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    Envelope gene-deficient Paramyxovirus vector
    1.
    发明授权
    Envelope gene-deficient Paramyxovirus vector 有权
    包膜基因缺陷型副粘病毒载体

    公开(公告)号：US07226786B2

    公开(公告)日：2007-06-05

    申请号：US10316535

    申请日：2002-12-10

    申请人： Kaio Kitazato ,  Tsugumine Shu ,  Hidekazu Kuma ,  Yasuji Ueda ,  Makoto Asakawa ,  Mamoru Hasegawa ,  Akihiro Iida ,  Fumino Tokito ,  Takahiro Hirata ,  Tsuyoshi Tokusumi ,  Makoto Inoue ,  Yumiko Tokusumi

    发明人： Kaio Kitazato ,  Tsugumine Shu ,  Hidekazu Kuma ,  Yasuji Ueda ,  Makoto Asakawa ,  Mamoru Hasegawa ,  Akihiro Iida ,  Fumino Tokito ,  Takahiro Hirata ,  Tsuyoshi Tokusumi ,  Makoto Inoue ,  Yumiko Tokusumi

    IPC分类号： C12N15/00 ,  C12N15/86

    CPC分类号： C12N15/86 ,  A61K48/00 ,  A61K2039/5254 ,  A61K2039/5256 ,  C12N7/00 ,  C12N2760/18823 ,  C12N2760/18843 ,  C12N2760/18845 ,  C12N2760/18861 ,  C12N2800/30 ,  C12N2810/6081

    摘要： F gene-deficient virus virions are successfully recovered by using an F gene-deficient Sendai virus genomic cDNA. Further, F gene-deficient infectious viral particles are successfully constructed by using F-expressing cells as helper cells. Also, F gene and HN gene-deficient virus virions are successfully recovered by using a virus genomic cDNA deficient in both F gene and HN gene. Further, F gene and HN gene-deficient infectious viral particles are successfully produced by using F- and HN-expressing cells as helper cells. A virus deficient in F gene and HN gene and having F protein is constructed by using F-expressing cells as helper cells. In addition, M gene-deficient infectious virus particles were produced using helper cells expressing M protein. From cells infected with M gene-deficient viruses, release of virus-like particles was inhibited. Further, a VSV-G pseudo type virus is successfully constructed by using VSV-G-expressing cells. Techniques for constructing these deficient viruses contribute to the development of vectors of Paramyxoviridae usable in gene therapy.

    摘要翻译： 通过使用F基因缺陷的仙台病毒基因组cDNA成功地回收了F基因缺陷型病毒粒子。 此外，通过使用F表达细胞作为辅助细胞，成功构建了F基因缺陷型感染性病毒颗粒。 此外，通过使用F基因和HN基因缺陷的病毒基因组cDNA，成功地回收了F基因和HN基因缺陷型病毒粒子。 此外，通过使用F-和HN表达细胞作为辅助细胞，F基因和HN基因缺陷型感染性病毒颗粒成功产生。 通过使用F表达细胞作为辅助细胞构建F基因和HN基因并具有F蛋白的病毒。 此外，使用表达M蛋白的辅助细胞产生M基因缺陷型感染性病毒颗粒。 从感染M基因缺陷病毒的细胞中，病毒样颗粒的释放被抑制。 此外，通过使用表达VSV-G的细胞成功地构建了VSV-G伪型病毒。 用于构建这些缺陷病毒的技术有助于可用于基因治疗的副粘病毒科的载体的发育。

    Methods of examining (-) strand rna virus vectors having lowered ability to form grains and method of constructing the same
    3.
    发明申请
    Methods of examining (-) strand rna virus vectors having lowered ability to form grains and method of constructing the same 审中-公开
    检查具有降低形成颗粒能力的（ - ）链rna病毒载体的方法及其构建方法

    公开(公告)号：US20050130123A1

    公开(公告)日：2005-06-16

    申请号：US10489384

    申请日：2002-09-18

    申请人： Makoto Inoue ,  Yumiko Tokusumi ,  Akihiro Iida ,  Mamoru Hasegawa

    发明人： Makoto Inoue ,  Yumiko Tokusumi ,  Akihiro Iida ,  Mamoru Hasegawa

    IPC分类号： A61K48/00 ,  C12N7/01 ,  C12N7/04 ,  C12N15/45 ,  C12N15/86 ,  C12Q1/70 ,  C12N15/867 ,  C12Q1/68

    CPC分类号： C12N7/00 ,  A61K48/00 ,  C12N15/86 ,  C12N2760/18843 ,  C12N2760/18861 ,  C12N2800/30

    摘要： The present invention provides methods for testing and producing (−) strand RNA virus vectors with reduced or eliminated particle formation ability or cytotoxicity. It was revealed that a deficiency in M protein localization in cells introduced with such a (−) strand RNA virus vector could result in the suppression of virus-like particle (VLP) formation in the cells. The present invention provides methods for testing and screening for a (−) strand RNA virus vector in which particle formation ability has been reduced or eliminated, and methods for producing a recombinant (−) strand RNA virus vector in which particle formation ability has been reduced or eliminated. Such a vector, in which VLP formation has been reduced or eliminated, is extremely useful as a vector for gene therapy, since it neither induces cytotoxicity nor immune response due to the secondary release of viruses from cells in which it has been introduced.

    摘要翻译： 本发明提供了测试和生产（ - ）链RNA病毒载体的方法，其具有降低或消除的颗粒形成能力或细胞毒性。 显示出用这种（ - ）链RNA病毒载体引入的细胞中M蛋白定位的缺陷可导致细胞中病毒样颗粒（VLP）形成的抑制。 本发明提供了用于测定和筛选其中减少或消除了颗粒形成能力的（ - ）链RNA病毒载体的方法，以及用于生产其中颗粒形成能力已经降低的重组（ - ）链RNA病毒载体的方法 或消除。 已经减少或消除VLP形成的这种载体作为用于基因治疗的载体是非常有用的，因为它既不引起细胞毒性也不诱导免疫应答，这是由于病毒从其引入细胞的二次释放引起的。

    VECTORS WITH MODIFIED PROTEASE-DEPENDENT TROPISM
    5.
    发明申请
    VECTORS WITH MODIFIED PROTEASE-DEPENDENT TROPISM 审中-公开
    具有修饰的蛋白依赖性TROPISM的载体

    公开(公告)号：US20100297732A1

    公开(公告)日：2010-11-25

    申请号：US12712905

    申请日：2010-02-25

    申请人： Hiroaki KINOH ,  Makoto Inoue ,  Yasuji Ueda ,  Akihiro Iida ,  Mamoru Hasegawa ,  Masanori Kobayashi

    发明人： Hiroaki KINOH ,  Makoto Inoue ,  Yasuji Ueda ,  Akihiro Iida ,  Mamoru Hasegawa ,  Masanori Kobayashi

    IPC分类号： C12N7/01 ,  C12N15/45

    CPC分类号： C07K14/005 ,  A61K38/00 ,  A61K48/00 ,  C07K2319/50 ,  C12N15/86 ,  C12N2760/18022 ,  C12N2760/18822 ,  C12N2760/18843 ,  C12N2800/30

    摘要： The present invention provides cell fusogenic vectors having replicative ability, whose protease-dependent tropism has been modified. M gene-deficient viral vectors encoding modified F proteins, in which the cleavage site of the F protein of paramyxovirus is modified to be cleaved by different proteases, were produced. In cells transfected with these vectors, the genomic RNA present in the vectors is replicated, and cell fusogenic infection spreads to neighboring cells depending on the presence of other proteases; however, no viral particles are released. The vectors of this invention, encoding the F proteins which are cleaved by proteases whose activity is enhanced in cancer, show cancer growth suppressive effect in vivo.

    摘要翻译： 本发明提供具有复制能力的细胞融合载体，其蛋白酶依赖性向性已被修饰。 产生编码修饰的F蛋白的M基因缺陷型病毒载体，其中修饰了被不同蛋白酶切割的副粘病毒的F蛋白的切割位点。 在用这些载体转染的细胞中，载体中存在的基因组RNA被复制，并且根据其它蛋白酶的存在，细胞融合感染扩散到相邻细胞; 然而，没有病毒颗粒被释放。 本发明的编码由蛋白酶切割的F蛋白的载体，其活性在癌症中增强，在体内显示出癌症生长抑制作用。

    Vectors with modified protease-dependent tropism
    6.
    发明授权
    Vectors with modified protease-dependent tropism 有权
    具有修饰的蛋白酶依赖性向量的载体

    公开(公告)号：US07709621B2

    公开(公告)日：2010-05-04

    申请号：US12140715

    申请日：2008-06-17

    申请人： Hiroaki Kinoh ,  Makoto Inoue ,  Yasuji Ueda ,  Akihiro Iida ,  Mamoru Hasegawa ,  Masanori Kobayashi

    发明人： Hiroaki Kinoh ,  Makoto Inoue ,  Yasuji Ueda ,  Akihiro Iida ,  Mamoru Hasegawa ,  Masanori Kobayashi

    IPC分类号： C07H21/04 ,  C12N15/45 ,  C12N7/00 ,  A61K48/00

    CPC分类号： C07K14/005 ,  A61K38/00 ,  A61K48/00 ,  C07K2319/50 ,  C12N15/86 ,  C12N2760/18022 ,  C12N2760/18822 ,  C12N2760/18843 ,  C12N2800/30

    摘要： The present invention provides cell fusogenic vectors having replicative ability, whose protease-dependent tropism has been modified. M gene-deficient viral vectors encoding modified F proteins, in which the cleavage site of the F protein of paramyxovirus is modified to be cleaved by different proteases, were produced. In cells transfected with these vectors, the genomic RNA present in the vectors is replicated, and cell fusogenic infection spreads to neighboring cells depending on the presence of other proteases; however, no viral particles are released. The vectors of this invention, encoding the F proteins which are cleaved by proteases whose activity is enhanced in cancer, show cancer growth suppressive effect in vivo.

    摘要翻译： 本发明提供具有复制能力的细胞融合载体，其蛋白酶依赖性向性已被修饰。 产生编码修饰的F蛋白的M基因缺陷型病毒载体，其中修饰了被不同蛋白酶切割的副粘病毒的F蛋白的切割位点。 在用这些载体转染的细胞中，载体中存在的基因组RNA被复制，并且根据其它蛋白酶的存在，细胞融合感染扩散到相邻细胞; 然而，没有病毒颗粒被释放。 本发明的编码由蛋白酶切割的F蛋白的载体，其活性在癌症中增强，在体内显示出癌症生长抑制作用。

    VECTORS WITH MODIFIED PROTEASE-DEPENDENT TROPISM
    7.
    发明申请
    VECTORS WITH MODIFIED PROTEASE-DEPENDENT TROPISM 有权
    具有修饰的蛋白依赖性TROPISM的载体

    公开(公告)号：US20080299642A1

    公开(公告)日：2008-12-04

    申请号：US12140715

    申请日：2008-06-17

    申请人： Hiroaki Kinoh ,  Makoto Inoue ,  Yasuji Ueda ,  Akihiro Iida ,  Mamoru Hasegawa ,  Masanori Kobayashi

    发明人： Hiroaki Kinoh ,  Makoto Inoue ,  Yasuji Ueda ,  Akihiro Iida ,  Mamoru Hasegawa ,  Masanori Kobayashi

    IPC分类号： C12N7/01 ,  C07H21/04 ,  C12N15/63

    CPC分类号： C07K14/005 ,  A61K38/00 ,  A61K48/00 ,  C07K2319/50 ,  C12N15/86 ,  C12N2760/18022 ,  C12N2760/18822 ,  C12N2760/18843 ,  C12N2800/30

    摘要： The present invention provides cell fusogenic vectors having replicative ability, whose protease-dependent tropism has been modified. M gene-deficient viral vectors encoding modified F proteins, in which the cleavage site of the F protein of paramyxovirus is modified to be cleaved by different proteases, were produced. In cells transfected with these vectors, the genomic RNA present in the vectors is replicated, and cell fusogenic infection spreads to neighboring cells depending on the presence of other proteases; however, no viral particles are released. The vectors of this invention, encoding the F proteins which are cleaved by proteases whose activity is enhanced in cancer, show cancer growth suppressive effect in vivo.

    摘要翻译： 本发明提供具有复制能力的细胞融合载体，其蛋白酶依赖性向性已被修饰。 产生编码修饰的F蛋白的M基因缺陷型病毒载体，其中修饰了被不同蛋白酶切割的副粘病毒的F蛋白的切割位点。 在用这些载体转染的细胞中，载体中存在的基因组RNA被复制，并且根据其它蛋白酶的存在，细胞融合感染扩散到相邻细胞; 然而，没有病毒颗粒被释放。 本发明的编码由蛋白酶切割的F蛋白的载体，其活性在癌症中增强，在体内显示出癌症生长抑制作用。

    Vectors with modified protease-dependent tropism
    8.
    发明授权
    Vectors with modified protease-dependent tropism 有权
    具有修饰的蛋白酶依赖性向量的载体

    公开(公告)号：US07402427B2

    公开(公告)日：2008-07-22

    申请号：US10513094

    申请日：2003-04-30

    申请人： Hiroaki Kinoh ,  Makoto Inoue ,  Yasuji Ueda ,  Akihiro Iida ,  Mamoru Hasegawa ,  Masanori Kobayashi

    发明人： Hiroaki Kinoh ,  Makoto Inoue ,  Yasuji Ueda ,  Akihiro Iida ,  Mamoru Hasegawa ,  Masanori Kobayashi

    IPC分类号： C12N15/86 ,  A01N63/00 ,  A61K39/155

    CPC分类号： C07K14/005 ,  A61K38/00 ,  A61K48/00 ,  C07K2319/50 ,  C12N15/86 ,  C12N2760/18022 ,  C12N2760/18822 ,  C12N2760/18843 ,  C12N2800/30

    摘要： The present invention provides cell fusogenic vectors having replicative ability, whose protease-dependent tropism has been modified. M gene-deficient viral vectors encoding modified F proteins, in which the cleavage site of the F protein of paramyxovirus is modified to be cleaved by different proteases, were produced. In cells transfected with these vectors, the genomic RNA present in the vectors is replicated, and cell fusogenic infection spreads to neighboring cells depending on the presence of other proteases; however, no viral particles are released. The vectors of this invention, encoding the F proteins which are cleaved by proteases whose activity is enhanced in cancer, show cancer growth suppressive effect in vivo.

    摘要翻译： 本发明提供具有复制能力的细胞融合载体，其蛋白酶依赖性向性已被修饰。 产生编码修饰的F蛋白的M基因缺陷型病毒载体，其中修饰了被不同蛋白酶切割的副粘病毒的F蛋白的切割位点。 在用这些载体转染的细胞中，载体中存在的基因组RNA被复制，并且根据其它蛋白酶的存在，细胞融合感染扩散到相邻细胞; 然而，没有病毒颗粒被释放。 本发明的编码由蛋白酶切割的F蛋白的载体，其活性在癌症中增强，在体内显示出癌症生长抑制作用。