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Leech extract and preparation method and use thereof

A leech extract and leech technology, applied in the direction of drug combination, medical raw materials derived from leeches/worms, extracellular fluid diseases, etc., can solve the problems of unclear active ingredients, loss of active ingredients, complex ingredients, etc., to improve activity and quality controllability, improved utilization, and strong physiological activity

Inactive Publication Date: 2010-09-29
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the leech preparations with definite clinical curative effect are Chinese patent medicines made of raw powder of dried medicinal materials, such as Huoxue Tongmai Capsules, Anti-Thrombotic Tablets, etc. These preparations have many adverse reactions due to the stench of leech medicinal materials. , it is not easy to control the quality of medicinal materials, and the efficacy of the medicine fluctuates greatly
[0004] As a traditional Chinese medicine, leeches are widely used clinically, but the active ingredients are not clear, and the literature reports that proteins and polypeptides are the main active ingredients.
The ambiguity of active ingredients is an inherent problem of animal medicinal materials. Traditional extraction and concentration methods may cause some ingredients to decompose, destroy, or fail to extract, which will easily cause the loss of active ingredients, so it is not suitable for animal medicinal materials.

Method used

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  • Leech extract and preparation method and use thereof
  • Leech extract and preparation method and use thereof
  • Leech extract and preparation method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Preparation of leech extract, the steps are as follows:

[0031] (1) Take the leeches and pulverize them into 100 mesh powder, add 8 times the amount of water, and homogenize;

[0032] (2) Take the homogenate, adjust the pH to 8.5, add trypsin at the ratio of 5000U enzyme per 1g substrate, control the temperature at 50°C, enzymolyze for 6 hours, and cool to room temperature;

[0033] (3) Add 2 times the volume of acetone to the enzymatic hydrolysis solution for precipitation under refrigeration, filter, take the filtrate to recover under reduced pressure below 60°C, dissolve the residue with water, and filter with a 0.45 micron microporous membrane;

[0034] (4) Pass the above-mentioned liquid medicine through an ultrafiltration membrane with a molecular cut-off of 5000 Da, and collect the part with a molecular weight of less than 5000 Da;

[0035] (5) Adjust the pH of the ultrafiltrate to 8.0, pass through Q Sepharose resin, first rinse with water, discard the rinsing...

Embodiment 2

[0036] Example 2: Preparation of leech extract, the steps are as follows:

[0037] (1) Take fresh and live whole leeches, add 2.5 times the amount of water, and homogenize;

[0038] (2) Take the homogenate, adjust the pH to 8.5, add trypsin and pancreatin at the ratio of 5000U enzyme per 1g of substrate, where trypsin provides protease activity accounting for 2% of the total protease activity, control the temperature at 50 ℃, enzymatic hydrolysis 8 hours, cool to room temperature;

[0039] (3) Add 3 times the volume of ethanol to the enzymatic hydrolysate for precipitation in cold storage, filter, take the filtrate under reduced pressure and recover to no alcohol, add 2 times the volume of acetone for refrigerated precipitation, filter, take the filtrate under 60℃ and recover under reduced pressure. Dissolve in water and filter with 0.45 micron microporous membrane;

[0040] (4) Pass the above-mentioned liquid medicine through an ultrafiltration membrane with a molecular cut-off of 5...

Embodiment 3

[0042] Example 3: Preparation of leech extract, the steps are as follows:

[0043] (1) Take fresh and live whole leeches, add 2.5 times the amount of water, and homogenize;

[0044] (2) Take the homogenate, adjust the pH to 8.0, add trypsin at the ratio of 10000U enzyme per 1g substrate, control the temperature at 50°C, enzymolyze for 4 hours, and cool to room temperature;

[0045] (3) Add 3 times the volume of ethanol to the enzymatic hydrolysis solution for precipitation in cold storage, filter, take the filtrate and recover under reduced pressure below 60°C, dissolve the residue with water, and filter with a 0.45 micron microporous membrane;

[0046] (4) Pass the above-mentioned liquid medicine through an ultrafiltration membrane with a molecular cut-off of 5000 Da, and collect the part with a molecular weight of less than 5000 Da;

[0047] (5) Adjust the pH of the ultrafiltrate to 7.9, pass through DE-52 resin, first rinse with pH 7.9, 0.02mol / L Tris-hydrochloric acid buffer, discar...

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Abstract

The present invention discloses a preparation method of bloodsucker extract, in which the bloodsucker is added with water and homogenized, and then the bloodsucker extract is obtained by controllable enzymolysis, organic solvent precipitation, ultrafiltration and anion-exchange chromatography. The bloodsucker extract can be adopted to prepare a medicine with anticoagulation, thrombolytic ability,inhibition on platelet aggregation and neuroprotection effect. The present invention, according to the modern technological requirements of Chinese medicine, adopts the technology of controllable enzymolysis for extracting the peptide component, the separating technology of ultrafiltration and gel chromatography for purification further, and the biological pharmacy technology, thus obtaining the extract with higher pureness and stronger activity than the traditional bloodsucker extract. The method breaks through the bottleneck of the traditional animal medicine in extraction and separation, solves the problem of the traditional animal medicine that the amount of raw powder is large and the taste is fishy, improves the activity and quality controllability of the animal medicine extract further and greatly improves the utilization rate of medicinal materials.

Description

Technical field [0001] The invention relates to a leech extract and its preparation method and application. Background technique [0002] The effect of leech is to break blood, expel blood stasis, and eliminate symptoms. It can be used to treat lumps, blood stasis, amenorrhea, bruises, thrombocytosis, and cerebral hemorrhage. Leech was first published in the "Shen Nong's Materia Medica": "The master chasing evil blood, blood stasis, moon closure, bleeding accumulation, childlessness, and conducive waterway." "Compendium of Materia Medica": "Salt to kill blood, bitterness wins blood. Of leech Salty and bitter, in addition to blood accumulation, it is the liver meridian blood to divide medicine, so it can pass the liver meridian and gather blood." [0003] As a traditional Chinese medicine for promoting blood circulation and removing blood stasis, leeches have the functions of breaking blood, expelling blood stasis, and dredging menstruation. The anticoagulant effect of leeches has...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K35/62A61P7/02A61P25/00
Inventor 吉爱国武建卓梁浩
Owner SHANDONG UNIV
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