公开(公告)号：US5674729A

公开(公告)日：1997-10-07

申请号：US268679

申请日：1994-06-30

申请人： Eckard Wimmer ,  Akhteruzzaman Molla ,  Aniko V. Paul

发明人： Eckard Wimmer ,  Akhteruzzaman Molla ,  Aniko V. Paul

IPC分类号： C12N7/00

CPC分类号： C12N7/00 ,  C12N2770/32051

摘要： A process for the de novo synthesis of a picornavirus using a cell-free medium has been developed. The cell-free medium is prepared from a lysate of mammalian cells from which the nuclei and mitochondria has been removed, endogenous mRNA deactivated and supplemented with nucleoside triphosphates, tRNA, amino acids and precursors necessary for generating proteins. The genomic RNA of the picornavirus or rhinovirus is added to the cell-free medium and after incubation at about 30.degree. C. to 40.degree. C. of from about 4 to 24 hours infectious, mature virus particles are formed.

摘要翻译： 已经开发了使用无细胞培养基从头合成小核糖核酸病毒的方法。 从细胞核和线粒体被去除的哺乳动物细胞的裂解物制备无细胞培养基，内源mRNA失活并补充有产生蛋白质所必需的三磷酸核苷，tRNA，氨基酸和前体。 将微小RNA病毒或鼻病毒的基因组RNA加入到无细胞培养基中，并且在约30至40℃孵育约4至24小时感染性的成熟病毒颗粒。

公开(公告)号：US06194141B1

公开(公告)日：2001-02-27

申请号：US09282351

申请日：1999-03-31

申请人： Aniko V. Paul ,  Eckard Wimmer ,  Elizabeth Rieder

发明人： Aniko V. Paul ,  Eckard Wimmer ,  Elizabeth Rieder

IPC分类号： C12Q100

CPC分类号： C12Q1/48 ,  G01N2333/105 ,  G01N2333/9125 ,  G01N2500/20

摘要： The present invention provides methods of inhibiting picornavirus genome replication in a subject. In particular, methods for interfering with VPg uridylylation and elongation are provided. The methods comprise administering to a subject an effective amount of at least one of VPg, VPg analog, VPg homology or biologically active fragment thereof as well as oligonucleotides, divalent cations, ribonucleotide or deoxyribonucleotide. Also provided are methods of identifying an inhibitor of picornaviral replication which comprise adding a potential inhibitor of picornaviral replication to an in vitro assay and analyzing levels of VPg uridylylation reaction products.

摘要翻译： 本发明提供了在受试者中抑制小RNA病毒基因组复制的方法。 特别地，提供了用于干扰VPg尿苷酸化和延长的方法。 所述方法包括向受试者施用有效量的VPg，VPg类似物，VPg同源物或其生物活性片段以及寡核苷酸，二价阳离子，核糖核苷酸或脱氧核糖核苷酸中的至少一种。 还提供了鉴定微小RNA病毒复制抑制剂的方法，其包括将潜在的微小RNA病毒复制抑制剂添加到体外测定中并分析VPg尿苷酸化反应产物的水平。

公开(公告)号：US06689559B2

公开(公告)日：2004-02-10

申请号：US09998900

申请日：2001-11-29

申请人： Eckard Wimmer ,  Chengyu Liang ,  Sung Key Jang ,  Bumsuk Hahm

发明人： Eckard Wimmer ,  Chengyu Liang ,  Sung Key Jang ,  Bumsuk Hahm

IPC分类号： C12Q170

CPC分类号： C07K14/005 ,  C12N7/00 ,  C12N15/86 ,  C12N2770/24222 ,  C12N2770/24243 ,  C12N2840/203 ,  G01N33/5767 ,  G01N2800/52

摘要： The present invention provides a Hepatitis C Virus (HCV) replicon that efficiently replicates in an eukaryotic cell. The HCV replicon includes a nucleic acid sequence encoding a subgenomic fragments of HCV of any genotype that confer on the RNA the ability to replicate, and a nucleic acid sequence encoding an acetyl transferase selectable marker, such as puromycin. Also provided is an HCV type 1a replicon that efficiently replicates in an eukaryotic cell and includes a nucleic acid sequence encoding subgenomic fragments of type 1a HCV that confer on the RNA the ability to replicate, and a nucleic acid sequence encoding a acetyl transferase selectable marker. Further provided are eukaryotic cell lines that include an HCV replicon or an HCV type 1a replicon which efficiently replicate in the eukaryotic cell. The present invention also provides screening methods for identifying candidate compounds that inhibit the propagation of HCV.

摘要翻译： 本发明提供在真核细胞中有效复制的丙型肝炎病毒（HCV）复制子。 HCV复制子包括编码赋予RNA复制能力的任何基因型的HCV亚基因组片段的核酸序列，以及编码乙酰转移酶选择标记如嘌呤霉素的核酸序列。 还提供了在真核细胞中有效复制的HCV 1a型复制子，并且包括编码赋予RNA复制能力的1a型HCV亚型基因组片段的核酸序列和编码乙酰转移酶选择性标记的核酸序列。 还提供了真核生物细胞系，其包括在真核细胞中有效复制的HCV复制子或HCV型1a复制子。 本发明还提供了用于鉴定抑制HCV繁殖的候选化合物的筛选方法。

公开(公告)号：US06264940B1

公开(公告)日：2001-07-24

申请号：US09129686

申请日：1998-08-05

申请人： Matthias Gromeier ,  Eckard Wimmer

发明人： Matthias Gromeier ,  Eckard Wimmer

IPC分类号： A01N6300

CPC分类号： C12N15/86 ,  A61K35/768 ,  A61K48/00 ,  C12N2770/32632 ,  C12N2770/32643 ,  C12N2840/203 ,  Y02A50/465

摘要： The present invention is directed to non-pathogenic, oncolytic, recombinant polioviruses for the treatment of various forms of malignant tumors. The recombinant polioviruses of the invention are those in which the internal ribosomal entry site (IRES) of the wild type poliovirus was exchanged with the IRES of other picornaviruses, and optionally P1, P3 or the 3′NTR thereof was exchanged with that of poliovirus Sabin type. More particularly, the present invention is directed to the administration of the non-pathogenic, oncolytic, recombinant poliovirus to the tumor directly, intrathecally or intravenously to cause tumor necrosis. The method of the present invention is particularly useful for the treatment of malignant tumors in various organs, such as: breast, colon, bronchial passage, epithelial lining of the gastrointestinal, upper respiratory and genito-urinary tracts, liver, prostate and the brain. Astounding remissions in experimental animals have been demonstrated for the treatment of malignant glioblastoma multiforme, an almost universally fatal neoplasm of the central nervous system.

摘要翻译： 本发明涉及用于治疗各种形式的恶性肿瘤的非致病性，溶瘤性重组脊髓灰质炎病毒。 本发明的重组体脊髓灰质炎病毒是其中野生型脊髓灰质炎病毒的内部核糖体进入位点（IRES）与其他微小核糖核酸病毒的IRES交换的，并且任选的P1，P3或其3'NTR与脊髓灰质炎病毒Sabin 类型。 更具体地，本发明涉及直接，鞘内或静脉内给予肿瘤非致病性，溶瘤性重组体脊髓灰质炎病毒以引起肿瘤坏死。 本发明的方法特别可用于治疗各种器官中的恶性肿瘤，例如：乳腺，结肠，支气管传代，胃肠道，上呼吸道和生殖泌尿道，肝脏，前列腺和脑部的上皮层。 实验动物的惊人缓解已被证明用于治疗多形性恶性胶质母细胞瘤，这是几乎普遍致命的中枢神经系统肿瘤。

公开(公告)号：US06464972B1

公开(公告)日：2002-10-15

申请号：US09566581

申请日：2000-05-08

申请人： Matthias Gromeier ,  Eckard Wimmer

发明人： Matthias Gromeier ,  Eckard Wimmer

IPC分类号： A01N6300

CPC分类号： C12N15/86 ,  A61K35/768 ,  A61K48/00 ,  C12N2770/32632 ,  C12N2770/32643 ,  C12N2840/203 ,  Y02A50/465

摘要： The present invention is directed to non-pathogenic, oncolytic, recombinant polioviruses for the treatment of various forms of malignant tumors. The recombinant polioviruses of the invention are those in which the internal ribosomal entry site (IRES) of the wild type poliovirus was exchanged with the IRES of other picornaviruses, and optionally P1, P3 or the 3′NTR thereof was exchanged with that of poliovirus Sabin type. More particularly, the present invention is directed to the administration of the non-pathogenic, oncolytic, recombinant poliovirus to the tumor directly, intrathecally or intravenously to cause tumor necrosis. The method of the present invention is particularly useful for the treatment of malignant tumors in various organs, such as: breast, colon, bronchial passage, epithelial lining of the gastrointestinal, upper respiratory and genito-urinary tracts, liver, prostate and the brain. Astounding remissions in experimental animals have been demonstrated for the treatment of malignant glioblastoma multiforme, an almost universally fatal neoplasm of the central nervous system.

摘要翻译： 本发明涉及用于治疗各种形式的恶性肿瘤的非致病性，溶瘤性重组脊髓灰质炎病毒。 本发明的重组体脊髓灰质炎病毒是其中野生型脊髓灰质炎病毒的内部核糖体进入位点（IRES）与其他微小核糖核酸病毒的IRES交换的，并且任选的P1，P3或其3'NTR与脊髓灰质炎病毒Sabin 类型。 更具体地，本发明涉及直接，鞘内或静脉内给予肿瘤非致病性，溶瘤性重组体脊髓灰质炎病毒以引起肿瘤坏死。 本发明的方法特别可用于治疗各种器官中的恶性肿瘤，例如：乳腺，结肠，支气管传代，胃肠道，上呼吸道和生殖泌尿道，肝脏，前列腺和脑部的上皮层。 实验动物的惊人缓解已被证明用于治疗多形性恶性胶质母细胞瘤，这是几乎普遍致命的中枢神经系统肿瘤。

公开(公告)号：US09476032B2

公开(公告)日：2016-10-25

申请号：US12594173

申请日：2008-03-31

申请人： Eckard Wimmer ,  Steve Skiena ,  Steffen Mueller ,  Bruce Futcher ,  Dimitris Papamichail ,  John Robert Coleman ,  Jeronimo Cello

发明人： Eckard Wimmer ,  Steve Skiena ,  Steffen Mueller ,  Bruce Futcher ,  Dimitris Papamichail ,  John Robert Coleman ,  Jeronimo Cello

IPC分类号： C12N7/04 ,  A61K39/00 ,  C12N7/00

CPC分类号： C12N7/00 ,  A61K39/13 ,  A61K39/145 ,  A61K2039/5254 ,  C07K14/005 ,  C12N2720/12361 ,  C12N2740/15061 ,  C12N2760/16061 ,  C12N2760/16121 ,  C12N2760/16134 ,  C12N2760/16162 ,  C12N2770/20061 ,  C12N2770/24161 ,  C12N2770/32061 ,  C12N2770/32621 ,  C12N2770/32634 ,  C12N2770/32661 ,  C12N2770/32662 ,  C12N2770/32671 ,  C12N2770/32721 ,  C12N2770/32761 ,  C12N2770/32762 ,  Y02A50/386 ,  Y02A50/388 ,  Y02A50/466

摘要： This invention provides an attenuated virus which comprises a modified viral genome containing nucleotide substitutions engineered in multiple locations in the genome, wherein the substitutions introduce synonymous deoptimized codons into the genome. The instant attenuated virus may be used in a vaccine composition for inducing a protective immune response in a subject. The invention also provides a method of synthesizing the instant attenuated virus. Further, this invention further provides a method for preventing a subject from becoming afflicted with a virus-associated disease comprising administering to the subject a prophylactically effective dose of a vaccine composition comprising the instant attenuated virus.

摘要翻译： 本发明提供了一种减毒病毒，其包含经修饰的病毒基因组，其含有在基因组中多个位置工程改造的核苷酸取代，其中所述取代引入同义脱酮密码子进入基因组。 本发明的减毒病毒可用于疫苗组合物中以在受试者中诱导保护性免疫应答。 本发明还提供了合成即时减毒病毒的方法。 此外，本发明进一步提供了一种防止受试者患有病毒相关疾病的方法，包括向受试者施用预防有效剂量的包含即时减毒病毒的疫苗组合物。

公开(公告)号：US08921101B2

公开(公告)日：2014-12-30

申请号：US13269213

申请日：2011-10-07

申请人： Eckard Wimmer ,  Jeronimo Cello ,  Aniko Paul ,  Hidemi Toyoda ,  Jiang Yin

发明人： Eckard Wimmer ,  Jeronimo Cello ,  Aniko Paul ,  Hidemi Toyoda ,  Jiang Yin

IPC分类号： A61K39/12 ,  C12N7/00 ,  A61K35/76

CPC分类号： C12N7/00 ,  A61K35/768 ,  C12N2770/32332 ,  C12N2770/32361 ,  C12N2800/30 ,  C12N2840/203

摘要： A novel and stable attenuated poliovirus, which replicates in neuroblastoma cells, is produced by engineering an indigenous replication element (cre), into the 5′ non-translated genomic region and inactivating the native cre element located in the coding region of 2C (mono-crePV). The stably attenuated poliovirus replicates in a neuroblastoma model (Neuro-2aCD155 tumors) expressing CD155, the poliovirus receptor, and is effective for oncolytic treatment and cure of solid tumors, such as neuroblastoma.

摘要翻译： 在神经母细胞瘤细胞中复制的新颖且稳定的减毒脊髓灰质炎病毒通过将本地复制元件（cre）工程化成5'非翻译的基因组区域并使位于2C编码区的天然cre元件失活而产生， crePV）。 稳定减毒的脊髓灰质炎病毒在表达CD155（脊髓灰质炎病毒受体）的神经母细胞瘤模型（Neuro-2aCD155肿瘤）中复制，并且对于固体肿瘤如神经母细胞瘤的溶瘤治疗和治愈是有效的。

公开(公告)号：US08066983B2

公开(公告)日：2011-11-29

申请号：US12405068

申请日：2009-03-16

申请人： Eckard Wimmer ,  Jeronimo Cello ,  Aniko Paul ,  Hidemi Toyoda ,  Jiang Yin

发明人： Eckard Wimmer ,  Jeronimo Cello ,  Aniko Paul ,  Hidemi Toyoda ,  Jiang Yin

IPC分类号： A61K39/12 ,  C12P19/33

CPC分类号： C12N7/00 ,  A61K35/768 ,  C12N2770/32332 ,  C12N2770/32361 ,  C12N2800/30 ,  C12N2840/203

摘要： A novel and stable attenuated poliovirus, which replicates in neuroblastoma cells, is produced by engineering an indigenous replication element (cre), into the 5′ non-translated genomic region and inactivating the native cre element located in the coding region of 2C (mono-crePV). The stably attenuated poliovirus replicates in a neuroblastoma model (Neuro-2aCD155 tumors) expressing CD155, the poliovirus receptor, and is effective for oncolytic treatment and cure of solid tumors, such as neuroblastoma.

摘要翻译： 在神经母细胞瘤细胞中复制的新颖且稳定的减毒脊髓灰质炎病毒通过将本地复制元件（cre）工程化成5'非翻译的基因组区域并使位于2C编码区的天然cre元件失活而产生， crePV）。 稳定减毒的脊髓灰质炎病毒在表达CD155（脊髓灰质炎病毒受体）的神经母细胞瘤模型（Neuro-2aCD155肿瘤）中复制，并且对于固体肿瘤如神经母细胞瘤的溶瘤治疗和治愈是有效的。

公开(公告)号：US07147848B2

公开(公告)日：2006-12-12

申请号：US10175247

申请日：2002-06-19

申请人： Matthias Gromeier ,  Eckard Wimmer

发明人： Matthias Gromeier ,  Eckard Wimmer

IPC分类号： A01N63/00 ,  C12N7/00 ,  C12N7/01 ,  C12N7/02 ,  C12N7/04 ,  C07H21/00

CPC分类号： C12N15/86 ,  A61K35/768 ,  A61K48/00 ,  C12N2770/32632 ,  C12N2770/32643 ,  C12N2840/203 ,  Y02A50/465

摘要： The present invention is directed to non-pathogenic, oncolytic, recombinant polioviruses for the treatment of various forms of malignant tumors. The recombinant polioviruses of the invention are those in which the internal ribosomal entry site (IRES) of the wild type poliovirus was exchanged with the IRES of other picornaviruses, and optionally P1, P3 or the 3′NTR thereof was exchanged with that of poliovirus Sabin type. More particularly, the present invention is directed to the administration of the non-pathogenic, oncolytic, recombinant poliovirus to the tumor directly, intrathecally or intravenously to cause tumor necrosis. The method of the present invention is particularly useful for the treatment of malignant tumors in various organs, such as: breast, colon, bronchial passage, epithelial lining of the gastrointestinal, upper respiratory and genito-urinary tracts, liver, prostate and the brain. Astounding remissions in experimental animals have been demonstrated for the treatment of malignant glioblastoma multiforme, an almost universally fatal neoplasm of the central nervous system.

公开(公告)号：US20120269849A1

公开(公告)日：2012-10-25

申请号：US13501045

申请日：2010-10-11

申请人： Eckard Wimmer ,  Steve Skiena ,  Steffen Mueller ,  Bruce Futcher ,  Dimitris Papamichail ,  John Robert Coleman ,  Jeronimo Cello

发明人： Eckard Wimmer ,  Steve Skiena ,  Steffen Mueller ,  Bruce Futcher ,  Dimitris Papamichail ,  John Robert Coleman ,  Jeronimo Cello

IPC分类号： A61K39/12 ,  C12N7/04 ,  A61P31/12 ,  C12N15/54

CPC分类号： C07K14/005 ,  A61K39/12 ,  A61K39/145 ,  A61K2039/5254 ,  A61K2039/543 ,  C12N7/00 ,  C12N2760/16122 ,  C12N2760/16134 ,  C12N2760/16162

摘要： The present provides attenuated influenza viruses comprising a modified viral genome containing a plurality of nucleotide substitutions. The nucleotide substitutions result in the rearrangement of preexisting codons of one or more protein encoding sequences and changes in codon pair bias. Substitutions of non-synonymous and synonymous codons may also be included. The attenuated influenza viruses enable production of improved vaccines and are used to elicit protective immune responses.

摘要翻译： 本发明提供减毒流感病毒，其包含含有多个核苷酸取代的经修饰的病毒基因组。 核苷酸取代导致一个或多个蛋白质编码序列的预先存在的密码子的重排以及密码子对偏差的变化。 也可以包括非同义和同义密码子的取代。 减毒的流感病毒能够生产改进的疫苗，并用于引发保护性免疫应答。