KR20190129322A - Composition for anti-virus Comprising Nano-Sized Lactic Acid Bacteria from Kimchi - Google Patents

Abstract

The present invention relates to an anti-influenza virus pharmaceutical composition using kimchi lactobacillus and a health functional food for alleviating and preventing influenza virus infectious diseases. More specifically, the present invention relates to a cosmetic composition using nano-sized dead cells of lactobacillus plantarum, excellent in an inhibitory effect of viruses.

Description

Composition for anti-virus Comprising Nano-Sized Lactic Acid Bacteria from Kimchi}

The present invention relates to an antiviral composition using kimchi lactic acid bacteria, and more specifically, to pharmaceutical composition and health functional composition using nano-organized microorganisms of Lactobacillus plantarum lactic acid bacteria, characterized in that it effectively controls influenza virus. It is about food.

Kimchi is a traditional Korean food fermented by lactic acid bacteria, and the lactic acid bacteria classified in kimchi are Leuconostoc mesenteroides, Leuconostoc dextranicum, Lactobacillus brevis, Lactobacillus plantarum, Pediococcus pentosacues, and the like.

Lactic acid bacteria (lactic acid bacteria, lactobacillus) is a bacterium that breaks down sugars such as glucose to produce lactic acid. Also called lactic acid bacteria. Lactic acid produced by lactic acid fermentation prevents the growth of pathogens and harmful bacteria. It is used in food production, such as brewed foods. In addition, lactic acid bacteria are used as intestinal agents by preventing abnormal fermentation caused by various bacteria by living in the intestine of mammals. The lactic acid bacteria are Gram-positive bacteria, have a characteristic of anaerobic or anaerobic, and are known in the genus Lactobacillus and Streptococcus.

Patents related to lactic acid bacteria isolated from kimchi include Lactobacillus Lactococcus lactis BH5 (Korean Patent No. 413335), which produces antibacterial peptide substances, Lactobacillus Plantarum NC (Korean Patent No. 771209), Bactero, which has excellent antioxidant activity. Lactic acid bacteria (Korean Patent No. 1068531) that produce gods.

However, conventionally disclosed kimchi lactic acid bacteria are all related to probiotics (probiotics), strains that can be grown in a certain volume of culture is limited, there is a problem that management and storage is difficult to change the morphology by activation of the bacteria.

Influenza viruses, on the other hand, create serious public health problems, ranging from school and work to serious illness. Influenza type A H1N1 (A / H1N1) and H3N2 (A / H3N2) subtypes and A, B, C, and D influenza viruses of the influenza B Yamagata and Victoria (B / Victoria) strains are human Induces seasonal respiratory diseases. Vaccines against these influenza viruses are provided, but vaccines are only 10-60% effective (CDC, 2017) and require many preventive measures for many adolescents (Palese and Shaw, 2013). (WHO, 2012). Influenza pandemic (eg swine parasite H1N1 pandemic in 2009) has a delay of at least 6 months before vaccination for newly emerging and antigen-transferred strains (Partridge et al., 2010). In this situation, antiviral agents (Park et al., 2012) and / or dietary measures (Park et al., 2013a) have the advantage of inhibiting viral spread and subsequently reducing the prevalence and mortality of influenza. Diet for influenza is beneficial for people who need an alternative to vaccination or medication due to allergies during the influenza season.

 Kimchi's Lactic Acid Bacteria (LAB) is known for its many health benefits, including immune response-related effects (Jung et al., 2014) .Lactobacillus of several species has proven protective against influenza viruses. Reported. The protective effect of oral Lactobacillus brevis KB290 has been demonstrated in mice and unvaccinated humans. (Waki et al., 2014), Lactobacillus pentosus strains are well known for their efficacy against influenza virus (Izumo et al., 2010; Kobayashi et al., 2011; Kiso et al., 2013). Much research is still needed on antiviral efficacy.

Korean Patent Registration No. 10-1287120

Accordingly, an object of the present invention is to provide a pharmaceutical composition having excellent virus suppression ability by using nanonized microorganisms of lactic acid bacteria isolated from kimchi, and excellent antiviral efficacy by using nanonized microorganisms of lactic acid bacteria isolated from kimchi. It is another object to provide a dietary supplement.

And, the present invention containing the nano Kimchi lactic acid bacteria use the cells as an active ingredient, wherein the nano Kimchi lactic acid bacteria is Lactobacillus Planta room (Lactobacillusplantarum) nF1 strain (accession number NITE P-1462) In order to achieve the above object, the Nanostructured kimchi lactic acid bacteria provides a pharmaceutical composition for the anti-influenza virus, characterized in that it is in the form of cocci.

The influenza virus is one influenza virus selected from the group consisting of H1N1 subtype and H3N2 subtype A influenza virus or one type B influenza virus selected from the group B influenza virus consisting of Victoria lineage and Yamagata lineage. It is preferable to be

More preferably, the influenza virus is A / Korea / 01/2009 (2009 pandemic A / H1N1; provided by Korean Centers for Disease Control and Prevention, KCDC, Osong, Republic of Korea), A / Philippines / 02/1982 ( A / H3N2; previously generated by reverse genetics using commercially constructed genetic plasmids) or B / Wisconsin / 01/2010 (B / Yamagata; provided by KCDC).

The nano- kimchi lactic acid bacteria microorganisms are cultured Lactobacillus plantarum ( Lactobacillus plantarum) nF1 strain (Accession No. NITE P-1462) in a condition of pH 5-6; Heat sterilizing the culture medium of the strain at 75 ° C. to 85 ° C. for 8 to 10 minutes; Adding the same amount of dextrin as the cell weight to the culture solution and dispersing it with 140 to 160 kgf / cm ² high pressure homogenizer; And lyophilizing and dispersing the dispersed culture solution.

In addition, the nano-ized kimchi lactic acid bacteria microorganisms may have a mode of the particle size distribution of 0.5 ~ 1.0 ㎛.

In addition, the nano-ized kimchi lactic acid bacteria microorganisms may include 100 to 5 trillion Lactobacillus plantarum nF1 strain per gram of lyophilized powder.

In accordance with still another aspect of the present invention, the present invention containing the nano Kimchi lactic acid bacteria use the cells as an active ingredient, wherein the nano Kimchi lactic acid bacteria is Lactobacillus Planta room (Lactobacillusplantarum) nF1 strain (accession number NITE P-1462 ), The nano-ized kimchi lactic acid bacteria microorganisms provide a health functional food for the prevention or improvement of viral infection diseases, characterized in that the form of the cocci.

Diseases caused by viral infections include, but are not limited to, diseases caused by influenza virus infections such as flu, cold, sore throat, bronchitis, pneumonia, bird flu, swine flu, and goat flu.

The antiviral composition containing nano-ized kimchi lactic acid bacteria microorganism according to the present invention as an active ingredient has an effect of reducing the replication of influenza A (H1N1 and H3N2 subtypes) and influenza B (Yamagata strain) virus at regular oral administration. have. In addition, the composition according to the present invention has the advantage that the aggregation of the cosmetic composition can be suppressed, the absorption rate can be improved by having the form of the nanonized kimchi lactic acid bacteria microorganism has a cocciform form, that is, a spherical form, In the case of the form of the cocci, the form of cocci has an advantage that can provide a composition having an improved antiviral effect.

1 is a (a) colony observation image and (b) gram staining result image of kimchi lactic acid bacteria of the present invention.
Figure 2 is a molecular diagram showing the analysis of the 16S rRNA sequence of the kimchi lactic acid bacteria (SIID11558) of the present invention (the lower left line is the scale bar, the number located at the branch of the branch is the bootstrap value, the end of the main name T Indicates the type strain.)
Figure 3 is a micrograph of the kimchi lactic acid bacteria of the present invention observed before and after the nanonization treatment. (a) before nanonization: rod-shaped; (b) after nanonization: spherical.
4 is a summary of mouse experiments to confirm antiviral efficacy.
5 is a weight and survival analysis of the mouse.
Figure 6 shows the results of experiments on the weight and survival rate of the mouse at the time of H1N1 inoculation.
7 shows the results of experiments on the weight and survival rate of mice upon inoculation of B / yamagata.
8 shows virus titer analysis in mouse lungs, in which (a) is H1N1, and (b) is experimental results for B / yamagata.

EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated in detail.

One aspect of the invention, and containing the nano Kimchi lactic acid bacteria use the cells as an active ingredient, wherein the nano Kimchi lactic acid bacteria is Lactobacillus Planta room (Lactobacillusplantarum) nF1 strain (accession number NITE P-1462), and wherein the nano the Kimchi Lactobacillus microorganisms are antiviral pharmaceutical composition, characterized in that the form of cocci.

Another aspect of the invention, and containing the nano Kimchi lactic acid bacteria use the cells as an active ingredient, wherein the nano Kimchi lactic acid bacteria is Lactobacillus Planta room (Lactobacillusplantarum) nF1 strain (accession number NITE P-1462), and wherein the nano the Kimchi Lactobacillus microorganisms are antiviral pharmaceutical composition, characterized in that the form of cocci.

The present inventors screened the microorganisms by separating lactic acid bacteria from kimchi, it was found that the Lactobacillus plantarum ( Lactobacillus plantarum). The Lactobacillus plantarum strain was designated as "nF1" as of November 8, 2012, at the National Institute of Technology and Evaluation (NITE; International Patent Organism Depositary, IPOD), a Japanese independent administrative corporation. Deposited (NITE P-1462).

Nanostructured kimchi lactic acid bacteria microorganism according to the present invention is preferably in the form of cocci. In general, lactic acid bacteria are known to change shape due to the stress when the growth environment during culture is not constant or poor. Thus, in the present invention, by controlling the culture and processing conditions constantly, the lactic acid bacteria are propagated so that the cocci form of lactic acid bacteria is kept constant. In other words, the lactic acid bacteria were in the form of rods (rods) before the nanonization treatment, but they had a coarse form (spherical) through the nanonization treatment, whereby the aggregation phenomenon could be suppressed and the absorption rate could be improved.

Nanostructured kimchi lactic acid bacteria microorganisms according to the present invention is the step of culturing Lactobacillus plantarum ( Lactobacillus plantarum) nF1 strain (Accession No. NITE P-1462) at the condition of pH 5-6; Heat sterilizing the culture medium of the strain at 75 ° C. to 85 ° C. for 8 to 10 minutes; Adding the same amount of dextrin as the cell weight to the culture solution and dispersing it with 140 to 160 kgf / cm ² high pressure homogenizer; And lyophilizing and dispersing the dispersed culture solution.

In one embodiment according to the present invention, the nano-ized kimchi lactic acid bacteria microorganisms are heat sterilized for 8 to 10 minutes at 75 ℃ ~ 85 ℃ for the bacteria to kill. Since live bacteria may cause a change in shape during delivery or display after the manufacture of the product, it is preferable that they are dead bacteria that do not cause any further change in shape.

Nanostructured kimchi lactic acid bacteria microorganism of the present invention is subjected to a dispersion treatment step. For dispersal treatment, the sterilized culture medium is wet-dispersed with a high pressure homogenizer of about 150 kgf / cm ² (1.5 MPa). In this case, it is preferable to add a well-known dispersing agent or excipient to a culture liquid in advance, and by this, reaggregation of a microbial cell can be prevented efficiently. Suitable dispersants and excipients can be used as well as dextrins, as well as trehalose, dextrins, scheme milk and the like.

In addition, in order to finally obtain the nanonized kimchi lactic acid bacteria microorganism of the present invention as a powder, it is preferable to add a known dispersant, excipient, etc. to the culture solution, and to disperse the cells so as not to reaggregate, and then lyophilize. As a result, a cell powder having excellent dispersibility in water can be obtained.

The mode of the strain particle size distribution of the nanonized kimchi lactic acid bacteria microorganisms according to the present invention is preferably 0.5 ~ 1.0 ㎛. "Mode of particle size distribution" is a value which is an index indicating the size of bacteria, and refers to the particle diameter at which the relative frequency in the particle size distribution when the particle diameter of the cells is measured is maximum. The nano-ized kimchi lactic acid bacteria microorganism of the present invention is micronized to a nanometer (nm) size having a particle size of 1.0 μm or less. If the mode of the strain particle size distribution of the nano-kimchi lactic acid bacteria microorganisms satisfy the above range, it can have a better anti-viral effect.

The nano-ized kimchi lactic acid bacteria microorganisms may include 100 million to 5 trillion Lactobacillus plantarum nF1 strain per gram of the lyophilized powder.

The concentration of the kimchi lactic acid bacteria Lactobacillus plantarum strain per 1 g of dry powder may be adjusted according to the amount of dispersant or excipient added during the production of the nanonized kimchi lactic acid bacteria microorganism of the present invention. The present invention is characterized in that the kimchi lactic acid bacteria strains per nanogram of kimchi lactic acid bacteria microbial powder containing 100 million to 5 trillion.

The pharmaceutical composition may preferably be formulated in an oral formulation, for example in formulations for oral administration such as liquids, suspensions, powders, granules, tablets, capsules, pills or extracts. And, when formulated into each formulation, it can be prepared by adding a pharmaceutically acceptable carrier or additive necessary for the preparation of each formulation. Typical formulations for oral administration include one or more of diluents, lubricants, binders, disintegrants, sweeteners, stabilizers, and preservatives as carriers, and one or more of fragrances, vitamins, and antioxidants. The above can be selected and used.

The carrier and the additive may be any pharmaceutically acceptable, and specifically, lactose, corn starch, soybean oil, microcrystalline cellulose, or mannitol as a diluent, magnesium stearate or talc as a lubricant, and polyvinylpyrrolidone as a binder. Or hydroxypropyl cellulose is preferred. In addition, as a disintegrant, calcium carboxymethyl cellulose, sodium starch glycolate, potassium polyacrylic acid, or crospovidone, sweetener as white sugar, fructose, sorbitol, or aspartame, stabilizer as carboxymethyl cellulose sodium, beta-cyclodextrin, As lead, or xanthan gum, and preservative, methyl paraoxybenzoate, propyl paraoxybenzoate, or potassium sorbate is preferable.

In addition to the above ingredients, in order to enhance the taste as a known additive, natural flavors such as plum flavor, lemon flavor, pineapple flavor, herbal flavor, natural pigments such as natural fruit juice, chlorophyllin, flavonoid, fructose, honey, sugar alcohol, sugar Sweetening ingredients such as, or may be used by mixing an acidulant such as citric acid, sodium citrate. Such formulation methods and the carriers and additives required for formulation are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

The preferred dosage of the pharmaceutical composition of the present invention depends on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, it is preferable to administer the Lactobacillus plantarium DSR M2 of the present invention at 0.01 mg / kg to 10 g / kg per day, preferably at 1 mg / kg to 1 g / kg. Administration can be done once a day or divided into several times.

Functional food for antiviral in one embodiment of the present invention is a meat, sausage, bread, chocolate, candy, confectionary, snacks, pizza, ramen, other noodles, gums, dairy products, including ice cream, various soups, drinks, tea It provides a functional food for antiviral, characterized in that any one of a drink, health drinks, alcoholic beverages and vitamin complexes.

When adding the nanonized kimchi lactic acid bacteria microorganism of the present invention to food, the kimchi-derived lactic acid bacteria can be added as it is or used with other foods or food ingredients, and can be appropriately used according to a conventional method. In this case, unlike the conventional lactic acid bacteria lactic acid bacteria kimchi lactic acid bacteria do not harm the original flavor of the food as well as a small amount can be added as needed can satisfy a variety of consumer preferences.

The nano-ized kimchi lactic acid bacteria microorganism of the present invention may be commercialized as it is, but in general, by adding and blending various additives or flavors (flavor) to increase the flavor, or can be produced in the required shape to make the final product.

Specifically, the present invention provides a food additive or food comprising the nanonized kimchi lactic acid bacteria microorganisms.

Specifically, various sugars, emulsifiers, sweeteners, acidulants, fruit juices, etc. may be added to the mixed kimchi lactic acid bacteria microorganism of the present invention and mixed. More specifically, sugars such as glucose, sucrose, fructose and honey; Sugar alcohols such as sorbitol, xylitol, erythritol, lactitol and paratinite; And emulsifiers such as sucrose fatty acid ester, glycerin sugar fatty acid ester, and lecithin. In addition, even if various vitamins such as vitamin A, vitamin B, vitamin C, and vitamin E, herbal extracts, grain components, vegetable components, milk components, and the like are blended, excellent flavor-flavored nano kimchi lactic acid bacteria microbial composition can be obtained. .

In addition, as a flavor, yoghurt, berry, orange, quince, cranberry, citrus, apple, mint, grape, fair, custard cream, peach, melon, banana, tropical, herbal, tea, coffee Flavours, such as type | system | group, are mentioned, These can be used 1 type or in combination or 2 or more types. Although the addition amount of a flavor is not specifically limited, 0.05-0.5 mass%, especially about 0.1-0.3 mass% is preferable in a cell from a flavor viewpoint.

In addition, the nano-ized kimchi lactic acid bacteria microorganism of the present invention can be used as a food composition, in particular can be used for health food. The health food is more active than a normal food, and means a food for the purpose of health, health maintenance and promotion. The form may be any of liquid, semi-solid and solid, and specifically, cookies, confectionery, jelly, yokan, sweets, bread, cakes and the like; Beverages such as yoghurt, soft drink, nutritional drink, sikhye, fruit drink; Soups, ice cream, powdered milk, kimchi, and ramen.

Hereinafter, the present invention will be described in more detail with reference to Examples.

However, it is apparent to those skilled in the art that the following examples are merely illustrative of the present invention and various changes and modifications can be made within the scope and spirit of the present invention, and it is obvious that such modifications and modifications belong to the appended claims. will be.

Example 1 Identification of New Kimchi Lactic Acid Bacteria

In order to develop a novel kimchi lactic acid bacteria, the inventors were diluted kimchi extract and incubated for 48 hours at 30 ℃, aerobic conditions in MRS agar (Oxoid, Hampshire, UK) medium. After cultivation, the milky white colonies were bred in the form of a typical lactic acid bacterium, and then separated, and the selected colonies were separated by pure culture by transferring them to fresh medium for three times. Among the pure cultured lactic acid bacteria, the most excellent strain (SIID11558) of intestinal environmental improvement effect was selected and analyzed.

As shown in Figure 1, the SIID11558 strain forms a milky white colony, gram-positive in form, it was confirmed that the bacillus having a size of 0.8 ~ 0.9 × 1.2 ~ 1.5 ㎛.

Technos Co., Ltd. Lu wrap (TechnoSuruga Laboratory Co., Ltd.) 16S rRNA of the strains isolated from the gene sequence were performed for determination and analysis, a strain isolated by this, Lactobacillus Planta room (Lactobacillusplantarum) and 99.9% of the high It was identified as a novel microorganism with same sex. The analyzed 16S rRNA nucleotide sequence of the isolated strain is shown in SEQ ID NO: 1, the phylogenetic tree showing the molecular systematic location (phylogenetics tree) is shown in FIG. PCR amplification of 16S rDNA using forward primer (5'- ga gtt tga tcc tgg ctc ag-3 ': SEQ ID NO: 2) and reverse primer (5'- tcg taa caa ggt agc c-3': SEQ ID NO: 3) The operation from PCR amplification to cycle sequence was based on each protocol below.

-DNA Extraction: Acromodoctidase (Wakojunyaku, Osaka)

PCR: PrimeSTAR HS DNA Polymerase (Takara Bio, Shiga)

*-Cycle Sequencing: BigDye Terminator v3.1 Cycle Sequencing Kit (Life Technologies, CA, USA)

Sequencing: ABI PRISM 3130 x1 Genetic Analyzer System (Life Technologies, CA, USA)

-Sequencing: Chromas Pro 1.5 (Technelysium Pty Ltd., Tewantin, AUS)

-BLAST homology search and simple molecular system analysis: Apollon 2.0 software (Techno Suruga Lab, Shizuoka)

-Database: Apollon DB-BA 7.0 Database (Techno Suruga Lab, Shizuoka), International Base Sequence Database (GeneBank / DDBJ / EMBL)

The new microorganism of the present invention identified by the above method is the November 8, 2012 to the National Institute of Technology and Evaluation (NITE; International Patent Organism Depositary, IPOD) Deposited as "nF1" (NITE P-1462).

Example 2 Preparation of Nano-ized Kimchi Lactobacillus nF1 Microbial Powder

The nF1 strain isolated and identified in Example 1 was cultivated in a known nutrient medium to which 5 mass% glucose was added, in an aqueous solution of 20 g% of sodium hydroxide, but was cultured under a neutral pH and a temperature of 36.5 ° C. This consumed time point was used as the culture end point. The pH range of preparing nanonized kimchi lactic acid bacteria microbial powder of the present invention is preferably in the neutral region of pH 5-7, most preferably pH 6.5.

After the completion of the culture, the culture solution was heat sterilized at 80 ° C. for 10 minutes, and the cells were washed with PBS, and the same amount of dextrin was added as an excipient based on the weight of the cells, followed by dispersion by a mixer. Next, the powder sample was prepared by freeze spray drying, and it was confirmed that the kimchi lactic acid bacteria nF1 strain of the present invention per 100 g of the prepared powder contained more than 100 million.

The nF1 powder was suspended in PBS again adjusting to a cell concentration of 10 mg / ml. The pH during the process was maintained at 6.5. As described above, the results of the nano-processing of the strain were confirmed by photographing under a microscope. As shown in FIG. 3, the nF1 strain extracted from kimchi was rod-shaped bacilli before the nano-ization treatment, but after the nano-ization treatment, the colony showed a form close to the cocci. It can be seen that the properties do not reaggregate with each other. The nanonized nF1 strain exhibited a mode of 0.5 ~ 1.0 μm in particle size distribution and has a property of not reaggregating, thereby obtaining lactic acid bacteria having excellent dispersibility in water or other solvents.

On the other hand, when prepared by adding the same amount of dextrin as the weight of the cell as an excipient, it was confirmed that 5 trillion of kimchi lactic acid bacteria per 1g of the powder produced.

Example 3. Antiviral Effect Experiment

1. Test substance

(1) Test substance: nF1 concentration of lactic acid bacterium extracted from Korean mature kimchi and subjected to heat sterilization nanostructured: 10 mg / kg / day, 0.05 mg / kg / day

(2) Control I: Phosphate buffer saline (PBS) from Lonza

(2) Control substance II: Oseltamivir phosphate concentration: 10 mg / kg / day Place of purchase: Toronto Research Chemicals Inc, Canada (Cat No. O701000)

2. Test method

2.1 Oral Administration

Two weeks before influenza virus infection, test and control substances were administered to mice (BALB / c, 6weeks, female; Nara-Biotec, Korea) as zonde. A dose of 200 μl was administered at the same time point (quaque die) once a day for a total of 4 weeks including 2 weeks after infection (see FIG. 4).

2.2 Vaccination

 Intramuscular anesthetics are injected to inoculate the mouse nasal cavity with seasonal influenza virus A / H1N1 and B / Yamagata lineage virus. Each virus was inoculated at a dose of 30 μl per mouse.

2.3 Virus titer analysis in mouse lungs

 Lung tissues were isolated by opening the abdominal cavity of mice on days 1, 3 and 6 after influenza virus infection. The isolated lung tissues were subjected to plaque assay, which is the best method for analyzing virus titers in our laboratory.

Experimental Example

(1) Weight change and survival rate analysis

1) uninfected group

The pathogenicity of the experimental group administered only nF1 without infecting the virus was analyzed. As a result, it was analyzed that nF1 had no effect on mice. Therefore, pathogenicity observed after viral infection can be judged to be caused by virus (see Fig. 5).

2) A / H1N1 inoculation

  Influenza virus of the H1N1 subtype was challenged to evaluate the antiviral efficacy of nF1 lactic acid bacteria. The survival rate of the experimental group administered with nF1 lactic acid bacteria at 0.05 mg / kg / day was not different from that of the experimental group not administered at all. However, the survival rate of 30% was observed in the experimental group in which the nF1 lactobacillus dose was increased to 10 mg / kg / day. In other words, as the concentration of nF1 increases, the survival rate also increases, and it is judged that nF1 lactic acid bacteria have the effect of suppressing H1N1 subtype influenza virus. And compared to the control Tamiflu nF1 lactic acid bacteria, despite being a dietary supplement, not a treatment, the survival rate of the mouse was about 30%, it can be seen that the continued administration of nF1 lactic acid bacteria can suppress some of the infectious diseases (Fig. 6). Reference).

3) B / Yamagata inoculation

   The antiviral efficacy of nF1 lactic acid bacteria was evaluated by inoculating the influenza B virus. In the experimental group administered with nF1 lactic acid bacteria at 0.05 mg / kg / day, the survival rate was increased by 10% compared to the control group, and in the experimental group which increased the dose to 10 mg / kg / day, the survival rate was increased by 20% compared to the control group. Was observed. As a result, as the concentration of nF1 is increased, the survival rate also increases, and it is determined that nF1 lactic acid bacteria have an effect of suppressing influenza B virus (see FIG. 7).

(2) virus titer analysis (lungs of the mouse)

Lungs of the experimental group challenged with the H1N1 subtype virus were extracted and analyzed for virus titers. The pulmonary viral titer of the experimental group to which the nF1 lactic acid bacteria was administered decreased more than the lung virus titer of the experimental group to which the nF1 lactobacillus was not administered (see FIG. 8A).

In the experimental group challenged with influenza B virus, the lung virus titer of the experimental group to which the nF1 lactobacillus was administered 6 days after the virus infection was further reduced compared to the experimental group to which the nF1 was not administered (see FIG. 8B).

Therefore, the increase in survival rate of mice by nF1 lactic acid bacteria administration is considered to be due to the decrease in the titer of virus present in the lungs of the nF1 lactic acid bacteria administration group. The decrease in viral titers in the lungs is evidence that mice are recovering from diseases caused by viral infections.

 Taken together, the results suggest that nF1 lactic acid bacteria are effective in protecting mice from influenza virus challenge.

(3) sintering

In the experimental group administered daily nanonized nF1 lactic acid bacteria, the survival rate was increased compared to the control group. In addition, as the concentration of nF1 is increased, the survival rate also increases, and it is judged that nF1 lactic acid bacteria have the effect of inhibiting H1N1 subtype and influenza B virus. Pulmonary viral titers in the experimental group treated with nF1 lactic acid bacteria were further reduced compared to the experimental group not administered nF1. Therefore, the increase in survival rate of mice by nF1 lactic acid bacteria administration is considered to be due to the decrease in the titer of virus present in the lungs of the nF1 lactic acid bacteria administration group.

Patent Microbial Deposit Center of Product Evaluation Technology Infrastructure nF1NITEP-1462 20121108

Claims (14)

And containing the nano Kimchi lactic acid bacteria use the cells as an active ingredient, wherein the nano Kimchi lactic acid bacteria is Lactobacillus Planta room (Lactobacillusplantarum) nF1 strain (accession number NITE P-1462), and wherein the nano Kimchi lactic acid bacteria used cell is a Streptococcus form Pharmaceutical composition for anti-influenza virus, characterized in that.
The method of claim 1,
The nano- kimchi lactic acid bacteria microorganisms are cultured Lactobacillus plantarum ( Lactobacillus plantarum) nF1 strain (Accession No. NITE P-1462) in a condition of pH 5-6;
Heat sterilizing the culture medium of the strain at 75 ° C. to 85 ° C. for 8 to 10 minutes;
Adding an excipient to the culture solution and dispersing it with 140 to 160 kgf / cm ² high pressure homogenizer; And
Pharmaceutical composition for anti-influenza virus, characterized in that it was prepared through; lyophilized powdered dispersion cultured.
The method of claim 1,
The nano-ized kimchi lactic acid bacteria microorganisms are anti-influenza virus pharmaceutical composition, characterized in that the mode of strain particle size distribution is 0.5 ~ 1.0 ㎛.
The method of claim 1,
The nano-kimchi lactic acid bacteria microorganisms are anti-influenza virus pharmaceutical composition, characterized in that containing 1 to 5 trillion Lactobacillus plantarum nF1 strain per gram of lyophilized powder.
The method of claim 1,
The virus is an influenza A virus or influenza B virus, characterized in that the pharmaceutical composition for anti-influenza virus.
The method of claim 5,
The influenza A virus is a H1N1 or H3N2 subtype pharmaceutical composition for anti-influenza virus, characterized in that.
The method of claim 5,
The influenza B virus is a pharmaceutical composition for anti-influenza virus, characterized in that the B / Yamagata lineage.
And containing the nano Kimchi lactic acid bacteria use the cells as an active ingredient, wherein the nano Kimchi lactic acid bacteria is Lactobacillus Planta room (Lactobacillusplantarum) nF1 strain (accession number NITE P-1462), and wherein the nano Kimchi lactic acid bacteria used cell is a Streptococcus form Health functional food for influenza virus suppression, characterized in that.
The method of claim 8,
The nano- kimchi lactic acid bacteria microorganisms are cultured Lactobacillus plantarum ( Lactobacillus plantarum) nF1 strain (Accession No. NITE P-1462) in a condition of pH 5-6;
Heat sterilizing the culture medium of the strain at 75 ° C. to 85 ° C. for 8 to 10 minutes;
Adding an excipient to the culture solution and dispersing it with 140 to 160 kgf / cm ² high pressure homogenizer; And
Freeze-drying the dispersed culture medium to powdered; functional foods for inhibiting influenza virus, characterized in that produced through.
The method of claim 8,
The nano- kimchi lactic acid bacteria microorganisms, the functional value for the prevention or improvement of influenza virus infectious disease, characterized in that the mode of the strain particle size distribution is 0.5 ~ 1.0 ㎛.
The method of claim 8,
The nano-ized kimchi lactic acid bacteria microbial microbial microbial lyophilized Lactobacillus plantarum (Lactobacillus plantarum ) nF1 strain characterized in that it comprises 100 million ~ 5 trillion health functional foods for the prevention or improvement of influenza virus infectious diseases.
The method of claim 8,
The virus is an influenza A virus or an influenza B virus, characterized in that the health functional food for the prevention or improvement of influenza virus infectious diseases.
The method of claim 12,
The influenza A virus is a H1N1 or H3N2 subtype health functional food for the prevention or improvement of influenza virus infection diseases, characterized in that.
The method of claim 12,
The influenza B virus is a B / Yamagata lineage health functional food for the prevention or improvement of influenza virus infection diseases, characterized in that.

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