CN1857358A - Use of atractylone, atractylone-containing plant and atractylone extract in preparing influenza virus resisting medicine - Google Patents
Use of atractylone, atractylone-containing plant and atractylone extract in preparing influenza virus resisting medicine Download PDFInfo
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- CN1857358A CN1857358A CN 200510034393 CN200510034393A CN1857358A CN 1857358 A CN1857358 A CN 1857358A CN 200510034393 CN200510034393 CN 200510034393 CN 200510034393 A CN200510034393 A CN 200510034393A CN 1857358 A CN1857358 A CN 1857358A
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Abstract
The present invention discloses the effective cold virus resisting effective component, atractylone, and features that atractylone and Chinese medicine or extract containing atractylone have effect of resisting influenza A and B virus. Extracorporeal experiments show that atractylone in concentration of 1 mg/ml has the effect of killing influenza A virus H3N2 subtype, high pathogenic bird flu virus H5N1 subtype and influenza B virus strain. All the said atractylone, Chinese medicine or extract containing atractylone may be used as the medicine for human body and animal to prevent and treat influenza and bird flu, and may be prepared into tablet, capsule, granule, oral liquid, injection, inhalant and spray.
Description
Technical field
The present invention relates to atractylone, contain the plant of atractylone and the purposes of atractylone extract in the preparation anti-influenza virus medicament.
Background technology
Influenza (Influenza) is called for short influenza, is the acute respiratory infectious disease that is caused by influenza virus.Main by the air droplet transmission.Influenza virus has first, second, the third three types, because the first type morphs easily, its caused influenza pandemic the most extensively and serious.B-mode often causing breaks out, and third type often causes that children's distributes more.Influenza is in a single day popular, propagates soon, involves widely, people ' s health and productivity are had a significant impact, and also very big to the threat of old many patients and infant, Chang Yin causes complication and death.At present, influenza is China's emphasis monitoring of diseases, classifies the Class C infectious disease as and manage in China's Law on the Prevention and Control of Infectious Diseases.
(Avian Influenza AI), is a kind of syndrome from respiratory tract disease to multiple symptoms such as seriousness septicemia that is caused poultry and wild fowl by influenza A virus to bird flu.China Ministry of Agriculture classifies bird flu as category A infectious disease.Popular H5N1 type can be passed on a skill of craft to others by avian in the Asia, the people is had very high pathogenic, and the people infects back case fatality rate height.Infect high pathogenic avian influenza, in China's Law on the Prevention and Control of Infectious Diseases, classify Category B notifiable disease as and manage.
The influenza patient generally adopts symptomatic treatment, uses antipyretic analgesic and antiallergic agent mostly.Old many patients and infant are noted preventing complication such as pneumonia.Patient is partitioned to and brings down a fever back 2 days.Present available resisiting influenza virus medicine only has amantadine and rimantadine, and is only effective to the first type, and causes the untoward reaction of nervous system and digestive system easily, and life-time service easily produces drug resistance.Develop have zanamivir (Zanamivir) and Austria of entering clinical research and taking charge of its Wei (Oseltamivir), be neuraminidase inhibitor, though it is effective to first, influenza B, but curative effect is relatively poor, systemic infection to influenza is invalid, side effect such as nauseating, vomiting are arranged, and approval is used for the prevention that influenza is subjected to as yet.The anti-influenza virus medicament of researching and developing at present suppresses the Pu Kenali (Pleconaril) and the protease inhibitor Lu Pu woods Qu Wei (Ruprinhivir) of virus uncoating in addition, and this equal end of two medicines enters clinical practice.
Chinese medicine has unique curative effect to the treatment of influenza.The effective ingredient of screening resisiting influenza virus from Chinese medicine is a kind of new trial of Tamiflu exploitation.
Summary of the invention
In order from the Chinese medicine Rhizoma Atractylodis, to filter out a kind of effective ingredient of influenza, the object of the present invention is to provide the effective ingredient of an anti-cold virus---atractylone, it is characterized in that: atractylone and contain the Chinese medicine and the extract thereof of atractylone all has the effect of anti-first type, Influenza B virus.Prove according to experiment in vitro: atractylone all has killing action to influenza A virus H3N2 hypotype, highly pathogenic bird flu virus H 5 N 1 hypotype and Influenza B virus strain when 1mg/ml concentration.
As containing atractylone (Atractylon, CAS:6989-21-5) plant, comprise: Compositae Atractylodes lancea (Thunb.) DC. Atractylodes lancea (Thunb.) DC., Atractylis chinensis Atractylodes chinensis (DC.) Koidz., atractylodes japonica Atractylodes japonica Koidz.ex Kitam., Atractylodes koreana Atractylodes koreana Nakai, full leaf Rhizoma Atractylodis Atractylodes chinensis Koidz.var.simplicifolia Kitag., the dry rhizome of Rhizoma Atractylodis Atractylodeschinensis var.quinqueloba Koidz. of Chifeng and Rhizoma Atractylodis Macrocephalae Atractylodes macrocephala Koidz..
Preferred as the Chinese medicine that contains atractylone, comprising: the dry rhizome of Atractylis chinensis Atractylodes chinensis (DC.) Koidz. or atractylodes japonica Atractylodes japonica Koidz.ex Kitam..
In order to realize obtaining containing from above-mentioned Chinese medicine the method for the extract of atractylone: the Chinese medicine powder that contains atractylone is broken into 10-30 order powder, 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 ℃, behind the extraction certain hour, obtains extract, for containing the extract of atractylone.
Method for optimizing as the extract that contains atractylone (3) extracting method: Atractylis chinensis or atractylodes japonica are ground into 14-20 order powder, put in the supercritical extraction device, use CO
2Carry out supercritical extraction, extraction conditions is: extracting pressure 20-22Mpa; Extraction temperature is 45-50 ℃, resolves pressure 6-7Mpa; Resolution temperature is 40-45 ℃, extracts after 3 hours, is rich in the extract of atractylone in the gained extract.
Above-mentioned atractylone and Chinese medicine thereof, Chinese medicine extract all can be used as human or brutish use medicine, also can make the disinfectant anti-epidemic product, can oral administration and externally, be used for the prevention and the treatment of people and bird flu.The dosage form of making comprises: tablet, capsule, granule, oral liquid, injection, inhalant, spray.
Description of drawings
Fig. 1 is the chemical structural drawing of atractylone of the present invention.
The specific embodiment
Embodiment one: the prevention of bird flu:
The prevention of influenza: the Atractylis chinensis powder includes atractylone 3.0%.Itself and chicken feedstuff are mixed thoroughly, and every day, consumption was Atractylis chinensis powder 5g/, divided sooner or later and took for twice, obeyed continuously 3 days.
Embodiment two: the prevention of influenza:
The Atractylis chinensis supercritical extract is made into the Emulsion that includes atractylone 2.0%.During the influenza epidemic situation, be used to spray disinfectant anti-epidemic.
Embodiment three: the treatment of influenza:
50 years old male, body weight is 60 kilograms, influenza; Medication: one day three time, continuously obey 3 day with containing atractylone capsule 450mg every day.
Experimental example
The screening of Rhizoma Atractylodis resisiting influenza virus effective ingredient
1 material and instrument
1.1 reagent and instrument: Atractylodes lancea (Thunb.) DC., purchase peaceful medical material market in Guangzhou, through evaluation the dry rhizome of Atractylodes lancea (Thunb.) DC. Atractylodeslancea (Thunb.) DC..Purchasing in the Inner Mongol and exhale city medical material company, is the dry rhizome of Atractylis chinensis Atractylodes chinensis (DC.) Koidz. through evaluation.1-5 rises supercritical carbon dioxide extraction apparatus, purchases in Guangzhou light industry institute; High pressure sharp separation chromatographic column (Flash-Chromatograghie-S ule) is purchased the company in German machery-nagel.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/H1N1 H5N1), Influenza B virus strain, cell strain MDCK, section provides by the Center of Diseases Prevention ﹠ Control, Shenzhen City Micro biological Tests.
2 methods and result
2.1 the preparation of sample: Atractylodes lancea (Thunb.) DC., Atractylis chinensis pulverize separately are become coarse powder, prepare Atractylodes lancea (Thunb.) DC. water extract, Atractylis chinensis water extract, Atractylodes lancea (Thunb.) DC. SFE-CO2 extract, Atractylis chinensis SFE-CO2 extract by the preparation method and the SFE-CO2 preparation method of extract of water extract.
The preparation method of water extract: get Rhizoma Atractylodis coarse powder 100g, decoct 3 times with the heating of 600ml water, each 1 hour, filter, merge extractive liquid, is concentrated into 100ml, filters, and puts in the clean container standby.
The SFE-CO2 preparation method of extract: Rhizoma Atractylodis are ground into 10-30 order powder, get 2000g, put in the supercritical extraction device, carry out supercritical extraction with CO2, and extraction conditions is: extracting pressure 21Mpa; Extraction temperature is 50 ℃, resolves pressure 6.5Mpa; Resolution temperature is 45 ℃, extracts 3 hours, gets extract 115g, puts in the clean container standby.
Sample is analyzed through GC:: chromatographic column: the U.S. ZB-WAX of Phenomenex company capillary column (30m * 0.25mm * 0.25um), fixative Polyethylene Glycol (Polyethylene Glycol), coating concentration is 100%; Post flow: 1.0mL.min-1; Carrier gas: N2:25mL.min-1; Detector: FID, H2:30mL.min-1, Air:300mL.min-1, detector temperature are 250 ℃; Injector temperature: 210 ℃, sample size 2 μ L, split sampling, split ratio is 20: 1; Temperature programming: 70 ℃ of (0min) → 15 ℃ min-1 of initial temperature → 145 ℃ of (4min) → 20 ℃ min-1 → 190 ℃ of (5min) → 10 ℃ min-1 → 230 ℃ (10min).Measurement result is calculated content by normalization method.The result shows, does not contain atractylone and atisine chloride atractydin in Atractylodes lancea (Thunb.) DC. water extract, the Atractylis chinensis water extract; Atractylodes lancea (Thunb.) DC. SFE-CO2 extract contains 0.07% atractylone and 3.4% atisine chloride atractydin; Atractylis chinensis SFE-CO2 extract contains 50.4% atractylone, 0.07% atisine chloride atractydin.
The extract part atractylone and the atisine chloride atractydin content of the different samples of table 1. Rhizoma Atractylodis
| The sample title | Atractylone | Atisine chloride atractydin |
| Atractylodes lancea (Thunb.) DC. water extract | - | - |
| Atractylis chinensis water extract | - | - |
| Atractylodes lancea (Thunb.) DC. SFE-CO2 extract | 0.07％ | 3.4％ |
| Atractylis chinensis SFE-CO2 extract | 50.4％ | 0.07％ |
| Atisine chloride atractydin | - | 98.9％ |
| Atractylone | 97.8％ | - |
2.2 the separation of atisine chloride atractydin, atractylone:
Get Atractylodes lancea (Thunb.) DC. carbon dioxide supercritical fluid extraction thing and carry out column chromatography for separation: thin layer, is cooled to after the room temperature with petroleum ether (30-60 ℃) wet method dress post at 110 ℃ of activation 1h with silica gel H, is eluent with the petroleum ether, collects the flow point of thin layer checking R f=0.7.The water bath method petroleum ether, atisine chloride atractydin, low temperature (18 ℃), keep in Dark Place.。
Get Atractylis chinensis carbon dioxide supercritical fluid extraction thing and carry out column chromatography for separation: thin layer, is cooled to after the room temperature with petroleum ether (30-60 ℃) wet method dress post at 110 ℃ of activation 1h with silica gel H, is eluent with the petroleum ether, collects the flow point of thin layer checking R f=0.4.The water bath method petroleum ether, atractylone, low temperature (18 ℃), keep in Dark Place.
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 Rhizoma Atractylodis, 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 ℃, and 5%CO2 incubator environment overnight incubation adds the 0.2mL1% red cell suspension in every hole, and room temperature is placed 30-60min. behind the mix homogeneously.Observe the blood clotting result.
After SFE-CO2 extract, atisine chloride atractydin, the atractylone sample of Rhizoma Atractylodis made the sample liquid of 100mg/ml with DMSO, with the PBS dilution (1: 2 of opposing doubly, 1: 8,1: 16,1: 32,1: 64,1: 128) totally 6 manage, the dilution back adds 50 μ l/ holes in the mdck cell that has grown up to monolayer, and 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 ℃, and 5%CO2 incubator environment overnight incubation adds the 0.2mL1% red cell suspension in every hole, and 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 extract parts of table 1. Rhizoma Atractylodis
| The sample title | H3N2 | H5N1 | Influenza B virus |
| Atractylodes lancea (Thunb.) DC. water extract | - | - | - |
| Atractylis chinensis water extract | - | - | - |
| Atractylodes lancea (Thunb.) DC. SFE-CO2 extract | 1∶20 | 1∶20 | 1∶20 |
| Atractylis chinensis SFE-CO2 extract | 1∶640 | 1∶640 | 1∶640 |
| Atisine chloride atractydin | - | - | - |
| Atractylone | 1∶1280 | 1∶1280 | 1∶1280 |
The result shows, atractylone 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 atisine chloride atractydin, Atractylodes lancea (Thunb.) DC. water extract, Atractylis chinensis water extract all do not 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, and Atractylodes lancea (Thunb.) DC. SFE-CO2 extract contains that atractylone is few, a little less than the resisiting influenza virus effect; And Atractylis chinensis SFE-CO2 extract contains the atractylone height, and the resisiting influenza virus effect is strong.It is high more to contain the atractylone amount in the sample, and the greatest dilution of resisiting influenza virus effect is big more.
2.4 the toxic mensuration of atractylone pair cell: adopt micro-culture method.In advance mdck cell is incubated in 96 well culture plates, puts 35 ℃, cultivate in the 5%CO2 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: 640,1: 1280) 100 μ l atractylone sample liquid, add respectively and grown up in the mdck cell of monolayer, 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 ℃, and 5%CO2 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 that the unlimited poison amount of atractylone is 1: 160, is 8 times of greatest dilution 1: 1280 of resisiting influenza virus effect.
3. brief summary and discussion
Atractylone all has the toxic action of going out for three types to influenza virus, and atisine chloride atractydin does not then have the toxic action of going out; The water extract that does not contain atractylone does not all have the toxic action of going out for three types to influenza virus, and Atractylodes lancea (Thunb.) DC. SFE-CO2 extract contains that atractylone is few, a little less than the resisiting influenza virus effect; And Atractylis chinensis SFE-CO2 extract contains the atractylone height, and the resisiting influenza virus effect is strong.Contain atractylone amount and the resisiting influenza virus effect relation of being proportionate in the sample.The unlimited poison amount of atractylone is 1: 160, be the resisiting influenza virus effect 8 times of greatest dilution 1: 1280.Can confirm that thus atractylone is the effective ingredient of Rhizoma Atractylodis 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.Atractylone can be killed H3N2, H5N1 influenza A virus and Influenza B virus, and the prompting atractylone can be used for the control of people's influenza A and influenza B, also can be used for bird flu.
The plant that contains atractylone has: Compositae Atractylodes lancea (Thunb.) DC. Atractylodes lancea (Thunb.) DC., Atractylis chinensis Atractylodes chinensis (DC.) Koidz., atractylodes japonica Atractylodes japonica Koidz.ex Kitam., Atractylodes koreana Atractylodes koreana Nakai, full leaf Rhizoma Atractylodis Atractylodes chinensis Koidz.var.simplicifolia Kitag., the dry rhizome of Rhizoma Atractylodis Atractylodes chinensis var.quinqueloba Koidz. of Chifeng and Rhizoma Atractylodis Macrocephalae Atractylodes macrocephala Koidz..This experimentation shows that it is few that Atractylodes lancea (Thunb.) DC. contains atractylone, and a little less than the resisiting influenza virus effect of its SFECO2 extract, and that Atractylis chinensis contains atractylone is many, and the resisiting influenza virus effect of its SFECO2 extract is strong.Prompting contains the Chinese medicine of atractylone, and the resisiting influenza virus effect is all arranged, and the content of atractylone and resisiting influenza virus effect are proportionate.
Claims (10)
- Atractylone, the plant that contains atractylone or atractylone extract the preparation influenza emit virus medicine in purposes.
- 2. atractylone, the plant that contains atractylone or atractylone extract are killed purposes 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.
- 3. purposes according to claim 1 and 2, it is characterized in that the plant that contains atractylone comprises: Compositae Atractylodes lancea (Thunb.) DC. Atractylodes lancea (Thunb.) DC., Atractylis chinensis Atractylodes chinensis (DC.) Koidz., atractylodes japonica Atractylodes japonica Koidz.ex Kitam., Atractylodes koreana Atractylodes koreana Nakai, full leaf Rhizoma Atractylodis Atractylodes chinensis Koidz.var.simplicifolia Kitag., the dry rhizome of Rhizoma Atractylodis Atractylodes chinensisvar.quinqueloba Koidz. of Chifeng or Rhizoma Atractylodis Macrocephalae Atractylodes macrocephala Koidz..
- 4. purposes according to claim 3 is characterized in that the plant that contains atractylone comprises: the dry rhizome of Atractylis chinensis Atractylodeschinensis (DC.) Koidz. or atractylodes japonica Atractylodes japonica Koidz.Ex Kitam..
- 5. purposes according to claim 1 and 2 is characterized in that the extract that contains atractylone prepares as follows: the plant that contains atractylone 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 ℃, behind the extraction certain hour, obtains containing the extract of atractylone.
- 6. purposes according to claim 5, it is characterized in that the extract that contains atractylone prepares as follows: Atractylis chinensis or atractylodes japonica are pulverized, put in the supercritical extraction device, carry out supercritical extraction with CO2, extraction conditions 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 atractylone.
- 7. purposes according to claim 1 is characterized in that described medicine is that human or birds and beasts are emitted viral medicine with influenza, or the disinfectant anti-epidemic product, is for oral administration or exterior-applied formulation.
- 8. purposes according to claim 7 is characterized in that described medicine makes tablet, capsule, granule, oral liquid, injection, inhalant, spray or drop.
- 9. an extract that contains atractylone is characterized in that preparing by the following method: contain the plant pulverizing of atractylone, put in the supercritical extraction device, use 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 ℃, behind the extraction certain hour, obtains containing the extract of atractylone.
- 10. extract according to claim 9 is characterized in that preparing as follows: Atractylis chinensis or atractylodes japonica are pulverized, put in the supercritical extraction device, use CO 2Carry out supercritical extraction, extraction conditions 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 atractylone.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104225651A (en) * | 2014-08-26 | 2014-12-24 | 赵兰 | Air freshener for preventing influenza and preparation method thereof |
| CN106942288A (en) * | 2017-03-16 | 2017-07-14 | 广东省中医院 | A kind of traditional medicine volatile oil composition and air sterilizing disinfectant acted on sterilizing |
| CN109157533A (en) * | 2018-10-29 | 2019-01-08 | 上海市浦东新区传染病医院 | Application of the atractylone in preparation anti-liver cancer and anti-molecular target drug |
-
2005
- 2005-04-30 CN CNB2005100343939A patent/CN100551366C/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104225651A (en) * | 2014-08-26 | 2014-12-24 | 赵兰 | Air freshener for preventing influenza and preparation method thereof |
| CN106942288A (en) * | 2017-03-16 | 2017-07-14 | 广东省中医院 | A kind of traditional medicine volatile oil composition and air sterilizing disinfectant acted on sterilizing |
| CN106942288B (en) * | 2017-03-16 | 2019-09-24 | 广东省中医院 | A kind of traditional medicine volatile oil composition and air sterilizing disinfectant with sterilizing effect |
| CN109157533A (en) * | 2018-10-29 | 2019-01-08 | 上海市浦东新区传染病医院 | Application of the atractylone in preparation anti-liver cancer and anti-molecular target drug |
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| CN100551366C (en) | 2009-10-21 |
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