KR970706292A - 평행 SELEX(Parall Selex) - Google Patents

평행 SELEX(Parall Selex) Download PDF

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Publication number
KR970706292A
KR970706292A KR1019970701738A KR19970701738A KR970706292A KR 970706292 A KR970706292 A KR 970706292A KR 1019970701738 A KR1019970701738 A KR 1019970701738A KR 19970701738 A KR19970701738 A KR 19970701738A KR 970706292 A KR970706292 A KR 970706292A
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South Korea
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product
nucleic acid
reactant
target
perform
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KR1019970701738A
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KR100457015B1 (ko
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래리 골드
브루스 이튼
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코크란 아담
넥스스타 파머수티컬스, 인크.
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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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/547—Chelates, e.g. Gd-DOTA or Zinc-amino acid chelates; Chelate-forming compounds, e.g. DOTA or ethylenediamine being covalently linked or complexed to the pharmacologically- or therapeutically-active agent
    • A—HUMAN NECESSITIES
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/549—Sugars, nucleosides, nucleotides or nucleic acids
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
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    • B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
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    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
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    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
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    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
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Abstract

본 발명은 2개 이상의 반응물질로부터의 생성물들을 공진화시키는 방벅과 함께, 생성물을 제조하는 반응을 촉진할 수 있는 핵산을 개시한다. 본 발명은 또한 생성물 및 상기 방법에 의하여 제조된 촉진성 핵산을 개시한다.

Description

평행 SELEX(Parallel Seles)
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음

Claims (34)

  1. 표적에, 미리 선택된 기능을 수행하는 능력을 기준으로 선별되는 생성물을, 생성물 라이브러리로부터 식별하는, 하기 (a)-(c)단계를 포함하는 방법: (a) 각각이 랜덤화 배열의 영역을 지니며, 제1반응물질과 회합되어 있는 힉산들을 포함하는 핵산-반응물질 시험혼합물의 제조; (b) 상기 핵산-반응물질 시험 혼합물을 유리 반응물질과 반응시켜, 상기 제1 및 유리 반응물질의 반응에 의하여 형성된 생성물을 포함하는 생성물 라이브러리를 형성하는데, 여기서 상기 반응은 상기 제1반응물질과 회합되어 있는 헥산에 의하여 촉진된다; 및 (c) 표적에 미리 선택된 기능을 수행하는 상대적 능력을 기준으로 상기 생성물 라이브러리의 멤버 사이를 분리하여 미리 선택된 기능을 수행할 수 있는 능력을 지닌 상기 생성물을 식별.
  2. 제1항에 있어서, 표적에 대하여 미리 선택된 기능을 수행하는 상기 능력이, 상기 표적에 결합하는 것인 방법.
  3. 제1항에 있어서, 표적에 대하여 미리 선택된 기능을 수행하는 상기 능력이, 상기 표적과 반응하여 상기 표적의 특징을 변화시키는 것인 방법.
  4. 제1항에 있어서, 상기 핵산 시험 혼합물이 보존 배열의 영역 및 랜덤화 배열의 영역을 지닌 핵산을 포함하는 방법.
  5. 제1항에 있어서, 상기 핵산이 단일-나선 RNA, 단일-나선 DNA 및 2중-나선 DNA로 구성되어 있는 군으로부터 선택되는 방법.
  6. 제1항에 있어서, 상기 핵산 시험 혼합물이 수식된 뉴클레오티드를 포함하는 방법.
  7. 제6항에 있어서, 상기 수식된 뉴클레오티드가 리보오스 및/또는 인산염 및/또는 염기 위치에서 화학적으로 수식되는 방법.
  8. 제6항에 있어서, 상기 수식된 뉴클레오티드가 2′- 또는 5-위치에서 수식된 피리미딘인 방법.
  9. 제6항에 있어서, 상기 수식된 뉴클레오티드가 8-위치에서 수식된 푸린인 방법.
  10. 제6항에 있어서, 상기 수식된 뉴클레오티드가, 뉴클레오티드의 전하, 분극성, 수소결합, 정전기적 상호 작용 또는 유동성을 증진시키는 화학기로 수식되는 방법.
  11. 제10항에 있어서, 상기 화학기가 소수성 성분, 친수성 성분, 다양한 산화상태의 금속원자, 강제구조물, 이미다졸, 1차 알코올, 카르복실산염, 구아니듐 그룹, 아미노 그룹, 티올 및 유기금속 촉매로 구성되어 있는 군으로부터 선택되는 방법.
  12. 제10항에 있어서, 상기 화학기가 아니노산 측쇄 또는 그 유사체를 포함하는 방법.
  13. 제1항에 있어서, 상기 핵산 시험혼합에 첨합되는 유기금속 촉매를 더 포함하는 방법.
  14. 제1항에 있어서, 상기 제1반응물질 및 상기 핵산 사이에 결합된 링커 그룹을 더 포함하는 방법.
  15. 제14항에 있어서, 상기 링커 그룹의 크기가 10-1000Å의 범위인 방법.
  16. 제15항에 있어서, 상기 링커 그룹이, PEG, 폴리비닐 알코올, 폴리아크릴레이트 및 폴리펩티드로 구성되어 있는 군으로부터 선택되는 방법.
  17. 제1항에 있어서, 상기 제1반응물질이 디에노필, 상기 유리 반응물질이 디엔, 및 상기 생성물이 시클로헥센 유도체인 방법.
  18. 제1항의 방법에 의하여 제조된 생성물.
  19. 제18항에 있어서, 비대칭 중심을 갖는 생성물.
  20. 제18항에 있어서, 분자량이 80-2,000범위인 생성물.
  21. 2개의 반응물질 사이의 반응 생성물을 포함하는 생성물 라이브러리로부터, 표적에 대여 미리 선택된 기능을 수행할 수 있는 능력을 지닌 생성물을 식별하는 방법으로서, 각 생성물이 상기 생성물의 형성을 촉진하는 핵산과 회합되어 있으머, 상기 방법이 미리 선택된 기능을 수행하는 상대적 능력을 기준으로 상기 생성물 라이브러리의 멤버 사이를 분리시킴으로써, 상기 미리 선택된 기능을 수행할 수 있는 능력을 지닌 상기 생성물이 식별될 수 있는 단계를 포함하는 방법.
  22. 생성물 라리브러리로부터 생성물을 식별하는 방법으로서, 상기 생성물이 표적 분자에 결합하는 능력을 기준으로 선별되며, 상기 방법이 하기 (a)-(e)단계를 포함하는 방법.
    (a) 각각이 랜덤화 배열의 영역을 지니며, 제1반응물질과 회합되어 있는 핵산들을 포함하는 핵산-반응물질 시험혼합물의 제조; (b) 상기 핵산-반응물질 시험 혼합물과 유리 반응물질을 반응시켜서, 상기 제1 및 제2반응물질의 반응에 의하여 형성된 생성물과 연합되어 있는 핵산을 포함하는 생성물 라이브러리를 제조; (c) 생성물 라이브러리와 표적을 접촉, 여기서 생성물 라이브러리에 비하여 상대적으로 표적에 대한 친화성이 증가된 생성물이, 잔류하는 생성물 라리브러리로부터 분리될 수 있다; (d) 잔류하는 생성물 라이브러리로부터 표적에 대한 친화성이 증가된 상기 생성물을 분리; 및 (e) 표적에 대한 친화성이 증가된 상기 생성물과 회합된 핵산을 증폭시킴으로써, 상기 생성물이 식별될 수 있다.
  23. 제22항에 있어서, (b) 및 (c)단계 사이에, 생성물 라이브러리를 비-표적과 접촉시키고, 상기 비-표적에 결합하는 생성물을 분리해버리는 단계를 더 포함하는 방법.
  24. 하기 (a)-(d)단계를 포함하는, 표적에 대하여 미리 선택된 기능을 수행하는 능력을 지닌 생성물의 제조 방법; (a) 핵산 시험혼합물의 각각의 멤버를 제1반응물질과 결합시켜서 핵산-반응물질 시험 혼합물을 형성; (b) 제1 및 유리 반응물질간의 결합형성에 유리한 조건하에서, 상기 핵산-반응물질 시험 혼합물을 유리 반응물질과 접촉시킴으로써 생성물 라이브러리를 형성하는데, 여기서 상기 결합 형성은 상기 제1반응물질에 결합된 촉진성 핵산에 의하여 매개된다; (c) (b)단계의 생성물 라이브러리를 표적과 접촉. 여기서 생성물 라이브러리에 비하여 상대적으로, 상기 표적에 대한 미리 선택된 기능을 수행할 수 있는 능력이 증가된 생성물이, 잔류하는 생성물 라이브러리로부터 분리될 수 있다; 및 (d) 잔류하는 생성물 라이브러리로부터 상기 표적에 대하여 미리 선택된 기능을 수행할 수 있는 능력을 지닌 상기 생성물을 분리시킴으로써 상기 생성물이 식별될 수 있다.
  25. 미리 선택된 기능을 수행할 수 있는 능력을 지닌 생성물을 제조하기 위하여 복수의 반응물질의 반응을 촉진할 수 있는 능력을 기준으로 핵산이 선별되는, 하기 (a)-(c)단계를 포함하는 촉진성 핵산의 식별방법. (a) 각각이 랜덤화 배열의 영역을 지니며, 제1반응물질과 회합되어 있는 핵산들을 포함하는 핵산-반응물질 시험혼합물의 제조; (b) 상기 생성물을 제조하기 위한 반응물질들간의 핵산 촉진 반응에 유리한 조건하에서, 상기 핵산-반응물질 시험혼합물을 복수의 유리 반응물질과 혼합; 및 (c) 생성물과 회합된 상기 핵산-반응물질 시험 혼합물의 멤버를 유리시킴으로써, 상기 촉진성 핵산이 식별될 수 있다.
  26. 생성물 라이브러리와 동시에, 생성물의 형성을 촉진하는 촉진성 핵산류를 공진화시키는 단계를 포함하는, 미리 선택된 기능을 수행할 수 있는 능력을 기준으로 선별된 생성물을 식별하는 방법.
  27. 동시에 촉진성 핵산을 식별하고 생성물 라이브러리를 제조하는, 하기 (a) 및 (b) 단계를 포함하는 방법. 여기서 상기 촉진성 핵산은 상기 생성물 라이브러리를 제조하기 위하여 적어도 제1 및 유리 반응물질 사이의 반응을 촉진한다.
    (a) 각각이 랜덤화 배열의 영역을 지니며, 제1반응물질과 회합되어 있는 핵산들을 포함하는 핵산-반응물질 시험혼합물의 제조; 및 (b) 생성물 라이브러리를 형성하기 위하여 상기 핵산-반응물질 시험혼합물과 유리 반응물질을 반응시킴으로써, 상기 촉진성 핵산이 식별될 수 있다.
  28. 하기 (a)-(d)단계를 포함하는 촉진성 핵산과, 표적에 대하여 미리 선택된 기능을 수행할 수 있는 능력을 지닌 생성물을, 함께 제조하는 방법.
    (a) 제1반응물질을 핵산 시험혼합물의 각 멤버에 결합시켜 핵산-반응물질 시험혼합물을 형성; (b) 결합형성에 유리한 조건하에서, 상기 핵산-반응물질 시험 혼합물과 유리 반응물질의 혼합물을 접촉시켜 생성물 라이브러리를 형성, 여기서 상기 결합형성은, 상기 제1반응물질에 결합하는 촉진성을 지닌 상기 핵산에 의하여 매개된다; (c) 상기 생성물 라이브러리를 표적과 접촉, 여기서 생성물 라이브러리에 비하여 상대적으로 표적에 대한 미리 선택된 기능을 수행할 수 있는 능력이 증가된 생성물이, 잔류하는 생성물 라이브러리로부터 분리될 수 있다; 및 (d) 생성물과 회합되어 있는 핵산을 증폭하여, 촉진성이 강화된 핵산의 혼합물을 산출.
  29. 제28항에 있어서, 하기 (e) 및 (f)단계를 더 포함하는 방법; (e) 촉진성-강화 핵산을 상기 제1반응물질과 결합; 및 (f) 상기 표적에 대하여 상기 미리 선택된 기능을 수행할 수 있는 상기 능력을 지닌 생성물이,식별되기에 충분할 정도의 양으로 제조될 수 있을 때까지 (b)-(e)단계를 반복.
  30. 적어도 결합된 반응물질 및 유리 반응물질 사이의 반응 결과인 생성물의 혼합물을 포함하는 생성물 라이브러리, 여기서 상기 결합된 반응물질은 상기 반응물질들 사이의 반응을 촉진한 핵산에 부착된다.
  31. 제1 및 유리 반응물질 사이의 결합 형성에 유리한 조건하에서, 각각이 핵산시험 혼합물의 멤버에 결합된 제1반응물질의 혼합물을, 유리 반응물질의 혼합물과 접촉시키는 단계를 포함하는 생성물 라리브러리의 제조방법, 여기서 상기 결합 형성 반응은 상기 제1반응물질에 결합된 핵산에 의하여 매개된다.
  32. 반응물질들 사이의 반응중에 제1반응물질이 핵산과 회합되어 있을 때, 적어도 제1 및 유리 반응물질 사이의 반응을 촉진하여 생성물을 형성하는 촉진성 핵산.
  33. 화학 반응을 촉진할 수 있는 비-자연 발생 확산.
  34. 제1 및 유리 반응물질 사이의 결합 형성에 유리한 조건하에서, 상기 제1반응물질과 상기 유리 반응물질을 접촉시키는 단계를 포함하는 생성물의 제조방법.
    여기서 상기 결합 형성 반응은 핵산에 의하여 매개된다.
    ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
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EP0782580A1 (en) 1997-07-09
RU2132853C1 (ru) 1999-07-10
US5723592A (en) 1998-03-03
JPH10508465A (ja) 1998-08-25
NZ294127A (en) 1999-06-29
DE69535365D1 (de) 2007-02-22
US5789160A (en) 1998-08-04
WO1996009316A1 (en) 1996-03-28
EP0782580A4 (en) 2002-01-16
US5858660A (en) 1999-01-12
CA2196286C (en) 2010-02-02
KR100457015B1 (ko) 2005-05-17
DE69535365T2 (de) 2007-11-15
CA2196286A1 (en) 1996-03-28
AU3679595A (en) 1996-04-09
JP4153030B2 (ja) 2008-09-17
US5723289A (en) 1998-03-03
AU714469B2 (en) 2000-01-06

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