公开(公告)号：US20100158890A1

公开(公告)日：2010-06-24

申请号：US12282695

申请日：2007-03-15

申请人： Kenichi Tanaka ,  Enrico Di Cera ,  Andras Gruber ,  Stephen Raymond Hanson

发明人： Kenichi Tanaka ,  Enrico Di Cera ,  Andras Gruber ,  Stephen Raymond Hanson

IPC分类号： A61K38/48 ,  G01N33/53 ,  A61P7/04

CPC分类号： A61K38/4833 ,  G01N33/86 ,  G01N2333/7452

摘要： The present invention provides methods for inhibiting the anticoagulation effect of a thrombin inhibitor in a patient in need thereof comprising administration of a therapeutically effective amount of a variant prothrombin or thrombin that is capable of binding the thrombin inhibitor and that has reduced procoagulant activity. Variant prothrombins or thrombins of use in the methods of the present invention include thrombin mutants W215A, W215A/E217A, or variants thereof in which the amino acids at positions 215 and/or 217 are alanine. Methods are also provided in which the thrombin mutants are administered with an additional active agent, particularly hemostatic agents such as activated factor VII or activated prothrombin complex concentrate. In one embodiment of the invention, the methods are useful in the treatment of patients in which a direct thrombin inhibitor has been administered, particularly argatroban. The present invention further provides a method for quantifying the concentration of an anticoagulant in the plasma or whole blood of a patient using a variant prothrombin or thrombin titration assay.

摘要翻译： 本发明提供了抑制凝血酶抑制剂在有需要的患者中的抗凝效果的方法，其包括施用治疗有效量的能够结合凝血酶抑制剂并降低促凝血活性的变体凝血酶原或凝血酶。 在本发明的方法中使用的变体前血栓素或血栓素包括其中215位和/或217位的氨基酸为丙氨酸的凝血酶突变体W215A，W215A / E217A或其变体。 还提供了其中凝血酶突变体与另外的活性剂，特别是止血剂如活化因子VII或活化的凝血酶原复合物浓缩物一起施用的方法。 在本发明的一个实施方案中，所述方法可用于治疗已经施用直接凝血酶抑制剂的患者，特别是阿加曲班。 本发明还提供了使用变体凝血酶原或凝血酶滴定测定来定量患者血浆或全血中抗凝血剂浓度的方法。

公开(公告)号：US08940297B2

公开(公告)日：2015-01-27

申请号：US13331109

申请日：2011-12-20

申请人： Enrico Di Cera ,  Andras Gruber ,  Prafull Gandhi ,  Leslie Pelc ,  Nicola Pozzi ,  David Collier Wood

发明人： Enrico Di Cera ,  Andras Gruber ,  Prafull Gandhi ,  Leslie Pelc ,  Nicola Pozzi ,  David Collier Wood

IPC分类号： C12N9/74 ,  A61K38/48

CPC分类号： C12N9/6429 ,  A61K38/4833 ,  C12Y304/21005 ,  Y02A50/473

摘要： One aspect of the invention contemplates a mutant E-WE thrombin precursor that contains the SEQ ID NO:1 amino acid residue sequence. Another aspect contemplates a thrombin precursor that contains the amino acid residue sequence Asp/Glu-Gly-Arg at positions 325, 326 and 327 based on the preprothrombin sequence. A third aspect contemplates a thrombin precursor that contains the SEQ ID NO:1 amino acid residue sequence as well as the amino acid residue sequence Asp/Glu Gly Arg at positions 325, 326 and 327 based on the preprothrombin sequence. Also contemplated is a composition that contains an effective amount of mutant thrombin dissolved or dispersed in a pharmaceutically acceptable carrier. A method is also disclosed for enhancing treating and preventing thrombosis in a mammal in need using that composition.

摘要翻译： 本发明的一个方面考虑了含有SEQ ID NO：1氨基酸残基序列的突变E-WE凝血酶前体。 另一方面考虑了基于前凝血酶原序列在325,326和327位含有氨基酸残基序列Asp / Glu-Gly-Arg的凝血酶前体。 第三方面考虑了基于前凝血酶原序列的含有SEQ ID NO：1氨基酸残基序列以及位置325,326和327的氨基酸残基序列Asp / Glu Gly Arg的凝血酶前体。 还考虑了包含溶解或分散在药学上可接受的载体中的有效量的突变凝血酶的组合物。 还公开了一种用于增强使用该组合物治疗和预防需要的哺乳动物血栓形成的方法。

公开(公告)号：US20120164129A1

公开(公告)日：2012-06-28

申请号：US13331109

申请日：2011-12-20

申请人： Enrico Di Cera ,  Andras Gruber ,  Prafull Gandhi ,  Leslie Pelc ,  Nicola Pozzi ,  David Collier Wood

发明人： Enrico Di Cera ,  Andras Gruber ,  Prafull Gandhi ,  Leslie Pelc ,  Nicola Pozzi ,  David Collier Wood

IPC分类号： A61K38/48 ,  C12N9/74 ,  A61P7/02

CPC分类号： C12N9/6429 ,  A61K38/4833 ,  C12Y304/21005 ,  Y02A50/473

摘要： One aspect of the invention contemplates a mutant E-WE thrombin precursor that contains the SEQ ID NO:1 amino acid residue sequence. Another aspect contemplates a thrombin precursor that contains the amino acid residue sequence Asp/Glu-Gly-Arg at positions 325, 326 and 327 based on the preprothrombin sequence. A third aspect contemplates a thrombin precursor that contains the SEQ ID NO:1 amino acid residue sequence as well as the amino acid residue sequence Asp/Glu Gly Arg at positions 325, 326 and 327 based on the preprothrombin sequence. Also contemplated is a composition that contains an effective amount of mutant thrombin dissolved or dispersed in a pharmaceutically acceptable carrier. A method is also disclosed for enhancing treating and preventing thrombosis in a mammal in need using that composition.

摘要翻译： 本发明的一个方面考虑了含有SEQ ID NO：1氨基酸残基序列的突变E-WE凝血酶前体。 另一方面考虑了基于前凝血酶原序列在325,326和327位含有氨基酸残基序列Asp / Glu-Gly-Arg的凝血酶前体。 第三方面考虑了基于前凝血酶原序列的含有SEQ ID NO：1氨基酸残基序列以及位置325,326和327的氨基酸残基序列Asp / Glu Gly Arg的凝血酶前体。 还考虑了包含溶解或分散在药学上可接受的载体中的有效量的突变凝血酶的组合物。 还公开了一种用于增强使用该组合物治疗和预防需要的哺乳动物血栓形成的方法。

公开(公告)号：US20130309753A1

公开(公告)日：2013-11-21

申请号：US13473566

申请日：2012-05-16

申请人： Nicola Pozzi ,  Enrico Di Cera ,  Sergio Barranco-Medina

发明人： Nicola Pozzi ,  Enrico Di Cera ,  Sergio Barranco-Medina

IPC分类号： C12N9/50

CPC分类号： C12N9/6429 ,  C12N9/6408 ,  C12Y304/21005

摘要： A recombinant serine protease precursor that auto-activates in an aqueous buffer to form a mature active enzyme is disclosed. A contemplated precursor contains 1 to about 10 heterologous amino acid residues that function to enhance by at least ten-fold the room temperature rate of auto-lytic bond cleavage to form the active enzyme relative to the auto-lytic cleavage rate of the native enzyme precursor when each precursor is dispersed in an aqueous buffer at an optimal pH value for the proteolytic activity of the protease. Illustrative active enzymes include serine proteases such as thrombin and protein C. A method of preparing and using an enzyme precursor is also disclosed.

摘要翻译： 公开了一种在水性缓冲液中自动激活以形成成熟活性酶的重组丝氨酸蛋白酶前体。 预期的前体含有1至约10个异源氨基酸残基，其功能相对于天然酶前体的自溶裂解速率增强自溶裂键切割的室温速率至少10倍以形成活性酶 当每种前体分散在蛋白酶的蛋白水解活性的最佳pH值的水性缓冲液中时。 说明性的活性酶包括丝氨酸蛋白酶如凝血酶和蛋白C.还公开了制备和使用酶前体的方法。