Dublin City University, School of Biotechnology, Glasnevin, Dublin 9, Ireland.
Expert Opin Ther Pat. 2011 Jun;21(6):967-70. doi: 10.1517/13543776.2011.577070. Epub 2011 Apr 9.
The application (WO2005001025) details recombinant fusion proteins attached to the constant region of heavy chains of immunoglobulins. They are found to be particularly useful for the treatment of hemostatic disorders, such as hemophilia B. It aims at engineering chimeric proteins comprising of a single molecule of human factor IX (FIX) and the constant region (Fc domain) of one or two heavy chain(s) of human IgG (rFIXFc). cDNA for rFIXFc was generated by a PCR. rFIXFc protein was isolated and purified from stably transfected mammalian cells. The concentration and clotting activity of rFIXFc were assessed in mice, rats, monkeys, and FIX-deficient mice and dogs, after intravenous administration. The half-life of rFIXFc activity is prolonged by three to fourfold, compared with rFIX, when administered intravenously in all animals. The generation of chimeric proteins, comprised of FIX fused to the Fc domain of IgG, extends the clotting activity of the recombinant molecule. rFIXFc represents a promising candidate for the treatment of patients with hemophilia B. The application claims the methods of making recombinant chimeric proteins comprising of one biologically active molecule fused to the Fc region of the heavy chain(s) of immunoglobulins and their use for therapy.
该申请（WO2005001025）详细介绍了附着在免疫球蛋白重链恒定区的重组融合蛋白。它们被发现特别适用于治疗止血紊乱，如乙型血友病。其目的是设计嵌合蛋白，包括单个分子的人凝血因子 IX（FIX）和一个或两个重链（s）的人 IgG 的恒定区（Fc 结构域）（rFIXFc）。通过 PCR 生成 rFIXFc 的 cDNA。rFIXFc 蛋白从稳定转染的哺乳动物细胞中分离和纯化。在静脉给药后，在小鼠、大鼠、猴子和 FIX 缺乏的小鼠和狗中评估 rFIXFc 的浓度和凝血活性。与静脉内给药时的 rFIX 相比，rFIXFc 的活性半衰期延长了三到四倍。由 FIX 融合到 IgG 的 Fc 结构域组成的嵌合蛋白的产生延长了重组分子的凝血活性。rFIXFc 是治疗乙型血友病患者的有前途的候选药物。该申请要求提供了一种制备包含一个生物活性分子融合到免疫球蛋白重链的 Fc 区域的重组嵌合蛋白的方法及其在治疗中的用途。
