CN101450084A - Ethyl acetate extract of Chinese caterpillar fungus, preparation method and use thereof in antiviral medicine - Google Patents
Ethyl acetate extract of Chinese caterpillar fungus, preparation method and use thereof in antiviral medicine Download PDFInfo
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- CN101450084A CN101450084A CNA2008100517243A CN200810051724A CN101450084A CN 101450084 A CN101450084 A CN 101450084A CN A2008100517243 A CNA2008100517243 A CN A2008100517243A CN 200810051724 A CN200810051724 A CN 200810051724A CN 101450084 A CN101450084 A CN 101450084A
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Abstract
The invention provides a cordycep militaris ethyl acetate extract and preparing process thereof, further discloses a purpose of the extract for preparing antiviral remedies. The preparing process comprises: smashing the dry cordyceps militaris, adding ethyl acetate, extracting and filtering in back flow manner by a Soxhlet extractor, and getting a filtrate; eluting the residue by ion liquid, filtering and get another filtrate; combining the tow filtrates, decompressing and concentrating to a constant weight to obtain the final extract. Extracting the effective constituents of the cordycep militaris by using ethyl acetate can increase the extraction rates of multiple effective constituents to higher limit, thereby improve medicine effect. The cordycep militaris ethyl acetate extract is a product obtained from natural resources, achieves excellent antivirus capacity, and advantages of no hemolytic activity and no plasma clkotting activity verified by experiment, can be used for preparing medicine for preventing AIDS, hepatitis B and virus form flu. The invention provides a new selection to the medicine for preventing AIDS, hepatitis B and virus form flu utilizing the abundant natural resources in our country.
Description
Technical field:
The present invention relates to a kind of ethyl acetate extract of Chinese caterpillar fungus, preparation method and the application in antiviral drugs thereof, belong to Chinese medicine extracts active ingredients and applied technical field, also belong to the biochemical pharmacy field.
Background technology:
Cordyceps militaris (L.) Link. (Cordyceps militaris), another name Cordyceps militaris (L.) Link., Cordyceps militaris belong to Ascomycotina (Ascomycotina), Clavicipitaceae (Clavicipitaceae), Cordyceps (Cordyceps).Under the Cordyceps situation that supply falls short of demand, the Cordyceps militaris (L.) Link. equally celebrated for their achievements with Cordyceps is subjected to people's attention in the world day by day, and its exploitation and production have been developed new nutrient source and medicine source for body-care and field of medicaments." Xinhua's book on Chinese herbal medicine outline " record Cordyceps militaris (L.) Link. effects such as " property are flat, and sweet in the mouth has the lung benefiting kidney, mends marrow, hemostasis and phlegm " is used for diseases such as pulmonary tuberculosis, old man's weakness and anemia, is the Chinese crude drug of preciousness.Cordyceps militaris (L.) Link. has multiple pharmacological effect: can blood fat reducing, prevent arteriosclerosis, and the protection heart, cerebral tissue, tranquilizing soporific strengthens macrophage activity, anticancer, antiinflammatory, antibiotic, anti-hypoxia etc.; Cordyceps militaris (L.) Link. can be used for treating emphysema, prostate hyperplasia; Cordyceps militaris (L.) Link. can also human body immunity improving power, and auxiliary renal transplant recipients is got well, and increases sexual function etc.
Former Cordyceps militaris (L.) Link. extracts active ingredients fabricating technology characteristics are the Cordyceps militaris (L.) Link. fungus filament to be adopted the method for decocting in water precipitate with ethanol, extract multiple proteins and polysaccharide effective ingredient in the Cordyceps militaris (L.) Link. thalline, and effective ingredient often extracts not exclusively.In addition, only limit to aspects such as human body immunity improving power about the pharmacological action of effective ingredient in the Cordyceps militaris (L.) Link., it is less report aspect the research of antivirus action.
Summary of the invention:
The object of the present invention is to provide a kind of ethyl acetate extract of Chinese caterpillar fungus, have tangible medicinal activity.
The invention also discloses the preparation method of ethyl acetate extract of Chinese caterpillar fungus, be applicable to suitability for industrialized production.
The present invention further discloses the application of ethyl acetate extract of Chinese caterpillar fungus in the preparation medicine.
Ethyl acetate extract of Chinese caterpillar fungus of the present invention obtains by the following method:
(1) exsiccant Cordyceps militaris (L.) Link. fungus filament is pulverized,,, filtered, get filtrate A with reflux, extract, 8～12h under 77～00 ℃ of temperature of apparatus,Soxhlet's in the ratio adding ethyl acetate of 1/ (10～15) m/v;
(2) filtering residue is extracted 3～5h in the ratio of 1/ (10～15) m/v for 100 ℃ with deionized water, filter, get liquor B;
(3) filtrate A and liquor B are merged, be evaporated to constant weight, promptly get extract.
This extract is a kind of mixture of being made up of organic acid, ciclosporin, sugar alcohol and sterols material.
Below experiment shows that ethyl acetate extract of Chinese caterpillar fungus is at antiviral pharmacologically active:
One, the experiment of ethyl acetate extract of Chinese caterpillar fungus AIDS virus resisting
On 96 porocyte culture plates, ethyl acetate extract of Chinese caterpillar fungus is carried out 2 times of doubling dilutions with complete medium, totally 6 dilution factors, every hole 100vl is provided with the normal cell contrast simultaneously, the hiv infected cell contrast.Every experimental port drips 3 * 10
4Individual C8166 cell and 200TCID
50HIV (human immunodeficiency virus) (Human Immunodeficiency Virus), the cumulative volume in every hole is 200vL.Put 37 ℃, 5%CO
2Cultivate in the incubator.Infect the back and observed the influence that testing compound forms syncytium on the 3rd day.Under 100 times inverted microscope, choose 5 nonoverlapping visuals field, the counting HIV (human immunodeficiency virus) (Human Immunodeficiency Virus) is induced the plastidogenetic syncytium number of C8166.IC50 reaches 50% o'clock drug level for suppressing syncytium formation.EC50 is the drug level when 50% host cell is produced cytotoxicity, and therapeutic index (SI) is the ratio of IC50/EC50.
Table 1: ethyl acetate extract of Chinese caterpillar fungus suppresses the effect of HIV (human immunodeficiency virus) (Human Immunodeficiency Virus) propagation:
| Embodiment one | IC50(vg/ml) | EC50(vg/ml) | SI |
| Ethyl acetate extract of Chinese caterpillar fungus | 11.7 | 1.8 | 6.5 |
| Embodiment two | IC50(vg/ml) | EC50(vg/ml) | SI |
| Ethyl acetate extract of Chinese caterpillar fungus | 12.0 | 1.9 | 6.3 |
| Embodiment three | IC50(vg/ml) | EC50(vg/ml) | SI |
| Ethyl acetate extract of Chinese caterpillar fungus | 12.7 | 1.9 | 6.7 |
| Embodiment four | IC50(vg/ml) | EC50(vg/ml) | SI |
| Ethyl acetate extract of Chinese caterpillar fungus | 12.6 | 1.9 | 6.6 |
Two, the experiment of ethyl acetate extract of Chinese caterpillar fungus anti-hepatitis B virus
On 96 porocyte culture plates, ethyl acetate extract of Chinese caterpillar fungus is carried out 2 times of doubling dilutions with complete medium, totally 6 dilution factors, every hole 100vl is provided with the normal cell contrast simultaneously, hepatitis b virus infected cell contrast.Every experimental port drips 3 * 10
4Individual C8166 cell and 300TCID
50Hepatitis B virus, the cumulative volume in every hole is 200vL.Put 37 ℃, 5%CO
2Cultivate in the incubator.Infect the back and observed the influence that testing compound forms syncytium on the 3rd day.Under 100 times inverted microscope, choose 5 nonoverlapping visuals field, the counting hepatitis B virus is induced the plastidogenetic syncytium number of C8166.IC50 reaches 50% o'clock drug level for suppressing syncytium formation.EC50 is the drug level when 50% host cell is produced cytotoxicity, and therapeutic index (SI) is the ratio of IC50/EC50.
Table 2: ethyl acetate extract of Chinese caterpillar fungus suppresses the effect of hepatitis B virus propagation:
| Embodiment one | IC50(vg/ml) | EC50(vg/ml) | SI |
| Ethyl acetate extract of Chinese caterpillar fungus | 11.9 | 1.6 | 7.4 |
| Embodiment two | IC50(vg/ml) | EC50(vg/ml) | SI |
| Ethyl acetate extract of Chinese caterpillar fungus | 11.5 | 1.5 | 7.7 |
| Embodiment three | IC50(vg/ml) | EC50(vg/ml) | SI |
| Ethyl acetate extract of Chinese caterpillar fungus | 11.8 | 1.5 | 7.9 |
| Embodiment four | IC50(vg/ml) | EC50(vg/ml) | SI |
| Ethyl acetate extract of Chinese caterpillar fungus | 12.8 | 1.7 | 7.5 |
Three, the experiment of ethyl acetate extract of Chinese caterpillar fungus resisiting influenza virus
On 96 porocyte culture plates, ethyl acetate extract of Chinese caterpillar fungus is carried out 2 times of doubling dilutions with complete medium, totally 6 dilution factors, every hole 100vl is provided with the normal cell contrast simultaneously, the contrast of influenza infection cell.Every experimental port drips 3 * 10
4Individual C8166 cell and 200TCID
50Influenza virus, the cumulative volume in every hole is 200vL.Put 37 ℃, 5%CO
2Cultivate in the incubator.Infect and observed the influence that testing compound forms syncytium in back second day.Under 100 times inverted microscope, choose 5 nonoverlapping visuals field, the counting influenza virus is induced the plastidogenetic syncytium number of C8166.IC50 reaches 50% o'clock drug level for suppressing syncytium formation.EC50 is the drug level when 50% host cell is produced cytotoxicity, and therapeutic index (SI) is the ratio of IC50/EC50.
Table 3: ethyl acetate extract of Chinese caterpillar fungus suppresses the effect of proliferation of influenza virus:
| Embodiment one | IC50(vg/ml) | EC50(vg/ml) | SI |
| Ethyl acetate extract of Chinese caterpillar fungus | 12.1 | 1.7 | 7.1 |
| Embodiment two | IC50(vg/ml) | EC50(vg/ml) | SI |
| Ethyl acetate extract of Chinese caterpillar fungus | 12.5 | 1.6 | 7.8 |
| Embodiment three | IC50(vg/ml) | EC50(vg/ml) | SI |
| Ethyl acetate extract of Chinese caterpillar fungus | 12.3 | 2.1 | 5.9 |
| Embodiment four | IC50(vg/ml) | EC50(vg/ml) | SI |
| Ethyl acetate extract of Chinese caterpillar fungus | 12.3 | 1.9 | 6.5 |
Above-mentioned experiment shows: ethyl acetate extract of Chinese caterpillar fungus of the present invention has significant inhibition HIV (human immunodeficiency virus) (Human Immunodeficiency Virus), hepatitis B virus and influenza virus, also has no hemolytic activity simultaneously, advantages such as no blood plasma coagulation activity can be used for preparing the medicine for the treatment of acquired immune deficiency syndrome (AIDS), hepatitis B and virus type influenza.
Good effect of the present invention is: take the effective ingredient in the ethyl acetate extraction Cordyceps militaris (L.) Link., can improve the extraction ratio of plurality of active ingredients in the Cordyceps militaris (L.) Link. to large extent, to improve drug effect.Ethyl acetate extract of Chinese caterpillar fungus is the product that obtains from natural resources, experimental results show that and have good anti-virus ability, also have advantages such as no hemolytic activity, no blood plasma coagulation activity simultaneously, can be used for preparing the medicine of anti-AIDS, hepatitis B and virus type influenza.Utilize the natural resources of China's abundant, for preparation anti-AIDS, hepatitis B and virus type influenza medicine provide new selection.
Specific implementation method:
Following embodiment is used for further specifying the present invention, but content of the present invention is not limited thereto.
Embodiment 1
The preparation of ethyl acetate extract of Chinese caterpillar fungus:
(1) exsiccant Cordyceps militaris (L.) Link. fungus filament is pulverized, got 20g mycelium powder, add the 200ml ethyl acetate, at 77 ℃ of reflux, extract, 8h, filter, get filtrate 1 with apparatus,Soxhlet's.
(2) filtering residue 25.8g is extracted 3h for 100 ℃ with the 258ml deionized water, filter, get filtrate 2.
(3) filtrate 1 and filtrate 2 are merged, be evaporated to constant weight, promptly obtain ethyl acetate extract of Chinese caterpillar fungus.
Embodiment 2:
The preparation of ethyl acetate extract of Chinese caterpillar fungus:
(1) exsiccant Cordyceps mycelium is pulverized, got 35g mycelium powder, add the 350ml ethyl acetate, at 93 ℃ of reflux, extract, 11h, filter, get filtrate 1 with apparatus,Soxhlet's.
(2) filtering residue 40.5g is extracted 3h for 100 ℃ with the 405ml deionized water, filter, get filtrate 2.
(3) filtrate 1 and filtrate 2 are merged, be evaporated to constant weight, promptly obtain ethyl acetate extract of Chinese caterpillar fungus.
Embodiment 3:
The preparation of ethyl acetate extract of Chinese caterpillar fungus:
(1) exsiccant Cordyceps militaris (L.) Link. fungus filament is pulverized, got 43g mycelium powder, add the 430ml ethyl acetate, extract 9h at 86 ℃ of streams, filter, get filtrate 1 with apparatus,Soxhlet's.
(2) filtering residue 48.7g is extracted 4.5h for 100 ℃ with the 487ml deionized water, filter, get filtrate 2.
(3) filtrate 1 and filtrate 2 are merged, be evaporated to constant weight, promptly obtain ethyl acetate extract of Chinese caterpillar fungus.
Embodiment 4:
The preparation of ethyl acetate extract of Chinese caterpillar fungus:
(1) exsiccant Cordyceps militaris (L.) Link. fungus filament is pulverized, got 34g mycelium powder, add the 340ml ethyl acetate, at 100 ℃ of reflux, extract, 12h, filter, get filtrate 1 with apparatus,Soxhlet's.
(2) filtering residue 40.1g is extracted 5h for 100 ℃ with the 401ml deionized water, filter, get filtrate 2.
(3) filtrate 1 and filtrate 2 are merged, be evaporated to constant weight, promptly obtain ethyl acetate extract of Chinese caterpillar fungus.
Claims (3)
1, a kind of ethyl acetate extract of Chinese caterpillar fungus prepares through following steps:
(1) exsiccant Cordyceps militaris (L.) Link. fungus filament is pulverized,,, filtered, get filtrate A with reflux, extract, 8～12h under 77～00 ℃ of temperature of apparatus,Soxhlet's in the ratio adding ethyl acetate of 1/ (10～15) m/v;
(2) filtering residue is extracted 3～5h in the ratio of 1/ (10～15) m/v for 100 ℃ with deionized water, filter, get liquor B;
(3) filtrate A and liquor B are merged, be evaporated to constant weight, promptly get extract.
2, the preparation method of the described ethyl acetate extract of Chinese caterpillar fungus of claim 1 prepares through following steps:
(1) exsiccant Cordyceps militaris (L.) Link. fungus filament is pulverized,,, filtered, get filtrate A with reflux, extract, 8～12h under 77～00 ℃ of temperature of apparatus,Soxhlet's in the ratio adding ethyl acetate of 1/ (10～15) m/v;
(2) filtering residue is extracted 3～5h in the ratio of 1/ (10～15) m/v for 100 ℃ with deionized water, filter, get liquor B;
(3) filtrate A and liquor B are merged, be evaporated to constant weight, promptly get extract.
3, the application of the described ethyl acetate extract of Chinese caterpillar fungus of claim 1 in the preparation antiviral drugs.
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| Application Number | Priority Date | Filing Date | Title |
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| CNA2008100517243A CN101450084A (en) | 2008-12-31 | 2008-12-31 | Ethyl acetate extract of Chinese caterpillar fungus, preparation method and use thereof in antiviral medicine |
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| CNA2008100517243A CN101450084A (en) | 2008-12-31 | 2008-12-31 | Ethyl acetate extract of Chinese caterpillar fungus, preparation method and use thereof in antiviral medicine |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102048770B (en) * | 2009-11-06 | 2013-02-13 | 华中科技大学 | Artificially-cultured cordceps militaris sporocarp extract and preparation process thereof |
| WO2015100579A1 (en) * | 2013-12-31 | 2015-07-09 | 王忠林 | Pharmaceutical composition for treating acquired immune deficiency syndrome and method for preparing same |
-
2008
- 2008-12-31 CN CNA2008100517243A patent/CN101450084A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102048770B (en) * | 2009-11-06 | 2013-02-13 | 华中科技大学 | Artificially-cultured cordceps militaris sporocarp extract and preparation process thereof |
| WO2015100579A1 (en) * | 2013-12-31 | 2015-07-09 | 王忠林 | Pharmaceutical composition for treating acquired immune deficiency syndrome and method for preparing same |
| US9623053B2 (en) | 2013-12-31 | 2017-04-18 | Zhonglin Wang | Pharmaceutical composition for treating AIDS and preparation method thereof |
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Open date: 20090610 |