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：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：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：US20030040100A1

公开(公告)日：2003-02-27

申请号：US09911020

申请日：2001-07-23

Applicant: GenVec, Inc.

Inventor： Douglas E. Brough ,  Imre Kovesdi

IPC: C12N007/02 ,  C12N005/02 ,  C07H021/04 ,  C12N005/00 ,  C12N007/01 ,  C12N007/00

CPC classification number: C12N15/86 ,  C12N7/00 ,  C12N2710/10321 ,  C12N2710/10343 ,  C12N2710/10352

Abstract: The invention provides a cell and a method of using the cell for the propagation of a replication-deficient adenoviral vector, wherein the cellular genome comprises a nucleic acid sequence whose expression produces a gene product that complements a replication-deficient adenoviral vector. The nucleic acid sequence is operatively linked to a chimeric expression control sequence comprising at least a functional portion of a CMV immediate early promoter/enhancer region and/or at least a functional portion of an adenoviral promoter, wherein the chimeric expression control sequence is upregulated by one or more viral proteins not produced by the nucleic acid sequence.

Abstract translation: 本发明提供了一种细胞和使用该细胞用于增殖复制缺陷型腺病毒载体的方法，其中所述细胞基因组包含其表达产生补充复制缺陷型腺病毒载体的基因产物的核酸序列。 核酸序列可操作地连接到包含CMV立即早期启动子/增强子区的至少一个功能部分和/或腺病毒启动子的至少功能部分的嵌合表达控制序列，其中嵌合表达控制序列被上调 不是由核酸序列产生的一种或多种病毒蛋白。

公开(公告)号：US20020004242A1

公开(公告)日：2002-01-10

申请号：US09905758

申请日：2001-07-13

Applicant: GenVec, Inc.

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

IPC: C12N015/86 ,  C07H021/04

CPC classification number: C12N15/86 ,  C12N15/65 ,  C12N2710/10343 ,  C12N2800/30 ,  C12N2830/001 ,  C12N2830/15 ,  C12N2830/55

Abstract: The present invention provides a dual selection cassette (DSC) comprising first and second DNA segments having homology to a eukaryotic viral vector, positive and negative selection genes, each operably linked to their own promoter, and one or more unique restriction enzyme sites (URES) or sitey-directed homologous recombination sites. The present invention also provides a plasmid, pN/P, comprising an independent positive selection marker gene, an origin of replication, and a dual selection cassette. The dual selection cassette and pN/P plasmid can be used to produce eukaryotic gene transfer vectors without requiring temporally-linked double recombination events or the use of specialized bacterial strains that allow the replication of plasmids comprising defective origins of replication. This method usefully increases the ratio of desired to undesired plasmid and vector constructs. Additionally, this invention provides a method for the creation of eukaryotic viral vector libraries.

Abstract translation: 本发明提供了一种双重选择盒（DSC），其包含与真核病毒载体具有同源性的第一和第二DNA片段，每个与其本身的启动子可操作地连接的阳性和阴性选择基因，以及一个或多个独特的限制酶位点（URES） 或定点同源重组位点。 本发明还提供了包含独立的阳性选择标记基因，复制起点和双选择盒的质粒pN / P。 双选择盒和pN / P质粒可用于产生真核基因转移载体，而不需要时间连接的双重重组事件或使用允许复制包含有缺陷复制起点的质粒的专门细菌菌株。 该方法有效地增加了期望的和不需要的质粒和载体构建体的比例。 此外，本发明提供了一种用于产生真核病毒载体文库的方法。

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：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：US20030087438A1

公开(公告)日：2003-05-08

申请号：US10001097

申请日：2001-11-02

Applicant: GenVec, Inc.

Inventor： Douglas E. Brough ,  Imre Kovesdi

IPC: A61K048/00 ,  C12N015/861 ,  C12N007/02 ,  C12N007/00

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：US20030045498A1

公开(公告)日：2003-03-06

申请号：US10211701

申请日：2002-08-02

Applicant: GenVec, Inc.

Inventor： Imre Kovesdi ,  Douglas E. Brough ,  Lisa Wei ,  Duncan L. McVey

IPC: A61K048/00 ,  C12N015/85

CPC classification number: C12N15/86 ,  A61K48/00 ,  A61K48/005 ,  A61K48/0075 ,  C07K14/71 ,  C07K14/811 ,  C12N2710/10343 ,  C12N2799/022

Abstract: The present invention is directed to a method of prophylactically or therapeutically treating an animal for at least one ocular-related disorder, e.g., ocular neovascularization or age-related macular degeneration. The method comprises contacting an ocular cell with an expression vector comprising a nucleic acid sequence encoding an inhibitor of angiogenesis and the same or different nucleic acid sequence encoding a neurotrophic agent. The method also can comprise contacting an ocular cell with different expression vectors, each comprising a nucleic acid sequence encoding an inhibitor of angiogenesis and/or a nucleic acid sequence encoding a neurotrophic agent. In addition, the present invention provides a viral vector comprising a nucleic acid sequence encoding pigment epithelium-derived factor (PEDF) or a therapeutic fragment thereof.

Abstract translation: 本发明涉及一种预防性或治疗性地治疗动物至少一种与眼部相关的疾病，例如眼睛新生血管形成或年龄相关性黄斑变性的方法。 该方法包括使眼细胞与包含编码血管生成抑制剂的核酸序列和编码神经营养剂的相同或不同核酸序列的表达载体接触。 该方法还可以包括使眼细胞与不同的表达载体接触，每种表达载体包含编码血管生成抑制剂的核酸序列和/或编码神经营养剂的核酸序列。 此外，本发明提供了包含编码色素上皮衍生因子（PEDF）的核酸序列或其治疗片段的病毒载体。

公开(公告)号：US20030017595A1

公开(公告)日：2003-01-23

申请号：US09911011

申请日：2001-07-23

Applicant: GenVec, Inc.

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

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

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。