公开(公告)号：EP0954591A2

公开(公告)日：1999-11-10

申请号：EP97928961.0

申请日：1997-05-30

Applicant: BAXTER INTERNATIONAL INC.

Inventor： ZHANG, Wei-Wei ,  ALEMANY, Ramon ,  DAI, Yifan ,  JOSEPHS, Steven ,  BALAGUE, Cristina ,  AYARES, David ,  SCHNEIDERMAN, Richard

IPC: A01K67 ,  A61K31 ,  A61K35 ,  A61K38 ,  A61K48 ,  A61P7 ,  C07K14 ,  C12N5 ,  C12N7 ,  C12N15 ,  C12Q1 ,  G01N21 ,  G01N33

CPC classification number: C12N15/8509 ,  A01K67/0278 ,  A01K2207/15 ,  A01K2217/00 ,  A01K2217/05 ,  A01K2217/075 ,  A01K2217/20 ,  A01K2267/0306 ,  A61K38/00 ,  A61K48/00 ,  C07K14/755 ,  C12N15/86 ,  C12N2710/10343 ,  C12N2800/108 ,  C12N2800/30 ,  C12N2810/859 ,  C12N2830/003 ,  C12N2830/008 ,  C12N2830/38 ,  C12N2830/85 ,  C12N2840/20 ,  C12N2840/203 ,  C12N2840/206

Abstract: This invention is related to adenoviral (Ad) vectors and their applications in the field of genetic medicine, including gene transfer, gene therapy, and gene vaccination. More specifically, this invention is related to the Ad vectors that carry the minimal cis-element of the Ad genome (mini-Ad vector) and are capable of delivering transgenes and/or heterologous DNA up to 36 kb. The generation and propagation of the mini-Ad vectors require trans-complementation of a packaging-attenuated and replication-defective helper Ad (helper) in an Ad helper cell line. This invention further comprises a methodology for generating a mini-adenoviral (mini-Ad) vector for use in gene therapy of hemophilia and animal test systems for in vivo evaluation of the Ad vectors. More specifically, this invention describes factor VIII (FVIII) Ad vectors that only contain minimal cis-element of the Ad genome (so-called mini-Ad) and comprise a human FVIII cDNA with other supporting DNA elements up to 36 kb. The FVIII mini-Ad can be generated and preferentially amplified through the assistance of a packaging-attenuated helper Ad and a helper cell line. This invention also reports designs and methods for producing transgenic mouse models that can be used for in vivo testing the mini-Ad.

公开(公告)号：EP0973866A1

公开(公告)日：2000-01-26

申请号：EP98908661.6

申请日：1998-02-23

Applicant: BAXTER INTERNATIONAL INC.

Inventor： AYARES, David ,  ALEMANY, Ramon ,  ZHANG, Wei-Wei

IPC: C12N5/00 ,  C12N15/63 ,  C12N1/21 ,  C12N15/64

CPC classification number: C12N7/00 ,  C12N15/86 ,  C12N2710/10343 ,  C12N2710/10345 ,  C12N2710/10352 ,  C12N2750/14143

Abstract: A new series of helper cell lines for the complementation, amplification, and controlled attenuation of E1-deleted adenovirus are disclosed in the present invention. These cell lines are advantageous because they can complement adenovirus E1 genedeletions without production of replication competent adenovirus (RCA), thus making them safer for the large-scale production of adenovirus stock for use in human genetherapy trials. A preferred embodiment is an A549E1 cell line that contains only the Ad5 E1 DNA sequences sufficient for complementation of E1-deleted adenoviral vectors without sequences that overlap with the adenovirus vector. In another aspect, the present invention embodies methods for the production of second generation A549-E1 complementing cell lines that, in addition to producing E1, also produce proteins required for further manipulation of adenoviral vectors. A preferred embodiment is an A549E1 cell line with DNA sequences that encode a polypeptide sufficient for packaging attenuation of E1-deleted helper virus, in order to enrich for packaging of mini-adenovirus.

Abstract translation: 本发明公开了一系列用于E1缺失型腺病毒的互补，扩增和受控减毒的辅助细胞系。 这些细胞系是有利的，因为它们可以补充腺病毒E1基因缺失而不产生具有复制能力的腺病毒（RCA），从而使它们更安全地用于人类基因疗法试验中的腺病毒储存的大规模生产。 一个优选的实施方案是A549E1细胞系，其仅包含Ad5E1 DNA序列，其足以补偿E1缺失的腺病毒载体而没有与腺病毒载体重叠的序列。 另一方面，本发明体现了用于产生第二代A549-E1补体细胞系的方法，所述第二代A549-E1补充细胞系除了产生E1之外还产生进一步操作腺病毒载体所需的蛋白质。 一个优选的实施方案是具有编码多肽的DNA序列的A549E1细胞系，所述多肽足以包装E1缺失的辅助病毒的减毒，以便富集微小腺病毒的包装。

公开(公告)号：EP0968281A2

公开(公告)日：2000-01-05

申请号：EP98904658.6

申请日：1998-01-23

Applicant: BAXTER INTERNATIONAL INC.

Inventor： ALEMANY, Ramon ,  FANG, Xiangming ,  ZHANG, Wei-Wei

IPC: C12N15/00

CPC classification number: C12N7/00 ,  A61K35/761 ,  A61K38/1774 ,  A61K38/217 ,  A61K48/00 ,  C07K14/4715 ,  C12N15/86 ,  C12N2710/10332 ,  C12N2710/10343 ,  C12N2830/008 ,  C12N2830/60 ,  C12N2840/20 ,  A61K2300/00

Abstract: This invention encompasses a composition for killing target cells, such as tumor cells. The composition comprises a first and a second adenoviral vector that have complementary function and are mutually dependent on each other for replication in a target cell. One of said adenoviral vectors has a target cell-activated promoter or a functional deletion that controls and limits propagation of the adenoviral vectors in the target cells which directly or indirectly kills the target cells. One of the adenoviral vectors comprises a gene encoding a protein which is expressed in the target cells and can induce anticancer immune responses. The target cells may be hepatoma, breast cancer, melanoma, colon cancer, or prostate cancer cells, for example. The vectors of this invention may also be utilized to treat other diseases such as restenosis, in which case the target cell may be a vascular smooth muscle cell, for example.