Nucleoside derivative or salt thereof, polynucleotide synthesis reagent, polynucleotide production method, polynucleotide, and binding nucleic acid molecule production method

A technology of nucleoside derivatives and polynucleotides, which is applied in the field of nucleoside derivatives or their salts, and can solve problems such as inability to obtain binding nucleic acid molecules

Active Publication Date: 2019-07-19
NEC SOLUTION INNOVATORS LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, using known natural nucleosides and their derivatives, there are also targets for which binding nucleic acid molecules with sufficient binding ability cannot be obtained

Method used

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  • Nucleoside derivative or salt thereof, polynucleotide synthesis reagent, polynucleotide production method, polynucleotide, and binding nucleic acid molecule production method
  • Nucleoside derivative or salt thereof, polynucleotide synthesis reagent, polynucleotide production method, polynucleotide, and binding nucleic acid molecule production method
  • Nucleoside derivative or salt thereof, polynucleotide synthesis reagent, polynucleotide production method, polynucleotide, and binding nucleic acid molecule production method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] IA8 was prepared by the following Synthesis Example.

[0089] Electrospray ionization mass spectrometry (ESI-MS) was performed using a mass spectrometer (API2000, supplier: Applied Biosystems) in positive or negative ion mode. Obtained using a nuclear magnetic resonance apparatus (JNM-ECS400, manufactured by JEOL) 1 H NMR spectrum. Chemical shifts are expressed relative to the internal standard tetramethylsilane (Me 4 δ (ppm) of Si). Ion exchange chromatography was performed using a chromatography system (ECONO system, manufactured by Bio-Rad). In ion exchange chromatography, a glass column packed with diethylaminoethyl (DEAE) A-25-Sephadex is used (manufactured by Amershambiosciences).

[0090] (Synthesis Example 1) Synthesis of TB4

[0091]

[0092] 2-Bromoethylamine hydrobromide (501 mg, 2.28 mmol) was weighed and suspended in 1,4-dioxane (5 ml), and the mixture thus obtained was stirred under ice bath for 30 minutes. To the mixture thus obtained was added...

Embodiment 2

[0147] This example examines whether sIgA-binding and amylase-binding nucleic acid molecules can be obtained using IA8.

[0148] (1) Binding to nucleic acid molecules

[0149] Binding nucleic acid molecules binding to the respective targets were obtained by the SELEX method except that, in addition to deoxyribonucleotides containing adenine, guanine and cytosine (dATP, dGTP and dCTP, respectively), IA8 was used as the deoxyribose Nucleotides to prepare candidate polynucleotides. Specifically, a binding nucleic acid molecule is obtained in the following manner. According to the protocol attached to the product, sIgA (product of MP Biomedicals, LLC-Cappel) or human salivary amylase (manufactured by LeeBioSolutions Corporation) was bound as a target to beads (Dynabeads MyOne Carboxylic Acid, manufactured by Invitrogen). After binding the target, use selection buffer (SB buffer: 40mmol / L HEPES, 125mmol / L NaCl, 5mmol / L KCl, 1mmol / L MgCl 2 , 0.01%Tween pH 7.5) to wash the beads...

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Abstract

Provided are a novel nucleoside derivative or salt thereof, polynucleotide synthesis reagent, polynucleotide production method, polynucleotide, and binding nucleic acid molecule production method. This nucleoside derivative or salt thereof is characterized in being represented by chemical formula (1). In chemical formula (1): Su is an atom group with a sugar backbone in the nucleoside residue or an atom group with a sugar phosphate backbone in the nucleotide residue, and may or may not have a protecting group; each of L1 and L2 independently is a straight or branched saturated or unsaturated C2-10 hydrocarbon group; X1 is an imino group (-NR1-), ether group (-O-) or thioether group (-S-); and R1 is a hydrogen atom or straight or branched saturated or unsaturated C2-10 hydrocarbon group.

Description

technical field [0001] The present invention relates to nucleoside derivatives or salts thereof, polynucleotide synthesis reagents, polynucleotide preparation methods, polynucleotides and nucleic acid-binding molecule preparation methods. Background technique [0002] To analyze a target in a sample, a binding molecule that binds to the target is used. In addition to antibodies, binding nucleic acid molecules such as aptamers that bind to targets are also used as binding molecules that bind to targets (Patent Document 1). [0003] As a method for obtaining binding nucleic acid molecules, SELEX (Systematic Evolution of Ligands by Exponential Enrichment method) is known, in which a target is brought into contact with a large number of candidate polynucleotides, and then selected from the Among the candidate polynucleotides, a polynucleotide that binds to the target is selected as a binding nucleic acid molecule. When a binding nucleic acid molecule is obtained by the SELEX m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H19/073C12N15/09C12N15/11
CPCY02P20/55C07H19/073C12N15/09C07H21/00C07H21/04C07H1/00C12N15/1048C12N15/115C07H19/20C07H21/02
Inventor 皆川宏贵堀井克纪秋富穰金子直人和贺岩桑原正靖
Owner NEC SOLUTION INNOVATORS LTD
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