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    • 4. 发明授权
    • Bivalent thrombin inhibitors
    • 二价凝血酶抑制剂
    • US6127337A
    • 2000-10-03
    • US636698
    • 1996-06-28
    • Yasuo KonishiZbigniew SzewczukYuko Tsuda
    • Yasuo KonishiZbigniew SzewczukYuko Tsuda
    • A61K38/00C07K14/815A61K38/03C07K7/00
    • C07K14/815A61K38/00
    • Hirudin is the most potent and specific thrombin inhibitor and is derived from the medicinal leech. It is reported to inhibit thrombin with an equilibrium dissociation constant (K.sub.i) value of 2.2.times.10.sup.-14 M. synthetic thrombin inhibitors have been designed based on the hirudin sequence but with a dramatically reduced size. The bulky active site inhibitor segment, hirudin.sup.1-48, has been substituted by small non-substrate type active site inhibitors of thrombin, e.g., dansyl-Arg-(D-pipecolic acid). The linker segment has also been modified using a combination of .omega.-amino acids to reduce the molecular weight but retaining sufficient length to span the two principal binding domains. Among the inhibitors designed, dansyl-Arg-(D-pipecolic acid)-(12-aminododecanoic acid)-4-aminobutyric acid)-Asp-Tyr-Glu-Pro-Ile-Pro-Glu-Glu-Ala-(L-.beta.-cyclohexylalamine)-(D-Glu)-OH (SEQ ID NO:45) showed the highest affinity and displays a competitive-type inhibition. The incorporation of the non-substrate type active site inhibitor segment and the linker of .omega.-amino acids into the bivalent thrombin inhibitors not only improved in vitro thrombin inhibitory activity to the pM level, overcame proteolytic susceptibility at the level of the "normal" scissile bond and confered high in vivo activity.
    • PCT No.PCT / CA94 / 00585 Sec。 371日期:1996年6月28日 102(e)日期1996年6月28日PCT 1994年10月25日PCT PCT。 第WO95 / 11921号公报 日期1995年5月4日水蛭素是最有效和特异性的凝血酶抑制剂,来源于药用水蛭。 据报道,以平均解离常数(Ki)值为2.2×10-14 M抑制凝血酶。已经基于水蛭素序列设计合成的凝血酶抑制剂,但其显着减小的大小。 大体积的活性位点抑制物片段,水蛭素1-48已经被小的非底物型的凝血酶活性位点抑制剂例如丹磺酰-Arg-(D-哌啶酸)所取代。 接头片段也已经使用ω-氨基酸的组合进行了修饰,以降低分子量,但保留足够的长度来跨越两个主要的结合结构域。 在设计的抑制剂中,丹磺酰-Arg-(D-哌啶酸) - (12-氨基十二烷酸)-4-氨基丁酸)-Asp-Tyr-Glu-Pro-Ile-Pro-Glu-Glu-Ala-(L- β-环己胺) - (D-Glu)-OH(SEQ ID NO:45)显示出最高的亲和力并显示出竞争性抑制。 将非底物型活性位点抑制物区段和ω-氨基酸接头加入到二价凝血酶抑制剂中不仅提高了对pM水平的体外凝血酶抑制活性,克服了在“正常”剪切水平下的蛋白水解敏感性 结合并提出高体内活性。