WO2005092909A1 - Procédés de fabrication d'analogue ribonucléotide de forte stéréorégularité et analogue désoxyribonucléotide - Google Patents

Procédés de fabrication d'analogue ribonucléotide de forte stéréorégularité et analogue désoxyribonucléotide Download PDF

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WO2005092909A1
WO2005092909A1 PCT/JP2005/003812 JP2005003812W WO2005092909A1 WO 2005092909 A1 WO2005092909 A1 WO 2005092909A1 JP 2005003812 W JP2005003812 W JP 2005003812W WO 2005092909 A1 WO2005092909 A1 WO 2005092909A1
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group
carbon atoms
represented
general formula
hydroxyl
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PCT/JP2005/003812
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Japanese (ja)
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Kazuhiko Saigo
Takeshi Wada
Satoshi Fujiwara
Terutoshi Sato
Naoki Iwamoto
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Toudai Tlo, Ltd.
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Priority to JP2006511417A priority Critical patent/JP4865544B2/ja
Publication of WO2005092909A1 publication Critical patent/WO2005092909A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
    • C07H21/02—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with ribosyl as saccharide radical
    • 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
    • C07H19/20—Purine 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
    • C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
    • C07H21/04—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with deoxyribosyl as saccharide radical
    • 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 a ribonucleotide analog having high stereoregularity and an oligodoxyribonucleotide analog.
  • the antisense method is a method in which a nucleic acid having a nucleotide sequence complementary to a target mRNA is selectively bound to the mRNA to inhibit protein translation.
  • the properties required as an antisense molecule include (1) the ability to recognize and specifically bind to the base sequence of the target mRNA, (2) the ability to form a stable duplex, and (3) the nuclease. (4) high cell membrane permeability.
  • Phosphate moiety-modified dinucleotides have an asymmetric center on the phosphorus atom and differ in their antisense effects due to differences in their absolute configuration.
  • properties of phosphate-modified dinucleotides such as their ability to form duplexes with DNA and RNA, nuclease resistance, and RNase H activity, are affected by chirality on the phosphorus atom.
  • DNA analogs in which two non-bridging oxygen atoms on the phosphorus atom of natural DMA internucleotides are variously substituted that is, inter-nucleotide modified analogs, have both nuclease resistance and cell membrane permeability. It is known to increase (Lev in, AA B i ochem.
  • the DNA analog will have chirality on the phosphorus atom. It is known that the properties and functions of these DNA relatives differ depending on the chirality (Yu, D .; Kanduma lla, ER; Rosky, A .; Zhao, Q .; Chen, J .; Agrawal , S. Bioorg. Med. Ghem., 2000, 8, 275-284.)).
  • phosphorothioate DNA an internucleotide-modified DNA analog in which one of two non-bridging oxygen atoms has been replaced with a sulfur atom, has a double-stranded structure that forms with complementary RNA, a nuclease.
  • H-phosphonate DNA is a DNA analog in which one of the two non-bridging oxygen atoms on the phosphorus atom of the internucleotide of natural DNA is replaced with a hydrogen atom. Having. In addition, it can be converted into various inter-nucleotide-modified DMA analogs by a stereospecific conversion reaction. Thus, if stereochemically pure H-phosphonate DNA is obtained, it will be possible to obtain an internucleotide-modified DMA analog whose stereochemistry is controlled as it is, using the DNA as it is. As described above, H-phosphonate DNA has various three-dimensionally controlled proteins. It is a useful synthetic intermediate that can be converted into a single nucleotide modified DNA analog.
  • stereochemically pure H-phosphonate DMA has only been reported at the dimer level ((a) See la, F .; Kretschner, UJ Org. Chem. 1991, 56, 3861-3869. (B) Loshmer, T .; Engels, JW Nucleic Acids Res. 1990, 18, 5143). Moreover, even when the dimer of H-phosphonate DNA is optically resolved, H-phosphonate DNA is unstable on silica gel column chromatography, and there is a polar difference between the two diastereomers.
  • H-phosphonate DNA Since there is no significant difference, the optical division is extremely inefficient. Considering stereochemically pure H-phosphonate DNA as a synthetic intermediate that can be applied to nucleic acid medicine, H-phosphonate DNA with a steric control at the oligomer level is required. In that case, the number of diastereomers increases exponentially, making optical resolution of H-phosphonate DMA oligomers virtually impossible. Therefore, if a stereoselective synthesis reaction of H-phosphonate DMA can be developed, it will be possible to obtain H-phosphonate DMA with steric control at the oligomer level.
  • RNAi was first reported in 1998 in a study using nematodes by Fire and Mel lo (Fire, A .; Xu, S .; Montgomery, ⁇ . ⁇ .; Kostas, SA; Driver, SE; Mel lo , CG Nature. 1998, 391, 806-811.), It has been revealed that it is a gene silencing system that is conventionally provided among various species such as insects, plants, and fungi. . In 2001, Tuschl et al. Showed that RNAi was applicable to mammalian cells (Elbashir, SM; Harborth, J .; Lendeckel,.; Yale in, A .; Weber, K .; Tuschl, T. Nature. 2001, 411, 494-498.).
  • RNAi has been attracting attention as a powerful method for gene therapy and gene function analysis, as an excellent gene suppression method with high gene suppression effect.
  • RNAi is characterized by its sequence specificity, which can knock out the target gene accurately, and the use of gene silencing mechanisms inherent in living organisms. This makes RNAi a promising new gene therapy with fewer side effects. ing.
  • the effect of RNAi cannot be maintained for a long time at present, because RNA strands of about 21 bases such as siRNA are gradually degraded by nucleases in the living body. Disclosure of the invention
  • An object of the present invention is to provide a method for producing a ribonucleotide analog and a deoxysilipnucleotide analog having a high stereoregularity, which can be used for the antisense method or RNA interference and has a controlled stereo structure on a phosphorus atom. Is to do.
  • the present invention provides a compound represented by the following general formula (I):
  • R 1 and R ′ may be the same or different and represent a hydrogen atom, an alkyl group having 1 to 3 carbon atoms, or an aryl group having 6 to 14 carbon atoms,
  • R 2 and R may be the same or different and represent a hydrogen atom, an alkyl group having 1 to 3 carbon atoms or an aryl group having 6 to 14 carbon atoms;
  • R 3 represents an alkyl group having 1 to 3 carbon atoms
  • R 4 is a hydroxyl-protecting group, is OR 5 (where R 5 is a hydroxyl-protecting group), a hydroxyl group or a hydrogen atom,
  • R 2 and R 3 may form a monocyclo structure or a bicyclo structure together with the nitrogen atom.
  • X— is BF 4 —, PF 6 —, T f O— (T f is CF 3 S 0 2 —; the same applies hereinafter), T f 2 N ⁇ A s F 6 — or S b F 6 — is indicated.
  • the cyclic structure A represents a monocyclo or bicyclo structure having 3 to 16 carbon atoms formed together with a nitrogen atom.
  • a method for producing a nucleotide analog, and a method for producing an oligoribonucleotide analog and an oligodoxyribonucleotide analog having high stereoregularity are provided.
  • B s has the same meaning as
  • first reaction step a condensation reaction
  • second reaction step a reaction with an electrophile
  • first and second divisions are for convenience of explanation only and are not limiting. If necessary, known processing steps such as purification processing can be added.
  • nucleoside (I) An optically active nucleoside 3'-phosphoramidite represented by the general formula (I) [hereinafter referred to as “phosphoramidite (I)”] and a nucleoside represented by the general formula (II) [hereinafter referred to as “nucleoside (II)”] Is condensed in the presence of an activating agent represented by the general formula (III) [hereinafter referred to as “activating agent (III)”].
  • activating agent (III) activating agent
  • the phosphoramidite (I) can be produced from an appropriate 1,2-amino alcohol by a known method as described below (for example, see Tetrahedron: Asy et al. 1995, 6, 1051-1054).
  • the general formula (VII) obtained by reacting an optically active 1,2-amino alcohol (hereinafter referred to as “amino alcohol (VI)”) represented by the general formula (VI) with phosphorus trichloride is used. It can be obtained by reacting the optically active phosphitylating agent represented by [hereinafter referred to as “phosphitylating agent (VII) J”] with the nucleoside represented by the general formula (VIII).
  • the amino alcohols (VI) include (S)-and (R) -2-methylamino-1-phenylethanol, (1R, 2S) -ephedrine, (1R, 2S) 1-2-methylamino-1 , 2-diph Xnylethanol and the like.
  • prolinol derivatives for example, (Of R, 2S)-(pyrrolidine-12-yl) benzyl alcohol, (aS, 2R)-(pyrrolidine-12-yl) benzyl alcohol Amino alcohols that can be converted to H-phosphonates, such as (S) -a, of-diphenyl (pyrrolidine-1-yl) methanol and (2S) -monomethyl (pyrrolidine-l-2-yl) ethanol Ethyl, (2R) -bimethyl (pyrrolidine-1-yl) ethanol, (oiR, 2S)-monomethyl (pyrrolidine-12-yl) benzyl alcohol, (S, 2R)-monomethyl (Pyrrolidine-1-yl)
  • Bs represents a group derived from peracyl, adenine, cytosine, guanine or thymine or a derivative thereof.
  • R 7 has the same meaning as above, and R 8 represents an alkyl group having 1 to 15 carbon atoms, an aryl group, an aralkyl group, an aryloxyalkyl group, among which a methyl group, an isopropyl group, A phenyl group, a benzyl group and a phenoxymethyl group are preferred, and a phenyl group is particularly preferred.
  • R 9 and R 1C) each represent an alkyl group having 1 to 4 carbon atoms, and a methyl group is particularly preferable.
  • R represents a protecting group at position 06 of guanine, preferably 2-cyanoethyl group, p-nitrophenylethyl group, phenylsulfonylethyl group, benzyl group, 2-trimethylsilylethyl group or the like.
  • Nucleoside (VIII) is Urashiru, adenosine, which was protected cytidine, guanosine, the hydroxyl group of thymine or 5 'position of their derivatives, the protecting group (R 4), tert - butyl diphenyl silyl group (TBDPS) Alkylsilyl groups such as tert-butyldimethylsilyl group (TBDMS), trityl groups such as 4,4'-dimethyoxytrityl group (DMT r) and 4-methoxytrityl group (MMT r), and a protecting group represented by the following formula: And the like.
  • Bs When is in the nucleoside (VIII) is a hydrogen atom, Bs is preferably thymine or a derivative thereof. When using a nucleoside other than thymine or a derivative thereof, it is desirable to introduce a protecting group into the base, since side reactions to the base may be feared.
  • Adenine and guanine can use the phenoxyacetyl (Pac) group, and cytosine can use the isobutyl (iBu) group.
  • R 1 and R 2 one of R 1 and R 2 is a hydrogen atom and the other is a phenyl group, one of R 1 and R 2 is a methyl group and the other is a phenyl group, or R 1 and R 2 R 2 is preferably a combination of phenyl groups, R 1 is a phenyl group, and R 2 is more preferably a combination of hydrogen atoms.
  • R 3 is preferably a methyl group. Further, it is preferable that R 1 forms a phenyl group and R 2 and R 3 form a pyrrolidine skeleton together with a nitrogen atom.
  • R 4 and R 5 are OR 5 , R 5 is preferably TBDP S or TBDMS, and more preferably TBD PS.
  • R ′ and R ′′ can be selected from a hydrogen atom, an alkyl group having 1 to 3 carbon atoms or an aryl group having 6 to 14 carbon atoms.
  • R 1 and R ′ may be the same or different and each may be an alkyl group having 1 to 3 carbon atoms or an alkyl group having 6 to 14 carbon atoms.
  • both R 1 and R are not hydrogen atoms (that is, R 1 and R ′ are The carbon atom to be bonded is a tertiary carbon), and the substituent of the tertiary carbon does not include an aryl group.
  • Nucleoside (II) protects the hydroxyl groups at the 2- and 3-positions of peridine, adenosine, cytidine, guanosine or their derivatives, and peracyl, adenine, cytosine, guanine, thymine or their derivatives represented by Bs
  • the group derived from is exemplified by the nucleoside (VIII).
  • the Bs of the nucleoside (II) and the nucleoside (VI) may be the same or different.
  • R 6 is the same as above, and when E, is 10 R 7 , the protecting group for the hydroxyl group represented by R 7 is TBDPS, TBDMS, acetyl group (Ac), phenoxy Asechiru group (PA c), benzyl group (B z), DMT r, MM Ding r etc. mentioned et been, R 6, R 7 is PA c is preferred.
  • the activator (III) has a capability of supplying a proton to the nitrogen atom of the phosphoramidite (I) and does not act as a nucleophile.
  • X— is preferably BF 4 —, PF 6 —, T f O—, or T f 2 N—.
  • the cyclic structure A represents a monocyclo or bicyclo structure having 3 to 16 carbon atoms formed with a nitrogen atom, and particularly preferably has a monocyclo structure represented by the formula (IU-1).
  • N represents a number of 3 to 7, preferably 4 or 5.
  • the activator (III) has the formula (IX)
  • the reaction between the phosphoamidite (I) and the nucleoside (M) is preferably carried out in a solvent such as acetonitrile.
  • ) are reacted with nucleoside (II) at a ratio of 5 to 1.0 equivalent times to phosphoramidite (I).
  • Activator (III) reacts with phosphoramidite (I)
  • the reaction temperature is preferably 0 to 40 ° C, and the reaction pressure is preferably 1 atm.
  • the phosphite (XI) obtained in the first reaction step is acylated with acetic anhydride, trifluoroacetic anhydride or the like, and then reacted with an electrophilic reagent such as a sulfurizing agent, a selenating agent, or a boranolating agent. Then, the asymmetric auxiliary group of the compound of the general formula (XII) is treated with 1,8-diazabicyclo [5.4.0] pendecar 7-ene (DBU) and the like to remove the compound.
  • DBU 1,8-diazabicyclo [5.4.0] pendecar 7-ene
  • E 13 ⁇ 4 B s and Y have the same meaning as described above.
  • a protected diphosphate site-modified dinucleotide represented by
  • the type of electrophile used for example, 1,2,4-dithiazolidine-1,3,5-dione, 3-ethoxy-1,2,4-dithiazoline-5-one, 3-methyl
  • the preceding acylation step may be omitted.
  • oligomer represented by the general formula (XIII) [hereinafter referred to as “oligomer”
  • the carbon to which R 1 of the monomer represented by the general formula (I) is bonded is a tertiary carbon (both R 1 and R ′ are not hydrogen atoms), and the substituent of the tertiary carbon is
  • the phosphite (XI) obtained in the first reaction step does not contain an aryl group
  • the phosphite (XI) obtained in the first reaction step is acylated with anhydrous acetic acid, trifluoroacetic anhydride, or the like, and then is acidified with an acid such as a 1% trifluoroacetic acid dichloromethane solution of methane.
  • the acylation step can be omitted in the second reaction step, but in order to reduce the carbocation formed, It is necessary to add a reducing agent such as triethylsilane or borane-pyridine complex.
  • an oligomer When an oligomer is synthesized by this method, a monomer having a DMTr group as a protecting group for the 5 ′ hydroxyl group is used, and the phosphite intermediate obtained by the above method is subjected to an acid treatment to form an asymmetric auxiliary group and a 5 ′ hydroxyl group.
  • the protecting group, DMTr is removed at the same time, and the resulting dimer having a hydroxyl group at the 5'-position is condensed with a monomer.
  • an oligomer having an H-phosphonate bond is subjected to a conversion reaction in the same manner as in the case of a dimer, and is guided to a desired phosphorus atom-modified DNA, followed by deprotection, whereby a target nucleic acid analog is obtained. Obtainable.
  • n represents an integer of 1 to 150, a preferred range is 5 to 50, a more preferred range is 10 to 30, and a still more preferred range is 15 to 22.
  • D 2 and E 2 represent a hydroxyl group or a hydrogen atom.
  • the oligomer (XII I) can be produced by applying an oligomer synthesis method by a solid phase method.
  • a solid phase method Specifically, a commercially available automatic synthesizer (Expedite, manufactured by ABI, or ABI Model 394, DNA / RNA Synthesizer ABI) Or a manual method using a solid-phase synthesis vessel equipped with a glass filter.
  • Solid-phase carriers used in the solid-phase method include aminoalkylated porous glass (control led pore glass: CPG) and aminoalkylated highly cross-linked polystyrene (HCP). ) Is preferred, and a polymer carrier that is as swellable as possible and can easily remove excess reagents by washing.
  • Either the 3 'or 2' hydroxyl groups of the ribonucleoside and the solid support are bound via a linker such as succinate, oxalate, or phthalate. May be.
  • a linker such as succinate, oxalate, or phthalate. May be.
  • Protecting groups for 2 'or 3' hydroxyl groups to which the solid phase carrier is not bound include acetyl, benzoyl, 2- (cyanoethoxy) ethyl, t-butyldimethylsilyl, and other RNA and DNA synthesis groups. The protecting groups used can be mentioned.
  • the highly stereoregular ribonucleotide analogs and deoxyribonucleotide derivatives obtained by the production method of the present invention can be used for antisense and RNA interference, which are one of the methods that have attracted attention in the field of gene therapy. Can be used.
  • ribonucleotide analogs and deoxyribonucleotide analogs having high stereoregularity and effective as antisense molecules can be obtained in high yield.
  • Production Example 1-1 Production of N-cyanomethylpyrrolidinium tetrafluoroborate
  • Production Example 1-2 Production of ⁇ -cyanomethylpyrrolidinium hexafluorophosphate
  • Production Example 2-2 Production of (5 R) —2-chloro-1--3-methyl-1-phenyl-1,1,3,2-oxazaphospholidine
  • Triethylamine (1.05 ml, 7.5 inmol) was added to the mixture, and the mixture was cooled to 78 ° G. Then, under an argon atmosphere, a 0.22 M THF solution of (5S) -18d shown in the following formula and Table 1 was added dropwise. After the reaction mixture was stirred at room temperature for 30 minutes, a saturated aqueous solution of sodium hydrogencarbonate (75 ml) and chloroform (75 ml) were added.
  • trans- 19b (0.0520 g, 50 mo I) and 2 ', 3' - 0 - off enoki Xia cetyl ⁇ lysine (0.0256 £, 50 ⁇ ⁇ ) 12 hours in a vacuum drying at [rho 2 0 5 on ⁇ — (cyanomethyl) pyrrolidinium trifluorofluoroester dried for 8 hours with MS 3 ⁇ A 0.25 M solution of methanesulfonate (27a) (400, 100 mol) in acetonitrile and CD 3 CN (100 ju I) were added under an argon atmosphere.
  • Beaucage reagent (0.0120 g, 0.06 mmo I) was added to this solution to sulfide the compound 7.
  • reaction solution was transferred from the NMR sample tube to a 50 ml narrow-necked eggplant flask, washed with 3 ml of pyridine, and then added with 20 ml of a mixed solution of ammonia water / ethanol (3: 1, v / v). In addition, it was sealed and heat-treated at 60 ° C for 4 hours.
  • Example 2 Oligomer (XIII) was produced by the following reactions (1) to (4) and (5) (the following reaction formula).
  • the ribonucleotide bound to the solid support was treated with 50 equivalents of Beaucage reagent (0.5 M) in acetonitrile solution for 60 seconds to sulfide the phosphite intermediate. After the reaction was completed, the substrate was washed with acetonitrile.
  • the ribonucleotide bound to the solid support was treated with a solution of trichloromouth acetate in dichloromethane for 60 seconds to remove the DMTr group at the 5 'end. After the completion of the reaction, the resultant was washed with dichloromethane and then with acetonitrile.
  • the solid support is reacted with 25% aqueous ammonia: ethanol (3: 1, v / v) at 60 ° C for 15 hours.
  • aqueous ammonia: ethanol 3: 1, v / v
  • the protecting groups at the base moiety and the phosphate moiety were removed.
  • the 3'-terminal hydroxyl-protecting group and the extraction of the oligomer from the solid support also proceeded at the same time.
  • a saturated aqueous solution of ammonium chloride (50 ml) and a saturated aqueous solution of sodium chloride (50 ml) were added, and the mixture was extracted with chloroform (50 mix 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Then, 30 ml of hexane was added, and the mixture was vigorously stirred, filtered by suction, and dried in vacuo to obtain 3a (7.22 g, 88%). Colorless amorphous.
  • N-ethy I carbamate- (2S)-a? -Methyl (pyrrol idi ⁇ -2-y I) ethano I 3b (9.60 g, 48.7 mmol), add methanol (50 ml), cool to 0 ° C, With stirring, potassium hydroxide (27.0 g, 481.1 mmol) was added. After heating and refluxing for 4 hours while stirring, methanol was distilled off under reduced pressure, 50 ml of water was added, concentrated hydrochloric acid was added until the pH became 1 and the mixture was washed with ether (100 ml x 2) to produce The aqueous phase was recovered from the precipitate.
  • NMR sample tube, 7b (35.1 mg, 55 jumol ) and 9 (17.8 mg, 50 ⁇ Mol) was vacuum-dried for 12 hours over P 2 0 5, 8 and dried for 8 hours at MS 3A (400 il, 1O0 ⁇ mol) of 0.25 M acetonitrile and CD 3 CN (100 jw I) were added under an Ar atmosphere.
  • the collected residue was dissolved in water (0.2 ml) and analyzed by reverse phase HPLG.

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Abstract

Il est prévu un procédé de fabrication d'analogue ribonucléotide de forte stéréorégularité. Le procédé de fabrication d'analogue ribonucléotide de forte stéréorégularité représentée par la formule (IV) ou (V) est caractérisé par la condensation d’un nucléoside 3'-phosphoroamidite actif optiquement avec un nucléoside, à l’aide d’un activateur, puis soumission du condensat à la sulfuration et l'élimination de groupe de protection. (IV) (V) [Dans les formules, Y+ représente un alkyle linéaire ou à ramification C1-3, etc. ; et Bs représente l’urasile, etc., sous réserve que les deux Bs dans chaque formule soient identiques ou différents.]
PCT/JP2005/003812 2004-03-25 2005-02-28 Procédés de fabrication d'analogue ribonucléotide de forte stéréorégularité et analogue désoxyribonucléotide WO2005092909A1 (fr)

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WO2011108682A1 (fr) * 2010-03-05 2011-09-09 国立大学法人 東京大学 Procédé de fabrication de ribonucléoside phosphorothioate
JP2012510460A (ja) * 2008-12-02 2012-05-10 株式会社キラルジェン リン原子修飾核酸の合成方法
WO2014012081A2 (fr) 2012-07-13 2014-01-16 Ontorii, Inc. Contrôle chiral
WO2014010250A1 (fr) * 2012-07-13 2014-01-16 Chiralgen, Ltd. Groupe auxiliaire asymétrique
WO2015107425A2 (fr) 2014-01-16 2015-07-23 Wave Life Sciences Pte. Ltd. Conception chirale
WO2017015575A1 (fr) 2015-07-22 2017-01-26 Wave Life Sciences Ltd. Compositions d'oligonucléotides et méthodes associées
US9605019B2 (en) 2011-07-19 2017-03-28 Wave Life Sciences Ltd. Methods for the synthesis of functionalized nucleic acids
US9617547B2 (en) 2012-07-13 2017-04-11 Shin Nippon Biomedical Laboratories, Ltd. Chiral nucleic acid adjuvant
US9744183B2 (en) 2009-07-06 2017-08-29 Wave Life Sciences Ltd. Nucleic acid prodrugs and methods of use thereof
WO2018177825A1 (fr) * 2017-03-29 2018-10-04 Roche Innovation Center Copenhagen A/S Groupes protecteurs orthogonaux pour la préparation d'oligonucléotides phosphorothioate stéréodéfinis
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JP2020525442A (ja) * 2017-06-21 2020-08-27 ウェイブ ライフ サイエンシズ リミテッドＷａｖｅ Ｌｉｆｅ Ｓｃｉｅｎｃｅｓ Ｌｔｄ． 合成のための化合物、組成物、及び方法
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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000056749A1 (fr) * 1999-03-24 2000-09-28 The Government Of The United States Of America, Represented By The Secretary, Department Of Health And Human Services N-acylphosphoramidites et leur utilisation dans la synthese d'oligonucleotides
JP2003238586A (ja) * 2002-02-12 2003-08-27 Daicel Chem Ind Ltd 立体規則性の高いジヌクレオシドホスホロチオエートの製造法

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000056749A1 (fr) * 1999-03-24 2000-09-28 The Government Of The United States Of America, Represented By The Secretary, Department Of Health And Human Services N-acylphosphoramidites et leur utilisation dans la synthese d'oligonucleotides
JP2003238586A (ja) * 2002-02-12 2003-08-27 Daicel Chem Ind Ltd 立体規則性の高いジヌクレオシドホスホロチオエートの製造法

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
OKA N. ET AL: "An Oxazaphospholidine Approach for the Stereocontrolled Synthesis of Oligonucleoside Phosphorothioates.", JOURNAL OF THE AMERICAN CHEMICAL SOCIETY., vol. 125, no. 27, 2003, pages 8307 - 8317, XP002315448 *

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Publication number Priority date Publication date Assignee Title
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