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    • 2. 发明申请
    • PROCESS FOR PURIFYING FACTOR VIII
    • 净化因子八
    • WO1996015150A1
    • 1996-05-23
    • PCT/SE1995001351
    • 1995-11-14
    • PHARMACIA ABALMSTEDT, AnnelieSANDBERG, HelenaSMEDS, Anna-LisaWRANGEL, MariaÖSTLIN, Anna
    • PHARMACIA AB
    • C07K14/755
    • C07K14/755Y10S530/83Y10S530/831
    • Proteases generally tend to reduce the activity of coagulation factor VIII by degrading the molecule. The present invention relates to a process for reducing the detrimental influence of metal-dependent proteases upon recombinant factor VIII molecules, by adding an inhibitor of metal-dependent proteases to a solution after harvest. The solution is suitably the harvest solution or any aqueous solution being fed to or leaving the primary isolation, being the initial step of the purifying sequence. The inhibitor is selected from: i) complexing agents with a stronger affinity for the metal ion of the protease than for the ion or ions stabilizing the factor VIII molecule and ii) compounds structurally related to the natural substrate of the protease and containing an electronegative moiety. The presence of an inhibitor, preferably a complexing agent, after harvest allows for a prolonged harvest period and considerably higher yield with essentially retained factor VIII activity. The invention further relates to an aqueous solution containing recombinant factor VIII which has been purified according to the present process and use of such an aqueous solution, for the manufacture of a medicament for administration to a patient having the symptoms of hemophilia. Also, the invention relates to a method for treatment of hemophilia by administration of a therapeutically effective amount of recombinant factor VIII which has been purified according to the present process.
    • 蛋白酶通常倾向于通过降解分子来降低凝血因子VIII的活性。 本发明涉及通过在收获后向溶液中加入金属依赖性蛋白酶抑制剂来减少金属依赖性蛋白酶对重组因子VIII分子的不利影响的方法。 该溶液适当地是收集溶液或被加入或离开初级分离的任何水溶液,作为纯化序列的初始步骤。 抑制剂选自:i)对蛋白质的金属离子具有更强的亲和力的络合剂,而不是稳定因子VIII分子的离子或离子,和ii)结构上与蛋白酶天然底物相关并含有电负性部分的化合物 。 在收获之后抑制剂,优选络合剂的存在允许延长的收获期和相当高的产率,基本保留因子VIII活性。 本发明还涉及含有根据本发明方法纯化的重组因子VIII的水溶液和这种水溶液的用途,用于制备用于给患有血友病症状的患者施用的药物。 此外,本发明还涉及通过给予治疗有效量的根据本方法纯化的重组因子VIII治疗血友病的方法。