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1. 发明申请

WO0034497A3 ENHANCED PACKAGING OF HERPES VIRUS AMPLICONS AND GENERATION OF RECOMBINANT VIRUS VECTORS 审中-公开
标题翻译：增强的病毒片段的包装和重组病毒载体的产生
公开(公告)号：WO0034497A3
公开(公告)日：2000-11-09
申请号：PCT/US9929120
申请日：1999-12-09
申请人： BREAKEFIELD XANDRA O , CHIOCCA E ANTONIO , SAEKI YOSHINAGA , FRAEFEL CORNEL , TOBLER KURT , ACKERMANN MATHIAS , SUTER MARK , ADEMA GOSSE J , SHORTMAN KEN
发明人： BREAKEFIELD XANDRA O , CHIOCCA E ANTONIO , SAEKI YOSHINAGA , FRAEFEL CORNEL , TOBLER KURT , ACKERMANN MATHIAS , SUTER MARK , ADEMA GOSSE J , SHORTMAN KEN
IPC分类号： A61K39/00 , A61K39/245 , A61K48/00 , C12N15/33 , C12N15/38 , C12N15/869
CPC分类号： C12N15/86 , A61K39/00 , A61K39/12 , A61K39/245 , A61K48/00 , A61K2039/5256 , A61K2039/53 , A61K2039/545 , C12N2710/16634 , C12N2710/16643 , C12N2800/204
摘要： The present invention relates to an enhanced and simplified herpes virus amplicon packaging system and method of generating recombinant herpes virus vectors by "pac rescue". In one embodiment of this packaging system, the HSW-1 genome is cloned and maintained as a single-copy, F-plasmid-based BAC in E.coli. Such a plasmid, containing the HSV-1 genome deleted for the pac signals, did not generate replication-competent progeny virus upon transfection into mammalian cells, but rather, supported the packaging of co-transfected amplicon DNA which contained a functional pac signal. In another embodiment, the HSV genome cloned in BAC has an intact pac signal, however, the helper HSV genome is made packaging defective by increasing or decreasing the size of the BAC. This method involves the co-introduction of DNA fragments containing a pac site flanked by homologous HSV sequences and pac-deleted helper virus genome to permissive cells. In another embodiment, any kind of modification can be introduced into the viral genome through homologous recombination in bacteria.
摘要翻译：本发明涉及一种增强和简化的疱疹病毒扩增子包装系统和通过“pac拯救”产生重组疱疹病毒载体的方法。在该包装系统的一个实施方案中，将HSW-1基因组克隆并维持为大肠杆菌中的基于单拷贝，F-质粒的BAC。含有针对pac信号缺失的HSV-1基因的这种质粒在转染哺乳动物细胞后没有产生复制能力的后代病毒，而是支持包含功能性pac信号的共转染的扩增子DNA的包装。在另一个实施方案中，克隆在BAC中的HSV基因组具有完整的pac信号，然而，辅助HSV基因组通过增加或减少BAC的大小而被制成包装有缺陷的。该方法涉及将含有同源HSV序列和pac缺失的辅助病毒基因组的Pac位点的DNA片段共同引入允许的细胞。在另一个实施方案中，可以通过细菌中的同源重组将任何种类的修饰引入到病毒基因组中。

2. 发明申请

WO0065077A8 TRIPLE HYBRID AMPLICON VECTOR SYSTEMS TO GENERATE RETROVIRAL PACKAGING LINES 审中-公开
标题翻译：三重混合放大器矢量系统生成回流包装线
公开(公告)号：WO0065077A8
公开(公告)日：2001-10-11
申请号：PCT/US0010669
申请日：2000-04-21
申请人： GEN HOSPITAL CORP , SENA ESTEVES MIGUEL , BREAKEFIELD XANDRA O , SAEKI YOSHINAGA
发明人： SENA-ESTEVES MIGUEL , BREAKEFIELD XANDRA O , SAEKI YOSHINAGA
IPC分类号： A61K48/00 , C07K14/15 , C12N5/10 , C12N15/38 , C12N15/867 , C12N15/864 , C12N15/63 , C12N15/64 , C12N15/869
CPC分类号： C12N7/00 , A61K48/00 , C07K14/005 , C12N15/86 , C12N2740/13022 , C12N2740/13043 , C12N2740/13052 , C12N2799/021
摘要： The present invention relates to a triple hybrid vector amplicon system comprising genetic elements derived from Herpes Simplex Virus (HSV), Epstein-Barr Virus (EBV) or Adeno-Associated Virus (AAV), and a retrovirus. The vector was developed to stably transform cells, both in culture or in vivo, into retrovirus packaging cells in a single step. This step can be accomplished both by transfection using liposomes, electroporation, calcium phosphate, or any other methodology used to transfer naked or complexed DNA into cells or by infection when the vector is packaged as an amplicon vector in HSV virions. In one embodiment, the system takes advantage of the host range and retention properties of HSV/EBV hybrid amplicons to efficiently convert cells to retrovirus vector producer cells after single-step transduction. Retrovirus genes gag-pol and env (GPE) and retroviral vector sequences were modified to minimize sequence overlap and cloned into an HSV/EBV hybrid amplicon. In a second embodiment, retrovirus gag-pol and env genes and a retrovirus vector carrying lacZ, were cloned into HSV/AAV hybrid amplicons. These hybrid amplicon vector systems hold great potential for the generation of new retrovirus packaging lines derived from cells that due to their migratory, tumor or tissue targeting properties, can expand the spatial distribution of gene delivery by retrovirus vectors in vivo.
摘要翻译：本发明涉及包含衍生自单纯疱疹病毒（HSV），爱泼斯坦 - 巴尔病毒（EBV）或腺相关病毒（AAV））和逆转录病毒的遗传元件的三重杂交载体扩增子系统。开发该载体以在单一步骤中将培养物或体内的细胞稳定地转化成逆转录病毒包装细胞。该步骤可以通过使用脂质体，电穿孔，磷酸钙或用于将裸露或复合DNA转移到细胞中的任何其它方法或当将载体作为扩增子载体包装在HSV病毒粒子中时通过感染来实现。在一个实施方案中，该系统利用HSV / EBV杂交扩增子的宿主范围和保留性质，以在单步转导后有效地将细胞转化为逆转录病毒载体生产细胞。逆转录病毒基因gag-pol和env（GPE）和逆转录病毒载体序列被修饰以最小化序列重叠并克隆到HSV / EBV杂交扩增子中。在第二个实施方案中，将逆转录病毒gag-pol和env基因和携带lacZ的逆转录病毒载体克隆到HSV / AAV杂交扩增子中。这些杂交扩增子载体系统具有巨大的潜力，用于产生由细胞衍生的新的逆转录病毒包装线，由于其迁移，肿瘤或组织靶向特性，可以扩大反转录病毒载体在体内的基因递送的空间分布。

3. 发明申请

WO2009052426A3 ONCOLYTIC VIRUS 审中-公开
公开(公告)号：WO2009052426A3
公开(公告)日：2009-07-30
申请号：PCT/US2008080367
申请日：2008-10-17
申请人： UNIV OHIO STATE , KAUR BALVEEN , CHIOCCA ANTONIO , SAEKI YOSHINAGA
发明人： KAUR BALVEEN , CHIOCCA ANTONIO , SAEKI YOSHINAGA
IPC分类号： C12N15/38
CPC分类号： C07K14/4703 , A61K35/763 , A61K38/00 , A61K38/177 , A61K48/00 , C12N7/00 , C12N15/89 , C12N2710/16632 , C12N2710/16643 , C12N2830/60 , A61K2300/00
摘要： Malignant tumors that are intrinsically resistant to conventional therapies are significant therapeutic challenges. An embodiment of the present invention provides an oncolytic virus capable of killing target cells, such as a tumor cells. In various embodiments presented herein, the oncolytic virus is armed or encodes a therapeutic polypeptide. In at least one embodiment, a recombinant oncolytic virus has been generated that can specifically replicate in cancer cells leading to their destruction and at the same time secrete robust amounts of an angiostatic factor to inhibit the regrowth of residual disease. Compositions and methods disclosed herein have broad therapeutic applicability.
摘要翻译：对传统疗法本质抵抗的恶性肿瘤是显着的治疗挑战。本发明的一个实施方案提供了能够杀死靶细胞如肿瘤细胞的溶瘤病毒。在本文提出的各种实施方案中，溶瘤病毒是配制或编码治疗性多肽的。在至少一个实施方案中，已经产生重组溶瘤病毒，其可以在导致其破坏的癌细胞中特异性复制，并且同时分泌强大量的血管生成抑制因子以抑制残留疾病的再生长。本文公开的组合物和方法具有广泛的治疗适用性。

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