di-substituted monoalkoxyphosphines examples include bis (diethylamino) cyanoethoxyphosphine, bis (getylamino) methansulfonylethoxyphosphine, bis (diisopropylamino) (2,2, Examples thereof include phosphines such as 2-tricycloethoxy) phosphine and bis (diisopropylamino) (4-chlorophenylmethoxy) phosphine, and preferably bis (getylamino).
  • It It is cyanoethoxyphosphine.
  • an acid is used, and in this case, the acid used is preferably tetrazole, acetic acid or P-toluenesulfonic acid.
  • reaction temperature is not particularly limited, it is generally ⁇ 50 to 50 ° C., and preferably room temperature.
  • the reaction time varies depending on the starting materials, reagents, temperature and the like to be used, but is usually 5 minutes to 30 hours, preferably 30 minutes when reacted at room temperature.
  • the target compound (13) of this reaction is collected from the reaction mixture according to a conventional method.
  • the reaction mixture is appropriately neutralized, and if insolubles are present, after removal by filtration, an immiscible organic solvent such as water and ethyl acetate is added. It is obtained by separating the layer, drying with anhydrous sodium sulfate or the like, and distilling off the solvent.
  • the obtained target compound can be further purified, if necessary, by a conventional method, for example, recrystallization, reprecipitation, or silica gel column chromatography.
  • the compound (12) produced by C-11 in an inert solvent preferably, a halogenated hydrocarbon such as methylene chloride
  • an inert solvent preferably, a halogenated hydrocarbon such as methylene chloride
  • the reaction temperature is not particularly limited, usually an 2 0 to 1 0 0 ° C, preferably from 1 0 to 4 0 D C.
  • the reaction time varies depending on the starting materials, reagents, temperature and the like to be used, but is usually 5 minutes to 30 hours, preferably 30 minutes when reacted at room temperature.
  • the target compound (13) of this reaction is collected from the reaction mixture according to a conventional method.
  • the reaction mixture is appropriately neutralized, and if insolubles are present, after removal by filtration, an immiscible organic solvent such as water and ethyl acetate is added. It is obtained by separating the layer, drying with anhydrous sodium sulfate or the like, and distilling off the solvent.
  • the obtained target compound can be further purified by a conventional method, for example, recrystallization, reprecipitation or chromatography.
  • At least one compound (1d) produced by C-12 and a commercially available phosphoramidite reagent necessary for producing an oligonucleotide having a desired nucleotide sequence are used. And the like, using an ordinary method to produce an oligonucleotide analog containing the desired structure (1a) or (1b) on an automatic DNA synthesizer.
  • Oligonucleotide analogs having a desired nucleotide sequence can be obtained from a DNA synthesizer, for example, using a phosphoramidite model 3922 of PerkinElmer Inc. (Nucleic Acids Research, 12, 4539 (1984)). ) It can be manufactured according to the method described.
  • TETD tetraethyl uram disulfide
  • Beau cage reagent Millipore
  • a reagent for forming a thioate by the method described in the literature (Tetarhedron Letters, 32, 3005 (1991), J. Am. Chem. Soc, 112, 1253 (1990)). Can be manufactured.
  • the resulting crude oligonucleotide can be purified using an Oligopack (reverse-phase chromatography), and the purity of the purified product can be confirmed by HPLC analysis.
  • 8 1 and scale 8 is the same as defined above, and the B 1 and B 1 of the formula (1 3) of the formula (1 d), may be the same or different.
  • R 1 7 shows such as succinic acid CPG (succinyl Controlled Pore Glass) or TEN Tageru (Tentagel), that are usually used in the synthesis of cage Gonuku Reochido resin.
  • CEO refers to the 2-cyanoethoxy group.
  • This step is a step of producing the compound (14) by reacting the compound (1e) with the compound (13), and is described in the literature (Nucleic Acids Research, Vol. 23, No. 14, pp. 2661-). 2668, 1995).
  • This step is a step of producing an oligonucleotide analog from the compound (14) obtained in the step D_1.
  • the protecting group R 1 for the hydroxyl group is deprotected according to the usual method, and then phosphorylated by the method described in the above-mentioned literature.
  • the reaction is repeated, and oligo nucleotides are synthesized by repeating this cycle.
  • the chain length of the obtained oligonucleotide nucleoside is usually 2 to 50, preferably 10 to 30, as nucleoside units.
  • the obtained oligonucleotide analog is not easily degraded by various nucleases and can exist in the living body for a long time after administration to the living body.
  • a stable duplex is formed with mRNA to inhibit the biosynthesis of a pathogenic protein, or a triplex is formed with a double-stranded DNA in the genome to form It can also inhibit transcription into RNA and inhibit the growth of viruses that infect cells.
  • the bicyclonucleoside derivative of the present invention can continuously enhance the action of interferon by being used as a 2-5A analog, and is a drug that regulates various biological phenomena involving 2-5A.
  • 2-5 A analogs and various antisense oligonucleotides can be combined with certain linkers. The conjugation can also enhance the activity of the antisense oligonucleotide (Proc. Natl. Sci. USA Vol. 95, pp. 8874-8879, July 1998 Medical Sciences).
  • oligonucleotide analogues of the present invention are expected to be used as pharmaceuticals for inhibiting diseases such as antitumor agents and antiviral agents, as well as for treating diseases.
  • oligonucleotide derivatives of the present invention can be formulated into a parenteral administration preparation or a ribosome preparation, for example, by blending a conventional auxiliary agent such as a buffer and / or a stabilizer.
  • a conventional auxiliary agent such as a buffer and / or a stabilizer.
  • a common pharmaceutical carrier can be compounded to prepare an ointment, cream, solution, salve or the like.
  • the dosage varies depending on symptoms, age, administration method, etc., for example, once per dose, 0.001 mg / kg body weight (preferably 0.01 mg / kg body weight) as the lower limit, and 100 mg as the upper limit / kg body weight (preferably 10 mg / kg body weight) should be used once or several times daily depending on the symptoms.
  • the oligosaccharide is cut from the support by concentrated ammonia water treatment according to a conventional method. In both cases, the protecting group on the phosphorus atom, the cyanoethyl group, was removed, and the protecting group on the nucleobase was also removed.
  • the obtained native oligonucleotides and modified oligonucleotide analogs were purified by reverse phase HPLC to obtain the target oligonucleotides. According to this synthesis method, the following sequence:
  • oligonucleotide (1) 5'-ttttttttttnt-3 '(SEQ ID NO: 1 in the sequence listing), having a base number of 11 and having an n force S 3'-amino-3'-doxy-3'-N, 4'- An oligonucleotide analog which is C-methylene-5-methylperidine (hereinafter referred to as “oligonucleotide (1)”) was obtained.
  • Acetic anhydride (850 ⁇ l) and concentrated sulfuric acid (25 ⁇ l) were added to an acetic acid solution (8 ml) of the compound of Reference Example 7 (525 mg, 0.98 mmol) under a nitrogen stream, and the mixture was stirred at room temperature for 2 hours. Under ice-cooling, the reaction solution was neutralized by adding a saturated aqueous solution of sodium bicarbonate, extracted with ethyl acetate, and washed with saturated saline.
  • the reaction solution was neutralized (weakly acidic) by adding saturated aqueous sodium bicarbonate under ice-cooling, and then added with saturated saline, extracted with ethyl acetate, and washed with saturated saline.
  • the desired product was obtained as a colorless oil (930 mg, 82%).
  • Buffer solution containing various oligonucleotides (80 ⁇ g) 2250 1 50 mM Tris (pH 8.0) and lOmM MgCl 2 ) in 3,-exonuclease (phosphodiesterase from Cro talus durissus (Boehringer Mannheim)
  • Add 0.3 / g and keep the mixture at 37 ° C to perform the reaction. After a certain period of time, remove a portion of the mixture and heat it at 90 ° C for 2 minutes to inactivate the enzyme and stop the reaction.
  • the novel bicyclonucleoside derivative of the present invention has excellent antisense or antigene activity and is an intermediate for producing an oligonucleotide analog that is stable in vivo. It is useful as an intermediate for the production of analogs of 2-5 A that are known as body and biological substances having antiviral activity.
  • novel oligonucleotide analogues of the present invention are stable in vivo and are useful as antisense or antigen drugs.

Abstract

Cette invention concerne des composés représentés par la formule (I) ou des sels pharmaceutiquement acceptables de ces composés convenant en tant qu'intermédiaires pour la préparation d'analogues oligonucléotidiques non naturels ou d'analogues 2-5A et qui présentent une excellente activité antisens ou antigène et sont stables in vivo. Dans la formule (I), R1 et R3 sont chacun indépendamment un hydrogène, un groupe protecteur hydroxyle pour la synthèse d'acides nucléiques ou analogues ; R2 est hydrogène, alkyle en C¿1?-C6 ou un groupe protecteur amino for la synthèse d'acides nucléiques : et B est un groupe purine-9-yle ou 2-oxo-1,2-dihydropyrimidine-1-yle qui peut avoir un substituant pris parmi halogéno, alkyle en C1-C6, hydroxyle, mercapto, amino, etc.
PCT/JP2000/004092 1999-06-25 2000-06-22 Nouveaux dérivés de bicyclonucléoside WO2001000641A1 (fr)

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Cited By (15)

* Cited by examiner, † Cited by third party
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WO2004037842A1 (fr) * 2002-10-23 2004-05-06 Sankyo Company, Limited Nouveaux acides nucleiques synthetiques dont les fractions saccharides sont a configuration s
EP1959012A2 (fr) 2004-12-29 2008-08-20 Exiqon A/S Nouvelles compositions oligonucléotides et séquences de sonde utiles pour la détection et l'analyse de micro ARN et leurs ARN cibles
EP2090665A2 (fr) 2006-10-20 2009-08-19 Exiqon A/S Nouveaux micro ARN humains associés au cancer
US8048998B2 (en) 2007-01-19 2011-11-01 Exiqon A/S Mediated cellular delivery of LNA oligonucleotides
WO2011134474A1 (fr) 2010-04-30 2011-11-03 Exiqon A/S Procédé et tampon d'hybridation in situ
WO2012061778A2 (fr) 2010-11-05 2012-05-10 Genalyte, Inc. Systèmes optiques de détection d'analytes et leurs procédés d'utilisation
US8188255B2 (en) 2006-10-20 2012-05-29 Exiqon A/S Human microRNAs associated with cancer
US8192937B2 (en) 2004-04-07 2012-06-05 Exiqon A/S Methods for quantification of microRNAs and small interfering RNAs
WO2013138251A1 (fr) 2012-03-12 2013-09-19 The Board Of Trustes Of The University Of Illinois Systèmes optiques de détection d'analyte avec amplification magnétique et procédés d'utilisation
WO2014134144A1 (fr) 2013-02-28 2014-09-04 The General Hospital Corporation Compositions de profilage de miarn et procédés d'utilisation
US9464106B2 (en) 2002-10-21 2016-10-11 Exiqon A/S Oligonucleotides useful for detecting and analyzing nucleic acids of interest
US9846126B2 (en) 2008-10-27 2017-12-19 Genalyte, Inc. Biosensors based on optical probing and sensing
US9983206B2 (en) 2013-03-15 2018-05-29 The Board Of Trustees Of The University Of Illinois Methods and compositions for enhancing immunoassays
US10365224B2 (en) 2007-12-06 2019-07-30 Genalyte, Inc. Label-free optical sensors
US11111535B2 (en) 2003-06-20 2021-09-07 Qiagen Gmbh Probes, libraries and kits for analysis of mixtures of nucleic acids and methods for constructing the same

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US9464106B2 (en) 2002-10-21 2016-10-11 Exiqon A/S Oligonucleotides useful for detecting and analyzing nucleic acids of interest
WO2004037842A1 (fr) * 2002-10-23 2004-05-06 Sankyo Company, Limited Nouveaux acides nucleiques synthetiques dont les fractions saccharides sont a configuration s
US11111535B2 (en) 2003-06-20 2021-09-07 Qiagen Gmbh Probes, libraries and kits for analysis of mixtures of nucleic acids and methods for constructing the same
US8383344B2 (en) 2004-04-07 2013-02-26 Exiqon A/S Methods for quantification of microRNAs and small interfering RNAs
US8192937B2 (en) 2004-04-07 2012-06-05 Exiqon A/S Methods for quantification of microRNAs and small interfering RNAs
EP1959012A2 (fr) 2004-12-29 2008-08-20 Exiqon A/S Nouvelles compositions oligonucléotides et séquences de sonde utiles pour la détection et l'analyse de micro ARN et leurs ARN cibles
US8188255B2 (en) 2006-10-20 2012-05-29 Exiqon A/S Human microRNAs associated with cancer
EP2090665A2 (fr) 2006-10-20 2009-08-19 Exiqon A/S Nouveaux micro ARN humains associés au cancer
US8048998B2 (en) 2007-01-19 2011-11-01 Exiqon A/S Mediated cellular delivery of LNA oligonucleotides
US10365224B2 (en) 2007-12-06 2019-07-30 Genalyte, Inc. Label-free optical sensors
US11041811B2 (en) 2008-10-27 2021-06-22 Genalyte, Inc. Biosensors based on optical probing and sensing
US9846126B2 (en) 2008-10-27 2017-12-19 Genalyte, Inc. Biosensors based on optical probing and sensing
US10023912B2 (en) 2010-04-30 2018-07-17 Exiqon A/S In situ hybridization method and buffer
WO2011134474A1 (fr) 2010-04-30 2011-11-03 Exiqon A/S Procédé et tampon d'hybridation in situ
US9212390B2 (en) 2010-04-30 2015-12-15 Exiqon A/S In situ hybridization method and buffer
US9921165B2 (en) 2010-11-05 2018-03-20 Genalyte, Inc. Optical analyte detection systems and methods of use
WO2012061778A2 (fr) 2010-11-05 2012-05-10 Genalyte, Inc. Systèmes optiques de détection d'analytes et leurs procédés d'utilisation
EP3266881A1 (fr) 2010-11-05 2018-01-10 Genalyte, Inc. Systèmes de détection d'analytes optiques et procédés d'utilisation
EP3733866A1 (fr) 2010-11-05 2020-11-04 Genalyte, Inc. Systèmes de détection d'analytes optiques et procédés d'utilisation
WO2013138251A1 (fr) 2012-03-12 2013-09-19 The Board Of Trustes Of The University Of Illinois Systèmes optiques de détection d'analyte avec amplification magnétique et procédés d'utilisation
EP3907506A1 (fr) 2012-03-12 2021-11-10 The Board of Trustees of the University of Illinois Systèmes optiques de détection d'analyte avec amplification magnétique et leurs procédés d'utilisation
WO2014134144A1 (fr) 2013-02-28 2014-09-04 The General Hospital Corporation Compositions de profilage de miarn et procédés d'utilisation
US9983206B2 (en) 2013-03-15 2018-05-29 The Board Of Trustees Of The University Of Illinois Methods and compositions for enhancing immunoassays
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