CN103638082B - Herba Centipedae CO 2supercritical extract is preparing the application in the medicine or food with resisiting influenza virus effect - Google Patents
Herba Centipedae CO 2supercritical extract is preparing the application in the medicine or food with resisiting influenza virus effect Download PDFInfo
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- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention discloses Herba Centipedae CO
2supercritical extract is preparing the application in the medicine or food with resisiting influenza virus effect.Described Herba Centipedae extract is for adopting CO
2the extract that obtains of supercritical extraction process extraction Herba Centipedae, or by described extract again through extract that petroleum ether, cyclohexane extraction, chloroform or benzene extracting and refining obtain.Described Herba Centipedae extract has good resisiting influenza virus effect, and its antivirus action and positive control drug ribavirin quite or more remarkable, have good medical applications and are worth.The present invention is that the plant resources of exploitation Herba Centipedae provides strong technical foundation.
Description
Technical field
The present invention relates to medical art, more specifically, relate to a kind of Herba Centipedae CO
2supercritical extract is preparing the application in the medicine or food with resisiting influenza virus effect.
Background technology
Influenza virus is that one of human health threatens greatly.Respiratory tract disease caused by influenza virus is called as influenza (influenza), has that popular wide, infectiousness is strong, sickness rate high.According to statistics, the annual flu cases in the whole world is 6 ~ 1,200,000,000, and wherein serious symptom influenza is 300 ~ 5,000,000 examples, and dead 25 ~ 500,000 people, the case fatality rate of serious symptom influenza can reach 10%.Influenza as a kind of viral infectious not only serious threat the health of the people, and bring white elephant to country and society, how effectively the especially highly pathogenic influenza of prevention and therapy influenza has become the important topic that a pendulum is urgently studied and solved in face of the medical personal of countries in the world.Due to influenza antigen variation, conventional vaccine at present still can not effectively flu-prevention break out with popular, therefore the research of anti-influenza virus medicament has great importance in treatment of influenza.
Herba Centipedae is feverfew Myriogyne minuta
centipedaminima(L.) A.Br.et.Aschers, its medicinal part is feverfew Myriogyne minuta
centipedaminima(L.) the wounded in the battle herb of A.Br.et.Aschers, its beginning of being used as medicine is loaded in Southern Tang Dynasty's " feeding habits book on Chinese herbal medicine ", and also on the books in Compendium of Material Medica.In China, Herba Centipedae mainly originates in the ground such as Zhejiang, Jiangsu, Guangdong, Hubei, and in Guangxi, also there is distribution on Henan, Jiangxi, Fujian, the ground such as Anhui.This medical herbs is recorded by 2000 editions and 2005 editions " Chinese Pharmacopoeias " respectively, can " clearing the nasal passage, cough-relieving ", for " headche due to wind-cold, cough with copious phlegm are had a stuffy nose obstructed, nasal sinusitis watery nasal discharge " etc.The chemical composition of Herba Centipedae mainly contains volatile oil, terpenoid, flavonoid and phytosterin compound." the resource of Herba Centipedae, extraction process and pharmacological research " (" Heilungkiang medicine ", December 6 volume 6 phase in 2009) in flavone compound in research summary Herba Centipedae extract there is antianaphylaxis activity, " research of Herba Centipedae alcohol extract bacteriostasis " (" China National folk medicine " the 14th phase in 2013, the antibacterial activity of ethanol extraction Li Jihua) summarized, " Herba Centipedae chemical composition and antitumor activity " (on May 30th, 2012, Zhu Yanping) systematically summarize the activity of ethanol extraction, comprise antiallergic, antitumor, antiinflammatory, antibacterial isoreactivity effect, about the ethanol extraction researching and proposing Herba Centipedae has the effect of relieving cough and asthma.But in many achievements in research, all do not relate to the effect correlational study achievement report whether Herba Centipedae extract has resisiting influenza virus.The resources of medicinal plant of Herba Centipedae needs further developmental research.
Summary of the invention
The technical problem to be solved in the present invention is the deficiency overcoming existing anti-influenza virus medicament, provides a kind of Herba Centipedae CO
2supercritical extract is preparing the application in the medicine or food with resisiting influenza virus effect.
Another technical problem that the present invention will solve is to provide the preparation method of described Herba Centipedae extract.
Object of the present invention is achieved by the following technical programs:
Herba Centipedae CO is provided
2supercritical extract is preparing the application in the medicine or food with resisiting influenza virus effect.
In order to play the effect of motherland's Chinese medicine in resisiting influenza virus further, the present inventor attempts effective component extracting from plurality of Chinese, but effect result badly, the extract that having only never appears in the newspapers has a Herba Centipedae of resisiting influenza virus demonstrates anti-influenza virus activity, obtain the active matter of resisiting influenza virus, further, inventor gropes through a large amount of tests, sums up and obtains at specific CO
2the Herba Centipedae extract obtained under supercritical extraction condition is better than the rnti-influenza virus effect of the related activity thing that other rule methods prepare.
Described specific CO
2supercritical extraction key condition is preferably: described Herba Centipedae CO
2the CO of supercritical extract
2supercritical extraction condition is: extracting pressure is 20 ~ 40 MPas, and extraction temperature is 30 ~ 50 DEG C, and flow velocity is 10 ~ 30 ls/h, and extraction time is 1 ~ 4 hour.
Preferred CO further
2supercritical extraction key condition is: extracting pressure is 20 ~ 30 MPas, and extraction temperature is 30 ~ 40 DEG C, and flow velocity is 15 ~ 25 ls/h, and extraction time is 1 ~ 3 hour.
More preferably, described Herba Centipedae CO
2supercritical extract is at employing CO
2condition during supercritical extraction is: extracting pressure is 25 MPas, and extraction temperature is 38 DEG C, and flow velocity is 20 ls/h, and extraction time is 2 hours.
The optimal technical scheme that the present invention further provides is, described Herba Centipedae CO
2supercritical extract is through CO
2cyclohexane extraction, petroleum ether, chloroform or benzene is used to extract after supercritical extraction more further.
As the further preferred version of one, described Herba Centipedae CO
2supercritical extract is through CO
2use cyclohexane extraction, petroleum ether, chloroform or benzene to extract after supercritical extraction more further 1 ~ 5 time, combining extraction liquid, concentrates, is drying to obtain.
As one most preferably scheme, described Herba Centipedae CO
2supercritical extract is through CO
2use cyclohexane extraction, petroleum ether, chloroform or benzene to extract after supercritical extraction more further 2 ~ 4 times, combining extraction liquid, concentrates, is drying to obtain.
Herba Centipedae CO of the present invention
2supercritical extract, preparing the application aspect in the medicine or food with resisiting influenza virus effect, is applied to medicine or the food aspect of anti-avian influenza virus more targetedly, has good application of active.
More preferably, described influenza virus is H1N1, H5N1 or H9N2 type influenza virus.
As a kind of preferred version, described Herba Centipedae is feverfew Myriogyne minuta
centipedaminima(L.) A.Br.et.Aschers.
As one most preferably scheme, described Herba Centipedae is feverfew Myriogyne minuta
centipedaminima(L.) the wounded in the battle herb of A.Br.et.Aschers.
The present invention provides a kind of medicine of resisiting influenza virus simultaneously, and described medicine contains the Herba Centipedae extract for the treatment of effective dose and pharmaceutically acceptable carrier.
Described medicine can be made into tablet, capsule, granule, powder, ointment, oral liquid, spray or injection.
The present invention provides a kind of food of resisiting influenza virus simultaneously, and described food is beverage or health food, the Herba Centipedae extract containing flu-prevention effective dose and food acceptable carrier.
Beneficial effect of the present invention:
The present invention breaks through prior art and limits about the application of Herba Centipedae, Herba Centipedae extract is applied to the medicine preparing resisiting influenza virus or food and health product aspect, develops the using value that Herba Centipedae is new.
The invention provides Herba Centipedae and prepare the application in anti-influenza virus medicament.The present invention adopts suitable CO
2supercritical extraction process, extracts in a large number by the active substance of the resisiting influenza virus in Herba Centipedae, obtains higher extraction efficiency.
Further, the present invention is through CO
2with the extract that petroleum ether, cyclohexane extraction, chloroform or benzene extracting and refining obtain, there is better resisiting influenza virus effect again after supercritical extraction process extraction Herba Centipedae, its antivirus action is better than positive control drug ribavirin far away, has good medical applications and is worth.
Extract energy specific aim of the present invention solves the treatment technology difficult problem bringing medical circle of bird flu virus particularly H1N1, H5N1 or H9N2 type influenza virus.
Detailed description of the invention
The present invention is further illustrated below in conjunction with specific embodiment, unless stated otherwise, the source chemicals that the present invention adopts is the conventional commercial source chemicals of the art, and unless stated otherwise, the method and apparatus of employing is the method and apparatus of the art routine.
Embodiment 1
300g Herba Centipedae (is picked up from Guangdong, is accredited as feverfew Myriogyne minuta by the Guangzhou medical material company senior engineer wheat ball that shakes
centipedaminima(L.) A.Br.et.Aschers, following embodiment with) dry herb or wounded in the battle herb (following embodiment with) pulverize, cross 60 mesh sieves, be placed in CO
2in supercritical extraction instrument, extracting pressure is 25 MPas, extraction temperature is 38 DEG C, and flow velocity is 20 ls/h, and extraction time is 2 hours, obtains Herba Centipedae CO
2supercritical extract 20.3g.
Embodiment 2
The dry herb of 300g Herba Centipedae (pick up from Guangdong, identified by the Guangzhou medical material company senior engineer wheat ball that shakes) is pulverized, crosses 60 mesh sieves, be placed in CO
2in supercritical extraction instrument, extracting pressure is 40 MPas, extraction temperature is 50 DEG C, and flow velocity is 30 ls/h, and extraction time is 4 hours, obtains Herba Centipedae CO
2supercritical extract 22.6g.
Embodiment 3
The dry herb of 300g Herba Centipedae (pick up from Guangdong, identified by the Guangzhou medical material company senior engineer wheat ball that shakes) is pulverized, crosses 60 mesh sieves, be placed in CO
2in supercritical extraction instrument, extracting pressure is 20 MPas, extraction temperature is 30 DEG C, and flow velocity is 10 ls/h, and extraction time is 1 hour, obtains Herba Centipedae CO
2supercritical extract 14.6g.
Embodiment 4
The dry herb of 300g Herba Centipedae (pick up from Guangdong, identified by the Guangzhou medical material company senior engineer wheat ball that shakes) is pulverized, crosses 60 mesh sieves, be placed in CO
2in supercritical extraction instrument, extracting pressure is 25 MPas, extraction temperature is 38 DEG C, and flow velocity is 20 ls/h, and extraction time is 2 hours, obtains Herba Centipedae CO
2supercritical extract, by Herba Centipedae CO
2supercritical extract is dissolved in water to 1L, then extracts with the cyclohexane extraction of 1L, repeats to extract 3 times with cyclohexane extraction, merges the Herba Centipedae CO that cyclohexane extraction extract must be refined
2supercritical extract.
Embodiment 5
The dry herb of 300g Herba Centipedae (pick up from Guangdong, identified by the Guangzhou medical material company senior engineer wheat ball that shakes) is pulverized, crosses 60 mesh sieves, be placed in CO
2in supercritical extraction instrument, extracting pressure is 25 MPas, extraction temperature is 38 DEG C, and flow velocity is 20 ls/h, and extraction time is 2 hours, obtains Herba Centipedae CO
2supercritical extract, by Herba Centipedae CO
2supercritical extract is dissolved in water to 1L, then uses the petroleum ether extraction of 1L, repeats to use petroleum ether extraction 3 times, merges the Herba Centipedae CO that petroleum ether extraction liquid must be refined
2supercritical extract.
Embodiment 6
The dry herb of 300g Herba Centipedae (pick up from Guangdong, identified by the Guangzhou medical material company senior engineer wheat ball that shakes) is pulverized, crosses 60 mesh sieves, be placed in CO
2in supercritical extraction instrument, extracting pressure is 25 MPas, extraction temperature is 38 DEG C, and flow velocity is 20 ls/h, and extraction time is 2 hours, obtains Herba Centipedae CO
2supercritical extract, by Herba Centipedae CO
2supercritical extract is dissolved in water to 1L, then uses the chloroform extraction of 1L, repeats to use chloroform extraction 3 times, the Herba Centipedae CO that combined chloroform extract must be refined
2supercritical extract.
Embodiment 7
The dry herb of 300g Herba Centipedae (pick up from Guangdong, identified by the Guangzhou medical material company senior engineer wheat ball that shakes) is pulverized, crosses 60 mesh sieves, be placed in CO
2in supercritical extraction instrument, extracting pressure is 25 MPas, extraction temperature is 38 DEG C, and flow velocity is 20 ls/h, and extraction time is 2 hours, obtains Herba Centipedae CO
2supercritical extract, by Herba Centipedae CO
2supercritical extract is dissolved in water to 1L, then extracts with the benzene of 1L, repeats to extract 3 times with benzene, the Herba Centipedae CO that combined benzene extract must be refined
2supercritical extract.
Comparative example 1
By 300g Herba Centipedae (pick up from Guangdong, by Guangzhou medical material company senior engineer wheat shake ball identify, be feverfew Myriogyne minuta
centipedaminima(L.) A.Br.et.Aschers) dry herb pulverize, be the alcohol heating reflux extraction of 75% with 1 liter of concentration of volume percent, extraction time is 1h, repeat extraction 3 times, merge extractive liquid, concentrate drying obtains Herba Centipedae ethanol (75%, v/v) extract.
Comparative example 2
By 300g Herba Centipedae (pick up from Guangdong, by Guangzhou medical material company senior engineer wheat shake ball identify, be feverfew Myriogyne minuta
centipedaminima(L.) A.Br.et.Aschers) dry herb pulverize, by 1 premium on currency heating and refluxing extraction, extraction time is 1h, and repeat extraction 3 times, merge extractive liquid, concentrate drying obtains Herba Centipedae water extract.
Embodiment 8 Herba Centipedae CO
2supercritical extract is tested the inhibitory action of influenza A virus in mdck cell
Get mdck cell (to derive from and large university's pharmaceutical college's Chinese medicine and researches on natural drugs institute, do not limit the present invention, those skilled in the art also can adopt other described cells of originating to test) grow to 96 porocyte culture plates of monolayer, abandoning supernatant, with PBS(pH7.2 ~ 7.4) wash twice, every hole adds by MEM fluid medium (brand: U.S. HyClone respectively, purchased from: Hui Ying bio tech ltd, Shanghai, article No. SH30024.01B) medicinal liquid of doubling dilution and virus liquid 100 μ L, establish cell controls group in addition, virus control group, be placed in 37 DEG C, saturated humidity, 5%CO
2hatch in incubator.After 72h, 0.5mMWST-1(brand: Roche, purchased from: Jia Meiniunuo bio tech ltd, Beijing, article No.: 11644807001) solution joins (concentration of WST-1 is 0.25mM) in every hole.After 4 hours, survey absorbance by microplate reader at 450nm.IC
50account form is as follows: cell survival rate=(cell control well OD value-medicine feeding hole OD value) × 100%; Take drug level as X-axis, cell survival rate is Y-axis, and in Origin mapping, obtaining concentration corresponding to 50% cell survival rate is then IC
50.
The Herba Centipedae CO that embodiment 1 ~ 3 is prepared
2the refining Herba Centipedae CO that supercritical extract and 4 ~ 7 prepares
2the product of supercritical extract and comparative example 1 and 2 carries out anti-influenza A virus active testing, the results are shown in Table 1, product (the Herba Centipedae CO that embodiment 1 ~ 3 prepares
2supercritical extract) to A/PR8/34(H1N1), A/Duck/Guangdong/99 (H5N1), AChicken/Guangdong/96(H9N2) these three kinds of influenza A viruss all have certain antivirus action, and their half-inhibition concentration (IC
50) there is no significant difference with positive control agents Ribavirin, their resisiting influenza virus effect and the suitable of ribavirin are described.Product (the refining Herba Centipedae CO that embodiment 4 ~ 7 prepares
2supercritical extract) Herba Centipedae extract is to A/PR8/34(H1N1), A/Duck/Guangdong/99 (H5N1), A/Chicken/Guangdong/96(H9N2) half-inhibition concentration (IC of these three kinds of influenza virus
50) much smaller than positive control agents Ribavirin, product (the refining Herba Centipedae CO that embodiment 4 ~ 7 prepares is described
2supercritical extract) antivirus action be significantly better than ribavirin.(A/PR8/34(H1N1), A/Duck/Guangdong/99 (H5N1), A/Chicken/Guangdong/96(H9N2) three kinds of strains of influenza viruses are the conventional conventional strains of influenza viruses of this area experiment, the present embodiment Strain derives from that College of Veterinary Medicine, South China Agricultural University is old builds recent studies on person).Experiment shows, Herba Centipedae CO of the present invention
2the application that supercritical extract has in the medicine of resisiting influenza virus effect or food in preparation has good application prospect.
Table 1 Herba Centipedae extract or extract suppress the IC of influenza infection mdck cell
50value
Claims (7)
1. Herba Centipedae CO
2supercritical extract, preparing the application in the medicine or food with resisiting influenza virus effect, is characterized in that, described Herba Centipedae CO
2supercritical extract is through CO by Herba Centipedae
2carry out extraction 1 ~ 5 time through cyclohexane extraction, petroleum ether, chloroform or benzene again after supercritical extraction, combining extraction liquid, concentrated, drying obtains; Wherein, described CO
2the condition of supercritical extraction is: extracting pressure is 20 ~ 40 MPas, and extraction temperature is 30 ~ 50 DEG C, and flow velocity is 10 ~ 30 ls/h, and extraction time is 1 ~ 4 hour; Described influenza virus is H
1n
1type, H
5n
1or H
9n
2influenza virus.
2. application according to claim 1, is characterized in that, described Herba Centipedae CO
2the Herba Centipedae raw material of supercritical extract is feverfew Myriogyne minuta
centipedaminima(L.) A.Br.et.Aschers.
3. application according to claim 1, is characterized in that, described Herba Centipedae CO
2the Herba Centipedae raw material of supercritical extract is feverfew Myriogyne minuta
centipedaminima(L.) the wounded in the battle herb of A.Br.et.Aschers.
4. the application according to any one of claims 1 to 3, is characterized in that, described Herba Centipedae CO
2the CO of supercritical extract
2supercritical extraction condition is: extracting pressure is 20 ~ 30 MPas, and extraction temperature is 30 ~ 40 DEG C, and flow velocity is 15 ~ 25 ls/h, and extraction time is 1 ~ 3 hour.
5. application according to claim 4, is characterized in that, described Herba Centipedae CO
2the CO of supercritical extract
2supercritical extraction condition is: extracting pressure is 25 MPas, and extraction temperature is 38 DEG C, and flow velocity is 20 ls/h, and extraction time is 2 hours.
6. an anti-H
1n
1the medicine of type influenza virus, is characterized in that, described medicine contain treatment effective dose claim 1 described in Herba Centipedae CO
2supercritical extract and pharmaceutically acceptable carrier.
7. an anti-H
1n
1the food of type influenza virus, is characterized in that, described food is beverage or health food, Herba Centipedae CO described in the claim 1 containing flu-prevention effective dose
2supercritical extract and food acceptable carrier.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
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| 超临界CO2萃取法与水蒸气蒸馏法提取鹅不食草油的化学成分研究;杨艳芳等;《中药材》;20070731;第30卷(第7期);第808-811页 * |
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