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    • 8. 发明授权
    • Process for manufacture of a modified collagen-induced platelet
aggregation inhibitor pallidipin
    • 制备改性胶原诱导的血小板聚集抑制剂帕拉地平的方法
    • US5882887A
    • 1999-03-16
    • US793294
    • 1997-03-11
    • Christiane Noeske-JungblutAndreas BeckerBernard Haendler
    • Christiane Noeske-JungblutAndreas BeckerBernard Haendler
    • C12N15/09A61K38/00A61K38/17A61P7/02A61P9/10A61P35/00C07K14/435C12N1/21C12N15/12C12N15/30C12N15/62C12P21/00C12P21/02C12R1/19C07K14/44C07K14/475
    • C07K14/435A61K38/00C07K2319/02Y10S514/822
    • The invention provides a process of manufacture of a recombinant protein called Asp-Pallidipin. Asp-Pallidipin inhibits the collagen-induced platelet aggregation of mammalian platelets. The Asp-Pallidipin comprises(i) a protein (Pallidipin) selected from the group of Pallidipin proteins, and(ii) the amino acid aspartic acid,wherein the aspartic acid is connected by a peptide bond with the N-terminal end of Pallidipin. The process comprises the stepsaa) transfecting at least one bacterium with an appropriate vector, wherein the vector comprises:(i) a DNA or cDNA coding for the recombinant Asp-Pallidipin(ii) a suitable signal peptide sequence which signal sequence is cleaved so that the amino acid aspartic acid is in the position +1 of the amino acid sequence seen from the position of the Pallidipin and(iii) a suitable promoter;bb) expressing the preprotein comprising Asp-Pallidipin and the signal sequence;cc) transporting the Asp-Pallidipin from the cytoplasm of the bacterium to the periplasm,cleavage of the preprotein by at least one protease during the transport, producing the Asp-Pallidipin,dd) isolating the Asp-Pallidipin by extracting the periplasm, andee) purifying the Asp-Pallidipin.The protein is used as a medicament for inhibiting of collagen-induced human platelet aggregation or of cancer with metastatic tumor cells.
    • PCT No.PCT / EP95 / 03573 371日期:1997年6月16日 102(e)日期1997年6月16日PCT提交1995年9月11日PCT公布。 公开号WO96 / 08563 日期:1996年3月21日本发明提供了称为Asp-Pallidipin的重组蛋白质的制造方法。 Asp-Pallidipin抑制胶原诱导的哺乳动物血小板血小板聚集。 Asp-Pallidipin包含(i)选自Pallidipin蛋白质的蛋白质(Pallidipin),和(ii)氨基酸天冬氨酸,其中天冬氨酸通过肽键与Pallidipin的N-末端连接。 该方法包括步骤aa)用合适的载体转染至少一种细菌,其中载体包括:(i)编码重组Asp-Pallidipin的DNA或cDNA(ii)信号序列被切割的合适的信号肽序列 氨基酸天冬氨酸位于从Pallidipin的位置看到的氨基酸序列的+1位,和(iii)合适的启动子; bb)表达包含Asp-Pallidipin和信号序列的前蛋白; cc)将Asp-Pallidipin从细菌的细胞质转运到周质,在运输期间由至少一种蛋白酶切割前蛋白,产生Asp-Pallidipin,dd)通过提取周质来分离Asp-Pallidipin,ee )纯化Asp-Pallidipin。 该蛋白质用作抑制胶原诱导的人血小板聚集或转移性肿瘤细胞癌的药物。