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将通过蛋白质工程获得的抗凝溶栓双功能蛋白水蛭素12肽-瑞替普酶(HV12p-rPA)制备成靶向脂质体,并考察其体内溶栓效果。用薄膜分散-超声法制备HV12p-rPA脂质体,通过正交设计优化处方,采用碳二亚胺法偶联抗纤维蛋白原单克隆抗体(McAbSZ-65)制备HV12p-rPA靶向脂质体,采用大鼠颈总动脉血栓模型,考察HV12p-rPA靶向脂质体的体内溶栓效果。结果:最佳处方中HV12p-rPA脂质体的粒径为(142.45±1.20)nm,Zeta电位为(-30.63±0.48)mV,平均包封率为(91.59±1.39)%。靶向脂质体组(0.48±0.083)mg在相同剂量下(80k IU/kg)与PBS空白对照组(2.04±0.114)mg、游离HV12p-rPA组(1.2±0.100)mg和普通HV12p-rPA脂质体组(0.74±0.089)mg分别比较,其血栓干重明显减轻(P<0.05);靶向脂质体组与5倍剂量(400k IU/kg)的游离HV12p-rPA组(0.52±0.084)mg比较,其血栓干重较之偏轻(P>0.05)。制备出的靶向HV12p-rPA脂质体具有体内靶向溶栓效果。
The anticoagulant-thrombolytic bifunctional hirudin 12 peptide-reteplase (HV12p-rPA) obtained by protein engineering was prepared into targeted liposomes and its in vivo thrombolytic effect was investigated. HV12p-rPA liposomes were prepared by thin-film dispersion-ultrasonic method. The optimal design of the liposomes was achieved by orthogonal design. Anti-fibrinogen monoclonal antibody (McAbSZ-65) The rat model of carotid artery thrombosis was used to investigate the in vivo thrombolytic effect of HV12p-rPA targeting liposomes. Results: The optimum formulation of HV12p-rPA liposomes was (142.45 ± 1.20) nm in size and (-30.63 ± 0.48) mV in zeta potential. The average encapsulation efficiency was (91.59 ± 1.39)%. Targeting liposomes (0.48 ± 0.083) mg at the same dose (80k IU / kg) with PBS blank control group (2.04 ± 0.114) mg, free HV12p-rPA group (1.2 ± 0.100) mg and ordinary HV12p-rPA (P <0.05). Compared with the free HV12p-rPA group (0.52 ± 0.05) in the liposome group (0.74 ± 0.089) mg, the thrombus dry weight was significantly reduced 0.084) mg, thrombosis lighter than the light (P> 0.05). The targeted HV12p-rPA liposomes were made to target the thrombolytic effect in vivo.