JP2009028040A - 非c群アデノウイルスベクター - Google Patents
非c群アデノウイルスベクター Download PDFInfo
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- JP2009028040A JP2009028040A JP2008179643A JP2008179643A JP2009028040A JP 2009028040 A JP2009028040 A JP 2009028040A JP 2008179643 A JP2008179643 A JP 2008179643A JP 2008179643 A JP2008179643 A JP 2008179643A JP 2009028040 A JP2009028040 A JP 2009028040A
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Abstract
【解決手段】A,B,D,EまたはF群に分類されるアデノウイルスから単離されるか、あるいはアデノウイルスに含まれるDNAセグメントと実質的に相同なアデノウイルスDNAセグメントを含有し、ウイルス複製に必要な領域の少なくとも1つに欠損のあるアデノウイルス遺伝子導入ベクター、および該ベクターの使用方法。
【選択図】図1
Description
本発明は、非C群アデノウイルスに見られるDNAセグメント、および通常は細胞内発現用の外来遺伝子を含有する、アデノウイルス遺伝子導入ベクターに関する。また、本発明はこのようなベクターの治療および診断への使用に関する。
1952〜1953年の冬から春にかけて、国立衛生研究所(NIH)のロウとその同僚らは、ワシントン特別区に住む幼児から外科的に切除されたアデノイド(腺様増殖組織)を取得し、組織培養を行った(Roweら,Proc.Soc.Exp.Biol.Med.,84,570-573(1953))。数週間後、該培養の多くが上皮細胞の破壊を特徴とする進行性の変性を示し始めた。この細胞変性効果は培養濾液によって、樹立されたヒト細胞株の組織培養に連続的に伝達することができた。該細胞変性剤は「アデノイド変性」(Ad)剤と呼ばれた。「アデノウイルス」という名前は、最終的にこれらの薬剤に一般的に用いられる名称となった。これら最初の発見に続いて、アデノウイルスの多くの原型株−その中の幾つかは呼吸疾患を引き起こす−が発見された(Roweら,上述;Dingle ら,Am.Rev.Respir.Dis.,97,1-65(1968);Horwitz,"Adenoviridae and Their Replication," in Fundamental Virology(Fieldsら編,Raven Press Ltd.,New York,NY,第2版,1991),pp.771-813)。
本発明は、A,B,D,EまたはF群に分類されるアデノウイルスから単離されるか、あるいはアデノウイルスに含まれるDNAセグメントと実質的に相同なアデノウイルスDNAセグメントを含有し、ウイルス複製に必要な領域の少なくとも1つに欠損のあるアデノウイルス遺伝子導入ベクターを提供する。本発明はまた、このようなベクターの使用方法を提供する。
本発明は、少なくとも部分的には、非C群アデノウイルスが複製欠損アデノウイルス遺伝子導入ベクターを調製するのに用いることができること、およびこのようなベクターが複製欠損C群アデノウイルスベクター用に設計された相補的な細胞株中で増殖し得ることの発見に基づいている。非C群アデノウイルスとC群アデノウイルスのE1AおよびE1B遺伝子産物の間の類似性が比較的小さいことに例示されるように、非C群アデノウイルスとC群アデノウイルスの間の実質的な相違を考慮すると、この発見は予期せぬものであった。
本実施例では、B群のAd7aウイルスのアデノウイルスDNAからE1A領域を欠失させることによる、ウイルスの大きなフラグメントの作製について説明する。
本実施例では、図2に示されるプラスミドpAd7aCMV-CATgDの作製について述べる。プラスミドpAd7aCMV-CATgDを3つの連続する工程におけるダイレクショナルクローニングにより調製した。
第二に、標準的な方法を用いて、該pBSベクターをNotI-SalI で開環し、サイトメガロウイルスプロモーター(CMV)をoriとpkgエレメントに連結し、その後に細菌のクロラムフェニコールアセチルトランスフェラーゼ配列(CAT)およびマウスのB−majグロビンポリ(A)部位を続けた。
本実施例では、組換えAd7aアデノウイルスの作製について述べ、該組換えAd7aアデノウイルスがHEK−293細胞内で複製され、該細胞によって相補され得ることを示す。
本実施例では、組換えAd7aアデノウイルスの生成および該組換えAd7aアデノウイルスがHEK−293細胞内で複製され、該細胞によって相補され得ることをさらに確認する。特に、本実施例では、CAT遺伝子発現に関する二次ウイルス溶解物の試験について述べる。
本実施例では、C群アデノウイルスに対する非C群アデノウイルスの類似性について評価する。
(1)一般情報:
(i)出願人:コーネル リサーチ ファウンデーション、インコーポレイティッド
(ii)発明の名称:非C群アデノウイルスベクター
(iii)配列の数:4
(iv)コンピューターリーダブル形式:
(A)媒体の種類:フロッピーディスク
(B)コンピューター:IBM PC互換機
(C)オペレーティングシステム:PC−DOS/MS−DOS
(D)ソフトウェア:パテントイン リリース #1.0, バージョン
#1.25
(v)出願の現在のデータ:
(A)出願番号:WO
(B)出願日:1996年9月24日
(C)分類:
(vi)先の出願のデータ:
(A)出願番号:US 537402
(B)出願日:1995年10月2日
(C)分類:
(2)配列番号1に関する情報
(i)配列の特徴:
(A)配列の長さ:30塩基対
(B)配列の型:核酸
(C)鎖の数:一本鎖
(D)トポロジー:直鎖状
(ii)配列の種類:DNA(ゲノミック)
(xi)配列:配列番号1:
GAGCTCACCG GTCTCTCTAT ATAATATACC 30
(2)配列番号2に関する情報
(i)配列の特徴:
(A)配列の長さ:35塩基対
(B)配列の型:核酸
(C)鎖の数:一本鎖
(D)トポロジー:直鎖状
(ii)配列の種類:DNA(ゲノミック)
(xi)配列:配列番号2:
TCTAGAGCGG CCGCAGCGAT CAGCTGACAC CTACG 35
(2)配列番号3に関する情報
(i)配列の特徴:
(A)配列の長さ:29塩基対
(B)配列の型:核酸
(C)鎖の数:一本鎖
(D)トポロジー:直鎖状
(ii)配列の種類:DNA(ゲノミック)
(xi)配列:配列番号3:
GCTAGCGTCG ACGAAGAAAT GCATGTTTG 29
(2)配列番号4に関する情報
(i)配列の特徴:
(A)配列の長さ:27塩基対
(B)配列の型:核酸
(C)鎖の数:一本鎖
(D)トポロジー:直鎖状
(ii)配列の種類:DNA(ゲノミック)
(xi)配列:配列番号4:
GGGCCCGGTA CCCAATGATC GAAACCG 27
Claims (19)
- アデノウイルスから単離されるか、あるいはアデノウイルスに含まれるDNAセグメントと実質的に相同なアデノウイルスDNAセグメントを含有するアデノウイルス遺伝子導入ベクター(ここで該アデノウイルスはA,B,D,EまたはF群アデノウイルスであり、該ベクターは複製欠損である)。
- 該アデノウイルスがB群アデノウイルスである請求の範囲1のベクター。
- 該アデノウイルスがAd7アデノウイルスである請求の範囲2のベクター。
- 該ベクターが、哺乳動物において治療または予防剤を発現し得る外来の核酸をさらに含む請求の範囲1のベクター。
- 該ベクターが、少なくともアデノウイルスゲノムのE1A領域を欠損したものである請求の範囲1のベクター。
- 該ベクターがHEK−293細胞株内で複製し得るものである請求の範囲5のベクター。
- 該ベクターが、哺乳動物において治療または予防剤を発現し得る外来の核酸をさらに含む請求の範囲5のベクター。
- 該ベクターが、少なくともアデノウイルスゲノムの別の領域をさらに欠損したものである請求の範囲5のベクター。
- 該ベクターが、哺乳動物において治療または予防剤を発現し得る外来の核酸をさらに含む請求の範囲8のベクター。
- 該ベクターが、少なくともウイルス複製に必要なアデノウイルスゲノムの別の領域をさらに欠損したものである請求の範囲5のベクター。
- 該ベクターが、哺乳動物において治療または予防剤を発現し得る外来の核酸をさらに含む請求の範囲10のベクター。
- 該ベクターが、アデノウイルスゲノムのE1,E2,E3およびE4領域からなる群より選択される領域をさらに欠損したものである請求の範囲5のベクター。
- 該ベクターが、アデノウイルスゲノムの後期領域からなる群より選択される領域をさらに欠損したものである請求の範囲5のベクター。
- 遺伝子治療を必要とするヒトに、治療上有効な量の請求の範囲4のベクターを投与することを含む遺伝子治療方法。
- 遺伝子治療を必要とするヒトに、治療上有効な量の請求の範囲7のベクターを投与することを含む遺伝子治療方法。
- 遺伝子治療を必要とするヒトに、治療上有効な量の請求の範囲9のベクターを投与することを含む遺伝子治療方法。
- 遺伝子治療を必要とするヒトに、治療上有効な量の請求の範囲11のベクターを投与することを含む遺伝子治療方法。
- 遺伝子治療を必要とするヒトの細胞における、外来核酸を含む請求の範囲4のベクターの発現または毒性を試験する方法であって、(a) 該ヒトから標的細胞のアリコートを取り出して培養し、(b) 該細胞に該ベクターを接触させ、(c)該外来核酸の発現と該培養標的細胞の生存性を測定することを含む方法。
- 標的細胞へのトランスフェクション後に、外来核酸を含むベクターの外来核酸発現を試験する方法であって、(a) 標的細胞を選択し、(b) 該標的細胞に外来核酸を含む請求の範囲4のベクターを接触させ、(c) 該標的細胞内での該外来核酸の発現を測定する方法。
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JPH07145079A (ja) * | 1993-08-11 | 1995-06-06 | American Home Prod Corp | 組換えアデノウイルス・ワクチン |
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ZA858044B (en) * | 1984-11-01 | 1987-05-27 | American Home Prod | Oral vaccines |
US5543328A (en) * | 1993-08-13 | 1996-08-06 | Genetic Therapy, Inc. | Adenoviruses having modified fiber proteins |
AU3551195A (en) * | 1994-09-23 | 1996-04-09 | Somatix Therapy Corporation | Chimeric adenovirus for gene delivery |
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1995
- 1995-10-02 US US08/537,402 patent/US5837511A/en not_active Expired - Lifetime
-
1996
- 1996-09-24 JP JP9514303A patent/JPH11514866A/ja not_active Withdrawn
- 1996-09-24 WO PCT/US1996/015274 patent/WO1997012986A2/en active IP Right Grant
- 1996-09-24 DE DE69636055T patent/DE69636055T2/de not_active Revoked
- 1996-09-24 AT AT96935925T patent/ATE323773T1/de not_active IP Right Cessation
- 1996-09-24 EP EP96935925A patent/EP0866873B1/en not_active Revoked
-
2008
- 2008-07-09 JP JP2008179643A patent/JP2009028040A/ja active Pending
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
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WO1994028152A1 (fr) * | 1993-05-28 | 1994-12-08 | Transgene S.A. | Adenovirus defectifs et lignees de complementation correspondantes |
JPH07145079A (ja) * | 1993-08-11 | 1995-06-06 | American Home Prod Corp | 組換えアデノウイルス・ワクチン |
Also Published As
Publication number | Publication date |
---|---|
DE69636055T2 (de) | 2007-04-12 |
JPH11514866A (ja) | 1999-12-21 |
EP0866873A2 (en) | 1998-09-30 |
WO1997012986A3 (en) | 1997-06-19 |
DE69636055D1 (de) | 2006-05-24 |
ATE323773T1 (de) | 2006-05-15 |
WO1997012986A2 (en) | 1997-04-10 |
US5837511A (en) | 1998-11-17 |
EP0866873B1 (en) | 2006-04-19 |
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