CN101843640B - Natural anti- virus and composition comprising thereof - Google Patents

Natural anti- virus and composition comprising thereof Download PDF

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Publication number
CN101843640B
CN101843640B CN2010101833264A CN201010183326A CN101843640B CN 101843640 B CN101843640 B CN 101843640B CN 2010101833264 A CN2010101833264 A CN 2010101833264A CN 201010183326 A CN201010183326 A CN 201010183326A CN 101843640 B CN101843640 B CN 101843640B
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virus
nutrient solution
lactobacillus
antiviral agent
composition
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CN2010101833264A
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Chinese (zh)
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CN101843640A (en
Inventor
李正华
张东逸
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高台有限公司
李正华
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Priority to KR10-2004-0056532 priority
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES, AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/10Animals; Substances produced thereby or obtained therefrom
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12Antivirals
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12Antivirals
    • A61P31/14Antivirals for RNA viruses
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12Antivirals
    • A61P31/14Antivirals for RNA viruses
    • A61P31/16Antivirals for RNA viruses for influenza or rhinoviruses

Abstract

Disclosed herein are a natural antiviral agent having high inhibitory activity against a broad spectrum of viruses without adverse side effects, obtainable using lactic acid bacteria isolated from kimchi, and a composition comprising the natural antiviral agent. The natural antiviral agent is safe to the body and can be applied to compositions, such as fodder, beverages, food, medicine, and cosmetics, imparting antimicrobial activity thereto.

Description

Natural anti-virus thing and the composition that comprises it

The application is international application no PCT/KR2005/002348, international filing date on July 20th, 2005, China's application number 200580024612.3 enters Chinese January 22 2007 date, and denomination of invention is divided an application for " natural anti-virus thing and the composition that comprises it ".

Technical field

The present invention relates to a kind of natural antiviral agent and the composition that comprises it.More specifically, the present invention relates to a kind of application from the natural antiviral agent of the milk-acid bacteria preparation of pickles (Kimchi) separation, its height that shows for viral growth suppresses active, and be free from side effects, and be safe to body, the invention still further relates to the composition comprising described natural antiviral agent, such as beverage, feed, food, medicine, makeup, etc.

Background technology

Causing the influenza virus of influenza pandemic once every year, is a kind of alkaline single strand RNA virus, and its size with the described RNA of parcel is the coating of 1/10,000mm.According to antigenicity, influenza virus is divided into A, B and C type.Among them, influenza type A and B virus cause prevailing disease.Be called hemagglutinin (HA) and neuraminidase (neuramidase) glycoprotein of two types (NA), outstanding from the surface as the projection, and 8 kinds of RNA fragments to be present in virus inner.Two kinds of surface glycoprotein HA of influenza virus and NA experience variation in identical hypotype usually, this causes the appearance of annual new antigenic mutant.

When cough, from lung's ejection, and enter in other body by nose or mouth together with the air of influenza virus and fierce release.In vivo, influenza virus is used viral glycoprotein h A projection, adheres to and penetrate the mucosal epithelium cell of the upper respiratory tract, and here breeds subsequently.The result of study of accumulation shows the definite mechanism of virus infection at present.Usually, on the target cell surface, the receptors bind that virus and sugar chain by glycoprotein form, and experience endocytosis and the fusion of peplos and endosome subsequently, and enter target cell.At cell interior, virus by transcribing and copy them gene and produce progeny virion from their host's plasma membrane and breed.The infection of influenza virus usually be accompanied by fever, headache, arthrodynia, whole body collapse, etc., and have respiratory symptom, comprise have a sore throat, have a running nose, have a stuffy nose, etc.Described influenza is different from flu on symptom and severity, and is characterised in that most evil bad infectivity, and this causes the fulminant prevailing disease of short period of time in the whole world.Because influenza virus has the annual HA structure that all makes a variation, the antibody that produces so reply previous infection can not be effectively for occurent prevailing disease, and this is why popular reason of influenza.

For the infection of flu-prevention virus, scientists planned and has been used for suppressing to be attached to epithelial cell, penetrates cell, transcribes, copies, translates and discharge from cell the method for filial generation, and all these is the target of resisiting influenza virus medicament.Under these methods, developed antiviral agent, such as amantadine, Rimantadine, zanamivir etc., but, owing to it is reported that they produce significant adverse side effect in central nervous system, Digestive tract and autonomic nervous system, such as hypersensitivity, so must use extremely carefully.

As indicated above, the mucosal epithelium of influenza infection respiratory tract, and breeding there.If a kind of Virus Type of vaccine is different from described popular influenza virus, so inoculation may not can flu-prevention Viral infection and propagation effectively.In fact, not for effectively treating in the treatment of influenza virus or the prevention medicament.Now, often brushing teeth, keep throat moistening, and have a rest fully and nutrition, is to be known as the prevention method of effective influenza infection.

Due to these reasons, there is the urgent need that suppresses active and usually can there is no the antiviral agent of adverse side effect ground application for having height for influenza virus.

Virus disease is one of dangerous factor with destroying poultry industry in the largest ground.Now, it is said that avian influenza virus becomes the threat of the universe and poultry industry.Recently, reported continually the outburst of H5N1 high pathogenic avian influenza (HPAI) in China and South East Asia bird.At brainstrust, the adaptation of avian influenza virus height suddenlys change, rather than by radiation, this causes the fearful problem to the mankind from present situation.

In Indonesia, have been found that for bird para-influenzal popular can not the measure of employing prevention property.In China, many H5N1 HPAI virus detected, and they with than its control speed faster speed propagate.Therefore, have no idea except inoculating.In fact, inoculation is just carried out in many areas.

Vietnam and Thailand are in more serious situation, have the report that the birds influenza that increases number has caused mankind's death.The possibility of influenza pandemic is not almost controlled in these countries in worse ground.Thailand, typical bird exporting country, the outburst of response bird flu is adhered to and will is in the policy that dangerous bird slaughters rather than inoculate.In order to eradicate bird flu, the Thai Government has been taked extreme measure, to without permission and immunity the personnel of poultry impose a fine.Reason is exactly, and can not eradicate bird flu in China because inoculation makes, and Thailand has to stop the bird outlet.

In nearly all country with bird industry, fowl source infectious bronchitis (IB) is greatly to destroy the representative disease of Poultry farming industry.In practice, even some countries have eradicated newcastle disease (ND) or bird flu (AI), also do not have up till now country to stop IB.

In Korea S, the inspection data of the animal doctor of country research and quarantine service organization (National Veterinary Researchand Quarantine Service) show, IB is most popular, and on the generation sum of poultry disease, it comprises 6% and comprised 5.2% in 2002 in calendar year 2001.IB virus is to study the most deep coronavirus, and has incredible simple structure.IB virus also is called removable virus, at present just in whole world experience sudden change.Simultaneously during infection host, described sudden change is mainly caused by the gene recombination of serotype IB virus differing from each other when them.When the IB of two or more types virus while infection host, they are accompanied by the restructuring of their gene at identical Intracellular growth, and this causes having the appearance of the new virus of the feature that is different from those viral features of origin.For example, if gene recombination occurs between attenuated vaccine virus and pernicious wild-type virus, there is the possibility that increases, namely, the advantage of described two-strain, that is, and higher proliferation and pathogenic, in connection with, and the progeny virus that therefore obtains becomes the acute illness initiator of fast breeding most probably.Due to this reason, vaccine, even attenuation, necessarily can not be introduced into domestic harmlessly.

The typical characteristics of IB virus comprises the lasting appearance of different serotypes and mutation, the ability (latent period of 18-36 hour) of propagating sooner than any Other diseases, and the survival ability (persistence in the farm) in bird colony.In addition, defence and the cross immunity between serotype for the serum antibody of described virus is limited.And, when two or more serotypes are attacked bird colony simultaneously, be difficult to only with the described virus of vaccine defence.

In Korea S, ND is caused by Avian pneumo-encephalitis virus, has been identified the communicable disease that the first is legal, and occur its every year in the hennery.Developed the various vaccines based on the ND inactivation of virus, yet the generation of ND does not reduce still.

Now, the hennery has 3 defence lines that suppress bird disease.Article one, the defence line depends on Biosafety, and second defence line depends on inoculation, and the 3rd immunity system of defending line to depend on bird self.Yet the generation of IB in the hennery proves that on the contrary IB virus crossed article one defence line of Biosafety, makes inoculation useless, and destroyed the health of chicken, destroyed immunity system fully.Therefore, there is urgent need for killing viral therapeutical agent or extremely strengthening immune multifunctional treating agent.

Summary of the invention

Technical problem

Therefore, the present invention keeps it in mind the problems referred to above that exist in the prior art, and an object of the present invention is to provide novel natural antiviral agent, its height that has for various virus infectiones effectively suppresses active, and be the disadvantageous side effect that do not have to the human body tight security, described virus comprises influenza virus, avian influenza virus, fowl source infectious bronchitis virus etc.

In addition, another object of the present invention is to provide and contains described natural antiviral agent as the composition of activeconstituents, and it can be contained in various products.

Technical scheme

According to an aspect of of the present present invention, purpose of the present invention can realize like this, namely, provide from a kind of of the group of selecting free following composition and the natural antiviral agent of preparation: the nutrient solution of the milk-acid bacteria that separates from pickles the pickle fermentation process, the enriched material of described nutrient solution, the dehydrate of described nutrient solution, and combination.

In described natural antiviral agent, the group that milk-acid bacteria can select free leuconos toc (Leuconostocspp.), lactobacillus (Lactobacillus spp.), Wei Si Bordetella (Weissella spp.) and combination thereof to form.

preferably, the leuconos toc that is used for natural antiviral agent of the present invention comprises lemon leukonid (Leuconostoc citreum), leuconostoc lactis (Leuconostoc lactis), Leuconostoc mesenteroides dextranicum (Leuconostoc mesenteroides subsp.Dextranicum), leuconostoc mesenteroides subsp mesenteroides (Leuconostoc mesenteroides subsp.Mesenteroides), Argentina leukonid (Leuconostoc argentinum), yellowish pink leukonid (Leuconostoc carnosum), hard leukonid (Leuconostoc gellidum), pickles leukonid (Leuconostoc kimchii), Leuconostoc inhae, Leuconostoc gasicomitatum, and their combination.

preferably, the lactobacillus that is used for natural antiviral agent of the present invention comprises short lactobacillus (Lactobacillus brevis), Lactobacterium acidophilum (Lactobacillus acidophilus), lactobacillus bulgaricus (Lactobacillus bulgaricus), plant lactobacillus (Lactobacillusplantarum), pickles Bacterium lacticum (Lactobacillus kimchii), class plant lactobacillus (Lactobacillus para-plantarum), lactobacillus curvatus subspecies (Lactobacillus curvatussubsp.Curvatus), Lactobacillus sakei subsp.Sakei, and their combination.

Preferably, the Wei Si Bordetella that is used for natural antiviral agent of the present invention comprises Weissellakoreens, Weissella hanii, pickles Wei Si Salmonella (Weissella kimchii), Weissella soli, merges Wei Si Salmonella (Weissella confusa), and their combination.

According to another aspect of the present invention, purpose of the present invention can comprise that natural antiviral agent realizes as the composition of activeconstituents by providing a kind of, described natural antiviral agent prepares from a kind of of the group of selecting free following composition: the nutrient solution of the milk-acid bacteria that separates from pickles the pickle fermentation process, the enriched material of described nutrient solution, the dehydrate of described nutrient solution, and combination.

In this, the described composition form of one of beverage, feed, food, medicine or makeup preferably.

Advantageous effects

The nutrient solution of pure lactobacillus, or their enriched material or dry substance have the fabulous antimicrobial acivity for wide spectrum virus, and can give food, feed, beverage, medicine and cosmetics safety effective antiviral activity.

Implement optimal mode of the present invention

About the present invention, the inventor carry out about foods and cosmetics preserve deeply and thorough research causes such discovery, namely, milk-acid bacteria (hereinafter referred to as pure lactobacillus) culture that separates from pickles can act as multi-functional natural antiviral agent, and it has for the effective inhibition of wide spectrum virus active.Known ground, pure lactobacillus is bred in the pickle fermentation process, it is usually by using the pickled Rhaphanus of salt, Caulis et Folium Brassicae capitatae and cucumber, described pickled prod is mixed with the seasoning composition that comprises red chilly powder, mashed garlic, green onion end, salted marine food etc., and described pickling vegetable is fermented to prepare.

The invention pattern

Better understanding of the present invention can obtain by following example, and described example proposes for illustrating, but can not be interpreted as restriction of the present invention.

Embodiment

Preparation pure lactobacillus nutrient solution

In pure lactobacillus, pickles leukonid (KCTC2386), leuconostoc mesenteroides subsp mesenteroides (KCTC3530), Leuconostoc citreum (KCTC3524), leuconostoc lactis (KCTC3528), plant lactobacillus (KCCM11322) and Weissella koreensis (KCTC3746) are for the preparation of described nutrient solution.

At first, 100ml MRS (Difco) meat soup 121 ℃ of autoclavings 15 minutes, and is inoculated into pure lactobacillus in meat soup, and 30 ℃ of wave and culture 16 hours.

, after the centrifugal 3min of 5000 * g supernatant liquid is filtered by the membrane filter with 0.22 oral pore size at the culture that will obtain, to obtain sterile solution.

Subsequently, this sterile solution is held back the film of size by having 10,000Da.Resulting filtrate is held back big or small membrane concentration with having 1,000Da.The enriched material that obtains is like this filtered by the membrane filter with 0.22 oral pore size again, in order to keep sterile state.In order to get rid of the pH-dependency effect of antimicrobial acivity, final nutrient solution is adjusted to pH 7.

-detect the pure lactobacillus nutrient solution for the antiviral activity of avian influenza virus-

The animal cell line (MDCK (Madin-Darby dog kidney) or CEF (chicken embryo fibroblast)) of monolayer growth is infected with avian influenza virus HK/220, then cultivated 3 days.Plaque Formation has proved virus infection.

To be diluted to 10 with PBS (pH 7.4) -3Avian influenza virus and with the mixture of the pure lactobacillus nutrient solution of identical buffering salt dilution, add on flat board with monolayer growth converged 80% or more animal cell line MDCK or CEF in, subsequently described clone was cultivated 3 days.Neutral red staining is fixed and used to the cell that infects, to observe the wherein formation of plaque.Result provides in table 1, and is as follows.

Table 1

Apparent from the data of table 1, do not allow to reach 1.3~1.6 * 10 although do not contain the contrast of pure lactobacillus nutrient solution 2Individual Plaque Formation, but add pickles dish lactic acid bacteria culture solution to reduce significantly the plaque number that forms.Therefore, the described pickles dish lactic acid bacteria culture solution of these result hints has very effective antiviral activity for avian influenza virus.

-detect the pure lactobacillus nutrient solution for the antiviral activity of influenza virus-

Replace avian influenza virus HK/220 except using influenza virus A 1H3N2, repeat and step identical in the antiviral activity for avian influenza virus detects.Described result provides in table 2, and is as follows.

Table 2

What there is no described pure lactobacillus nutrient solution although wherein only used the viral dilution thing has 1.3~1.6 * 10 to impinging upon each culture plate 2Individual plaque, as shown in table 2, but when having the pure lactobacillus nutrient solution, the infection of influenza virus has developed the plaque of remarkable minimizing number, and the described pure lactobacillus nutrient solution of this hint has very effective antiviral activity for influenza virus.

The animal detection of-pure lactobacillus nutrient solution-

1,100 hen of laying eggs of the egg productivity of minimizing is divided into 10 groups at random to occur experience significantly in the time of will be due to bird flu and newcastle disease, and every group comprises 110 hens.The pure lactobacillus nutrient solution of above preparation is diluted 10 times in the hen tap water, and observe the variation of egg productivity in the process of 17 days.Result provides in following table 3.

in table 3, sample 1 prepares based on pickles leukonid (KCTC2386) nutrient solution, sample 2 is based on leuconostoc mesenteroides subsp mesenteroides (KCTC3530) nutrient solution, sample 3 is based on Leuconostoccitreum (KCTC3524) nutrient solution, sample 4 is based on leuconostoc lactis (KCTC3528) nutrient solution, sample 5 is based on plant lactobacillus (KCCM11322) nutrient solution, sample 6 is based on Weissellakoreensis (KCTC3746) nutrient solution, sample 7 is based on the mixture of leuconostoc mesenteroides subsp mesenteroides (KCTC3530) nutrient solution and plant lactobacillus (KCCM11322) nutrient solution, sample 8 is based on the mixture of pickles leukonid (KCTC2386) nutrient solution and Weissella koreensis (KCTC3746) nutrient solution, sample 9 is based on the mixture of Weissella koreensis (KCTC3746) nutrient solution and plant lactobacillus (KCCM11322) nutrient solution and prepare.

Table 3

As shown in table 3, cause egg productivity to reduce, suffer from dysentery and show the bird inlay of unusual respiratory symptom owing to infecting virus, drink to them contain according to the water of nutrient solution of the present invention after 10 days, return to standard state, this determines by the level that egg productivity returns to normal hen.Therefore, also has fabulous interior resisting virus activity according to pure lactobacillus nutrient solution of the present invention.

In addition, because natural antiviral agent of the present invention is to prepare from eugonic pure lactobacillus nutrient solution between the food pickles yeast phase of Korean traditional, therefore, detect even without carrying out extra security, about described natural anti-virus product, there is not suspection in the security of body.

As in the tap water of bird inlay, even without carrying out concrete detection, the nutrient solution of antiviral activity of the present invention also can be included in food, feed, beverage, makeup and medicine.

When nutrient solution of the present invention is used for makeup, beverage, food, feed, medicine etc., it should be appreciated by those skilled in the art, according to the characteristic of described product and the embodiment of preparation, distribution, storage and the application of described product, various modifications, interpolation and replacement be can carry out in described nutrient solution, and the present invention scope of disclosure and spirit in the claim of enclosing do not deviated from.

Industrial usability

In sum, the data of acquisition show in an embodiment, and the pure lactobacillus nutrient solution has the fabulous antiviral activity for broad-spectrum micro-organisms, and can give food, feed, beverage, medicine and cosmetics safety and effective antiviral activity.

Claims (1)

1. the application of composition in the preparation antiviral agent, wherein said composition comprise from a kind of of the group of selecting free following composition and the natural antiviral agent of preparation as activeconstituents: Wei Si Bordetella species (Weissella spp.) nutrient solution that separates from pickles the pickle fermentation process, the enriched material of described nutrient solution, the dehydrate of described nutrient solution, and their combination, wherein said virus is influenza virus, avian influenza virus or Avian pneumo-encephalitis virus, wherein said Wei Si Bordetella species are Weissella koreensis.
CN2010101833264A 2004-07-20 2005-07-20 Natural anti- virus and composition comprising thereof CN101843640B (en)

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KR20050087742A (en) 2005-08-31
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CN101843640A (en) 2010-09-29
EP1781308A4 (en) 2009-07-01
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