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 biological activity

Inactive Publication Date: 2008-12-31
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, lee...

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] Embodiment 1: prepare leech extract, the steps are as follows:

[0031] (1) Get the dried leeches and grind them into 100 mesh powder, add 8 times the amount of water, and homogenate;

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

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

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

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

Embodiment 2

[0036] Embodiment 2: prepare leech extract, the steps are as follows:

[0037] (1) Get fresh and alive whole body of leeches, add 2.5 times the amount of water, and homogenate;

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

[0039] (3) Add 3 times the volume of ethanol refrigerated precipitation to the enzymolysis solution, filter, take the filtrate and recover it under reduced pressure until it has no alcohol smell, add 2 times the volume of acetone refrigerated precipitation, filter, take the filtrate below 60 °C and recover under reduced pressure, the residue Add water to dissolve, filter with 0.45 micron microporous membrane;

[0040] (4) passing the above-mentioned medicinal liquid through an u...

Embodiment 3

[0042] Embodiment 3: prepare leech extract, the steps are as follows:

[0043] (1) Get fresh and alive whole body of leeches, add 2.5 times the amount of water, and homogenate;

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

[0045] (3) Add 3 times the volume of ethanol to the enzymatic hydrolysis solution to refrigerate and precipitate, filter, take the filtrate and recover it under reduced pressure below 60°C, add water to dissolve the residue, and filter through a 0.45 micron microporous membrane;

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

[0047] (5) Adjust the pH of the ultrafiltrate to 7.9, pass through DE-52 resin, wash with pH7.9, 0.02mol / L Tris-h...

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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, a preparation method and application thereof. Background technique [0002] The effect of leech is to break blood, expel blood stasis, and eliminate symptoms. It can be used for lumps in the abdomen, blood stasis, amenorrhea, bruises, thrombocytosis, and cerebral hemorrhage. Leeches were first recorded in "Shen Nong's Materia Medica": "The main purpose is to drive away bad blood, blood stasis, moon closure, accumulation of broken blood, no children, and water passages." Salty and bitter, in order to remove blood accumulation, it is the medicine of the liver meridian blood, so it can dredge the liver meridian and gather blood." [0003] As a traditional traditional Chinese medicine for promoting blood circulation and removing blood stasis, leech has the functions of breaking blood, expelling blood stasis, and dredging menstruation. Since the anticoagulant effect of leeches has been affirmed, and in recent year...

Claims

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Application Information

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IPC IPC(8): A61K35/62A61P7/02A61P25/00
Inventor 吉爱国武建卓梁浩
Owner SHANDONG UNIV
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