The invention relates to the field of
biopharmaceutics, in particular to a
liposome preparation for
anticoagulant thrombolytic difunctional
fusion protein, namely
anticoagulant thrombolytic difunctional
fusion protein of 12 peptides of
hirudin and reteplase (HV12p-rPA) and a preparation method thereof. The
anticoagulant thrombolytic difunctional
fusion protein (HV12p-rPA) constructed in the laboratory has the dual effects of anticoagulation and
thrombolysis by structural identification, expression,
chromatography renaturation, purification and extracorporal and intracorporal pharmacodynamic experiments. In the preparation method, the
liposome preparation is prepared from the anticoagulant thrombolytic difunctional fusion
protein serving as a
raw material, and a
liposome is prepared by an optimized membrane dispersion-probe ultrasonic method; and the optimized membrane dispersion-probe ultrasonic method is characterized in that a prescription which is most suitable for improving the envelop rate of the HV12p-rPA liposome is selected by taking a ratio of
phospholipid to
cholesterol, the concentration of
protein medicaments, the volume of
buffer solution and water-bath ultrasonic time as influence factors, and the grain
diameter of the HV12p-rPA liposome is reduced and homogenized further by probe ultrasonic, so that the HV12p-rPA liposome of which the grain
diameter is between 140 and 145 nanometers and which is used for intravenous injection is prepared. By the preparation method, the envelop rate of the prepared HV12p-rPA liposome is over 90 percent, and extracorporal thrombolytic activity and extracorporal
anticoagulant activity are 23,810
international unit (IU) .mg<1> and 414
antithrombin unit (ATU) .mg<1> respectively.