公开(公告)号：US20060182733A1

公开(公告)日：2006-08-17

申请号：US11402388

申请日：2006-04-12

申请人： Fritz Bach ,  Simon Robson ,  Adrien Beaudoin ,  Jean Sevigny

发明人： Fritz Bach ,  Simon Robson ,  Adrien Beaudoin ,  Jean Sevigny

IPC分类号： A61K38/46

CPC分类号： C12N15/8509 ,  A01K2217/05 ,  A01K2217/20 ,  A01K2227/10 ,  A01K2227/108 ,  A01K2267/025 ,  A01K2267/03 ,  A01K2267/0368 ,  A61K35/44 ,  A61K38/00 ,  A61K48/00 ,  A61L33/0047 ,  C07K14/705 ,  C12N9/14 ,  C12Y306/01005

摘要： A method to render endothelial cells capable of inhibiting platelet and leukocyte-mediated injury and inflammation is described, comprising genetically modifying the cells by inserting DNA encoding ecto-ATP diphosphohydrolase or an oxidation-resistant analog thereof, and expressing a protein having functional ecto-ATP diphosphohydrolase activity, such as the human CD39 protein, by said cells under cellular activating conditions. The method, which can be carried out in vivo, ex vivo or in vitro, has use in allogeneic or xenogeneic transplantation as well as to treat systemic or local inflammatory conditions characterized by platelet aggregation leading to thrombus formation.

摘要翻译： 描述了一种能够抑制血小板和白细胞介导的损伤和炎症的内皮细胞的方法，包括通过插入编码ecto-ATP二磷酸水解酶或其抗氧化类似物的DNA，并且表达具有功能性ecto-ATP的蛋白质来遗传修饰细胞 在细胞活化条件下由所述细胞产生二磷酸水解酶活性，例如人CD39蛋白。 可以在体内，离体或体外进行的方法可用于同种异体或异种移植，以及治疗特征在血小板聚集导致血栓形成的全身或局部炎症状态。

公开(公告)号：US06800284B2

公开(公告)日：2004-10-05

申请号：US09781796

申请日：2001-02-12

申请人： Adrien R. Beaudoin ,  Jean Sevigny

发明人： Adrien R. Beaudoin ,  Jean Sevigny

IPC分类号： A61K3846

CPC分类号： C07K14/70596 ,  A61K38/00 ,  C12N9/14 ,  C12Y306/01005

摘要： The present invention relates to two ATP diphosphohydrolases (ATPDase enzymes) isolated from bovine aorta and pig pancreas, which enzymes have a molecular weight for their catalytic unit of about 78 and 54 Kilodaltons, respectively. A first process for obtaining a highly purified ATPDase is also an object of the present invention. This process has been successfully applied to the purification of both the pancreatic and the aorta enzymes and is deemed to work in the purification of any ATPDase. For both sources of enzymes, the process allows the specific activity of the enzyme to be increased by at least 10,000 fold when compared to the activity retrieved in the crude cell homogenates. The novel process involves an ion exchange chromatography step, a separation on an affinity column, followed by an electrophoresis under non-denaturing conditions. The two enzymes purified by this process (aortic and pancreatic) are glycosylated and, when deglycosylated, have molecular weights shifted to about 56 and 35 Kdaltons, respectively. Partial amino acid sequences have been obtained for each enzyme. The partial sequences appear highly homologous with a human lymphoid cell activation antigen named CD39. An antibody directed against the porcine pancreatic enzyme cross-reacts with a protein present in endothelial cell lines and in bovine aorta (78 KDa). The high degree of homology of the pancreatic and aortic enzymes with CD39 and their cross-reactivity are indications that both enzymes are related. The pancreatic enzyme completely lacks the first 200 amino acids of CD39, which means the ATPDase activity is comprised between residues 200 and 510 of CD39. Since this is the first time that a sequence is assigned to ATPDases, a second new process for producing ATPDases by recombinant technology can also be used. Therefore a second new process for producing an ATPDase using the CD39-encoding nucleic acid or part or variant thereof is also described.

公开(公告)号：US06287837B1

公开(公告)日：2001-09-11

申请号：US08930921

申请日：1998-01-02

申请人： Adrien R. Beaudoin ,  Jean Sevigny

发明人： Adrien R. Beaudoin ,  Jean Sevigny

IPC分类号： C12N1100

CPC分类号： C07K14/70596 ,  A61K38/00 ,  C12N9/14 ,  C12Y306/01005

摘要： The present invention relates to two ATP-diphosphohydrolases (ATPDase enzymes) isolated from bovine aorta and pig pancreas, which enzymes have a molecular weight for their catalytic unit of about 78 and 54 Kilodaltons, respectively. A first process for obtaining a highly purified ATPDase is also an object of the present invention. The process has been successfully applied in the purification of both the pancreatic and the aorta enzymes and is deemed to work in the purification of any ATPDase. For both sources of enzymes, the process allows the specific activity of the enzyme to be increased by at least 10,000 fold when compared to the activity retrieved in the crude cell homogenates. The novel process involves an ion exchange chromatography step, a separation on an affinity column, followed by an electrophoresis under non-denaturing conditions. The two enzymes purified by this process (aortic and pancreatic) are glycosylated and, when deglycosylated, have molecular weights shifted to about 56 and 35 Kdaltons, respectively. Partial amino acid sequences have been obtained for each enzyme. The partial sequences appear highly homologous with a human lymophoid cell activation antigen named CD39. An antibody directed against the porcine pancreatic enzyme cross-reacts with a protein present in endothelial cell lines and in bovine aorta (78 KDa). The high degree of homology of the pancreatic and aortic enzymes with CD39 and their cross-reactivity are indications that both enzymes are related. The pancreatic enzyme completely lacks the first 200 amino acids of CD39, which means the ATPDase activity is comprised between residues 200 and 510 of CD39. Since this is the first time that a sequence is assigned to ATPDase, a second new process for producing ATPDases by recombinant technology can also be used. Therefore, a second new process for producing an ATPDase using the CD39-encoding nucleic acid or part or variant thereof is also described.

公开(公告)号：US07129074B2

公开(公告)日：2006-10-31

申请号：US11076982

申请日：2005-03-11

申请人： Jean Sevigny

发明人： Jean Sevigny

IPC分类号： C12N9/14 ,  C12N1/20 ,  C12N15/00

CPC分类号： C12N9/14 ,  A61K38/00

摘要： The present invention relates to a new ectohydroxynucleotidase, namely the NTPDasse8, which allows to regulate platelet aggregation or activation involved in the formation of thrombosis and related diseases. The nucleic acid sequence and the corresponding amino acid sequence and uses thereof are described.

摘要翻译： 本发明涉及一种新的二羟基核苷酸酶，即NTPDasse8，其允许调节参与血栓形成和相关疾病形成的血小板聚集或活化。 描述了核酸序列及其相应的氨基酸序列及其用途。

公开(公告)号：US20050196798A1

公开(公告)日：2005-09-08

申请号：US11076982

申请日：2005-03-11

申请人： Jean Sevigny

发明人： Jean Sevigny

IPC分类号： A61K38/00 ,  A61K48/00 ,  C07H21/04 ,  C12N9/14 ,  C12N9/22 ,  C12N15/55 ,  C12Q1/68

CPC分类号： C12N9/14 ,  A61K38/00

摘要： The present invention relates to a new ectohydroxynucleotidase, namely the NTPDasse8, which allows to regulate platelet aggregation or activation involved in the formation of thrombosis and related diseases. The nucleic acid sequence and the corresponding amino acid sequence and uses thereof are described.

摘要翻译： 本发明涉及一种新的二羟基核苷酸酶，即NTPDasse8，其允许调节参与血栓形成和相关疾病形成的血小板聚集或活化。 描述了核酸序列及其相应的氨基酸序列及其用途。

公开(公告)号：US5798241A

公开(公告)日：1998-08-25

申请号：US777859

申请日：1996-12-31

申请人： Adrien R. Beaudoin ,  Jean Sevigny

发明人： Adrien R. Beaudoin ,  Jean Sevigny

IPC分类号： A61K38/00 ,  C07K14/705 ,  C12N9/14 ,  C12N11/00 ,  C12N9/96

CPC分类号： C07K14/70596 ,  C12N9/14 ,  C12Y306/01005 ,  A61K38/00

摘要： The present invention relates to two ATP diphosphohydrolases ATPDase enzymes isolated from bovine aorta and pig pancreas, which enzymes have a molecular weight for their catalytic unit of about 78 and 54 Kilodaltons, respectively. A novel process for obtaining a highly purified ATPDase is also an object of the present invention. This process has been successfully applied to the purification of both the pancreatic and the aorta enzymes and is deemed to work in the purification of any ATPDase. For both sources of enzymes, the process allows the specific activity of the enzyme to be increased by at least 10,000 fold when compared to the activity retrieved in the crude cell homogenates. The novel process involves an ion exchange chromatography step, a separation on an affinity column, followed by an electrophoresis under non-denaturing conditions. The two enzymes purified by this process (aortic and pancreatic) are glycosylated and, when deglycosylated, have molecular weights of about 56 and 35 Kdaltons, respectively.

摘要翻译： 本发明涉及从牛主动脉和猪胰腺分离的两种ATP二磷酸水解酶ATPDase酶，该酶分别具有约78和54千道尔顿的催化单位的分子量。 获得高度纯化的ATPDase的新方法也是本发明的目的。 该方法已成功应用于胰腺和主动脉酶的纯化，并被认为在纯化任何ATPDase中起作用。 对于两种酶的来源，当与粗细胞匀浆中回收的活性相比时，该过程允许酶的比活性增加至少10,000倍。 该新方法涉及离子交换层析步骤，在亲和柱上分离，然后在非变性条件下进行电泳。 通过该方法（主动脉和胰腺）纯化的两种酶被糖基化，并且当脱糖基化时，分别具有约56和35道尔顿的分子量。