Method for liquid-phase synthesis of nucleic acid

A compound, representative technology, applied in the field of liquid phase synthesis of nucleic acid

Active Publication Date: 2015-09-16
TAKEDA PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This solid-phase synthesis method allows easy preparation of nucleic acid oligomers

Method used

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  • Method for liquid-phase synthesis of nucleic acid
  • Method for liquid-phase synthesis of nucleic acid
  • Method for liquid-phase synthesis of nucleic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0607] Synthesis of a deoxythymine (dT) type nucleoside monomer compound (compound (E5)) with three octadecyloxy groups

[0608] [Formula 55]

[0609]

[0610] (1) Methyl benzoate substituted by three octadecyloxy groups (compound (E1))

[0611] [Formula 56]

[0612]

[0613] To methyl gallate (9.2 g) and potassium carbonate (103.7 g) was added 1,3-dimethyl-2-imidazolidinone (170 mL), and the mixture was stirred at 80° C. for 30 minutes. 1-Bromooctadecane (69.1 mL) was added thereto, and the mixture was stirred at 80°C for 12 hr. 40° C. hot water was added to the reaction liquid, suspended, and then the precipitate was collected by suction filtration, and the obtained solid was washed with acetonitrile, acetone and methanol to quantitatively obtain the compound represented by E1 (48.0 g).

[0614] 1 H NMR (500MHz, CDCl 3 )δ7.25(s,2H),3.99-4.03(m,6H),3.89(s,3H),1.78-1.84(m,4H),1.71-1.77(m,2H),1.44-1.50(m, 6H), 1.20-1.38(m, 84H), 0.88(t, J=7.0Hz, 9H)

[0615] (2) Sy...

Embodiment 2

[0636] Synthesis of uracil (U) nucleoside monomer compound (compound (E7)) with three octadecyloxy groups

[0637] [Formula 61]

[0638]

[0639] (1) Synthesis of U-shaped nucleosides (compound (E6))

[0640] [Formula 62]

[0641]

[0642] 1-((2R,3R,4R,5R)-5-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-3-((tert-butyldimethyl Silyl)oxy)-4-hydroxytetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H)-dione (1 g) was dissolved in tetrahydrofuran (5 mL). Add levulinic acid (0.272g), N,N'-dimethylaminopyridine (0.0185g) and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride in sequence Salt (0.444g), and the mixture was stirred at room temperature for 18 hours. At room temperature, 10% acetic acid / triethylamine (10 mL) and ethyl acetate aqueous solution (10 mL) were added to the reaction liquid, followed by separation and extraction. The obtained organic layer was concentrated under reduced pressure to obtain compound (E6) (1.21 g, yield percentage: 105.7%).

[0643] 1 ...

Embodiment 3

[0650] Synthesis of uracil (U) type nucleoside monomer compound (compound (E8)) with three tetradecyloxy groups

[0651] [Formula 64]

[0652]

[0653] (1) Methyl benzoate substituted by three tetradecyloxy groups (compound (E9))

[0654] [Formula 65]

[0655]

[0656] To methyl gallate (5.5 g) and potassium carbonate (62.2 g) was added 1,3-dimethyl-2-imidazolidinone (100 mL), and the mixture was stirred at 80° C. for 45 minutes. 1-Bromotetradecane (26.9 mL) was added thereto, and the mixture was stirred at 80°C for 12 hr. Water was added to the reaction liquid, suspended, and then the precipitate was collected by suction filtration, and the obtained solid was washed with a 50% aqueous solution of acetonitrile and acetonitrile to obtain compound (E9) (22.6 g).

[0657] 1 H NMR (500MHz, CDCl 3 )δ7.25(2H,s),3.99-4.03(6H,m),3.89(3H,s,COOMe),1.71-1.84(6H,m),1.44-1.50(6H,m),1.23-1.38( 60H, br), 0.88 (9H, t J = 7.0 Hz).

[0658] (2) Synthesis of benzoic acid (compound (...

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Abstract

In this method, an oligonucleotide is prepared by using, as a synthesis unit, a novel nucleoside monomer compound represented by formula (I) [wherein X, R 1 , Y, Base, Z, Ar, R 2 , R 3 and n are each as defined in Claim 1]. The novel nucleoside monomer compound is a nucleoside, the base moiety of which is substituted with an aromatic-hydrocarbon-ring-carbonyl or -thiocarbonyl group having at least one hydrophobic group. The method can dispense with column-chromatographic purification in every reaction, and enables base elongation not only in the 3'-direction but also in the 5'-direction, thus attaining efficient liquid-phase mass synthesis of an oligonucleotide.

Description

technical field [0001] The present invention relates to a liquid phase synthesis method of nucleic acid. More specifically, the present invention relates to novel nucleoside monomer compounds wherein the base portion of the nucleoside is substituted by (i) an aromatic hydrocarbon ring carbonyl having at least one hydrophobic group, or (ii) The aromatic hydrocarbon ring thiocarbonyl group having at least one hydrophobic group also relates to a method for preparing a nucleic acid oligomer using a nucleoside monomer compound as a synthetic unit. This preparation method can avoid the need for column chromatographic purification after each reaction, and can achieve efficient large-scale synthesis. The invention also relates to a method for preparing nucleic acid oligomers in liquid phase. Background of the invention [0002] In recent years, with the rapid development or progress of frontier research on bioengineering, including genomic drug development and genetic diagnosis or...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H19/073C07H19/067C07H21/04
CPCC07H21/00C07H19/067C07H21/04C07H19/073C07H1/00C07H19/167C07H19/207C07H19/06C07H19/16
Inventor 野野川满长田敏明斋藤英树安间常雄
Owner TAKEDA PHARMA CO LTD
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