公开(公告)号：US07666405B2

公开(公告)日：2010-02-23

申请号：US10935576

申请日：2004-09-07

Applicant: Andrea Amalfitano ,  Yuan Tsong Chen ,  Huimin Hu ,  Bradley Lowell Hodges

Inventor： Andrea Amalfitano ,  Yuan Tsong Chen ,  Huimin Hu ,  Bradley Lowell Hodges

IPC: A61K48/00 ,  C12N15/00 ,  A01N43/04

CPC classification number: C12N15/86 ,  A61K48/00 ,  C07K14/005 ,  C12N2710/10322 ,  C12N2710/10343 ,  C12N2800/30 ,  C12N2830/008 ,  C12N2830/42 ,  C12N2840/20

Abstract: The present invention provides deleted adenovirus vectors. The inventive adenovirus vectors carry one or more deletions in the IVa2, 100K, polymerase and/or preterminal protein sequences of the adenovirus genome. The adenoviruses may additionally contain other deletions, mutations or other modifications as well. In particular preferred embodiments, the adenovirus genome is multiply deleted, i.e., carries two or more deletions therein. The deleted adenoviruses of the invention are “propagation-defective” in that the virus cannot replicate and produce new virions in the absence of complementing function(s). Preferred adenovirus vectors of the invention carry a heterologous nucleotide sequence encoding a protein or peptide associated with a metabolic disorder, more preferably a protein or peptide associated with a lysosomal or glycogen storage disease, most preferably, a lysosomal acid α-glucosidase. Further provided are methods for producing the inventive deleted adenovirus vectors. Further provided are methods of administering the deleted adenovirus vectors to a cell in vitro or in vivo.

Abstract translation: 本发明提供了缺失的腺病毒载体。 本发明的腺病毒载体在腺病毒基因组的IVa2,100K，聚合酶和/或早产蛋白质序列中携带一个或多个缺失。 腺病毒还可以另外含有其它缺失，突变或其它修饰。 在特别优选的实施方案中，腺病毒基因组被多次缺失，即在其中携带两个或多个缺失。 本发明的缺失的腺病毒是“繁殖缺陷型”，因为在没有补体功能的情况下病毒不能复制并产生新的病毒粒子。 本发明优选的腺病毒载体携带编码与代谢紊乱相关的蛋白质或肽的异源核苷酸序列，更优选与溶酶体或糖原贮积病相关的蛋白质或肽，最优选溶酶体酸性α-葡糖苷酶。 还提供了用于产生本发明缺失的腺病毒载体的方法。 还提供了在体外或体内将缺失的腺病毒载体施用于细胞的方法。

公开(公告)号：US06451596B1

公开(公告)日：2002-09-17

申请号：US09562919

申请日：2000-05-02

Applicant: Jeffrey S. Chamberlain ,  Andrea Amalfitano ,  Michael A. Hauser ,  Rajendra Kumar-Singh ,  Denis J. Hartigan-O'Connor

Inventor： Jeffrey S. Chamberlain ,  Andrea Amalfitano ,  Michael A. Hauser ,  Rajendra Kumar-Singh ,  Denis J. Hartigan-O'Connor

IPC: C12N1586

CPC classification number: C12N15/86 ,  A61K48/00 ,  C07K14/4708 ,  C12N7/00 ,  C12N2710/10343 ,  C12N2800/30 ,  C12N2830/008 ,  C12N2830/30

Abstract: The present invention provides improved adenovirus vectors and packaging cell lines. One type of improved adenoviral vector comprises deletions within the E2b region of the adenoviral genome. These E2b-deleted virus are used in conjunction with novel cell lines that constitutively express E2b gene products. The present invention further provides adenoviral vectors deleted for all viral coding regions. These “gutted” vectors permit the transfer of large genes to cells as demonstrated herein by the transfer of the dystrophin gene to the muscle of mice. The E2b-deleted vectors and the gutted vectors provide improved adenoviral vectors useful for a wide variety of gene therapy applications.

Abstract translation: 本发明提供了改进的腺病毒载体和包装细胞系。 一种类型的改良的腺病毒载体包含在腺病毒基因组的E2b区域内的缺失。 这些E2b缺失的病毒与组成型表达E2b基因产物的新型细胞系一起使用。 本发明还提供了所有病毒编码区缺失的腺病毒载体。 这些“内含”载体允许将大基因转移到细胞中，如本文通过将肌营养不良蛋白基因转移到小鼠的肌肉所证明的。 E2b缺失的载体和内切的载体提供了可用于各种基因治疗应用的改良的腺病毒载体。

公开(公告)号：US20110294193A1

公开(公告)日：2011-12-01

申请号：US12976334

申请日：2010-12-22

Applicant: Andrea Amalfitano ,  Dwight D. Koeberl ,  Baodong Sun

Inventor： Andrea Amalfitano ,  Dwight D. Koeberl ,  Baodong Sun

IPC: C12N7/01

CPC classification number: C12N15/86 ,  A61K38/47 ,  A61K48/00 ,  A61K48/0066 ,  A61K48/0075 ,  A61K48/0083 ,  A61K2039/5256 ,  C12N9/2408 ,  C12N9/2411 ,  C12N2710/10344 ,  C12N2750/14143 ,  C12Y302/0102

Abstract: A recombinant hybrid virus, including: (a) a deleted adenovirus vector genome comprising the adenovirus 5′ and 3′ cis-elements for viral replication and encapsidation, and further comprising a deletion in an adenovirus genomic region selected from the group consisting of: (i) the polymerase region, wherein said deletion essentially prevents the expression of a functional polymerase protein from said deleted region and said hybrid virus does not otherwise express a functional polymerase protein, (ii) the preterminal protein region, wherein said deletion essentially prevents the expression of a functional preterminal protein from said deleted region, and said hybrid virus does not otherwise express a functional preterminal protein, and (iii) both the regions of (i) and (ii); and (b) a recombinant adeno-associated virus (AAV) vector genome flanked by the adenovirus vector genome sequences of (a), said recombinant AAV vector genome comprising (i) AAV 5′ and 3′ inverted terminal repeats, (ii) an AAV packaging sequence, and (iii) a heterologous nucleic acid sequence, wherein said heterologous nucleic acid sequence is flanked by the 5′ and 3′ AAV inverted terminal repeats of (i). Methods of making and using the recombinant hybrid virus are also disclosed.

Abstract translation: 一种重组杂合病毒，其包括：（a）缺失的腺病毒载体基因组，其包含用于病毒复制和壳化的腺病毒5'和3'顺式元件，并且还包含选自以下的腺病毒基因组区域中的缺失：（ i）聚合酶区域，其中所述缺失基本上防止来自所述缺失区的功能性聚合酶蛋白的表达，并且所述杂合病毒不另外表达功能性聚合酶蛋白，（ii）早产蛋白区，其中所述缺失基本上防止表达 的来自所述缺失区的功能性早产蛋白，并且所述杂交病毒不另外表达功能性早产蛋白，和（iii）（i）和（ii）的两个区域; 所述重组腺病毒载体基因组包含（i）AAV 5'和3'反向末端重复序列，（ii）一个或多个重组腺病毒载体基因组（AAV） AAV包装序列，和（iii）异源核酸序列，其中所述异源核酸序列侧翼为（i）的5'和3'AAV反向末端重复序列。 还公开了制备和使用重组杂交病毒的方法。

公开(公告)号：US07858367B2

公开(公告)日：2010-12-28

申请号：US10511980

申请日：2003-04-30

Applicant: Andrea Amalfitano ,  Dwight D. Koeberl ,  Baodong Sun

Inventor： Andrea Amalfitano ,  Dwight D. Koeberl ,  Baodong Sun

IPC: C12N15/00 ,  C12N7/00 ,  A61K48/00

CPC classification number: C12N15/86 ,  A61K38/47 ,  A61K48/00 ,  A61K48/0066 ,  A61K48/0075 ,  A61K48/0083 ,  A61K2039/5256 ,  C12N9/2408 ,  C12N9/2411 ,  C12N2710/10344 ,  C12N2750/14143 ,  C12Y302/0102

Abstract: A recombinant hybrid virus which includes: (a) a deleted adenovirus vector genome having the adenovirus 5′ and 3′ cis-elements for viral replication and encapsidation and a deletion in an adenovirus genomic region selected from the polymerase region and/or the preterminal protein region, wherein the deletion essentially prevents the expression of a functional polymerase and/or preterminal protein from the deleted region and the hybrid virus does not otherwise express a functional polymerase protein; and (b) a recombinant adeno-associated virus (AAV) vector genome flanked by the adenovirus vector genome sequences of (a), wherein the recombinant AAV vector genome includes an AAV packaging sequence and a heterologous nucleic acid sequence, wherein the heterologous nucleic acid sequence is flanked by 5′ and 3′ AAV inverted terminal repeats.

Abstract translation: 一种重组杂合病毒，其包括：（a）具有用于病毒复制和壳化的腺病毒5'和3'顺式元件的缺失的腺病毒载体基因组，以及在选自聚合酶区和/或早产蛋白的腺病毒基因组区域中的缺失 区，其中所述缺失基本上阻止来自缺失区的功能性聚合酶和/或早产蛋白的表达，并且所述杂合病毒不另外表达功能性聚合酶蛋白; 和（b）侧翼为（a）的腺病毒载体基因组序列的重组腺相关病毒（AAV）载体基因组，其中所述重组AAV载体基因组包含AAV包装序列和异源核酸序列，其中所述异源核酸 序列侧接5'和3'AAV反向末端重复序列。

公开(公告)号：US07129043B1

公开(公告)日：2006-10-31

申请号：US09830045

申请日：1999-10-21

Applicant: Rose-Mary N. Boustany ,  Wei-Xing Guo ,  Andrea Amalfitano

Inventor： Rose-Mary N. Boustany ,  Wei-Xing Guo ,  Andrea Amalfitano

IPC: C07H21/04 ,  C12Q1/68

CPC classification number: C12Q1/6886 ,  C12Q2600/118 ,  C12Q2600/136 ,  G01N2800/52

Abstract: A method of screening a subject for a proliferative disease risk factor comprises detecting the presence or absence of upregulation of the CLN3 gene in the subject. The upregulation of the CLN3 gene in the subject indicates the subject is at increased risk of developing a proliferative disease. Methods of screening compounds for the treatment of proliferative diseases based on the CLN3 gene and its product are also disclosed, along with methods of treating such diseases and vectors useful therefore.

Abstract translation: 筛选受试者增殖性疾病危险因素的方法包括检测受试者中CLN3基因上调的存在或不存在。 受试者中CLN3基因的上调表明受试者处于发展增殖性疾病的风险增加。 还公开了用于治疗基于CLN3基因及其产物的增殖性疾病的化合物的方法，以及用于治疗这样的疾病和载体的方法。

公开(公告)号：US20050019929A1

公开(公告)日：2005-01-27

申请号：US10935576

申请日：2004-09-07

Applicant: Andrea Amalfitano ,  Yuan Chen ,  Huimin Hu ,  Bradley Hodges

Inventor： Andrea Amalfitano ,  Yuan Chen ,  Huimin Hu ,  Bradley Hodges

IPC: C12N15/09 ,  A61K38/46 ,  A61K48/00 ,  A61P3/08 ,  A61P43/00 ,  C07K14/075 ,  C07K14/75 ,  C12N5/10 ,  C12N7/00 ,  C12N7/02 ,  C12N7/04 ,  C12N15/86 ,  C12N15/861 ,  C12R1/92

CPC classification number: C12N15/86 ,  A61K48/00 ,  C07K14/005 ,  C12N2710/10322 ,  C12N2710/10343 ,  C12N2800/30 ,  C12N2830/008 ,  C12N2830/42 ,  C12N2840/20

Abstract: The present invention provides deleted adenovirus vectors. The inventive adenovirus vectors carry one or more deletions in the IVa2, 100 K, polymerase and/or preterminal protein sequences of the adenovirus genome. The adenoviruses may additionally contain other deletions, mutations or other modifications as well. In particular preferred embodiments, the adenovirus genome is multiply deleted, i.e., carries two or more deletions therein. The deleted adenoviruses of the invention are “propagation-defective” in that the virus cannot replicate and produce new virions in the absence of complementing function(s). Preferred adenovirus vectors of the invention carry a heterologous nucleotide sequence encoding a protein or peptide associated with a metabolic disorder, more preferably a protein or peptide associated with a lysosomal or glycogen storage disease, most preferably, a lysosomal acid α-glucosidase. Further provided are methods for producing the inventive deleted adenovirus vectors. Further provided are methods of administering the deleted adenovirus vectors to a cell in vitro or in vivo.

Abstract translation: 本发明提供了缺失的腺病毒载体。 本发明的腺病毒载体在腺病毒基因组的IVa2,100K，聚合酶和/或早产蛋白质序列中携带一个或多个缺失。 腺病毒还可以另外含有其它缺失，突变或其它修饰。 在特别优选的实施方案中，腺病毒基因组被多次缺失，即在其中携带两个或多个缺失。 本发明的缺失的腺病毒是“繁殖缺陷型”，因为在没有补体功能的情况下病毒不能复制并产生新的病毒粒子。 优选的本发明的腺病毒载体携带编码与代谢紊乱相关的蛋白质或肽的异源核苷酸序列，更优选与溶酶体或糖原贮积病相关的蛋白质或肽，最优选溶酶体酸性α-葡糖苷酶。 还提供了用于产生本发明缺失的腺病毒载体的方法。 还提供了在体外或体内将缺失的腺病毒载体施用于细胞的方法。

公开(公告)号：US06063622A

公开(公告)日：2000-05-16

申请号：US244752

申请日：1999-02-05

Applicant: Jeffrey S. Chamberlain ,  Andrea Amalfitano

Inventor： Jeffrey S. Chamberlain ,  Andrea Amalfitano

IPC: A61K48/00 ,  C07K14/47 ,  C12N5/08 ,  C12N7/01 ,  C12N15/861 ,  C12N5/10 ,  C12N15/86

CPC classification number: C12N15/86 ,  C07K14/4708 ,  C12N7/00 ,  A61K48/00 ,  C12N2710/10343 ,  C12N2800/30 ,  C12N2830/008 ,  C12N2830/30

Abstract: The present invention provides improved adenovirus vectors and packaging cell lines. One type of improved adenoviral vector comprises deletions within the E2b region of the adenoviral genome. These E2b-deleted virus are used in conjunction with novel cell lines that constitutively express E2b gene products. The present invention further provides adenoviral vectors deleted for all viral coding regions. These "gutted" vectors permit the transfer of large genes to cells as demonstrated herein by the transfer of the dystrophin gene to the muscle of mice. The E2b-deleted vectors and the gutted vectors provide improved adenoviral vectors useful for a wide variety of gene therapy applications.

公开(公告)号：US08003327B2

公开(公告)日：2011-08-23

申请号：US12357750

申请日：2009-01-22

Applicant: Rose-Mary N. Boustany ,  Wei-Xing Guo ,  Andrea Amalfitano

Inventor： Rose-Mary N. Boustany ,  Wei-Xing Guo ,  Andrea Amalfitano

IPC: C07H21/04 ,  C12Q1/68

CPC classification number: C12Q1/6886 ,  C12Q2600/118 ,  C12Q2600/136 ,  G01N2800/52

Abstract: A method of screening a subject for a proliferative disease risk factor comprises detecting the presence or absence of upregulation of the CLN3 gene in the subject. The upregulation of the CLN3 gene in the subject indicates the subject is at increased risk of developing a proliferative disease. Methods of screening compounds for the treatment of proliferative diseases based on the CLN3 gene and its product are also disclosed, along with methods of treating such diseases and vectors useful therefore.

Abstract translation: 筛选受试者增殖性疾病危险因素的方法包括检测受试者中CLN3基因上调的存在或不存在。 受试者中CLN3基因的上调表明受试者处于发展增殖性疾病的风险增加。 还公开了用于治疗基于CLN3基因及其产物的增殖性疾病的化合物的方法，以及用于治疗这样的疾病和载体的方法。

公开(公告)号：US20070054302A1

公开(公告)日：2007-03-08

申请号：US11524618

申请日：2006-09-21

Applicant: Rose-Mary Boustany ,  Wei-Xing Guo ,  Andrea Amalfitano

Inventor： Rose-Mary Boustany ,  Wei-Xing Guo ,  Andrea Amalfitano

IPC: C12Q1/68

CPC classification number: C12Q1/6886 ,  C12Q2600/118 ,  C12Q2600/136 ,  G01N2800/52

Abstract: A method of screening a subject for a proliferative disease risk factor comprises detecting the presence or absence of upregulation of the CLN3 gene in the subject. The upregulation of the CLN3 gene in the subject indicates the subject is at increased risk of developing a proliferative disease. Methods of screening compounds for the treatment of proliferative diseases based on the CLN3 gene and its product are also disclosed, along with methods of treating such diseases and vectors useful therefore.

Abstract translation: 筛选受试者增殖性疾病危险因素的方法包括检测受试者中CLN3基因上调的存在或不存在。 受试者中CLN3基因的上调表明受试者处于发展增殖性疾病的风险增加。 还公开了用于治疗基于CLN3基因及其产物的增殖性疾病的化合物的方法，以及用于治疗这样的疾病和载体的方法。

公开(公告)号：US20050220766A1

公开(公告)日：2005-10-06

申请号：US10511980

申请日：2003-04-30

Applicant: Andrea Amalfitano ,  Dwight Koeberl ,  Baodong Sun

Inventor： Andrea Amalfitano ,  Dwight Koeberl ,  Baodong Sun

IPC: A61K38/47 ,  A61K38/55 ,  A61K48/00 ,  C12N7/00 ,  C12N9/26 ,  C12N15/861 ,  C12N15/864

CPC classification number: C12N15/86 ,  A61K38/47 ,  A61K48/00 ,  A61K48/0066 ,  A61K48/0075 ,  A61K48/0083 ,  A61K2039/5256 ,  C12N9/2408 ,  C12N9/2411 ,  C12N2710/10344 ,  C12N2750/14143 ,  C12Y302/0102

Abstract: A recombinant hybrid virus, including: (a) a deleted adenovirus vector genome comprising the adenovirus 5′ and 3′ cis-elements for viral replication and encapsidation, and further comprising a deletion in an adenovirus genomic region selected from the group consisting of: (i) the polymerase region, wherein said deletion essentially prevents the expression of a functional polymerase protein from said deleted region and said hybrid virus does not otherwise express a functional polymerase protein, (ii) the preterminal protein region, wherein said deletion essentially prevents the expression of a functional preterminal protein from said deleted region, and said hybrid virus does not otherwise express a functional preterminal protein, and (iii) both the regions of (i) and (ii); and (b) a recombinant adeno-associated virus (AAV) vector genome flanked by the adenovirus vector genome sequences of (a), said recombinant AAV vector genome comprising (i) AAV 5′ and 3′ inverted terminal repeats, (ii) an AAV packaging sequence, and (iii) a heterologous nucleic acid sequence, wherein said heterologous nucleic acid sequence is flanked by the 5′ and the 3′ AAV inverted terminal repeats of (i). Methods of making and using the recombinant hybrid virus are also disclosed.

Abstract translation: 一种重组杂合病毒，其包括：（a）缺失的腺病毒载体基因组，其包含用于病毒复制和壳化的腺病毒5'和3'顺式元件，并且还包含选自以下的腺病毒基因组区域中的缺失：（ i）聚合酶区域，其中所述缺失基本上防止来自所述缺失区的功能性聚合酶蛋白的表达，并且所述杂合病毒不另外表达功能性聚合酶蛋白，（ii）早产蛋白区，其中所述缺失基本上防止表达 的来自所述缺失区的功能性早产蛋白，并且所述杂交病毒不另外表达功能性早产蛋白，和（iii）（i）和（ii）的两个区域; 所述重组腺病毒载体基因组包含（i）AAV 5'和3'反向末端重复序列，（ii）一个或多个重组腺病毒载体基因组（AAV） AAV包装序列，和（iii）异源核酸序列，其中所述异源核酸序列侧翼为（i）的5'和3'AAV反向末端重复序列。 还公开了制备和使用重组杂交病毒的方法。