KR960705930A - 재조합 p53 아데노바이러스 제조방법 및 조성물(Recombinant p53 adenovirus methods and compositions) - Google Patents

재조합 p53 아데노바이러스 제조방법 및 조성물(Recombinant p53 adenovirus methods and compositions)

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KR960705930A
KR960705930A KR1019960702193A KR19960702193A KR960705930A KR 960705930 A KR960705930 A KR 960705930A KR 1019960702193 A KR1019960702193 A KR 1019960702193A KR 19960702193 A KR19960702193 A KR 19960702193A KR 960705930 A KR960705930 A KR 960705930A
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장 웨이-웨이
에이. 로트 잭
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레이 파라비
보드 오브 리전츠, 더 유니버시티 오브 텍사스 시스템
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Abstract

본 발명은 리포좀-매개된 공동 형질감염 및 형질감염된 세포에서 세포변성 효과(CPE)의 직접 관찰을 사용하여 재조합 아데노바이러스를 제조하는 단순화되고 효율적인 방법을 기술한다. 또한, 비정상적인 p53을 갖는 세포 및 동물에서 p53 작용 및 종양 억제를 회복하는 방법을 포함하여, 신규한 p53 아데노바이러스 작제물을 포함하는 조성물 및 방법을 기술한다.

Description

재조합 p53 아데노바이러스 제조방법 및 조성물(Recombinant p53 adenovirus methods and compositions)
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음
제 1도는 재조합 p53 아데노바이러스를 제조하기 위한 도식이다. 제 2A도는 Ad5CMV-p53 DNA의 구조 분석을 위해 사용된 지도를 도시한다, 제 2B도는 PCR 생성물을 아가로즈 겔 분석을 도시한다, 제 2C도는 Ad5CMV-p53 DNA의 제한 매핑을 도시한다, 제 3A, 3B, 3C 및 3D도는 재조합 아데노바이러스에 의한 293에 대한 세포변성 효과를 관찰한 것이다, 제 7A도는 세포계 H358의 바이러스적으로 감염시킨 사람 폐암 세포의 성장곡선을 도시한다, 제 7B도는 세포계 H322의 바이러스적으로 감염시킨 사람 폐암 세포의 성장 곡선을 도시한다, 제 7C도는 세포계 H460의 바이러스적으로 감염시킨 사람 폐암 세소의 성장 곡선을 도시한다, 제 8도는 동소 폐암 모델에서 Ad5CMV-p53를 시험한 순서도이다.

Claims (21)

  1. p53을 암호화하는 발현 영역을 포함하고, 사람 악성 종양 세포에서 p53을 발현시킬 수 있는 아데노바이러스 벡터 작제물을 갖는 재조합 아데노바이러스.
  2. 제 1항에 있어서, 벡터 작제물을 사이토메갈로바이러스 IE 프로모터의 조절하에 위치된 p53을 발현 영역을 포함하는 재조합 아데노바이러스.
  3. 제 2항에 있어서, 벡터 작제물이 p53 발현 영역, 사이토메갈로바이러스 IE 프로모터 및 SV40 초기 폴리아데닐화 시그날을 포함하는 재조합 아데노바이러스.
  4. 제 1항에 있어서, 아데노바이러스 복제에 필수적인 유전자가 벡터 작제물로부터 결실되고 p53 발현 영역이 그 위치에 도입되어 있는 재조합 아데노바이러스.
  5. 제 1항에 있어서, 아데노바이러스 벡터의 EIA 및 EIB 영역이 결실되고 p53 발현 영역이 그 위치에 도입되어 있는 재조합 아데노바이러스.
  6. 제 1항에 있어서, 제 1도의 게놈 구조를 가지는 재조합 아데노바이러스.
  7. 제 1항에 있어서, 약리학적으로 허용되는 제형에 분산되어 있는 재조합, 아데노바이러스.
  8. 제 1항에 따른 재조합 아데노바이러스로 감염된 재조합 숙주 세포.
  9. 제 1항의 재조합 아데노바이러스를 야생형 p53이 결여된 세포내에서 야생형 p53을 발현시키기에 효과적인 양으로 상기 세포와 접촉시킴을 특징으로 하여 야생형 p53이 결여된 세포에게 야생형 p53 기능을 회복시키는 방법.
  10. 제 9항에 있어서, 세포가 p53 유전자에서 돌연변이를 갖는 암세포인 방법.
  11. 제 10항에 있어서, 세포가 사람 폐암 세포인 방법.
  12. 제 10항에 있어서, 세포가 사람 유방암 세포인 방법.
  13. 제 9항에 있어서, 세포가 포유동물내에 위치하고 아데노바이러스가 약리학적으로 허용되는 형태로 포유동물에게 투여되는 방법.
  14. 아데노바이러스 플라스미드 및 발현 벡터를 리포좀-매개된 형질감염에 의해 적합한 숙주 세포내에 도입하고, 상동성 재조합 및 바이러스 생성을 나타내는 세포변성 효과의 존재에 대해 배양된 숙주 세포를 분석함을 특징으로 하여 재조합 아데노바이러스를 제조하는 방법.
  15. 제 14항에 있어서, 리포좀-매개된 형질감염 단계가 DOTAP-매게된 형질감염인 방법.
  16. 제 14항에 있어서, 아데노바이러스 플라스미드가 복제-결함 아데노바이러스 플라스미드이고 숙주 세포가 그 결함을 보충하는 방법.
  17. 제 16항에 있어서, 아데노바이러스 플라스미드가 작용성 EIA 및 EIB가 결여되고 숙주 세포가 E1-발현 세포인 방법.
  18. 제 17항에 있어서, 숙주 세포가 293 세포인 방법.
  19. 제 14항에 있어서, 숙주 세포가 MEM 배지에서 배양되는 방법.
  20. 제 14항에 있어서, 세포변성 효과를 나타내는 세포의 상등액으로부터 DNA를 수득하고, 발현 벡터-특이성 및 아데노아비러스 게놈-특이성 DNA 프라이머를 사용하는 PCR에 의해 DNA를 분석하여 재조합 아데노이바이러스의 존재를 확인함을 추가로 포함하는 방법.
  21. 제 14항에 있어서, 발현 벡터가 사람 전이성 세포내에서 p53 발현을 유도할 수 있는 프로모터의 조절하에 위치된 p53-암호화 DNA 단편을 포함하는 방법.
    ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
KR1019960702193A 1993-10-29 1994-10-28 재조합 p53 아데노바이러스 제조방법 및 조성물(Recombinant p53 adenovirus methods and compositions) KR960705930A (ko)

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US08/145,826 US6410010B1 (en) 1992-10-13 1993-10-29 Recombinant P53 adenovirus compositions
US08/145,826 1993-10-29
PCT/US1994/012401 WO1995012660A2 (en) 1993-10-29 1994-10-28 Recombinant p53 adenovirus methods and compositions

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HU222981B1 (hu) 2004-01-28
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US6905873B1 (en) 2005-06-14
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WO1995012660A3 (en) 1995-08-03
KR100812631B1 (ko) 2008-03-13
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CA2174556A1 (en) 1995-05-11
US20030012770A1 (en) 2003-01-16
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