一种溶栓酶及其制备方法和应用

摘要

本发明涉及一种新溶栓酶，使用溶栓酶已成为治疗心肌梗塞和脑血栓疾病的标准疗法。新溶栓酶来源于人类大脑，是经重组基因克隆所获得的新溶栓酶。该分子由大肠杆菌生产。新溶栓酶基因克隆于T7启动子和lac调控下表达。在DnaY的辅助下，新溶栓酶表达量为细胞总蛋白的35％以上。新溶栓酶具有组织型纤溶酶原激活剂tPA的优点，如血栓特异性的同时，在药理、药代和药效上优于tPA。新溶栓酶的半衰期为15至23分钟，而tPA只有3.5分钟。

主权项

1、一种溶栓酶，其特征在于所述的溶栓酶由395个氨基酸组成，其序列如下：Met Ala Ser Cys Arg Asp Glu Lys Thr Gln 10Met Ile Tyr Gln Gln His Gln Ser Trp Leu 20Arg Pro Val Leu Arg Ser Asn Arg Val Glu 30Tyr Csy Trp Cys Asn Ser Gly Arg Ala Gln 40Cys Ser Glu Gly Asn Ser Asp Cys Tyr Phe 50Gly Asn Gly Ser Ala Tyr Arg Gly Thr His 60Ser Leu Thr Glu Ser Gly Ala Ser Cys Leu 70Pro Trp Asn Ser Met Ile Leu Ile Gly Lys 80Val Tyr Thr Ala Gln Asn Pro Ser Ala Gln 90Ala Leu Gly Leu Gly Lys His Asn Tyr Cys 100Arg Asn Pro Asp Gly Asp Ala Lys Pro Trp 110Cys His Val Leu Lys Asp Arg Arg Leu Thr 120Trp Glu Tyr Cys Asp Val Pro Ser Cys Ser 130Thr Cys Gly Leu Arg Gln Tyr Ser Gln Pro 140Gln Phe Arg Ile Lys Gly Gly Leu Phe Ala 150Asp Ile Ala Ser His Pro Trp Gln Ala Ala 160Ile Phe Ala Lys His Glu Glu Ser Pro Gly 170Glu Arg Phe Leu Cys Gly Gly Ile Leu Ile 180Ser Ser Cys Trp Ile Leu Ser Ala Ala His 190Cys Phe Gln Glu Arg Phe Pro Pro His His 200Leu Thr Val Ile Leu Gly Leu The Tyr Arg 210Val Tyr Pro Gly Glu Glu Glu Gln Lys Phe 220Glu Val Glu Lys Tyr Ile Val His Lys Glu 230Phe Asp Asp Asp Thr Tyr Asp Asn Asp Ile 240Ala Leu Leu Gln Leu Lys Ser Asp Ser Ser 250Arg Cys Ala Gln Glu Ser Ser Val Val Arg 260Thr Val Cys Leu Pro Pro Ala Asp Leu Gln 270Leu Pro Asp Trp Thr Glu Cys Glu Leu Ser 280Gly Tyr Gly Lys His Glu Ala Leu Ser Pro 290Phe Tyr Ser Glu Arg Leu Lys Glu Ala His 300Val Arg Leu Tyr Pro Ser Ser Arg Cys Thr 310Ser Gln His Leu Leu Asn Arg Thr Val Thr 320Asp Asn Met Leu Cys Ala Gly Asp Thr Arg 330Ser Gly Gly Phe Gln Ala Asn Leu His Asp 340Ala Cys Gln Gly Asp Ser Gly Gly Pro Leu 350Val Cys Leu Asn Asp Gly Arg Met Thr Leu 360Val Gly Ile Ile Ser Trp Gly Leu Gly Cys 370Gly Gln Lys Asp Val Pro Gly Val Tyr Thr 380Lys Val Thr Asn Tyr Leu Asp Trp Ile Arg 390Asp Asn Met Arg Pro 395