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Method for producing nattokinase liposome from phytosterol

A technology of phytosterol and soybean kinase lipid, which is applied in the directions of liposome delivery, drug combination, pharmaceutical formulation, etc., can solve problems such as hypercholesterolemia, and achieve the effects of good biocompatibility, fast release speed and low toxicity

Inactive Publication Date: 2009-10-21
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when cholesterol is excessive in the body, it will lead to hypercholesterolemia, which will have an adverse effect on the body.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Utilize phytosterol to produce the method for nattokinase liposome, it comprises the steps:

[0028] 1) Preparation of blank liposomes: Weigh 180 mg of lecithin, add 90 mg of stigmasterol and 3 ml of diethyl ether, rotate to evaporate to form a uniform lipid film covering the bottom of the pear-shaped bottle, add 5 mL, 0.02 mol / L to the bottle , pH7.4 buffer solution (phosphate buffer solution 5mL, mannitol 360mg), add glass beads, then rotate and shake at 60°C for 60min, ultrasonicate in a water bath for 20min (frequency 40Hz, power 500W), and then freeze-dry for 15h (-10~ 0°C, 1mbar), to obtain liposome precursor material (blank liposome);

[0029] 2) Hydration of liposomes: Take 5ml of nattokinase solution (nattokinase activity 2500U / ml), add it to the liposome precursor substance, and hydrate under nitrogen flow for 10min to obtain nattokinase liposomes. The encapsulation rate of nattokinase liposome was determined to be 64.4%.

[0030] The nattokinase liposome pr...

Embodiment 2

[0034] Utilize phytosterol to produce the method for nattokinase liposome, it comprises the steps:

[0035] 1) Preparation of blank liposomes: Weigh 270 mg of lecithin, add 90 mg of sitosterol, and 5 ml of diethyl ether, rotate to evaporate to form a uniform lipid film covering the bottom of the pear-shaped bottle, add 10 mL, 0.02 mol / L , pH7.4 buffer solution (phosphate buffer solution 10mL, mannitol 180mg), add glass beads, then rotate and shake at 60°C for 45min, ultrasonicate in a water bath for 20min (frequency 40Hz, power 500W), and then freeze-dry for 10h (-10~ 0°C, 1mbar), to obtain liposome precursor material (blank liposome);

[0036] 2) Hydration of liposomes: Take 10ml of nattokinase solution (nattokinase activity 1000U / ml), add it to the liposome precursor substance, and hydrate under nitrogen flow for 15min to obtain nattokinase liposomes. The encapsulation rate of nattokinase liposome was determined to be 55.2%

Embodiment 3

[0038] Utilize phytosterol to produce the method for nattokinase liposome, it comprises the steps:

[0039] 1) Preparation of blank liposomes: Weigh 270 mg of lecithin, add 90 mg of stigmasterol and 3 ml of ether, rotate to evaporate to form a uniform lipid film covering the bottom of the pear-shaped bottle, add 8 mL, 0.02 mol / L , pH7.4 buffer solution (phosphate buffer solution 8mL, mannitol 270mg), add glass beads, then rotate and shake at 60°C for 60min, ultrasonicate in a water bath for 20min (frequency 40Hz, power 500W), and then freeze-dry for 15h (-10~ 0°C, 1mbar), to obtain liposome precursor material (i.e. blank liposome);

[0040] 2) Hydration of liposomes: Take 5ml of nattokinase solution (nattokinase activity 2500U / ml), add it to the liposome precursor substance, and hydrate under nitrogen flow for 10min to obtain nattokinase liposomes. The encapsulation rate of nattokinase liposome was determined to be 48.2%.

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Abstract

The invention relates to a method for producing a nattokinase liposome from phytosterol. The method for producing a nattokinase liposome from phytosterol is characterized by comprising: (1) a step of raw material selection, in which lecithin, phytosterol, ether, a buffer solution and a solution of nattokinase are selected in mass ratio of 100-300 mg:90-200 mg:2-5 mL:5-10 mL:5-10 mL for later use; 2) a step of blank liposome preparation, in which the lecithin, the phytosterol and the ether are mixed, the mixture is subjected to rotary evaporation to be formed into a uniform lipoid membrane to cover the internal bottom of a pear-shaped flask, the buffer solution is added into the flask, glass beads are added into the flask, the flask is rotated and shaken at 60 DEG C for 30 to 90 minutes, the mixture in the flask is treated by ultrasonic wave in a water bath for 10 to 20 minutes, and the materials in the flask is subjected to freeze drying to form a precursor substance of the liposome; and 3) a step of hydration, in which the solution of nattokinase is added into the precursor substance of the liposome for hydration in nitrogen flow for 10 to 20 minutes and thus the nattokinase liposome is obtained. The method is simple in process and can reduce the influences of cholesterol on human body.

Description

technical field [0001] The invention relates to a method for producing nattokinase liposome. Background technique [0002] Nattokinase is fermented by Bacillus natto, and its molecular weight is much smaller than that of urokinase, streptokinase, tissue-type plasminogen activator, and can be naturally absorbed by the intestinal tract. The in vitro and in vivo thrombolytic function of nattokinase has also been confirmed through experiments: the in vivo thrombolytic activity of nattokinase is four times that of plasmin and plasmin, and the duration of action in the body is long. The most important point is It can also activate tissue-type plasminogen activator in the human body, so that it can gently and continuously improve the fibrinolytic activity of blood. Nattokinase can reduce fibrin and platelet aggregation rate, prevent arteriosclerosis, and improve thrombotic diseases. In recent years, a lot of research has been done on the properties, mechanism of action, separatio...

Claims

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Application Information

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IPC IPC(8): A61K9/127A61K38/48A61K47/28A61P7/02
Inventor 董绪燕陈洪魏芳袁钢友李光明赵元弟
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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