CN1997378B - natural antiviral composition - Google Patents

Abstract

The present application discloses a natural antiviral agent and a composition comprising the natural antiviral agent. The natural antiviral agent has high inhibitory activity against a broad spectrum of viruses without adverse side effects and can be obtained using lactic acid bacteria isolated from kimchi. The natural antiviral agent is safe for the human body and can be used in compositions such as feed, beverages, food, medicine, and cosmetics to impart antimicrobial activity.

Description

natural antiviral composition

technical field

The present invention relates to a natural antiviral agent and compositions comprising the same. More specifically, the present invention relates to a natural antiviral agent prepared by using lactic acid bacteria isolated from Kimchi (Kimchi), which exhibits high inhibitory activity against virus growth without side effects and is safe to the body. It relates to compositions, such as beverages, feeds, foods, medicines, cosmetics, etc., in which said natural antiviral agents are included. the

Background technique

The influenza virus that causes the annual influenza epidemic is a basic single-stranded RNA virus that has an envelope with a size of 1/10,000mm enclosing the RNA. According to antigenicity, influenza viruses are divided into A, B and C types. Among them, influenza A and B viruses cause epidemics. Two types of glycoproteins, called hemagglutinin (HA) and neuramidase (NA), protrude like protrusions from the surface, and eight RNA segments exist inside the virus. The two surface glycoproteins HA and NA of influenza viruses often undergo variation within the same subtype, which leads to the emergence of new antigenic mutants each year. the

When coughing, the flu virus is expelled from the lungs with a violent release of air and enters the rest of the body through the nose or mouth. In vivo, influenza virus, using the viral glycoprotein HA protrusion, attaches to and penetrates the mucosal epithelium of the upper respiratory tract, where it subsequently proliferates. Accumulated findings so far indicate the exact mechanism of viral infection. Generally, on the surface of target cells, viruses bind to receptors composed of sugar chains of glycoproteins, and undergo endocytosis and subsequent fusion of viral envelopes with endosomes to enter target cells. Inside cells, viruses multiply by transcribing and replicating their genes and producing progeny virus particles from the plasma membrane of their host. Influenza virus infection is usually accompanied by fever, headache, joint pain, general prostration, etc., and respiratory symptoms including sore throat, runny nose, nasal congestion, etc. The influenza is different from colds in symptoms and severity, and is characterized by extremely bad infectivity, which leads to short-term outbreak-type epidemics all over the world. Because influenza viruses have an HA structure that mutates every year, antibodies produced in response to earlier infections are not effective against ongoing epidemics, which is why influenza circulates. the

To prevent influenza virus infection, scientists have devised methods for inhibiting attachment to epithelial cells, cell penetration, transcription, replication, translation, and release of progeny from cells, all of which are targets of anti-influenza virus agents. Under these approaches, antiviral agents such as amantadine, rimantadine, zanamivir, etc. have been developed, however, since they are reported to produce significant adverse side effects in the central nervous system, digestive system, and autonomic nervous system , such as hypersensitivity, so must be applied with extreme caution. the

As mentioned above, influenza viruses infect and multiply in the mucosal epithelium of the respiratory tract. If a vaccine is of a different virus type than the circulating influenza virus, vaccination may not be effective in preventing infection and multiplication of the influenza virus. In fact, there are no therapeutically effective therapeutic or prophylactic agents against influenza virus. Now, regular brushing of teeth, keeping your throat moist, and getting enough rest and nutrition are the most effective prevention methods known for flu infection. the

For these reasons, there is an urgent need for antiviral agents that have high inhibitory activity against influenza virus and can generally be applied without adverse side effects. the

Viral diseases are one of the factors that most endanger and destroy the poultry industry. Now, the avian influenza virus is said to be a threat to all human beings as well as the poultry industry. Recently, outbreaks of H5N1 highly pathogenic avian influenza (HPAI) in poultry in China and Southeast Asia have been frequently reported. From the current situation it seems to experts that bird flu viruses are highly adapted to undergo mutations, rather than being irradiated, which leads to dire problems for humans. the

In Indonesia, it has been found impossible to take preventive measures against the prevalence of avian influenza. In China, many H5N1 HPAI viruses have been detected and they are spreading faster than they can control. Therefore, there is no other way but to vaccinate. In fact, vaccinations are being carried out in many areas. the

Vietnam and Thailand are in a more serious situation, with increased numbers of reports of avian influenza that has caused human deaths. Worse, there is little possibility of controlling influenza epidemics in these countries. Thailand, a typical poultry exporting country, responded to an outbreak of bird flu by maintaining a policy of culling at-risk birds rather than vaccinating them. In an effort to eradicate bird flu, the Thai government has taken drastic measures, imposing fines on those who vaccinate poultry without permission. The reason is that Thailand had to stop poultry exports because vaccination made it impossible to eradicate bird flu nationwide. the

In almost all countries with a poultry industry, avian infectious bronchitis (IB) is the representative disease that greatly destroys the poultry farming industry. In practice, even though some countries have eradicated Newcastle disease (ND) or avian influenza (AI), no country has terminated IB so far. the

In Korea, inspection data from the National Veterinary Research and Quarantine Service show that IB is the most prevalent, comprising 6% in 2001 and 5.2% in 2002 of the total incidence of poultry diseases . The IB virus is the best studied coronavirus and has an incredibly simple structure. IB virus, also called mobile virus, is currently undergoing mutations all over the world. The mutation is mainly caused by genetic recombination of IB viruses whose serotypes are different from each other when they simultaneously infect a host. When two or more types of IB viruses simultaneously infect a host, they grow within the same cell, accompanied by recombination of their genes, which leads to the emergence of novel viruses with characteristics different from those of the virus of origin. For example, if genetic recombination occurs between an attenuated vaccine virus and a vicious wild-type virus, there is an increased likelihood that the advantages of the two viruses, namely high proliferative and pathogenicity, will combine, and thus The resulting progeny viruses are highly likely to be rapidly proliferating agents of acute disease. For this reason, vaccines, even attenuated ones, must not be introduced into the country without damage. the

Typical properties of IB viruses include the persistent emergence of different serotypes and variants, the ability to spread faster than any other disease (18-36 hour incubation period), and the ability to survive in poultry populations (persistence on farms). In addition, the defense of serum antibodies against the virus and cross-immunity between serotypes is limited. Also, when two or more serotypes attack a poultry population at the same time, it is difficult to protect against the virus with a vaccine alone. the

In Korea, ND, caused by Newcastle disease virus, has been recognized as the first notifiable infectious disease, which occurs annually in poultry farms. Various vaccines based on ND virus inactivation have been developed, however, the occurrence of ND has still not decreased. the

Poultry farms now have 3 lines of defense against bird disease. The first line of defense depends on biosecurity, the second on vaccination, and the third on the bird's own immune system. However, the occurrence of IB in poultry farms proves to the contrary that the IB virus crosses the first line of defense in biosecurity, rendering vaccination useless and destroying the body of the chicken, completely destroying the immune system. Therefore, there is an urgent need for therapeutic agents that kill viruses or multifunctional therapeutic agents that abnormally enhance the immune system. the

Contents of the invention

technical problem

Therefore, the present invention keeps the above-mentioned problems in the prior art in mind, and an object of the present invention is to provide novel natural antiviral agents which have highly effective inhibitory activity against various viral infections and are highly effective against human body. The safety has no adverse side effects, and the viruses include influenza virus, avian influenza virus, avian infectious bronchitis virus and the like. the

In addition, another object of the present invention is to provide a composition containing the natural antiviral agent as an active ingredient, which can be included in various products. the

Technical solutions

According to an aspect of the present invention, the object of the present invention can be achieved by providing a natural antiviral agent prepared from one selected from the group consisting of: cultivation of lactic acid bacteria isolated from Kimchi during fermentation of Kimchi solution, a concentrate of said culture solution, a dehydrated product of said culture solution, and combinations thereof. the

Among the natural antiviral agents, lactic acid bacteria may be selected from the group consisting of Leuconostoc spp., Lactobacillus spp., Weissella spp., and combinations thereof. the

Preferably, the Leuconostoc genus used in the natural antiviral agent of the present invention includes Leuconostoc citreum, Leuconostoc lactis, Leuconostoc mesenteroides subsp. .Dextranicum), Leuconostoc mesenteroides subsp. Mesenteroides, Leuconostoc argentinum, Leuconostoc carnosum, Leuconostoc gellidum, Kimchi Ming Leuconostoc kimchii, Leuconostoc inhae, Leuconostoc gasicomitatum, and combinations thereof. the

Preferably, the Lactobacillus genus used in the natural antiviral agent of the present invention includes Lactobacillus brevis, Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus plantarum, pickles Lactobacillus kimchii, Lactobacillus para-plantarum, Lactobacillus curvatus subsp. Curvatus, Lactobacillus sakei subsp. Sakei, and combinations thereof. the

Preferably, the Weissella genus used in the natural antiviral agent of the present invention includes Weissellakoreens, Weissella hanii, Weissella kimchii, Weissella soli, Weissella confusa, and their combination. the

According to another aspect of the present invention, the object of the present invention can be achieved by providing a composition comprising, as an active ingredient, a natural antiviral agent selected from one selected from the group consisting of Preparation: a culture solution of lactic acid bacteria isolated from pickles during the fermentation process of pickles, a concentrate of the culture solution, a dehydrated product of the culture solution, and combinations thereof. the

In this regard, the composition is preferably in the form of one of beverage, feed, food, medicine or cosmetic. the

beneficial effect

The culture solution of Kimchi lactic acid bacteria, or their concentrates or dried substances have excellent antimicrobial activity against a broad spectrum of viruses, and can impart safe and effective antiviral activity to foods, feeds, beverages, drugs and cosmetics. the

The best mode for implementing the present invention

With regard to the present invention, intensive and thorough studies on preservation of food and cosmetics conducted by the present inventors have led to the discovery that a culture of lactic acid bacteria isolated from Kimchi (hereinafter referred to as Kimchi lactic acid bacteria) can act as a multifunctional natural disease-resistant Toxic agent with potent inhibitory activity against a broad spectrum of viruses. It is known that Kimchi lactic acid bacteria proliferate during the fermentation process of Kimchi, which is generally obtained by pickling radish, cabbage, and cucumber with salt, combining the pickles with seasonings including chili powder, garlic paste, green onion powder, salted seafood, etc. mixture and fermenting the pickled vegetables. the

invention model

A better understanding of the invention may be gained through the following examples, which are presented for purposes of illustration and are not to be construed as limitations of the invention. the

Example

Preparation of Kimchi Lactic Acid Bacteria Culture Solution

Among the kimchi lactic acid bacteria, Leuconostoc kimchi (KCTC2386), Leuconostoc enterica subsp. enterica (KCTC3530), Leuconostoc citreum (KCTC3524), Leuconostoc lactis (KCTC3528), Lactobacillus plantarum (KCCM11322), and Weissella koreensis (KCTC3746) was used to prepare the culture medium. the

First, 100 ml of MRS (Difco) broth was autoclaved at 121° C. for 15 minutes, and Kimchi lactic acid bacteria were inoculated into the broth, and cultured with shaking at 30° C. for 16 hours. the

After the obtained culture was centrifuged at 5000×g for 3 min, the supernatant was filtered through a membrane filter having a pore size of 0.22 D to obtain a sterile solution. the

Subsequently, this sterile solution was passed through a membrane with a cut-off of 10,000 Da. The resulting filtrate was concentrated using a membrane with a 1,000 Da cut-off. The concentrate thus obtained was filtered again through a membrane filter having a pore size of 0.22 D in order to maintain sterility. To exclude pH-dependent effects of antimicrobial activity, the final culture broth was adjusted to pH 7. the

- Detection of antiviral activity of pickle lactic acid bacteria culture solution against avian influenza virus -

Monolayer-grown animal cell lines (MDCK (Madin-Darby canine kidney) or CEF (chicken embryonic fibroblasts)) were infected with avian influenza virus HK/220 and cultured for 3 days. Plaque formation evidenced viral infection. the

A mixture of avian influenza virus diluted to 10 ^-3 with PBS (pH 7.4) and culture solution of Kimchi lactic acid bacteria diluted with the same buffer salt was added to animals grown confluent by 80% or more in a monolayer on a plate In the cell lines MDCK or CEF, the cell lines were then cultured for 3 days. Infected cells were fixed and stained with neutral red to visualize plaque formation therein. The results are given in Table 1, below.

Table 1

As apparent from the data in Table 1, although the control without Kimchi lactic acid bacteria culture solution allowed as many as 1.3˜1.6×10 ² plaques to be formed, the addition of Kimchi lactic acid bacteria culture solution significantly reduced the number of plaques formed. Therefore, these results suggest that the kimchi vegetable lactic acid bacteria culture solution has very effective antiviral activity against avian influenza virus.

- Detection of the antiviral activity of Kimchi lactic acid bacteria culture fluid against influenza virus -

Except that influenza virus A1H3N2 was used instead of avian influenza virus HK/220, the same steps as in the detection of antiviral activity against avian influenza virus were repeated. The results are given in Table 2, as follows. the

Table 2

Although the control in which only the virus dilution was applied without the Kimchi lactic acid bacteria culture had 1.3 to 1.6× ¹⁰² plaques per culture plate, as shown in Table 2, in the presence of the Kimchi lactic acid bacteria culture, influenza Infection with the virus developed a significantly reduced number of plaques, suggesting that the Kimchi lactic acid bacteria culture solution has very potent antiviral activity against influenza virus.

-Animal detection of Kimchi lactic acid bacteria culture fluid-

1,100 laying hens that experienced significantly reduced egg production due to the co-occurrence of avian influenza and Newcastle disease were randomly divided into 10 groups, each group consisting of 110 hens. The kimchi lactic acid bacteria culture solution prepared above was diluted 10 times in the drinking water of hens, and the change of egg production was observed in the course of 17 days. The results are given in Table 3 below. the

In Table 3, sample 1 is prepared based on the culture solution of Leuconostoc kimchi (KCTC2386), sample 2 is based on the culture solution of Leuconostoc enterica subspecies (KCTC3530), sample 3 is based on the culture solution of Leuconostoccitreum (KCTC3524), and sample 4 is based Leuconostoc lactis (KCTC3528) culture fluid, sample 5 is based on the culture fluid of Lactobacillus plantarum (KCCM11322), sample 6 is based on the culture fluid of Weissellakoreensis (KCTC3746), and sample 7 is based on the culture fluid of Leuconostoc enterica subspecies (KCTC3530) and The mixture of Lactobacillus plantarum (KCCM 11322) culture fluid, sample 8 is based on the mixture of Kimchi Leuconostoc (KCTC2386) culture fluid and Weissella koreensis (KCTC3746) culture fluid, and sample 9 is based on Weissella koreensis (KCTC3746) culture fluid and Lactobacillus plantarum ( KCCM11322) culture medium mixture. the

table 3

As shown in Table 3, laying hens whose egg production decreased due to virus infection, suffered from dysentery and exhibited abnormal respiratory symptoms recovered 10 days after they were given water containing the culture solution according to the present invention. to normal, which is determined by the return of egg production to normal hen levels. Therefore, the Kimchi lactic acid bacteria culture solution according to the present invention also has excellent in vivo antiviral activity. the

In addition, since the natural antiviral agent of the present invention is prepared from the Kimchi lactic acid bacteria culture fluid that thrives during the fermentation of kimchi, a traditional Korean food, even without additional safety tests, the natural antiviral product has no effect on There is no doubt about the safety of the body. the

As in the drinking water of laying hens, the culture solution of the antiviral activity of the present invention can be contained in foods, feeds, beverages, cosmetics, and medicines even without specific testing. the

When the culture solution of the present invention is used for cosmetics, beverages, food, feed, medicine, etc., those skilled in the art should understand that according to the characteristics of the product and the embodiment of preparation, distribution, storage and application of the product, it can be Various modifications, additions and substitutions can be made in the culture medium without departing from the scope and spirit of the present invention as disclosed in the appended claims. the

Industrial Applicability

In summary, the data obtained in the examples show that Kimchi lactic acid bacteria culture fluid has excellent antiviral activity against a broad spectrum of microorganisms, and can impart safe and effective antiviral activity to food, feed, beverages, drugs and cosmetics . the

Claims (2)

1. Use of a composition for the preparation of an antiviral agent, wherein the composition comprises as an active ingredient a natural antiviral agent prepared from one selected from the group consisting of: Leuconostoc spp. culture fluid, a concentrate of said culture fluid, a dehydrated product of said culture fluid, and combinations thereof. 2. the application that claim 1 proposes, wherein said Leuconostoc comprises Leuconostoc citreum (Leuconostoc citreum), Leuconostoc lactis (Leuconostoc lactis), Leuconostoc mesenteroides subsp. dextranicum), Leuconostoc mesenteroides subsp. Mesenteroides, Leuconostoc argentinum, Leuconostoc carnosum, Leuconostoc gellidum, Leuconostoc kimchi (Leuconostoc kimchii), Leuconostoc inhae, Leuconostoc gasicomitatum, and combinations thereof.