CN1732999A - Method for preparing extracting solution of gnetum montanum which can inhibit human liver cancer cell strain - Google Patents
Method for preparing extracting solution of gnetum montanum which can inhibit human liver cancer cell strain Download PDFInfo
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- CN1732999A CN1732999A CN 200510027940 CN200510027940A CN1732999A CN 1732999 A CN1732999 A CN 1732999A CN 200510027940 CN200510027940 CN 200510027940 CN 200510027940 A CN200510027940 A CN 200510027940A CN 1732999 A CN1732999 A CN 1732999A
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Abstract
The invention provides a process for preparing Gnetopsida extract having suppressing effect to human hepatic carcinoma cell line, which comprises, airing vines of Gnetopsida, drying, disintegrating into 40 mesh particles, mixing disintegrated Gnetopsida substance with 10 times by weight of solvent containing ethanol and deionized water by the volume ratio of 7:3, heating for backflow extraction, vacuum concentrating, concentrating the mixed solvent extract into extract, dissolving grease form substance with deionized water, extracting with acetic acid ethyl ester of equal volume, vacuum concentrating the vinyl acetate extract, using polyamide as fixed phase, deionized water-ethanol as flow phase for gradient elution, vacuum concentrating the eluent, freeze drying, obtaining powdered deep brown substance, finally preparing suspension with RPMI1640 culture liquid of cow's serum.
Description
Technical field
What the present invention relates to is the method in a kind of Chinese drug preparation technique field, specifically, is a kind of preparation method that human hepatoma cell strain is had the Caulis Gneti extractive liquid that suppresses effect.
Background technology
Caulis Gneti mainly is distributed in the Yunnan Province of China south, and province and south east asia such as Guangdong are used with Caulis Gneti Parvifolii among the people, treatment rheumatic arthritis, lumbar muscle strain, soreness of bones and muscles, traumatic injury, ulcerative hemorrhage, diseases such as bronchitis.Confirm that through the pharmacology zoopery Caulis Gneti ethanol extraction has tangible antiinflammatory action to rat Ovum Gallus domesticus album and yeast arthritis, but do not have bibliographical information at anti-tumor aspect.
Find by prior art documents, Chen Hao, Lin Mao mention in " Chinese herbal medicine " the 8th phase in 1999 " Da Ye Caulis Gneti chemical constitution study " literary composition, it is immobile phase that Da Ye Caulis Gneti crude extract is adopted polyamide, silica gel, water-ethanol, chloroform-methanol, methanol-water are that mobile phase is carried out repeatedly column chromatography, obtain 15 chemical compounds, but it is not clear and definite as yet whether to have an antitumous effect with the prepared material of this method.
Summary of the invention
The objective of the invention is to overcome the deficiencies in the prior art, a kind of preparation method that human hepatoma cell strain is had the Caulis Gneti extractive liquid that suppresses effect is provided, after making it through heating and refluxing extraction and column chromatography drip washing, improve the inhibition effect of its eluent to human hepatoma cell strain BEL-7402.
The present invention is achieved by the following technical solutions, and the present invention is a raw material with the Caulis Gneti rattan, adopts solvent extraction method, vacuum concentration, solvent extraction, lyophilization and column chromatography process to be prepared from.
Concrete steps of the present invention are as follows:
(1) rattan of Caulis Gneti is dried after, 20-30 ℃ down dry, be ground into 40 order granules again;
(2) with 10 times of mixed solvents to Caulis Gneti rattan ground product weight, the ethanol in the mixed solvent: the volume ratio of deionized water is 7: 3;
(3) 50-90 ℃ of following heating and refluxing extraction is four times, and 1 time 2 hours, 45 ℃ of following vacuum concentration were condensed into extractum with the mixed solvent extract;
(4) utilize the ultrasound wave of frequency for 100KHZ, behind deionized water dissolving cream shape thing, equal-volume ethyl acetate extraction 6 times;
(5) with 45 ℃ of following vacuum concentration of acetic acid ethyl ester extract after do, polyamide (14-100 order) is an immobile phase, deionized water-ethanol is the eluent gradient eluting;
(6) 45 ℃ of following vacuum concentration of eluent ,-40 ℃ of following lyophilizations obtain the Powdered thing of dark brown;
(7) the RPMI1640 culture fluid with 5% hyclone is made into suspension.
Find from the Caulis Gneti rattan being carried out chemical constitution study, wherein contain isorhapontigenin, resveratrol, daucosterol, stearic acid, Caulis Gneti penta element, Caulis Gneti alcohol, cupreol, Caulis Gneti third element and isorhapontigenin-3-O-β-compositions such as D-glucoside.Wherein isorhapontigenin and resveratrol etc. belong to the stilbene monomeric compound, as everyone knows, that the stilbene monomeric compound has is antibiotic, blood fat reducing, blood pressure lowering, antioxidation, free radical resisting and protein kinase inhibitory action isoreactivity, anti-inflammatory activity is in various degree still arranged in addition, particularly isorhapontigenin and resveratrol have very strong leukotriene generation inhibition and receptor acting isoreactivity, and this may be that this eluent has the reason that suppresses the active effect of human hepatoma cell strain BEL-7402.
The present invention is a raw material with the rattan of perennial wild plant Caulis Gneti, utilizes its reproducibility advantage, and abundant plant resources can be provided; Extraction process is simple, need not special instrument; In vitro tests with human tumor cell line shows: the concentration of 0.063mg/ml has significant inhibition effect to human hepatoma cell strain BEL-7402.
The specific embodiment
Embodiment 1
Take by weighing the Caulis Gneti rattan powder 200g (fine powders of 40 order granularities) of 20 ℃ of crushed after being dried, choose dehydrated alcohol: deionized water=7: 3 mixed solvent 2000ml, 50 ℃ of following heating and refluxing extraction three times, each 2 hours, 45 ℃ of following vacuum concentration reclaim solvent, get paste 80.9g, use deionized water dissolving, equal-volume ethyl acetate extraction 6 times, concentrate 10.8g extractum.Getting 1g material wherein, is leacheate with the deionization water-ethanol, and polyamide (14 order) carries out column chromatography and gets active component, concentrates, and-40 ℃ of lyophilizations get 81.4mg.
External this component 0.063mg/ml adopts sulphonyl rhodamine B protein staining method, is 87.0% to the suppression ratio of human hepatoma cell strain BEL-7402.
Embodiment 2
Take by weighing the Caulis Gneti rattan powder 200g (fine powders of 40 order granularities) of 25 ℃ of following crushed after being dried, choose dehydrated alcohol: deionized water=7: 3 mixed solvent 2000ml, 70 ℃ of following heating and refluxing extraction three times, each 2 hours, 45 ℃ of following vacuum concentration reclaim solvent, get paste 80.9g, use deionized water dissolving, equal-volume ethyl acetate extraction 6 times, concentrate 10.8g extractum.Getting 1g material wherein, is leacheate with the deionization water-ethanol, and polyamide (100 order) carries out column chromatography and gets active component, concentrates, and-40 ℃ of lyophilizations get 62.3mg.
External this component 0.25mg/ml adopts sulphonyl rhodamine B protein staining method, is 63.7% to the suppression ratio of human hepatoma cell strain BEL-7402.
Embodiment 3
Take by weighing the Caulis Gneti rattan powder 200g (fine powders of 40 order granularities) of 30 ℃ of crushed after being dried, choose dehydrated alcohol: deionized water=7: 3 mixed solvent 2000ml, 90 ℃ of following heating and refluxing extraction three times, each 2 hours, 45 ℃ of following vacuum concentration reclaim solvent, get paste 80.9g, use deionized water dissolving, equal-volume ethyl acetate extraction 6 times, concentrate 10.8g extractum.Getting 1g material wherein, is leacheate with the deionization water-ethanol, and polyamide (40 order) carries out column chromatography and gets active component, concentrates, and-40 ℃ of lyophilizations get 71.5mg.
External this component 0.016mg/ml adopts sulphonyl rhodamine B protein staining method, is 63.1% to the suppression ratio of human hepatoma cell strain BEL-7402.
Claims (10)
1. one kind has the preparation method of the Caulis Gneti extractive liquid that suppresses effect to human hepatoma cell strain, it is characterized in that, may further comprise the steps:
(1) rattan of Caulis Gneti is dried after, drying is ground into 40 order granules again;
(2) with 10 times of mixed solvents to Caulis Gneti rattan ground product weight, the ethanol in the mixed solvent: the volume ratio of deionized water is 7: 3;
(3) heating and refluxing extraction, vacuum concentration is condensed into extractum with the mixed solvent extract;
(4) utilize ultrasound wave, behind deionized water dissolving cream shape thing, the equal-volume ethyl acetate extraction;
(5) with the acetic acid ethyl ester extract vacuum concentration after do, polyamide is an immobile phase, deionized water-ethanol is the eluent gradient eluting;
(6) eluent vacuum concentration, lyophilization obtains the Powdered thing of dark brown;
(7) the RPMI1640 culture fluid with hyclone is made into suspension.
2. according to claim 1 have the preparation method of the Caulis Gneti extractive liquid that suppresses effect to human hepatoma cell strain, it is characterized in that, and described step (1), its baking temperature is 20-30 ℃.
3. according to claim 1 have the preparation method of the Caulis Gneti extractive liquid that suppresses effect to human hepatoma cell strain, it is characterized in that described heating and refluxing extraction, its temperature are 50-90 ℃.
4. according to claim 3 have the preparation method of the Caulis Gneti extractive liquid that suppresses effect to human hepatoma cell strain, it is characterized in that described heating and refluxing extraction is four times, 1 time 2 hours.
5. according to claim 1 have the preparation method of the Caulis Gneti extractive liquid that suppresses effect to human hepatoma cell strain, it is characterized in that, and described vacuum concentration, its temperature is 45 ℃.
6. according to claim 1 have the preparation method of the Caulis Gneti extractive liquid that suppresses effect to human hepatoma cell strain, it is characterized in that described polyamide is the 14-100 order.
7. according to claim 1 have the preparation method of the Caulis Gneti extractive liquid that suppresses effect to human hepatoma cell strain, it is characterized in that, and described ultrasound wave, its frequency is 100KHZ.
8. according to claim 1 have the preparation method of the Caulis Gneti extractive liquid that suppresses effect to human hepatoma cell strain, it is characterized in that described extraction is 6 times.
9. according to claim 1 have the preparation method of the Caulis Gneti extractive liquid that suppresses effect to human hepatoma cell strain, it is characterized in that, and described step (6), its lyophilization temperature is-40 ℃.
10. according to claim 1 have the preparation method of the Caulis Gneti extractive liquid that suppresses effect to human hepatoma cell strain, it is characterized in that, and the RPMI1640 culture fluid of described hyclone, its concentration is 5%.
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| Application Number | Priority Date | Filing Date | Title |
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| CNB2005100279400A CN1315485C (en) | 2005-07-21 | 2005-07-21 | Method for preparing extracting solution of gnetum montanum which can inhibit human liver cancer cell strain |
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| CNB2005100279400A CN1315485C (en) | 2005-07-21 | 2005-07-21 | Method for preparing extracting solution of gnetum montanum which can inhibit human liver cancer cell strain |
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| CN1732999A true CN1732999A (en) | 2006-02-15 |
| CN1315485C CN1315485C (en) | 2007-05-16 |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1790239A1 (en) * | 2004-09-14 | 2007-05-30 | Hosoda SHC Inc. | Gnetum extract |
| CN110582289A (en) * | 2017-03-16 | 2019-12-17 | 株式会社山田养蜂场本社 | Enhancer of genome stability |
| CN112236136A (en) * | 2018-10-04 | 2021-01-15 | 株式会社细田Shc | Composition for improving intestinal flora |
| CN113981703A (en) * | 2021-11-16 | 2022-01-28 | 高梵(浙江)信息技术有限公司 | Antibacterial warm-keeping jean down pants |
| CN114052309A (en) * | 2021-11-16 | 2022-02-18 | 高梵(浙江)信息技术有限公司 | Sportswear with antibacterial and deodorant down fabric |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100342862C (en) * | 2003-06-27 | 2007-10-17 | 中国医学科学院药物研究所 | Resveratrol oligo cattail compounds, its manufacturing process, pharmaceutical combination and uses thereof |
-
2005
- 2005-07-21 CN CNB2005100279400A patent/CN1315485C/en not_active Expired - Fee Related
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1790239A1 (en) * | 2004-09-14 | 2007-05-30 | Hosoda SHC Inc. | Gnetum extract |
| EP1790239A4 (en) * | 2004-09-14 | 2010-03-31 | Hosoda Shc Inc | Gnetum extract |
| US8067036B2 (en) | 2004-09-14 | 2011-11-29 | Hosoda Shc Inc. | Gnetum extract |
| CN110582289A (en) * | 2017-03-16 | 2019-12-17 | 株式会社山田养蜂场本社 | Enhancer of genome stability |
| CN112236136A (en) * | 2018-10-04 | 2021-01-15 | 株式会社细田Shc | Composition for improving intestinal flora |
| CN112236136B (en) * | 2018-10-04 | 2023-12-01 | Ms健康科学实验室株式会社 | Composition for improving intestinal flora |
| CN113981703A (en) * | 2021-11-16 | 2022-01-28 | 高梵(浙江)信息技术有限公司 | Antibacterial warm-keeping jean down pants |
| CN114052309A (en) * | 2021-11-16 | 2022-02-18 | 高梵(浙江)信息技术有限公司 | Sportswear with antibacterial and deodorant down fabric |
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| CN1315485C (en) | 2007-05-16 |
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