WO2001016151A1 - Acide nucleique a photocouplage reversible et phosphoroamidite - Google Patents
Acide nucleique a photocouplage reversible et phosphoroamidite Download PDFInfo
- Publication number
- WO2001016151A1 WO2001016151A1 PCT/JP2000/005715 JP0005715W WO0116151A1 WO 2001016151 A1 WO2001016151 A1 WO 2001016151A1 JP 0005715 W JP0005715 W JP 0005715W WO 0116151 A1 WO0116151 A1 WO 0116151A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- nucleic acid
- reversible
- dna
- photocoupling
- photocleavage
- Prior art date
Links
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 18
- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 18
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 18
- 230000002441 reversible effect Effects 0.000 title claims abstract description 17
- SXADIBFZNXBEGI-UHFFFAOYSA-N phosphoramidous acid Chemical compound NP(O)O SXADIBFZNXBEGI-UHFFFAOYSA-N 0.000 title 1
- 238000010168 coupling process Methods 0.000 claims abstract description 10
- 230000008878 coupling Effects 0.000 claims abstract description 9
- 238000005859 coupling reaction Methods 0.000 claims abstract description 9
- 238000000034 method Methods 0.000 claims description 7
- 230000003287 optical effect Effects 0.000 claims description 7
- 150000008300 phosphoramidites Chemical class 0.000 claims description 6
- 230000001678 irradiating effect Effects 0.000 claims description 4
- 229920000642 polymer Polymers 0.000 claims description 4
- 229920001222 biopolymer Polymers 0.000 abstract 1
- 108020004414 DNA Proteins 0.000 description 12
- 238000003776 cleavage reaction Methods 0.000 description 8
- 108090000623 proteins and genes Proteins 0.000 description 7
- 230000007017 scission Effects 0.000 description 7
- 229940104302 cytosine Drugs 0.000 description 5
- 229920002554 vinyl polymer Polymers 0.000 description 5
- 230000015572 biosynthetic process Effects 0.000 description 4
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Natural products NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- -1 vinyl cytosine Chemical compound 0.000 description 4
- CEQJZGYGJSOEEL-UHFFFAOYSA-N 6-(ethenylamino)-1h-pyrimidin-2-one Chemical compound C=CNC1=CC=NC(=O)N1 CEQJZGYGJSOEEL-UHFFFAOYSA-N 0.000 description 3
- 108020004491 Antisense DNA Proteins 0.000 description 3
- 239000003816 antisense DNA Substances 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 239000002243 precursor Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 125000000391 vinyl group Chemical group [H]C([*])=C([H])[H] 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- NSPMIYGKQJPBQR-UHFFFAOYSA-N 4H-1,2,4-triazole Chemical compound C=1N=CNN=1 NSPMIYGKQJPBQR-UHFFFAOYSA-N 0.000 description 2
- 101001007348 Arachis hypogaea Galactose-binding lectin Proteins 0.000 description 2
- 230000000975 bioactive effect Effects 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 239000002777 nucleoside Substances 0.000 description 2
- 150000003833 nucleoside derivatives Chemical class 0.000 description 2
- 230000001443 photoexcitation Effects 0.000 description 2
- 238000006303 photolysis reaction Methods 0.000 description 2
- 230000015843 photosynthesis, light reaction Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 230000002194 synthesizing effect Effects 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 238000000862 absorption spectrum Methods 0.000 description 1
- 230000002862 amidating effect Effects 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000010533 azeotropic distillation Methods 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 239000007978 cacodylate buffer Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000004020 conductor Substances 0.000 description 1
- 238000007596 consolidation process Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 238000012482 interaction analysis Methods 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 239000012044 organic layer Substances 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- FAIAAWCVCHQXDN-UHFFFAOYSA-N phosphorus trichloride Chemical compound ClP(Cl)Cl FAIAAWCVCHQXDN-UHFFFAOYSA-N 0.000 description 1
- 125000006239 protecting group Chemical group 0.000 description 1
- 125000000714 pyrimidinyl group Chemical group 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- 238000010532 solid phase synthesis reaction Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 125000001424 substituent group Chemical group 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
Classifications
-
- 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
-
- 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
Definitions
- the invention of this application relates to a reversible photoligating nucleic acid and phosphoramidite. More specifically, the invention of this application is useful for applications such as antisense DNA, antigene DNA, optical gene diagnosis and treatment, analysis of DNA-protein interaction, and the formation and specificity of linked structures of bio-related molecules.
- New photoreversible nucleic acids capable of controlling chemical cleavage by light and phosphoramidites useful in their synthesis, and methods of photoligation and photocleavage using these. is there. Background art
- linked structures such as DNA-DNA, DNA-RNA, DNA-PNA, and DNA-protein
- the control of their cleavage are controlled by antisense DNA, antigene DNA, gene diagnostic therapy, DNA- It is extremely important for elucidating the interaction of proteins and the like, and controlling biomolecular structures based on this.
- the invention of this application solves the above-mentioned problems of the conventional technology, selectively provides a high coupling yield, and enables specific photocleavage of the coupling portion, thereby achieving optical coupling. It is an object of the present invention to provide a new technical means that can freely and reversibly control photolysis and photolysis. Disclosure of the invention
- the invention of the present application is characterized in that the system of the reversible photoligating nucleic acid and the bio-related polymer is irradiated with light to form a coupling structure therebetween.
- the present invention provides a photocleaving method characterized by cleaving the linking structure between a reversible photoligating nucleic acid and a bio-related polymer by irradiation with short-wavelength light.
- FIG. 1 is an absorption spectrum diagram showing an example of a reversible photocoupling and photocleavage reaction.
- the reversible photoligating nucleic acid of the formula (1) ie, the photoligating nucleoside-containing DNA of the present invention, has a vinyl group at the 5-position of the pyrimidine ring. It has a characteristic structure. Presence of vinyl group in this structure Light-control of the linkage and cleavage of biologically relevant macromolecules such as DNA, RNA, PNA, proteins, etc. is enabled. This reversible photoligating nucleic acid can also be called a functional nucleic acid containing vinylcytosine.
- the reversible photoligating nucleic acid may have other substituents permitted on the bilimidine ring-nucleoside moiety, and may have an amino group (NH 2 ), a hydroxyl group ( ⁇ H) may be protected by an acceptable protecting group.
- the invention of this application includes these conductors.
- the photoligating nucleic acid of the present invention represented by the formula (1) can be easily prepared, for example, by passing through the phosphoramidite represented by the formula (2) by the route of step 1 and step 2 of the following formula. Can be synthesized.
- step 1 5-cytosine amidite is reacted by reacting an amiditizing agent with a 5-vinyldoxydzidine DMT r form. It is carried out by synthesizing the precursor and then reacting with 1,2,4-triazole. The following are shown as amiditizing agents in this case.
- step 2 the reaction is carried out, for example, by synthesizing a DNA oligomer by a solid-phase synthesis method using a DNA synthesizer.
- Linkage by light can be achieved by irradiating light in the coexistence of a partner bio-related polymer, that is, DNA, RNA, PNA, or a protein.
- photocleavage is performed as a specific cleavage at the joint by irradiating light of shorter wavelength.
- optical coupling can be generated by photoexcitation having a longer wavelength than 330 nm, and cleavage can be generated by photoexcitation having a single wavelength than 320 nm.
- the method of light irradiation is selected from various means, and for example, a transilluminator can be used.
- a functional nucleic acid containing vinylcytosine as a reverse photoligating nucleic acid was synthesized.
- the CH 3 CN solution (0.1 M, 3 mL) of the vinyl cytosine amidite obtained in step 1 was attached to a fully automatic DNA synthesizer, and then DNA oligomers were synthesized by the phase synthesis method, excised, deprotected, and purified by high-performance liquid chromatography.
- the invention of this application gives a selectively high ligation yield, enables specific photocleavage of the linking part, and enables optical connection and photocleavage freely.
- New technological tools are provided that can be controlled reversibly.
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Saccharide Compounds (AREA)
Description
Claims
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP00954997A EP1125945B1 (en) | 1999-08-27 | 2000-08-24 | Reversible photocoupling nucleic acid and phosphoroamidite |
US09/830,401 US6593088B1 (en) | 1999-08-27 | 2000-08-24 | Reversible photocoupling nucleic acid and phosphoroamidite |
DE60002821T DE60002821T2 (de) | 1999-08-27 | 2000-08-24 | Reversibel photokuppelnde nukleinsäure und phosphoramidit |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP24068599 | 1999-08-27 | ||
JP11/240685 | 1999-08-27 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2001016151A1 true WO2001016151A1 (fr) | 2001-03-08 |
Family
ID=17063194
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/JP2000/005715 WO2001016151A1 (fr) | 1999-08-27 | 2000-08-24 | Acide nucleique a photocouplage reversible et phosphoroamidite |
Country Status (5)
Country | Link |
---|---|
US (1) | US6593088B1 (ja) |
EP (1) | EP1125945B1 (ja) |
DE (1) | DE60002821T2 (ja) |
TW (1) | TWI239337B (ja) |
WO (1) | WO2001016151A1 (ja) |
Families Citing this family (20)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2002102820A1 (en) | 2001-06-20 | 2002-12-27 | Nuevolution A/S | Nucleoside derivatives for library preparation |
DE10147074A1 (de) * | 2001-09-25 | 2003-05-08 | Beru Ag | Verfahren zum Betreiben einer aus mehreren Heizelementen bestehenden mehrstufigen elektrischen Heizung |
DE20200926U1 (de) * | 2002-01-23 | 2002-04-18 | Hegenscheidt Mfd Gmbh & Co Kg | Festwalzgerät einer Festwalzmaschine für Kurbelwellen |
AU2003214033A1 (en) * | 2002-03-15 | 2003-09-29 | Nuevolution A/S | A building block capable of transferring a functional entity |
AU2003218629A1 (en) * | 2002-03-15 | 2003-09-29 | Nuevolution A/S | A building block forming a c-c or a c-hetero atom bond uponreaction |
AU2003253069A1 (en) * | 2002-03-15 | 2003-09-29 | Nuevolution A/S | A building block forming a c-c bond upon reaction |
US7413854B2 (en) | 2002-03-15 | 2008-08-19 | Nuevolution A/S | Method for synthesising templated molecules |
US10730906B2 (en) * | 2002-08-01 | 2020-08-04 | Nuevolutions A/S | Multi-step synthesis of templated molecules |
EP3299463B1 (en) | 2002-10-30 | 2020-10-21 | Nuevolution A/S | Enzymatic encoding |
WO2004056994A2 (en) * | 2002-12-19 | 2004-07-08 | Nuevolution A/S | Quasirandom structure and function guided synthesis methods |
EP1597394A2 (en) * | 2003-02-21 | 2005-11-23 | Nuevolution A/S | A method for obtaining structural information about an encoded molecule |
WO2004074429A2 (en) * | 2003-02-21 | 2004-09-02 | Nuevolution A/S | Method for producing second-generation library |
DE602004019764D1 (de) * | 2003-03-20 | 2009-04-16 | Nuevolution As | Ligationsvermittelnde codierung von kleinen molekülen |
WO2005026387A1 (en) | 2003-09-18 | 2005-03-24 | Nuevolution A/S | A method for obtaining structural information concerning an encoded molecule and method for selecting compounds |
US20090239211A1 (en) * | 2004-02-17 | 2009-09-24 | Nuevolution A/S | Method For Enrichment Involving Elimination By Mismatch Hybridisation |
EP1730277B1 (en) * | 2004-03-22 | 2009-10-28 | Nuevolution A/S | Ligational encoding using building block oligonucleotides |
EP1956089B1 (en) * | 2005-11-17 | 2012-02-08 | Japan Science and Technology Agency | Method for detecting target nucleic acid with specific base sequence and set of nucleic acids for detection |
LT3305900T (lt) | 2005-12-01 | 2021-11-10 | Nuevolution A/S | Fermentiniai kodavimo būdai didelių bibliotekų efektyviai sintezei |
JP4814904B2 (ja) * | 2008-04-16 | 2011-11-16 | 国立大学法人北陸先端科学技術大学院大学 | 核酸類の配列選択的な精製方法 |
US11225655B2 (en) | 2010-04-16 | 2022-01-18 | Nuevolution A/S | Bi-functional complexes and methods for making and using such complexes |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0324616A2 (en) * | 1988-01-13 | 1989-07-19 | Amoco Corporation | Template-directed photoligation |
JPH1129591A (ja) * | 1997-07-08 | 1999-02-02 | Kansai Shin Gijutsu Kenkyusho:Kk | 人工ヌクレアーゼ機能を有するオリゴデオキシリボヌクレオチド、ならびに、その製造用の中間体、その製造方法および使用方法 |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4849513A (en) * | 1983-12-20 | 1989-07-18 | California Institute Of Technology | Deoxyribonucleoside phosphoramidites in which an aliphatic amino group is attached to the sugar ring and their use for the preparation of oligonucleotides containing aliphatic amino groups |
US5767264A (en) * | 1993-01-22 | 1998-06-16 | Mta Zozponti Kemiai Kutato Intezet | Oligodeoxynucleotides containing 5-alkyl, 5-(1-alkenyl)- and 5-(1-alkynl) pyrimidines |
HU212717B (en) * | 1993-01-22 | 1997-02-28 | Mta Koezponti Kemiai Kutato In | Oligonucleotides containing 5-alkyl-, 5-(1-alkenyl)- or 5-(1-alkynyl)-pyrimidine bases, pharmaceutical compositions containing them and process for their preparation |
-
2000
- 2000-08-24 DE DE60002821T patent/DE60002821T2/de not_active Expired - Fee Related
- 2000-08-24 EP EP00954997A patent/EP1125945B1/en not_active Expired - Lifetime
- 2000-08-24 WO PCT/JP2000/005715 patent/WO2001016151A1/ja active IP Right Grant
- 2000-08-24 US US09/830,401 patent/US6593088B1/en not_active Expired - Lifetime
- 2000-08-25 TW TW089117263A patent/TWI239337B/zh not_active IP Right Cessation
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0324616A2 (en) * | 1988-01-13 | 1989-07-19 | Amoco Corporation | Template-directed photoligation |
JPH1129591A (ja) * | 1997-07-08 | 1999-02-02 | Kansai Shin Gijutsu Kenkyusho:Kk | 人工ヌクレアーゼ機能を有するオリゴデオキシリボヌクレオチド、ならびに、その製造用の中間体、その製造方法および使用方法 |
Non-Patent Citations (3)
Title |
---|
FUJIMOTO KENZO ET AL.: "Template-directed photoreversible ligation of deoxyoligonucleotides via-5-vinyldeoxyuridine", J. AM. CHEM. SOC., vol. 122, no. 23, 14 June 2000 (2000-06-14), pages 5646 - 5647, XP002933956 * |
S.G. RAHIM ET AL.: "Synthesis and biological properties of 2'-deoxy-5-vinyluridine and 2'-deoxy-5-vinylcytidine", NUCLEID ACIDS RESEARCH, vol. 10, no. 17, 1982, pages 5285 - 5295, XP002933957 * |
See also references of EP1125945A4 * |
Also Published As
Publication number | Publication date |
---|---|
DE60002821T2 (de) | 2004-05-06 |
US6593088B1 (en) | 2003-07-15 |
TWI239337B (en) | 2005-09-11 |
DE60002821D1 (en) | 2003-06-26 |
EP1125945A4 (en) | 2002-01-02 |
EP1125945A1 (en) | 2001-08-22 |
EP1125945B1 (en) | 2003-05-21 |
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