CN101230086A - Technique for producing hirudin by using poecilobdella manillensis saliva as raw material - Google Patents
Technique for producing hirudin by using poecilobdella manillensis saliva as raw material Download PDFInfo
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- CN101230086A CN101230086A CNA2007100264616A CN200710026461A CN101230086A CN 101230086 A CN101230086 A CN 101230086A CN A2007100264616 A CNA2007100264616 A CN A2007100264616A CN 200710026461 A CN200710026461 A CN 200710026461A CN 101230086 A CN101230086 A CN 101230086A
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- hirudinaria manillensis
- hirudin
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Abstract
The invention discloses a hirudin manufacturing technique with the saliva of poecilobdella manillensis as the raw material, which belongs to the traditional Chinese medicine pharmacy field, relates to extraction of effective elements of animals, and aims at solving the problem that the traditional hirudin extraction technique is difficult to realize industrialized production. The technique adopts the saliva of poecilobdella manillensis to extract hirudin, and industrialized production can be realized. The saliva of poecilobdella manillensis is extracted through normal saline, the extracting solution is boiled and filtered, and the filter liquor is obtained. The solids is extracted through normal saline, and filtered, and the filter liquor is obtained. All the filter liquor produced in the two steps is combined together, Lyphgel is added to treat for twice, filtering is performed, and the filter liquor is removed. The solids is extracted through normal saline and filtered. After being boiled, the filter liquor is cooled down rapidly, and refrigerated over night. Centrifugation is performed, the supernatant is taken, and the filter liquor is frozen and dried into powder after decompression and condensation. The manufacturing technique is simple, the product activity is high, and has very strong thrombus dissolution function.
Description
Technical field
This technology relates to field of traditional Chinese medicine pharmacy, and being specifically related to a kind of is the production technique of raw-material extraction r-hirudin from Hirudinaria manillensis saliva.
Background technology
China has abundant leech resource to utilize.Leech is a kind of activating blood and removing stasis drug commonly used in traditional traditional Chinese medicine and pharmacy, based on many Chinese patent medicines of leech medicinal material in the market popularity.But this class medicine uses inconvenient, and curative effect is not obvious.
The related leech of this technology is a Hirudinaria manillensis, is the hirudinaria manillensis of doctor leech section Poecilobdella, extensively is distributed in the ground such as Hainan, Guangdong, Guangxi, Fujian and Yunnan of China.The application of Hirudinaria manillensis is the clinical medicine that effective prevention and treatment cardiovascular disorder are provided.The used Hirudinaria manillensis saliva of this technology be meant with the pig clot feed raise the Hirudinaria manillensis of propagating artificially after, the sorrel liquid of extruding in the Hirudinaria manillensis body through staff again.This liquid is-18 ℃ of freezing preservations.
The research of Hirudinaria manillensis biologically active substance, BIOPHARM company (EP6373767,1990) is purified into the hirudin class material of antithrombin.But this invention hirudin class material is acetone extraction with an organic solvent, separates, and the purifying process complexity is single-activity.Though active higher, yield is extremely low, is difficult to suitability for industrialized production.HOFMAN etc. (WO007221,2001) are purified into Unidasa.This invention material also is the single-activity composition, complicated process of preparation, and content is extremely low, can't carry out suitability for industrialized production.Zhou Weihai (03113566,2003) is a raw material with the saliva of Hirudinaria manillensis, and the further separating and filtering of its saliva is obtained the r-hirudin crude product, the r-hirudin crude product is carried out deep processing again and obtains meeting medical r-hirudin.The hirudin activity height that this method obtains, medicinal effect is good, and it is simple to extract the r-hirudin method, but yield is extremely low.Kang Qiang (02116727.3,2003) adopts Hirudinaria manillensis live body normal saline extraction, or the method for the melt and dissolved back of living body freezing normal saline extraction, obtains the lyophilized solid extract of biologically active.
Summary of the invention
The r-hirudin that this technology is extracted is starting material with Hirudinaria manillensis saliva, and it is extract and production method thereof starting material production, that be used to prevent and treat the cardiovascular and cerebrovascular diseases that thrombus causes by Hirudinaria manillensis saliva that purpose is to provide a kind of.This production method technology is simple, and cost is lower, but suitability for industrialized production, and also leech can extract saliva repeatedly, and raw-material source is more stable.This extract is easy to use, and curative effect is obvious.
Description of drawings
Fig. 1 is a technique for producing hirudin schema of the present invention.
Embodiment
The present invention realizes by following measure:
After refrigerated Hirudinaria manillensis saliva was thawed and liquefied, the sodium chloride solution that adds isopyknic 0.15mol/L mixed.240r/min stirred 2 hours, boiled, and was cooled to room temperature rapidly.120 order nylon net filters are collected filtrate.Solid substance adds the uniform suspension of 0.15mol/L sodium chloride solution furnishing of 1.5 times of Hirudinaria manillensis saliva volumes.180r/min stirred 1 hour, 150 order nylon net filters.Collect filtrate, solid substance repeats aforesaid operations and extracts 2 times.Discard solid substance, merge three times filtrate, be cooled to room temperature after boiling rapidly, merge whole filtrates and spend the night in refrigerator cold-storage.Filtrate after the refrigeration discards solid substance with 150 orders and 300 order nylon wires elder generation after-filtration, collects filtrate.Quality by filtrate added Lyphgel with 50: 1, and 180r/min stirred 2 hours, 150 order nylon net filters.Filtrate repeats to discard after aforesaid operations is handled once again.Merge twice solid substance, the sodium chloride solution 180r/min that presses 2 times of 0.15mol/L of quality adding of solid substance stirred 1 hour.150 order nylon net filters are collected filtrate, and solid substance repeats aforesaid operations and handles 2 times again.Solid substance is reusable after treatment.After merging three filtrates and boiling, be cooled to room temperature rapidly.After refrigerator cold-storage spends the night, centrifugal 10 minutes of 5000r/min.Collect supernatant liquor, be concentrated into 60AIU/ml with the rotatory evaporator that reduces pressure in 55 ℃, lyophilize becomes lyophilized powder again.
Claims (3)
1. one kind is that the starting material technique for producing hirudin is characterized in that with Hirudinaria manillensis saliva:
Raw materials used is Hirudinaria manillensis saliva, is meant that the Hirudinaria manillensis propagated artificially picks up after raising pig blood through feeding, again the sorrel liquid of extruding in the Hirudinaria manillensis body through staff.This liquid is-18 ℃ of freezing preservations.
2. one kind is that raw-material technique for producing hirudin is characterized in that with Hirudinaria manillensis saliva:
After this process using Hirudinaria manillensis saliva is thawed, produce the freeze-dry extract of biologically active with the method for normal saline extraction.
3. described according to right 2, adopting Hirudinaria manillensis saliva is raw material, by extract, concentrate, lyophilize makes this product, its feature adopts following steps and method:
1) after refrigerated Hirudinaria manillensis saliva was thawed liquefaction, the sodium chloride solution that adds isopyknic 0.15mol/L mixed.240r/min stirred 2 hours, boiled, and was cooled to room temperature rapidly.
2) 120 order nylon net filters are collected filtrate.Solid substance adds the uniform suspension of 0.15mol/L sodium chloride solution furnishing of 1.5 times of Hirudinaria manillensis saliva volumes.180r/min stirred 1 hour, 150 order nylon net filters.Collect filtrate, solid substance repeats aforesaid operations and extracts 2 times, discards solid substance.
3) merge three times filtrate, be cooled to room temperature after boiling rapidly, merge whole filtrates and spend the night in refrigerator cold-storage.
4) filtrate after the refrigeration discards solid substance with 150 orders and 300 order nylon wires elder generation after-filtration, collects filtrate.
5) press the quality of filtrate with 50: 1 adding Lyphgel.180r/min stirred 2 hours, 150 order nylon net filters.Filtrate repeats to discard after aforesaid operations is handled once again, merges twice solid substance.
6) the sodium chloride solution 180r/min that presses 2 times of 0.15mol/L of quality adding of solid substance stirred 1 hour.150 order nylon net filters are collected filtrate, and solid substance repeats aforesaid operations and handles 2 times again.Solid substance is reusable after treatment.
7) after three filtrates of merging are boiled, be cooled to room temperature rapidly.After refrigerator cold-storage spends the night, centrifugal 10 minutes of 5000r/min.
8) collect supernatant liquor, be concentrated into 60AIU/ml with the rotatory evaporator that reduces pressure in 55 ℃, lyophilize becomes lyophilized powder again.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007100264616A CN101230086A (en) | 2007-01-22 | 2007-01-22 | Technique for producing hirudin by using poecilobdella manillensis saliva as raw material |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
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| CNA2007100264616A CN101230086A (en) | 2007-01-22 | 2007-01-22 | Technique for producing hirudin by using poecilobdella manillensis saliva as raw material |
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| CN101230086A true CN101230086A (en) | 2008-07-30 |
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| CNA2007100264616A Pending CN101230086A (en) | 2007-01-22 | 2007-01-22 | Technique for producing hirudin by using poecilobdella manillensis saliva as raw material |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103417580A (en) * | 2013-09-04 | 2013-12-04 | 南宁市净雪皇生物工程有限公司 | Method for preparing poecilobdella manillensis medium-high activity anticoagulant crude product |
| CN104837495A (en) * | 2012-09-17 | 2015-08-12 | 比奥佩普解决方案有限公司 | A whole, leech saliva extract |
| CN111249170A (en) * | 2020-04-01 | 2020-06-09 | 广州今盛美精细化工有限公司 | Hirudin and application thereof in preparation of cosmetics |
| CN114699432A (en) * | 2022-05-19 | 2022-07-05 | 广西蛭郎中生物科技有限公司 | Method for extracting hirudo manillensis anticoagulant crude product |
| CN115386445A (en) * | 2022-09-23 | 2022-11-25 | 文泽沂 | Preparation method of natural hirudin wine |
-
2007
- 2007-01-22 CN CNA2007100264616A patent/CN101230086A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104837495A (en) * | 2012-09-17 | 2015-08-12 | 比奥佩普解决方案有限公司 | A whole, leech saliva extract |
| CN103417580A (en) * | 2013-09-04 | 2013-12-04 | 南宁市净雪皇生物工程有限公司 | Method for preparing poecilobdella manillensis medium-high activity anticoagulant crude product |
| CN103417580B (en) * | 2013-09-04 | 2015-07-22 | 南宁市净雪皇生物工程有限公司 | Method for preparing poecilobdella manillensis medium-high activity anticoagulant crude product |
| CN111249170A (en) * | 2020-04-01 | 2020-06-09 | 广州今盛美精细化工有限公司 | Hirudin and application thereof in preparation of cosmetics |
| CN114699432A (en) * | 2022-05-19 | 2022-07-05 | 广西蛭郎中生物科技有限公司 | Method for extracting hirudo manillensis anticoagulant crude product |
| CN114699432B (en) * | 2022-05-19 | 2024-02-23 | 广西蛭郎中生物科技有限公司 | Method for extracting anti-coagulation crude product of Poecilobdella manillensis |
| CN115386445A (en) * | 2022-09-23 | 2022-11-25 | 文泽沂 | Preparation method of natural hirudin wine |
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Open date: 20080730 |