公开(公告)号：US20090203077A1

公开(公告)日：2009-08-13

申请号：US12252024

申请日：2008-10-15

申请人： Randal J. Kaufman ,  Steven W. Pipe ,  Michael Griffith

发明人： Randal J. Kaufman ,  Steven W. Pipe ,  Michael Griffith

IPC分类号： C12P21/02 ,  C12N15/12

CPC分类号： C07K14/755

摘要： Provided herein are methods and compositions for producing Factor VIII proteins. Such methods include introducing into a cell a nucleic acid molecule encoding a Factor VIII protein operably linked to a promoter, wherein the promoter is characterized by the ability to produce commercially viable Factor VIII protein; and incubating the cell under conditions for producing commercially viable Factor VIII protein. Also provided are nucleic acid molecules which encode a Factor VIII protein operably linked to a Chinese hamster elongation factor 1-α (CHEF1) promoter, which may be used in the methods provided herein.

摘要翻译： 本文提供了用于产生因子VIII蛋白质的方法和组合物。 这样的方法包括向细胞中引入编码与启动子可操作连接的因子VIII蛋白的核酸分子，其中启动子的特征在于产生商业上可行的因子VIII蛋白的能力; 并在用于生产商业上可行的因子VIII蛋白的条件下孵育细胞。 还提供了编码与中国仓鼠延长因子1-α（CHEF1）启动子可操作地连接的因子VIII蛋白的核酸分子，其可用于本文提供的方法中。

公开(公告)号：US08183344B2

公开(公告)日：2012-05-22

申请号：US11455116

申请日：2006-06-16

申请人： Randal J. Kaufman ,  Steven W. Pipe

发明人： Randal J. Kaufman ,  Steven W. Pipe

IPC分类号： A61K38/37 ,  C07K14/755

CPC分类号： C07K14/755 ,  A61K38/00 ,  A61K38/37

摘要： The present invention provides novel purified and isolated nucleic acid sequences encoding procoagulant-active FVIII proteins. The nucleic acid sequences of the present invention encode amino acid sequences corresponding to known human FVIII sequences, wherein residue Phe309 is mutated. The nucleic acid sequences of the present invention also encode amino acid sequences corresponding to known human FVIII sequences, wherein the APC cleavage sites, Arg336 and Ile562, are mutated. The nucleic acid sequences of the present invention further encode amino acid sequences corresponding to known human FVIII sequences, wherein the B-domain is deleted, the von Willebrand factor binding site is deleted, a thrombin cleavage site is mutated, an amino acid sequence spacer is inserted between the A2- and A3-domains. Methods of producing the FVIII proteins of the invention, nucleotide sequences encoding such proteins, pharmaceutical compositions containing the nucleotide sequences or proteins, as well as methods of treating patients suffering from hemophilia, are also provided.

摘要翻译： 本发明提供了编码促凝血活性FVIII蛋白的新型纯化和分离的核酸序列。 本发明的核酸序列编码对应于已知人FVIII序列的氨基酸序列，其中残基Phe309被突变。 本发明的核酸序列还编码对应于已知人FVIII序列的氨基酸序列，其中APC切割位点Arg336和Ile562突变。 本发明的核酸序列进一步编码对应于已知人FVIII序列的氨基酸序列，其中缺失B结构域，缺失血管性血友病因子结合位点，突变凝血酶切割位点，氨基酸序列间隔物 插入在A2-和A3-域之间。 还提供了产生本发明的FVIII蛋白的方法，编码这种蛋白质的核苷酸序列，含有核苷酸序列或蛋白质的药物组合物，以及治疗患有血友病的患者的方法。

公开(公告)号：US06838437B2

公开(公告)日：2005-01-04

申请号：US09819098

申请日：2001-04-11

申请人： Randal J. Kaufman ,  Steven W. Pipe ,  Kagehiro Amano

发明人： Randal J. Kaufman ,  Steven W. Pipe ,  Kagehiro Amano

IPC分类号： C12N15/09 ,  A61K31/00 ,  A61K38/00 ,  A61K38/43 ,  A61K48/00 ,  A61P7/00 ,  A61P7/04 ,  C07K14/755 ,  C12N1/21 ,  C12P21/08

CPC分类号： C07K14/755 ,  A61K38/00

摘要： The present invention provides novel purified and isolated nucleic acid sequences encoding procoagulant-active FVIII proteins. The nucleic acid sequences of the present invention encode amino acid sequences corresponding to known human FVIII sequences, wherein residue Phe3O9 is mutated. The nucleic acid sequences of the present invention also encode amino acid sequences corresponding to known human FVIII sequences, wherein the APC cleavage sites, Arg336 and Ile562, are mutated. The nucleic acid sequences of the present invention further encode amino acid sequences corresponding to known human FVIII sequences, wherein the B-domain is deleted, the von Willebrand factor binding site is deleted, a thrombin cleavage site is mutated and an amino acid sequence spacer is inserted between the A2- and A3-domains. Methods of producing the FVIII proteins of the invention, nucleotide sequences encoding such proteins, pharmaceutical compositions containing the nucleotide sequences or proteins, as well as methods of treating patients suffering from hemophilia, are also provided.

摘要翻译： 本发明提供了编码促凝血活性FVIII蛋白的新型纯化和分离的核酸序列。 本发明的核酸序列编码对应于已知人FVIII序列的氨基酸序列，其中残基Phe3O9被突变。 本发明的核酸序列还编码对应于已知人FVIII序列的氨基酸序列，其中APC切割位点Arg336和Ile562突变。 本发明的核酸序列进一步编码对应于已知人FVIII序列的氨基酸序列，其中缺失B结构域，缺失血管性血友病因子结合位点，突变凝血酶切割位点，氨基酸序列间隔物为 插入在A2-和A3-域之间。 还提供了产生本发明的FVIII蛋白的方法，编码这种蛋白质的核苷酸序列，含有核苷酸序列或蛋白质的药物组合物，以及治疗患有血友病的患者的方法。

公开(公告)号：US07459534B2

公开(公告)日：2008-12-02

申请号：US10974534

申请日：2004-10-26

申请人： Randal J. Kaufman ,  Steven W. Pipe ,  Kagehiro Amano

发明人： Randal J. Kaufman ,  Steven W. Pipe ,  Kagehiro Amano

IPC分类号： A61K38/00 ,  C07K14/00

CPC分类号： C07K14/755 ,  A61K38/00

摘要： The present invention provides novel purified and isolated nucleic acid sequences encoding procoagulant-active FVIII proteins. The nucleic acid sequences of the present invention encode amino acid sequences corresponding to known human FVIII sequences, wherein residue Phe309 is mutated. The nucleic acid sequences of the present invention also encode amino acid sequences corresponding to known human FVIII sequences, wherein the APC cleavage sites, Arg336 and Ile562, are mutated. The nucleic acid sequences of the present invention further encode amino acid sequences corresponding to known human FVIII sequences, wherein the B-domain is deleted, the von Willebrand factor binding site is deleted, a thrombin cleavage site is mutated and an amino acid sequence spacer is inserted between the A2- and A3-domains. Methods of producing the FVIII proteins of the invention, nucleotide sequences encoding such proteins, pharmaceutical compositions containing the nucleotide sequences or proteins, as well as methods of treating patients suffering from hemophilia, are also provided.

摘要翻译： 本发明提供了编码促凝血活性FVIII蛋白的新型纯化和分离的核酸序列。 本发明的核酸序列编码对应于已知人FVIII序列的氨基酸序列，其中残基Phe309被突变。 本发明的核酸序列还编码对应于已知人FVIII序列的氨基酸序列，其中APC切割位点Arg336和Ile562突变。 本发明的核酸序列进一步编码对应于已知人FVIII序列的氨基酸序列，其中缺失B结构域，缺失血管性血友病因子结合位点，突变凝血酶切割位点，氨基酸序列间隔物为 插入在A2-和A3-域之间。 还提供了产生本发明的FVIII蛋白的方法，编码这种蛋白质的核苷酸序列，含有核苷酸序列或蛋白质的药物组合物，以及治疗患有血友病的患者的方法。

公开(公告)号：US20130072434A1

公开(公告)日：2013-03-21

申请号：US13542010

申请日：2012-07-05

申请人： Randal J. Kaufman ,  Steven W. Pipe

发明人： Randal J. Kaufman ,  Steven W. Pipe

IPC分类号： C07K14/755 ,  A61K38/37

CPC分类号： C07K14/755 ,  A61K38/00 ,  A61K38/37

摘要： The present invention provides novel purified and isolated nucleic acid sequences encoding procoagulant-active FVIII proteins. The nucleic acid sequences may encode amino acid sequences corresponding to known human FVIII sequences, wherein residue Phe309 is mutated. The nucleic acid sequences also may encode amino acid sequences corresponding to known human FVIII sequences, wherein the APC cleavage sites, Arg336 and Ile562, are mutated. The nucleic acid sequences of the present invention further encode amino acid sequences corresponding to known human FVIII sequences, wherein the B-domain is deleted, the von Willebrand factor binding site is deleted, a thrombin cleavage site is mutated, an amino acid sequence spacer is inserted between the A2- and A3-domains. Provided herein are methods of producing the FVIII proteins of the invention, nucleotide sequences encoding such proteins, pharmaceutical compositions containing the nucleotide sequences or proteins, as well as methods of treating patients suffering from hemophilia.

摘要翻译： 本发明提供了编码促凝血活性FVIII蛋白的新型纯化和分离的核酸序列。 核酸序列可以编码对应于已知人FVIII序列的氨基酸序列，其中残基Phe309被突变。 核酸序列还可以编码对应于已知人FVIII序列的氨基酸序列，其中APC切割位点Arg336和Ile562被突变。 本发明的核酸序列进一步编码对应于已知人FVIII序列的氨基酸序列，其中缺失B结构域，缺失血管性血友病因子结合位点，突变凝血酶切割位点，氨基酸序列间隔物 插入在A2-和A3-域之间。 本文提供了制备本发明的FVIII蛋白质，编码这种蛋白质的核苷酸序列，含有核苷酸序列或蛋白质的药物组合物以及治疗患有血友病患者的方法。

公开(公告)号：US20120190623A1

公开(公告)日：2012-07-26

申请号：US13015043

申请日：2011-01-27

申请人： Randal J. Kaufman ,  Steven W. Pipe

发明人： Randal J. Kaufman ,  Steven W. Pipe

IPC分类号： A61K38/37 ,  A61K31/7088 ,  C12N5/10 ,  A61P7/04 ,  C12N15/63 ,  C07H21/04 ,  C07K14/755

CPC分类号： C07K14/755 ,  A61K38/00 ,  A61K38/37

摘要： The present invention provides novel purified and isolated nucleic acid sequences encoding procoagulant-active FVIII proteins. The nucleic acid sequences of the present invention encode amino acid sequences corresponding to known human FVIII sequences, wherein residue Phe309 is mutated. The nucleic acid sequences of the present invention also encode amino acid sequences corresponding to known human FVIII sequences, wherein the APC cleavage sites, Arg336 and Ile562, are mutated. The nucleic acid sequences of the present invention further encode amino acid sequences corresponding to known human FVIII sequences, wherein the B-domain is deleted, the von Willebrand factor binding site is deleted, a thrombin cleavage site is mutated, an amino acid sequence spacer is inserted between the A2- and A3-domains. Methods of producing the FVIII proteins of the invention, nucleotide sequences encoding such proteins, pharmaceutical compositions containing the nucleotide sequences or proteins, as well as methods of treating patients suffering from hemophilia, are also provided.

摘要翻译： 本发明提供了编码促凝血活性FVIII蛋白的新型纯化和分离的核酸序列。 本发明的核酸序列编码对应于已知人FVIII序列的氨基酸序列，其中残基Phe309被突变。 本发明的核酸序列还编码对应于已知人FVIII序列的氨基酸序列，其中APC切割位点Arg336和Ile562突变。 本发明的核酸序列进一步编码对应于已知人FVIII序列的氨基酸序列，其中缺失B结构域，缺失血管性血友病因子结合位点，突变凝血酶切割位点，氨基酸序列间隔物 插入在A2-和A3-域之间。 还提供了产生本发明的FVIII蛋白的方法，编码这种蛋白质的核苷酸序列，含有核苷酸序列或蛋白质的药物组合物，以及治疗患有血友病的患者的方法。