Top > Search of International Patents > OLIGONUCLEOTIDE DERIVATIVE, OLIGONUCLEOTIDE CONSTRUCT USING SAME, AND METHODS FOR PRODUCING THESE

OLIGONUCLEOTIDE DERIVATIVE, OLIGONUCLEOTIDE CONSTRUCT USING SAME, AND METHODS FOR PRODUCING THESE

Foreign code F170008931
File No. (S2015-0845-N0)
Posted date Jan 19, 2017
Country WIPO
International application number 2016JP059398
International publication number WO 2016152980
Date of international filing Mar 24, 2016
Date of international publication Sep 29, 2016
Priority data
  • P2015-060689 (Mar 24, 2015) JP
Title OLIGONUCLEOTIDE DERIVATIVE, OLIGONUCLEOTIDE CONSTRUCT USING SAME, AND METHODS FOR PRODUCING THESE
Abstract [Problem] To provide a novel oligonucleotide derivative that is easy to synthesize and can be introduced into cells without using a lipofection reagent.
[Solution] The oligonucleotide derivative of the present invention is shown by, e.g., (1). This derivative is considered to bind an aminoglycan moiety to a ligand on the surface of a cell and be introduced into the cell, and can be expected to have selective drug delivery functions. In the formula, R1 and R2 each independently represent a hydrogen or phosphate group; a, b, and c independently represent integers of 0 or higher, at least one of a, b, and c being 1 or higher; A and B independently represent optionally modified oligonucleotides having a combined chain length of A and B of 3 or higher. However, A and B do not include hydroxyl groups at the 3' end and 5' end of the oligonucleotide. Furthermore, S in the formula indicates a sugar substituent, a peptide chain, or a tocopherol binding group. Additionally, an alkyl group may be bonded, in lieu of hydrogen, to the benzene ring in the formula.
Outline of related art and contending technology BACKGROUND ART
In recent years, various substrates such as DNA or RNA oligonucleotide treatment, such as a diagnosis applications has been to be used.For example, a nucleic acid of interest as a technology, interference RNA (RNAi) using, of a specific gene knock-down method can be used.RNAi is, by the action of the double-stranded RNA (dsRNA), such that sequences homologous to a phenomenon in which the action of the genes is suppressed.The medicament is a nucleic acid using RNAi, next generation as a therapeutic agent, are great expectations.
On the other hand, the chemical modification of oligonucleotide, are not present in the native, given a new function studies have also been carried out.The present inventors, ethynyl groups are introduced to the end of the oligonucleotide, further, the ethynyl groups, a benzene ring using the click reaction was new substituent modified to develop a technique (patent document 1).In addition, in this technique the resulting artificial oligonucleotides, natural type oligonucleotide is higher than the nuclease are resistant, hardly degraded inside the cell the result is obtained.
And have a negative charge at the nucleic acid itself is however, not able to pass through the cell membrane.For this reason, in a nucleic acid drug, such as a drug delivery system (DDS) nucleotides to pass through the cell membrane is required.The current, as a method for introducing nucleotides into cells, using liposomes are ribonucleic transfection method has been developed.The method, having a negative charge around DNA, cationic liposomes with a positive charge to form a complex that has bound, phenomenon from the cell surface by endocytosis into the cell DNA is incorporated into the method and the like.
However, lipofection reagent is used in the lipofection method, and toxicity to the liver and the kidney, has been a problem.In addition, lipofection method, and using a simple endocytosis DDS, is a lack of selectivity for the cells.
In order to solve such a problem, the oligonucleotide 3' - terminus of the peptide chain is introduced into the asialoglycoprotein, the asialoglycoprotein receptor (ASGPR) that introducing into the cell through, chemically modified oligonucleotides have been developed (Patent Document 2, 3).However, in this method, such as shown in Fig. 4, three asialoglycoprotein - peptide chain and a complex has a chemical structure, in order to synthesize a complex operation is required.
Scope of claims (In Japanese)[請求項1]
次式(1)で表されるオリゴヌクレオチド誘導体。
[化1]
[請求項2]
次式(2)で表されるオリゴヌクレオチド誘導体。
[化2]
[請求項3]
前記Sは下記化学構造式(3)又は(3´)で示される請求項1又は2に記載のオリゴヌクレオチド誘導体。
[化3]
[請求項4]
次式で表されるオリゴヌクレオチド誘導体。
[化4]
[請求項5]
R1及びR2はHであることを特徴とする請求項1乃至4のいずれか1項に記載のオリゴヌクレオチド誘導体。
[請求項6]
bは0であることを特徴とする請求項1乃至5のいずれか1項に記載のオリゴヌクレオチド誘導体。
[請求項7]
a及びbはともに0であることを特徴とする請求項1乃至6のいずれか1項に記載のオリゴヌクレオチド誘導体。
[請求項8]
cは1以上5以下であることを特徴とする請求項7に記載のオリゴヌクレオチド誘導体。
[請求項9]
A及びBは、所定の遺伝子mRNAの部分配列又はその相補配列を有する請求項1乃至8のいずれか1項に記載のオリゴヌクレオチド誘導体。
[請求項10]
A及びBを合わせた鎖長は、10以上35以下である請求項1乃至9のいずれか1項に記載のオリゴヌクレオチド誘導体。
[請求項11]
A及びBはオリゴリボヌクレオチドであることを特徴とする請求項1乃至10のいずれか1項に記載のオリゴヌクレオチド誘導体。
[請求項12]
遺伝子発現調節用オリゴヌクレオチド構築物であって、
請求項1乃至11のいずれか1項に記載のオリゴヌクレオチド誘導体を有する構築物。
[請求項13]
1本鎖及び2本鎖DNA、1本鎖及び2本鎖RNA、DNA/RNAキメラ並びにDNA/RNAハイブリッドから選択される遺伝子発現調節用オリゴヌクレオチド構築物であって請求項12に記載の構築物。
[請求項14]
アンチジーン、アンチセンス、アプタマー、siRNA、miRNA、shRNA及びリポザイムから選択される請求項12又は13に記載の構築物。
[請求項15]
ダングリングエンド部分に以下の式(4)又は(5)で表されるユニットを有する請求項12乃至14のいずれか1項記載の構築物。
[化5]
[請求項16]
siRNAであって、前記オリゴヌクレオチド誘導体において、a及びbは0であり、cは1又は2であり、3’末端ダングリングエンド部分に以下の式(4)又は(5)で表されるユニットを含む請求項15記載の構築物。
[化6]
[請求項17]
遺伝子診断用オリゴヌクレオチド構築物であって、請求項1乃至11のいずれかに記載のオリゴヌクレオチド誘導体を有する構築物。
[請求項18]
プローブであることを特徴とする請求項17に記載の構築物。
[請求項19]
以下の式(6)又は(7)で表される化合物。
[化7]
[請求項20]
siRNAのダングリングエンドユニット用である請求項19に記載の化合物。
[請求項21]
請求項19に記載の化合物から選択される1種又は2種以上を用いることを特徴とするオリゴヌクレオチドの修飾方法。
[請求項22]
オリゴヌクレオチドに、少なくとも1個の以下の式(4)又は(5)で表されるユニットを付加、置換及び挿入のいずれか又はこれらを組み合わせて導入することを特徴とするオリゴヌクレオチドの修飾方法。
[化8]
  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • GIFU UNIVERSITY
  • Inventor
  • KITADE, Yukio
  • SHIBATA, Aya
IPC(International Patent Classification)
Specified countries National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS JP KE KG KN KP KR KZ LA LC LK LR LS LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SA SC SD SE SG SK SL SM ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN ZA ZM ZW
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW KM ML MR NE SN ST TD TG
Please contact us by E-mail or facsimile if you have any interests on this patent.

PAGE TOP

close
close
close
close
close
close