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    Screening methods for identifying viral proteins with interferon antagonizing functions and potential antiviral agents
    1.
    发明授权
    Screening methods for identifying viral proteins with interferon antagonizing functions and potential antiviral agents 有权
    用干扰素拮抗功能鉴定病毒蛋白的筛选方法和潜在的抗病毒剂

    公开(公告)号：US07833774B2

    公开(公告)日：2010-11-16

    申请号：US12218280

    申请日：2008-07-14

    Applicant: Peter M. Palese ,  Adolfo Garcia-Sastre ,  Christopher Basler

    Inventor： Peter M. Palese ,  Adolfo Garcia-Sastre ,  Christopher Basler

    IPC: C12N7/02 ,  C12N5/10

    CPC classification number: G01N33/5008 ,  C12N15/1086 ,  C12Q1/18 ,  G01N33/502 ,  G01N33/6866 ,  G01N2333/005 ,  G01N2333/115 ,  G01N2333/145

    Abstract: The present invention relates, in general, to a screening method for identifying novel viral proteins with interferon antagonizing function using a transfection-based assay, and the use of such proteins in isolating various types of attenuated viruses for the development of vaccine and pharmaceutical formulations. The invention also relates to the use of viral interferon antagonists in screening assays to identify potential anti-viral agents. The invention further relates to protocols utilizing interferon antagonists, e.g., NS1, to enhance gene therapy or DNA vaccination based on their ability to increase gene expression.

    Abstract translation: 本发明一般涉及使用基于转染的测定法鉴定具有干扰素拮抗功能的新型病毒蛋白的筛选方法，以及使用这些蛋白质分离各种类型的减毒病毒以开发疫苗和药物制剂。 本发明还涉及病毒干扰素拮抗剂在筛选测定中鉴定潜在的抗病毒剂的用途。 本发明进一步涉及利用干扰素拮抗剂（例如NS1）基于其增加基因表达能力来增强基因治疗或DNA疫苗接种的方案。

    Recombinant negative strand RNA virus expression systems
    2.
    发明授权
    Recombinant negative strand RNA virus expression systems 失效
    重组负链RNA病毒表达系统

    公开(公告)号：US5840520A

    公开(公告)日：1998-11-24

    申请号：US316439

    申请日：1994-09-30

    Applicant: David Kirkwood Clarke ,  Peter M. Palese

    Inventor： David Kirkwood Clarke ,  Peter M. Palese

    IPC: A61K39/00 ,  C07K14/11 ,  C07K14/16 ,  C12N7/01 ,  C12N9/12 ,  C12N9/24 ,  C12N15/10 ,  C12N15/11 ,  C12N15/86 ,  C12P21/06 ,  A61K39/12 ,  C12P19/34

    CPC classification number: C07K14/005 ,  C12N15/10 ,  C12N15/11 ,  C12N15/86 ,  C12N9/127 ,  C12N9/2402 ,  C12Y302/01018 ,  A61K2039/5256 ,  A61K39/00 ,  C12N2740/16122 ,  C12N2760/16122 ,  C12N2840/20

    Abstract: Recombinant negative strand virus RNA templates which may be used to express heterologous gene products and/or to construct chimeric viruses are described. Influenza viral polymerase, which was prepared depleted of viral RNA, was used to copy small RNA templates prepared from plasmid-encoded sequences. Template constructions containing only the 3' end of genomic RNA were shown to be efficiently copied, indicative that the promoter lay solely within the 15 nucleotide 3' terminus. Sequences not specific for the influenza viral termini were not copied, and, surprisingly, RNAs containing termini identical to those from plus sense cRNA were copied at low levels. The specificity for recognition of the virus-sense promoter was further defined by site-specific mutagenesis. It was also found that increased level of viral protein were required in order to catalyze both the cap-endonuclease primed and primer-free RNA synthesis from these model templates as well as from genomic length RNAs. This indicated that this reconstituted system had catalytic properties very similar to those of native viral RNPs. High levels of expression of a heterologous gene was obtained using the constructs and methods described. The system was exemplified using Influenza and respiratory syncytial virus.

    Abstract translation: 描述了可用于表达异源基因产物和/或构建嵌合病毒的重组负链病毒RNA模板。 用于制备耗尽病毒RNA的流感病毒聚合酶用于复制由质粒编码序列制备的小RNA模板。 显示仅含有3'末端基因组RNA的模板结构被有效拷贝，表明启动子仅位于15个核苷酸3'末端内。 不具体针对流感病毒末端的序列没有复制，令人惊讶的是，含有与正义cRNA相同的末端的RNA被复制在低水平。 通过位点特异性诱变进一步定义识别病毒感应启动子的特异性。 还发现需要增加水平的病毒蛋白以便催化来自这些模型模板以及来自基因组长度RNA的封端核酸内切酶引物和无引物的RNA合成。 这表明这种重构系统具有与天然病毒RNP非常相似的催化性质。 使用描述的构建体和方法获得异源基因的高水平表达。 该系统使用流感和呼吸道合胞病毒来证明。

    Identification and use of antiviral compounds that inhibit interaction of host cell proteins and viral proteins required for viral replication
    3.
    发明授权
    Identification and use of antiviral compounds that inhibit interaction of host cell proteins and viral proteins required for viral replication 有权
    鉴定和使用抑制病毒复制所需的宿主细胞蛋白和病毒蛋白相互作用的抗病毒化合物

    公开(公告)号：US07632801B2

    公开(公告)日：2009-12-15

    申请号：US10337213

    申请日：2003-01-06

    Applicant: Peter M. Palese ,  Robert O'Neill ,  Ronald Harty

    Inventor： Peter M. Palese ,  Robert O'Neill ,  Ronald Harty

    IPC: A61K38/00 ,  A61K38/08 ,  A61K38/10 ,  C07K4/00 ,  C07K4/02 ,  C07K7/00

    CPC classification number: C07K14/005 ,  A61K38/00 ,  C12N15/1055 ,  C12N2760/16122 ,  G01N33/56983 ,  G01N2333/11

    Abstract: The present invention relates to the identification of host cell proteins that interact with viral proteins required for virus replication, and high throughput assays to identify compounds that interfere with the specific interaction between the viral and host cell protein. Interfering compounds that inhibit viral replication can be used therapeutically to treat viral infection. The invention is based, in part, on the Applicants' discovery of novel interactions between viral proteins and a human host cell proteins. One of these host cell proteins, referred to herein as NPI-1, interacts with influenza virus protein NP. Also, host cell proteins, referred to herein as NS1I-1 and NS1-BP interact with influenza virus protein NS1. In addition, host cell proteins containing WW domains that interact with viral proteins such as Rhabdoviral M protein are described. Compounds that interfere with the binding of the host cell and viral proteins, and inhibit viral replication can be useful for treating viral infection in vivo.

    Abstract translation: 本发明涉及鉴定与病毒复制所需的病毒蛋白相互作用的宿主细胞蛋白质，以及用于鉴定干扰病毒和宿主细胞蛋白质之间特异性相互作用的化合物的高通量测定。 抑制病毒复制的干扰化合物可用于治疗性治疗病毒感染。 本发明部分地基于申请人发现病毒蛋白和人宿主细胞蛋白之间的新的相互作用。 这些宿主细胞蛋白之一，本文称为NPI-1，与流感病毒蛋白NP相互作用。 此外，本文称为NS1I-1和NS1-BP的宿主细胞蛋白与流感病毒蛋白NS1相互作用。 此外，描述了含有与病毒蛋白如Rhabdoviral M蛋白相互作用的WW结构域的宿主细胞蛋白质。 干扰宿主细胞和病毒蛋白结合并抑制病毒复制的化合物可用于在体内治疗病毒感染。

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