公开(公告)号：US20040209247A1

公开(公告)日：2004-10-21

申请号：US10844133

申请日：2004-05-12

Applicant: GenVec, Inc.

Inventor： Douglas E. Brough ,  Imre Kovesdi

IPC: C12Q001/70

CPC classification number: C12N15/86 ,  A61K48/00 ,  C12N2710/10343

Abstract: The invention provides a composition comprising particles of an adenoviral vector comprising deficiencies in two or more gene functions required for viral replication, wherein at least one of the deficiencies is of a gene function of the E1 region of the adenoviral genome and (b) a carrier therefor, with relatively high ratios of (i) the number of particles of the adenoviral vectors to the number of particles of E1-revertant replication-deficient adenoviral vectors not comprising one or more of the deficiencies in gene functions of the E1 region of the adenoviral and (ii) the number of particles of the adenoviral vectors to the number of particles of replication-competent adenoviral vectors, as well as a method of preparing such a composition.

Abstract translation: 本发明提供包含腺病毒载体颗粒的组合物，其包含病毒复制所需的两个或更多个基因功能的缺陷，其中至少一个缺陷是腺病毒基因组的E1区域的基因功能，和（b）载体 因此，具有相对高的比例（i）腺病毒载体的颗粒数与E1复发缺失型复制缺陷型腺病毒载体的颗粒数量不包含腺病毒E1区的基因功能的一个或多个缺陷 和（ii）腺病毒载体的颗粒数与复制能力的腺病毒载体的颗粒数目，以及制备这种组合物的方法。

公开(公告)号：US20040161848A1

公开(公告)日：2004-08-19

申请号：US10778832

申请日：2004-02-13

Applicant: GenVec, Inc.

Inventor： Imre Kovesdi

IPC: A61K048/00 ,  C12N015/861

CPC classification number: C12N7/00 ,  A61K48/00 ,  C12N15/86 ,  C12N2710/10322 ,  C12N2710/10332 ,  C12N2710/10343 ,  C12N2710/10352 ,  C12N2830/002 ,  C12N2830/85

Abstract: An adenoviral vector comprising an adenoviral genome comprising (i) at least one deletion in a region of the adenoviral genome selected from the group consisting of E1, E2A and E4, (ii) (a) at least one deletion in the VAI gene of the adenoviral genome, alone or in further combination with at least one deletion in the VAII gene of the adenoviral genome, (b) a recombinant VAI gene, alone or in further combination with a recombinant VAII gene, wherein the recombinant gene comprises either of a regulatable promoter in place of the native promoter or a mutated native promoter and 5null to the mutated native promoter, a pol II promoter, or (c) a dominant negative, double-stranded, RNA-dependent protein kinase (PKR) and, optionally, (iii) a polymerase II (pol II) construct comprising a pol II promoter operably linked to a coding region and/or a polymerase III (pol III) construct comprising a pol III promoter operably linked to a coding region, as well as a system comprising such an adenoviral vector and a cell line that complements the adenoviral vector, and related systems and methods.

Abstract translation: 一种腺病毒载体，其包含腺病毒基因组，其包含（i）在选自E1，E2A和E4的腺病毒基因组的区域中的至少一个缺失，（ii）（a）在所述腺病毒基因的VAI基因中的至少一个缺失 腺病毒基因组单独或与腺病毒基因组的VAII基因中的至少一个缺失进一步组合，（b）单独的或与重组VAII基因进一步组合的重组VAI基因，其中重组基因包括可调节的 启动子代替天然启动子或突变的天然启动子和5'至突变的天然启动子，pol II启动子，或（c）显性失活的，双链的RNA依赖性蛋白激酶（PKR） （iii）包含可操作地连接到编码区的pol II启动子和/或包含可操作地连接到编码区的pol III启动子的聚合酶III（pol III）构建体的聚合酶II（pol II）构建体，以及系统 包括这样的腺病毒 ector和补充腺病毒载体的细胞系，以及相关系统和方法。

公开(公告)号：US20040063203A1

公开(公告)日：2004-04-01

申请号：US10695605

申请日：2003-10-28

Applicant: GenVec, Inc.

Inventor： Douglas E. Brough ,  Jason G. D. Gall ,  Imre Kovesdi

IPC: C12N005/08 ,  C12N015/861

CPC classification number: C12N7/00 ,  C12N2710/10051 ,  C12N2710/10052 ,  C12N2710/10351 ,  C12N2710/10352

Abstract: The invention provides cells and methods of using the cells for the propagation of replication-deficient adenoviral vectors. The cells comprise at least one heterologous nucleic acid sequence which upon expression produces at least one non-adenoviral gene product that complements in trans for a deficiency in at least one essential gene function of one or more regions of an adenoviral genome so as to propagate a replication-deficient adenoviral vector comprising an adenoviral genome deficient in the at least one essential gene function of the one or more regions when present in the cell.

Abstract translation: 本发明提供了细胞和使用细胞促进复制缺陷型腺病毒载体的方法。 所述细胞包含至少一种异源核酸序列，其在表达时产生至少一种非腺病毒基因产物，所述非腺病毒基因产物在腺病毒基因组的一个或多个区域的至少一个必需基因功能缺陷中互补互补，从而传播 复制缺陷型腺病毒载体，其包含存在于细胞中时所述一个或多个区域的至少一个必需基因功能缺陷的腺病毒基因组。

公开(公告)号：US20030027751A1

公开(公告)日：2003-02-06

申请号：US09832355

申请日：2001-04-10

Applicant: GenVec, Inc.

Inventor： Imre Kovesdi ,  Paul D. Kessler

IPC: A61K038/18 ,  C07K014/515

CPC classification number: C12N9/16 ,  A61K38/00 ,  C07K14/475 ,  C07K14/50 ,  C07K14/515 ,  C07K14/52 ,  C07K2319/00

Abstract: The invention provides therapeutic fusion proteins which include a first peptide portion comprising a first non-heparin binding VEGF peptide portion and a second non-VEGF peptide portion covalently associated with the first peptide portion, which first and second peptide portions separately promote angiogenesis, bone growth, wound healing, or any combination thereof. Further provided are polynucleotides encoding such fusion proteins, vectors including such polynucleotides, methods of making such proteins, and methods of promoting angiogenesis, bone growth, and/or wound healing using such proteins, polynucleotides, and vectors.

Abstract translation: 本发明提供治疗性融合蛋白，其包括第一肽部分，其包含与第一肽部分共价缔合的第一非肝素结合VEGF肽部分和第二非VEGF肽部分，所述第一和第二肽部分分别促进血管生成，骨生长 ，伤口愈合或其任何组合。 还提供了编码这种融合蛋白的多核苷酸，包括这种多核苷酸的载体，制备这种蛋白质的方法，以及使用这些蛋白质，多核苷酸和载体促进血管发生，骨生长和/或伤口愈合的方法。

公开(公告)号：US20020155602A1

公开(公告)日：2002-10-24

申请号：US10123886

申请日：2002-04-16

Applicant: GenVec, Inc.

Inventor： Joseph T. Bruder ,  Imre Kovesdi ,  Alena Lizonova

IPC: C12N007/01 ,  C12N005/08 ,  C12N015/861

CPC classification number: C12N15/86 ,  C07K14/4747 ,  C12N9/0075 ,  C12N2710/10343

Abstract: The present invention provides a method of in vitro propagation of a viral eukaryotic gene transfer vector comprising a deleterious, i.e., a cytostatic, cytotoxic, or apoptotic, gene in a eukaryotic, e.g., a mammalian, host-production cell, comprising a blocking gene. The blocking gene inhibits the adverse effects of the deleterious gene on the eukaryotic host-production cell. Vectors and cells useful in the context of the present inventive method are also provided.

Abstract translation: 本发明提供了一种在包含阻断基因的真核（例如哺乳动物宿主生产细胞）中包含有害的，即细胞抑制性，细胞毒性或凋亡基因的病毒真核基因转移载体的体外繁殖的方法， 。 阻断基因抑制有害基因对真核宿主生产细胞的不利影响。 还提供了在本发明方法的上下文中有用的载体和细胞。

公开(公告)号：US20010010933A1

公开(公告)日：2001-08-02

申请号：US09771832

申请日：2001-01-29

Applicant: GenVec, Inc.

Inventor： Douglas E. Brough ,  Imre Kovesdi

IPC: A61K048/00 ,  C12N015/09 ,  C12N007/01

CPC classification number: C12N15/86 ,  C07K14/005 ,  C12N15/63 ,  C12N2710/10322 ,  C12N2710/10345 ,  C12N2710/16622 ,  C12N2830/60 ,  C12N2840/60

Abstract: The present invention provides a method of modulating the persistence of expression of a transgene in an at least E4null adenoviral vector in a cell. In one embodiment, the method comprises contacting the cell with an at least E4null adenoviral vector comprising (i) a transgene and (ii) a gene encoding a trans-acting factor, which is not from the E4 region of an adenovirus and which modulates the persistence of expression of the transgene. In another embodiment, the method comprises contacting the cell simultaneously or sequentially with (i) an at least E4null adenoviral vector comprising a transgene and (ii) a viral vector comprising a gene encoding a trans-acting factor, which is not from the E4 region of an adenovirus and which modulates the persistence of expression of the transgene. In addition, the present invention provides a recombinant at least E4null adenoviral vector for use in the method and a composition comprising the vector and a carrier therefor. Also provided by the present invention is a system for modulation of a recombinant at least E4null adenoviral vector for use in the method and a composition comprising the system and a carrier therefor.

Abstract translation: 本发明提供了调节细胞中至少E4DELTA腺病毒载体中转基因表达的持久性的方法。 在一个实施方案中，该方法包括使细胞与至少E4DELTA腺病毒载体接触，所述载体包含（i）转基因和（ii）编码反式作用因子的基因，其不是来自腺病毒的E4区域，并且其调节 持续表达的转基因。 在另一个实施方案中，该方法包括使细胞同时或顺序地与（i）包含转基因的至少E4DELTA腺病毒载体接触，和（ii）包含编码反作用因子的基因的病毒载体，其不是来自E4区 的腺病毒，并且其调节转基因表达的持续性。 此外，本发明提供了用于该方法的重组至少E4DELTA腺病毒载体和包含载体及其载体的组合物。 本发明还提供了一种用于调节用于该方法的重组至少E4DELTA腺病毒载体的系统和包含该系统和其载体的组合物。

公开(公告)号：US20030170899A1

公开(公告)日：2003-09-11

申请号：US10162043

申请日：2002-06-03

Applicant: GenVec, Inc.

Inventor： Duncan L. McVey ,  Douglas E. Brough ,  Mohammed Zuber ,  Imre Kovesdi

IPC: C12N007/00 ,  C12N015/74 ,  C12N001/21

CPC classification number: C12N15/86 ,  C12N2710/10343 ,  C12N2710/10344 ,  C12N2795/10344 ,  C12N2810/405 ,  C12N2830/002 ,  C12N2830/005 ,  C12N2830/15 ,  C12N2830/55

Abstract: The present invention provides an improved method of making eukaryotic gene transfer vectors comprising homologous recombining lambdid vectors with a second DNA in a bacterium to generate novel recombinant eukaryotic viral gene transfer vectors as well as a novel lambdid vector used in the inventive method and an inventive system comprising the novel lambdid vector.

公开(公告)号：US20030158132A1

公开(公告)日：2003-08-21

申请号：US10053637

申请日：2002-01-22

Applicant: GenVec, Inc.

Inventor： Imre Kovesdi

IPC: A61K048/00 ,  C12N005/08

CPC classification number: C07K14/52 ,  A61K48/00 ,  C07K14/47 ,  C07K14/475 ,  C07K2319/00 ,  C07K2319/02 ,  C12N9/16 ,  C12N2799/022

Abstract: The invention pertains to a method for enhancing bone density or formation. In accordance with the method, a nucleic acid encoding a secreted alkaline phosphatase (SEAP) is administered to a cell in a region of a bone such that the nucleic acid is expressed to produce the SEAP, whereby bone density or formation is enhanced within the region. The method can be employed to produce a bone graft having a cell harboring an exogenous nucleic acid encoding a SEAP. To facilitate the inventive method, the invention provides a recombinant viral vector having a nucleic acid encoding a SEAP. Optionally, a nucleic acid encoding an angiogenic protein and/or a nucleic acid encoding an osteogenic protein is employed in conjunction with the nucleic acid encoding a SEAP.

Abstract translation: 本发明涉及一种增强骨密度或形成的方法。 根据该方法，将编码分泌的碱性磷酸酶（SEAP）的核酸施用于骨的区域中的细胞，使得所述核酸被表达以产生SEAP，从而在该区域内增强骨密度或形成 。 该方法可用于产生具有含有编码SEAP的外源核酸的细胞的骨移植物。 为了促进本发明的方法，本发明提供了具有编码SEAP的核酸的重组病毒载体。 任选地，编码血管生成蛋白和/或编码成骨蛋白的核酸的核酸与编码SEAP的核酸结合使用。

公开(公告)号：US20020004040A1

公开(公告)日：2002-01-10

申请号：US09797064

申请日：2001-03-01

Applicant: GenVec, Inc.

Inventor： Imre Kovesdi ,  Douglas E. Brough ,  Duncan L. McVey ,  Joseph T. Bruder ,  Alena Lizonova

IPC: A61K048/00

CPC classification number: C12N7/00 ,  A61K38/00 ,  A61K48/00 ,  A61K2039/5256 ,  C07K14/005 ,  C07K14/4712 ,  C12N15/86 ,  C12N2710/10322 ,  C12N2710/10343 ,  C12N2710/10352 ,  C12N2810/60 ,  C12N2810/6081 ,  C12N2830/002 ,  C12N2830/85 ,  C12N2840/20 ,  C12N2840/203 ,  C12N2840/44 ,  Y10S977/799

Abstract: The present invention provides multiply deficient adenoviral vectors and complementing cell lines. Also provided are recombinants of the multiply deficient adenoviral vectors and a therapeutic method, particularly relating to gene therapy, vaccination, and the like, involving the use of such recombinants.

公开(公告)号：US20030203480A1

公开(公告)日：2003-10-30

申请号：US10427759

申请日：2003-04-30

Applicant: GenVec, Inc.

Inventor： Imre Kovesdi ,  Duncan L. McVey

IPC: C12N015/861 ,  C12N007/00

CPC classification number: C12N15/86 ,  C12N2710/10043 ,  C12N2795/10043 ,  C12N2810/80

Abstract: The inventive method of producing a eukaryotic viral vector comprises contacting a eukaryotic cell, which comprises a unique enzyme that nicks or cleaves a DNA molecule, with a recombinant phage vector, or contacting a eukaryotic cell, which does not comprise a unique enzyme that nicks or cleaves a DNA molecule, simultaneously or sequentially, in either order, with (i) a unique enzyme that nicks or cleaves a DNA molecule, and (ii) a recombinant phage vector. The recombinant phage vector comprises the DNA molecule comprising (a) a eukaryotic viral vector genome comprising a coding sequence, (b) a phage packaging site that is not contained within the eukaryotic viral vector genome, and (c) a promoter that is operably linked to the coding sequence. Alternatively, the DNA molecule is not present within the recombinant phage vector. The eukaryotic cell is contacted with the first DNA molecule and a recombinant phage vector. The first DNA molecule comprises a replication deficient eukaryotic viral vector genome comprising at least one adenoviral inverted terminal repeat and a packaging signal. The recombinant phage vector comprises a second DNA molecule and a phage packaging site, wherein the second DNA molecule complements in trans the replication deficient eukaryotic viral vector genome. The DNA molecule(s) enter the eukaryotic cell, and the unique enzyme nicks or cleaves the DNA molecule in the eukaryotic cell in at least one region not contained within the eukaryotic viral vector genome, thereby inducing the production of and ultimately producing a eukaryotic viral vector.

Abstract translation: 产生真核病毒载体的本发明方法包括将包含切割或切割DNA分子的独特酶与重组噬菌体载体接触的真核细胞或接触真核细胞，所述真核细胞不包含缺陷或 以（i）切割或切割DNA分子的独特的酶，和（ii）重组噬菌体载体以任一顺序同时或顺序地切割DNA分子。 重组噬菌体载体包含DNA分子，其包含（a）真核病毒载体基因组，其包含编码序列，（b）不包含在真核病毒载体基因组内的噬菌体包装位点，和（c）可操作地连接的启动子 到编码序列。 或者，DNA分子不存在于重组噬菌体载体内。 将真核细胞与第一DNA分子和重组噬菌体载体接触。 第一DNA分子包含复制缺陷型真核病毒载体基因组，其包含至少一个腺病毒反向末端重复和包装信号。 重组噬菌体载体包含第二DNA分子和噬菌体包装位点，其中第二DNA分子与复制缺陷型真核病毒载体基因组相反。 DNA分子进入真核细胞，独特的酶在至少一个不包含在真核病毒载体基因组内的区域中切割或切割真核细胞中的DNA分子，由此诱导产生并最终产生真核病毒 向量。