CN100493532C - Traditional Chinese medicine effective part for treating influenza virus and its preparation process - Google Patents
Traditional Chinese medicine effective part for treating influenza virus and its preparation process Download PDFInfo
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- CN100493532C CN100493532C CNB2006100053822A CN200610005382A CN100493532C CN 100493532 C CN100493532 C CN 100493532C CN B2006100053822 A CNB2006100053822 A CN B2006100053822A CN 200610005382 A CN200610005382 A CN 200610005382A CN 100493532 C CN100493532 C CN 100493532C
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Abstract
The invention discloses an effective portion of traditional Chinese medicament for resisting influenza virus, which is prepared from selfheal 5-200 parts, mulberry leaf 5-1-00 parts, wild chrysanthemum flower 5-100 parts as raw material through disintegrating, and supercritical CO2 extraction. The effective portion can be prepared into tablets, capsules, granules, oral liquids, injections, inhalants, sprays or drop pills.
Description
Technical field
The present invention relates to the compound preparation medicine, particularly a kind of Chinese medicine preparation for the treatment of influenza virus and preparation method thereof.
Background technology
Chinese medicine has unique curative effect to the treatment of influenza." summer Sang Ju Yin " is made up of Spica Prunellae, Folium Mori, Flos Chrysanthemi Indici three flavor Chinese medicines, and " XIASANGJU KELI agent " that water is obtained through refining is the famous Chinese patent medicine of China, is usually used in the treatment of traditional Chinese medical science heat symptom-complex, is commonly used for the herbal tea influenza in the daily life and emits.But owing to use the water extraction crude drug usually, what obtain is water soluble ingredient only, has influenced the drug effect performance of Chinese medicine; On technology, exist long heating again, cause the thermal sensitivity effective ingredient destroyed, can not be used for influenza as a line medicine and emit; Simultaneously since the drug manufacture Technology fall behind, the scientific and technological content deficiency, dosage form is coarse, consumer is difficult to accept.
Summary of the invention
The objective of the invention is to overcome the deficiencies in the prior art, providing a kind of combines ancient prescription with modern preparation technique, not only attractive in appearance but also convenient, taking dose is little, can be used for treating the effective ingredient in Chinese and preparation method thereof of the resisiting influenza virus of acute influenza virus.
The object of the present invention is achieved like this, and its fat-soluble effective site by following crude drug is made: Spica Prunellae 5-200 part, Folium Mori 5-100 part, Flos Chrysanthemi Indici 5-100 part.
As a kind of prioritization scheme, its fat-soluble effective site by following crude drug is made: Spica Prunellae 50-150 part, Folium Mori 20-50 part, Flos Chrysanthemi Indici 15-100 part.
Most preferred scheme is: its fat-soluble effective site by following crude drug is made: 100 parts of Spica Prunellaes, 35 parts on Folium Mori, 16 parts of Flos Chrysanthemi Indicis.
This effective ingredient in Chinese can be made tablet, capsule, granule, oral liquid, injection, inhalant, spray or drop, can be used as to make human or brutish emit viral medicine with influenza, or the disinfectant anti-epidemic product.Can be applied to prepare the medicine of killing influenza A virus H3N2 hypotype, highly pathogenic bird flu virus H 5 N 1 hypotype and Influenza B virus strain.
The preparation method of the effective ingredient in Chinese of this resisiting influenza virus is: with aforementioned Spica Prunellae, and Folium Mori, Flos Chrysanthemi Indici is pulverized, and puts in the supercritical extraction device, uses CO
2Carry out supercritical extraction, extraction conditions is: extracting pressure 15-33Mpa; Extraction temperature is 30-50 ℃, resolves pressure 5-9Mpa; Resolution temperature is 30-55 ℃, extracts after 1-4 hour, can obtain containing the extract of fat-soluble effective site.
The extraction conditions of optimizing is: extracting pressure 20-22Mpa; Extraction temperature is 45-50 ℃, resolves pressure 6-7Mpa; Resolution temperature is 40-45 ℃, extracts after 3 hours, obtains containing the extract of fat-soluble effective site.(carbon dioxide flow is 25 ± 3L/h)
Drug effective region of the present invention can be used for the treatment of box prophylaxis of acute influenza, and effective ingredient is clear and definite, and drug effect is remarkable, and taking dose is little, and dosage form is various, and is easy to use.Further specify beneficial effect of the present invention below in conjunction with relevant test:
1, material and instrument
1.1, reagent and instrument: Spica Prunellae, Folium Mori, Flos Chrysanthemi Indici is purchased the peaceful medical material market in Guangzhou, is the kind that Chinese Pharmacopoeia version in 2005 is recorded through evaluation.
1-5 rises supercritical carbon dioxide extraction apparatus, purchases in Guangzhou light industry institute; 24,96 porocyte culture plates and 25cm3 Tissue Culture Flask are purchased the company in German Greiner bio-one.CO2 gas incubator 3210 types are purchased the Forma company in the U.S..U.S. VARIAN-3900 gas chromatograph.
1.2, strain and cell: influenza A virus strain (A/shenzhen/203/2001/H3N2, A/shenzhen/45/2001/H1N1H5N1), Influenza B virus strain, cell strain MDCK, section provides by the Center of Diseases Prevention ﹠ Control, Shenzhen City Micro biological Tests.
2, method and result
2.1, the preparation of sample: with 100 parts of Spica Prunellaes, 35 parts on Folium Mori, 16 parts of Flos Chrysanthemi Indicis are ground into coarse powder, press the preparation method and the SFE-CO of water extract
2Preparation method of extract prepares summer Sang Ju Yin water extract, summer Sang Ju Yin SFE-CO
2Extract.
The preparation method of water extract: get 100 parts of Spica Prunellaes, 35 parts on Folium Mori, 16 parts of Flos Chrysanthemi Indicis, the medical material gross weight is 151g, decocts 3 times with the heating of 1000ml water, each 1 hour, filter, merge extractive liquid, is concentrated into 151ml, filters, and puts in the clean container standby.
SFE-CO
2Preparation method of extract: get 100 parts of Spica Prunellaes, 35 parts on Folium Mori, 16 parts of Flos Chrysanthemi Indicis, the medical material gross weight is 453g, is ground into 10-30 order powder, puts in the supercritical extraction device, uses CO
2Carry out supercritical extraction, extraction conditions is: extracting pressure 21Mpa; Extraction temperature is 50 ℃, resolves pressure 6.5Mpa; Resolution temperature is 45 ℃, extracts 3 hours, gets extract 25g, puts in the clean container standby.2.3, the effect of vitro inhibition first type and Influenza B virus: adopt micro-culture method.
At first three kinds of viruses (H3N2, H5N1 and Influenza B virus) are diluted to test concentrations (100 TCID50) respectively.With the water extract of summer Sang Ju Yin, do not dilute 100 μ l/ holes in the mdck cell (Testis et Pentis Canis passage cell) that direct adding grown up to monolayer, add influenza virus liquid 50 μ l/ holes again, add the liquid of keeping in 100 μ l/ holes again.Cell control well is established in each concentration 4 hole simultaneously, puts 35 ℃, 5%CO
2Incubator environment overnight incubation adds the 0.2mL1% red cell suspension in every hole, room temperature is placed 30-60min behind the mix homogeneously.Observe the blood clotting result.
SFE-CO with the summer Sang Ju Yin
2The extract sample, make the sample liquid of 100mg/ml with DMSO after, with PBS oppose doubly dilution (1:2,1:8,1:16,1:32,1:64 1:128) 6 manages totally, and the dilution back adds 50 μ l/ holes in the mdck cell that has grown up to monolayer, every hole adds influenza virus liquid 50 μ l/ holes again, adds the liquid of keeping in 100 μ l/ holes, every hole again.Each sample concentration 4 hole is established DMSO hole and cell control well simultaneously, puts 35 ℃, 5%CO
2Incubator environment overnight incubation adds the 0.2mL1% red cell suspension in every hole, room temperature is placed 30-60min. behind the mix homogeneously.Observe the blood clotting result.The results are shown in Table 1.
The greatest dilution of the resisiting influenza virus of the different extraction method extracts of table 1. summer Sang Ju Yin
| The sample title | H3N2 | H5N1 | Influenza B virus |
| Summer Sang Ju Yin water extract | — | — | — |
| Summer Sang Ju Yin SFE-CO 2Extract | 1:320 | 1:320 | 1:320 |
The result shows, summer Sang Ju Yin SFE-CO
2The extract liposoluble constituent all has the toxic action of going out to influenza virus A type H3N2, influenza virus A type H5N1 (bird flu virus) and Influenza B virus, and summer Sang Ju Yin water extract does not all have the toxic action of going out to influenza virus A type H3N2, influenza virus A type H5N1 (bird flu virus) and Influenza B virus.
2.4, summer Sang Ju Yin SFE-CO
2The toxic mensuration of extract pair cell: adopt micro-culture method.In advance mdck cell is incubated in 96 well culture plates, puts 35 ℃, 5%CO
2Cultivate in the incubator environment, after about 24 hour cell stratification, discard culture fluid in the hole, will pass doubly dilution (1:20 again, 1:40,1:80,1:160,1:320,1:640,100 μ l liposoluble constituent sample liquid 1:1280), adding has grown up in the mdck cell of monolayer respectively, and every hole adds the liquid of keeping in 100 μ l/ holes again.Cell control well is established in each concentration 4 hole simultaneously, puts 35 ℃, 5%CO
2The incubator environment is cultivated, and writes down the cytopathy result every day, declares the result when the toxicity of medicine pair cell no longer continues to make progress.The minimum extension rate of cytopathic medicine not occur is unlimited poison amount.
The result shows, summer Sang Ju Yin SFE-CO
2The unlimited poison amount of extract is 1:16, is 20 times of greatest dilution 1:320 of resisiting influenza virus effect.
3, brief summary and discussion
Summer Sang Ju Yin SFE-CO
2Extract all has the toxic action of going out for three types to influenza virus; And the water extract does not all have the toxic action of going out for three types to influenza virus.Summer Sang Ju Yin SFE-CO
2The unlimited poison amount of extract is 1:16, is 20 times of greatest dilution 1:320 of resisiting influenza virus effect.Can confirm that thus the summer Sang Ju Yin adopts SFE-CO
2Extract the drug effect that just has influenza.
Strains of influenza viruses is selected H3N2, H5N1 strain and the Influenza B virus strain of first type for use, wherein the H3N2 of first type is the popular influenza morbidity virus in the Asia in the recent period, and being the people, H5N1 infects high pathogenic avian influenza virus, once causing 1997 and Hong Kong bird flu in 2003, is the Strain of the popular high pathogenic avian influenza in Asia.Summer Sang Ju Yin liposoluble constituent can be killed H3N2, H5N1 influenza A virus and Influenza B virus, prompting summer Sang Ju Yin SFE-CO
2The extract liposoluble constituent can be used for the control of people's influenza A and influenza B, also can be used for bird flu.
The specific embodiment:
Embodiment 1: get 100 parts of Spica Prunellaes, and 35 parts on Folium Mori, 16 parts of Flos Chrysanthemi Indicis, the medical material gross weight is 453g, is ground into 10-30 order powder, puts in the supercritical extraction device, uses CO
2Carry out supercritical extraction, extraction conditions is: extracting pressure 21Mpa; Extraction temperature is 50 ℃, resolves pressure 6.5Mpa; Resolution temperature is 45 ℃, extracts 3 hours, must contain the extract 25g of fat-soluble effective site, adds appropriate amount of auxiliary materials, makes the soft capsule that contains 0.1g/ grain supercritical extract, is used for birds flu-preventing, every day 3 times, one time one, took continuously 3 days.
Embodiment 2: get 5 parts of Spica Prunellaes, and 20 parts on Folium Mori, 100 parts of Flos Chrysanthemi Indicis, the medical material gross weight is 250g, is ground into 30 order powder, puts in the supercritical extraction device, uses CO
2Carry out supercritical extraction, extraction conditions is: extracting pressure 16Mpa; Extraction temperature is 35 ℃, resolves pressure 5Mpa; Resolution temperature is 30 ℃, extracts 2 hours, must contain the extract 14g of fat-soluble effective site, adds appropriate amount of auxiliary materials, makes the drop that contains supercritical extract, is used for the poultry birds flu-preventing.
Embodiment 3: get 200 parts of Spica Prunellaes, and 100 parts on Folium Mori, 10 parts of Flos Chrysanthemi Indicis, the medical material gross weight is 620g, is ground into 20 order powder, puts in the supercritical extraction device, uses CO
2Carry out supercritical extraction, extraction conditions is: extracting pressure 33Mpa; Extraction temperature is 50 ℃, resolves pressure 9Mpa; Resolution temperature is 55 ℃, extracts 4 hours, must contain the extract 35g of fat-soluble effective site, adds appropriate amount of auxiliary materials, makes the Emulsion that contains 2% supercritical extract, is used for during the bird flu epidemic situation, sprays disinfectant anti-epidemic.
Embodiment 4: get 150 parts of Spica Prunellaes, and 50 parts on Folium Mori, 100 parts of Flos Chrysanthemi Indicis, the medical material gross weight is 300g, is ground into 30 order powder, puts in the supercritical extraction device, uses CO
2Carry out supercritical extraction, extraction conditions is: extracting pressure 25Mpa; Extraction temperature is 48 ℃, resolves pressure 8Mpa; Resolution temperature is 40 ℃, extracts 3 hours, must contain the extract 17g of fat-soluble effective site, adds appropriate amount of auxiliary materials, makes the tablet that contains 0.1g/ grain supercritical extract, is used for birds flu-preventing, every day 3 times, one time one, took continuously 3 days.
Embodiment 5: get 180 parts of Spica Prunellaes, and 75 parts on Folium Mori, 80 parts of Flos Chrysanthemi Indicis, the medical material gross weight is 670g, is ground into 30 order powder, puts in the supercritical extraction device, uses CO
2Carry out supercritical extraction, extraction conditions is: extracting pressure 28Mpa; Extraction temperature is 40 ℃, resolves pressure 9Mpa; Resolution temperature is 50 ℃, extracts 4 hours, must contain the extract 40g of fat-soluble effective site, adds appropriate amount of auxiliary materials, makes the granule that contains supercritical extract, is used for birds flu-preventing, takes continuously 3 days.
Embodiment 6: get 20 parts of Spica Prunellaes, and 100 parts on Folium Mori, 5 parts of Flos Chrysanthemi Indicis, the medical material gross weight is 500g, is ground into 30 order powder, puts in the supercritical extraction device, uses CO
2Carry out supercritical extraction, extraction conditions is: extracting pressure 30Mpa; Extraction temperature is 45 ℃, resolves pressure 7Mpa; Resolution temperature is 40 ℃, extracts 4 hours, must contain the extract 27g of fat-soluble effective site, adds appropriate amount of auxiliary materials, makes the Emulsion that contains 2% supercritical extract, during the influenza epidemic situation, is used to spray disinfectant anti-epidemic.
Embodiment 7: get 100 parts of Spica Prunellaes, and 50 parts on Folium Mori, 10 parts of Flos Chrysanthemi Indicis, the medical material gross weight is 480g, is ground into 30 order powder, puts in the supercritical extraction device, uses CO
2Carry out supercritical extraction, extraction conditions is: extracting pressure 22Mpa; Extraction temperature is 50 ℃, resolves pressure 6.5Mpa; Resolution temperature is 40 ℃, extracts 3 hours, must contain the extract 27g of fat-soluble effective site, adds appropriate amount of auxiliary materials, makes the oral liquid that contains supercritical extract, during the influenza epidemic situation, is used for the treatment of influenza.
Embodiment 8, get 50 parts of Spica Prunellaes, 50 parts on Folium Mori, 50 parts of Flos Chrysanthemi Indicis, the medical material gross weight is 450g, is ground into 30 order powder, puts in the supercritical extraction device, uses CO
2Carry out supercritical extraction, extraction conditions is: extracting pressure 21Mpa; Extraction temperature is 45 ℃, resolves pressure 7Mpa; Resolution temperature is 40 ℃, extracts 3 hours, must contain the extract 25g of fat-soluble effective site, adds appropriate amount of auxiliary materials, makes the injection that contains supercritical extract, during the influenza epidemic situation, is used for the treatment of influenza.
Embodiment 9: get 40 parts of Spica Prunellaes, and 80 parts on Folium Mori, 20 parts of Flos Chrysanthemi Indicis, the medical material gross weight is 420g, is ground into 30 order powder, puts in the supercritical extraction device, uses CO
2Carry out supercritical extraction, extraction conditions is: extracting pressure 22Mpa; Extraction temperature is 50 ℃, resolves pressure 6.5Mpa; Resolution temperature is 40 ℃, extracts 3 hours, must contain the extract 23g of fat-soluble effective site, adds appropriate amount of auxiliary materials, makes the inhalant that contains supercritical extract, during the influenza epidemic situation, is used for the treatment of influenza.
Embodiment 10: get 120 parts of Spica Prunellaes, and 75 parts on Folium Mori, 65 parts of Flos Chrysanthemi Indicis, the medical material gross weight is 260g, is ground into 30 order powder, puts in the supercritical extraction device, uses CO
2Carry out supercritical extraction, extraction conditions is: extracting pressure 20Mpa; Extraction temperature is 40 ℃, resolves pressure 5Mpa; Resolution temperature is 40 ℃, extracts 2 hours, must contain the extract 14g of fat-soluble effective site, adds appropriate amount of auxiliary materials, makes the spray that contains supercritical extract, during the influenza epidemic situation, is used for the spraying epidemic prevention.
Embodiment 11: get 75 parts of Spica Prunellaes, and 20 parts on Folium Mori, 80 parts of Flos Chrysanthemi Indicis, the medical material gross weight is 350g, is ground into 30 order powder, puts in the supercritical extraction device, uses CO
2Carry out supercritical extraction, extraction conditions is: extracting pressure 22Mpa; Extraction temperature is 50 ℃, resolves pressure 6.5Mpa; Resolution temperature is 40 ℃, extracts 3 hours, must contain the extract 20g of fat-soluble effective site, adds appropriate amount of auxiliary materials, makes the drop that contains supercritical extract, during the influenza epidemic situation, is used for the treatment of influenza.
Embodiment 12: get 175 parts of Spica Prunellaes, and 10 parts on Folium Mori, 40 parts of Flos Chrysanthemi Indicis, the medical material gross weight is 550g, is ground into 30 order powder, puts in the supercritical extraction device, uses CO
2Carry out supercritical extraction, extraction conditions is: extracting pressure 20Mpa; Extraction temperature is 35 ℃, resolves pressure 5Mpa; Resolution temperature is 40 ℃, extracts 2 hours, must contain the extract 30g of fat-soluble effective site, adds appropriate amount of auxiliary materials, makes the tablet that contains 0.1g/ grain supercritical extract, is used for flu-prevention, takes continuously 3 days.
Claims (8)
1, a kind of effective ingredient in Chinese of resisiting influenza virus is characterized in that: its fat-soluble effective site by the carbon dioxide supercritical fluid extraction of following crude drug through not adding entrainer is made: Spica Prunellae 5-200 part, Folium Mori 5-100 part, Flos Chrysanthemi Indici 5-100 part.
2, the effective ingredient in Chinese of a kind of resisiting influenza virus according to claim 1, it is characterized in that: its fat-soluble effective site by the carbon dioxide supercritical fluid extraction of following crude drug through not adding entrainer is made: Spica Prunellae 50-150 part, Folium Mori 20-50 part, Flos Chrysanthemi Indici 15-100 part.
3, the effective ingredient in Chinese of a kind of resisiting influenza virus according to claim 2, it is characterized in that: its fat-soluble effective site by the carbon dioxide supercritical fluid extraction of following crude drug through not adding entrainer is made: 100 parts of Spica Prunellaes, 35 parts on Folium Mori, 16 parts of Flos Chrysanthemi Indicis.
4, a kind of effective ingredient in Chinese as claim 1,2 or 3 described resisiting influenza virus is killed application in the medicine of influenza A virus H3N2 hypotype, highly pathogenic bird flu virus H 5 N 1 hypotype and Influenza B virus strain in preparation.
5, according to the effective ingredient in Chinese of claim 1,2 or 3 described a kind of resisiting influenza virus, it makes tablet, capsule, granule, oral liquid, injection, inhalant, spray or drop.
6, according to the effective ingredient in Chinese of claim 1,2 or 3 described a kind of resisiting influenza virus, it makes human or birds and beasts are emitted viral medicine with influenza, or the disinfectant anti-epidemic product.
7, a kind of method for preparing the effective ingredient in Chinese of the described resisiting influenza virus of claim 1 is characterized in that: with aforementioned Spica Prunellae, and Folium Mori, Flos Chrysanthemi Indici is pulverized, and puts in the supercritical extraction device, uses CO
2Carry out supercritical extraction, extraction conditions is: do not add entrainer, extracting pressure 15-33Mpa; Extraction temperature is 30-50 ℃, resolves pressure 5-9Mpa; Resolution temperature is 30-55 ℃, extracts after 1-4 hour, obtains containing the extract of fat-soluble effective site.
8, preparation method according to claim 7 is characterized in that: with aforementioned Spica Prunellae, and Folium Mori, Flos Chrysanthemi Indici is pulverized, and puts in the supercritical extraction device, uses CO
2Carry out supercritical extraction, extraction conditions is: do not add entrainer, extracting pressure 20-22Mpa; Extraction temperature is 45-50 ℃, resolves pressure 6-7Mpa; Resolution temperature is 40-45 ℃, extracts after 3 hours, obtains containing the extract of fat-soluble effective site.
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| CNB2006100053822A CN100493532C (en) | 2006-01-19 | 2006-01-19 | Traditional Chinese medicine effective part for treating influenza virus and its preparation process |
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| CN100493532C true CN100493532C (en) | 2009-06-03 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101279028B (en) * | 2008-05-26 | 2011-01-19 | 雷清莲 | Method of preparing Oral liquid vaccine for preventing and controlling avian influenza of domestic animals and domestic birds |
| CN101361941B (en) * | 2008-09-28 | 2011-05-11 | 东莞广州中医药大学中医药数理工程研究院 | Medicine combination for preventing and treating cold and preparation method thereof |
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