WO2016159582A2 - Composition for innate immunity enhancement and antiviral activity containing schizonepeta tenuifolia extract as active ingredient - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for innate immunity enhancement and antiviral activity containing a Schizonepeta tenuifolia extract as an active ingredient, a health functional food for innate immunity enhancement and antiviral activity containing a Schizonepeta tenuifolia extract as an active ingredient, a feed composition for innate immunity enhancement and antiviral activity containing a Schizonepeta tenuifolia extract as an active ingredient, and a method for innate immunity enhancement and antiviral activity enhancement in an animal, the method comprising administering a Schizonepeta tenuifolia extract to an animal excluding humans, in need of innate immunity enhancement. The Schizonepeta tenuifolia-containing composition for innate immunity enhancement and antiviral activity according to the present invention exhibits antiviral activity and innate immunity enhancement efficacy, and thus can be applied for prevention and treatment of bacterial and viral infectious diseases, and can be widely used for a medicine or a health functional food for immunity enhancement and regulation in immunodeficient or immunosuppressed patients and for a feed for innate immunity enhancement and antiviral activity, for the purpose of anti-disease enhancement of animals. Furthermore, the composition for innate immunity enhancement and antiviral activity according to the present invention scarcely causes toxicity or side effects, and thus can be safely used even in the long-term use for prophylactic purposes.

Description

Congenital immunity enhancement and antiviral composition containing the extract of Hyeophyeong as an active ingredient

The present invention relates to an innate immunity enhancement and antiviral composition containing the extract as an active ingredient, and more particularly to an innate immunity containing the extract as an active ingredient, which can be used as a pharmaceutical, health food or feed composition. And antiviral compositions.

Immunity can be divided into innate immunity, which has been born since birth, and acquired immunity, which is obtained by adapting to life after being acquired. Innate immunity, also known as 'natural immunity', is characterized by non-specific responses to antigens and no special memory function.

Recently, the field of innate immunity induced by interferon (interferon) in the innate defense immune system in vivo, because the activation of interferon-mediated immunity can be a fundamental preventive method against various infectious disease pathogens. Therefore, studies on the mechanism of interferon activation and the development of immunomodulators capable of inducing interferon are being actively conducted.

On the other hand, as one of the defense mechanisms of innate immunity following infection of the pathogen, immune factors such as cytokines are secreted, thereby causing an immune response and defense against the pathogen. Therefore, by inducing an innate immune response can be a method of prevention and treatment of various infectious disease pathogens, and research on the enhancement of innate immunity that can induce it.

Virus is a Latin word for toxic substance and is a group of infectious pathogenic particles that pass through bacterial filter paper (0.22㎛). Viruses may be classified as bacteriophages, plant viruses, or animal viruses according to host cell types, and may be classified as DNA viruses or RNA viruses according to nucleic acid types. Recently, various viral diseases such as swine flu, AI and foot-and-mouth disease caused social problems, and consequently, anxiety about effective measures for viral diseases has attracted great social attention.

Currently, the best way to prevent viral diseases is vaccination, but in the case of viral diseases, the effectiveness of vaccines due to the large number of virus serotypes (subtypes) is raised. Development and dissemination of inhibitors for the prevention of viruses that can supplement the problems of such vaccines is an important matter. For this purpose, the preventive agents that enhance the immunity of individual animals by stimulating the innate immune system, which is the initial defense system against viruses, are particularly important. Discovery and development can be an important method of drug development.

Representative antiviral agents that inhibit the growth of influenza viruses include amantadine and rimantadine, but these two antiviral agents are effective only against serotype A influenza virus and serotype B without M2 protein. Influenza virus was found to be ineffective. In addition, amantadine and rimantadine have been found to have the disadvantage that the emergence of a mutant virus that does not affect the ion channel function of the influenza virus M2 protein when used. To alleviate this drawback, zanamivir and oseltamivir have been developed as effective antiviral agents for all 16 serotype A influenza viruses and serotype B influenza viruses. However, zanamivir has the disadvantage of inhalation and intravenous administration, while oseltamivir is orally available, but has been pointed out as a disadvantage due to side effects such as recent reports of the emergence of resistant virus and vomiting and dizziness upon oral administration.

In addition, since the main control method of Vesicular stomatitis virus is impossible to treat the disease completely, prevention and blocking prevention and the eradication of susceptible livestock in the developing area is the best method, like foot-and-mouth disease.

In addition, vaccines against Newcastle disease virus are largely divided into live poison vaccine and dead poison vaccine. The most widely used representative Newcastle disease live vaccine vaccines, B1 and La Sota (including clone), are known to induce vaccination response. Multivalent mixed poisoning oil vaccine, a globally used Newcastle disease poisoning vaccine, can prevent three or more diseases at the same time with a single vaccination. However, in case of laying hen farms, cases of newcastle disease are increasing due to reduced immunity.

There are also relatively small viruses among RNA viruses. They combine the word 'pico' which means small and 'RNA' to call piconaviruses, and the viruses belonging to them are collectively called the piconavirus family (Picona viridae). Enteroviruses belonging to the piconavirus family include about 70 serotypes that cause various clinical symptoms such as aseptic meningitis, hand and foot disease, herpes stenosis, dilated myocarditis, and acute hemorrhagic conjunctivitis. , Cossackie virus and Echo virus and other enteroviruses. Enteroviruses are about 20 to 30 nm in diameter and contain a single strand of RNA as a gene. Most of the infections in the digestive organs of vertebrates, as well as the respiratory tract and the central nervous system, but often do not show any obvious symptoms. Coxsackie virus (CXV) is a human enterovirus belonging to the family of piconaviruses. It is largely divided into types A and B (Pallansch MA and Roos RP, Fields Virology, 4th edi, pp723-775). , 2001).

In addition, recently, high-risk enteroviruses and mutant viruses (enterovirus type 71 (EV-71) and coxsackie A24 mutant strains) have been newly discovered and spread in various parts of the world. It is required.

Enteroviruses, including coxsackieviruses, infect the vertebrate digestive system, respiratory tract and central nervous system and cause various clinical symptoms. Therefore, it is urgently required to prepare measures at the national level, but the types and serotypes of viruses are very diverse. As such, no effective commercialized vaccines or therapeutics have been developed.

In addition, Herpes Simplex Virus (HSV) is a DNA virus belonging to the herpes virus, a relatively large virus having a size of about 175 nm, and is a widespread infectious agent in humans. Herpes simplex infection is an infection of HSV type 1 (hereinafter referred to as 'HSV-1') and HSV type 2 (hereinafter referred to as 'HSV-2'), which is an acute bullous disease involving the mucous membranes and skin. It is a common infectious disease in people. HSV-1 mainly forms around the mouth and eye, while HSV-2 forms blisters around the genitals. HSV infection is reported to be severe in patients with compromised immune function and also to cause cervical cancer (Melnick, JL, Adam, E. and Rawls, W., Concer, 34, 1355-1385, 1974). In addition, newborns and fetuses are not only able to produce HSV antibodies themselves, but because the antibodies of the mother are not passed on to the fetus, it is known that most infections have a fatal effect. Currently, about 500,000 cases of HSV-1-related eye disease occur in the United States alone, and more than 1,000 of them are undergoing ocular transplantation. About 95% of HSV-2 infections are known to be transmitted through sexual contact with a partner with active lesions. About 20-30% of US adults are infected with HSV-2, of which 20-50% relapse It has been reported to have genital herpes (Johnson RE et al., N. Engl. J. Med., 321, 7,1989).

HSV begins to replicate in the skin or mucosal epidermal cells after initial infection, migrates along the nerve tissue, lurks in the vertebral ganglion throughout life, and reactivates when the immune function is compromised. It is known to generate. Nucleoside derivative Acyclovir is known to be very effective for early infection as a therapeutic agent for HSV infection (Bryson, YJ et al., N. Engl. J. Med., 308, 916, 1983) In order to maintain the effectiveness of the drug, the drug must be continuously administered, and it is known that HSV relapses infectious diseases (Mindel A. et al., Lancet i: 926-928 (1988); Straus SE et al., N. Engl. J. Med., 310, 1545 (1984)). It has also been reported that mortality rates are high in patients with relapse after the initial infection of HSV (Nahmias, A. J. and Coleman, R. M., Immunobiology of Herpes Simplex Virus Infection CRC Press, Boca Raton, 92-102,1984).

Although vaccines have been developed to prevent such infections of HSV, live attenuated vaccines that use attenuated virus itself have been found to be directly related to the oncogenesis of HSV genomes. Use has been prohibited (Cappel, R., Sprecher, S., De Cuyper, F. and De Braekeleer, J., J. Med. Virol., 16, 137-145,1985).

Under these circumstances, many efforts have been made to overcome the shortcomings of existing antiviral agents in Korea and abroad, and among them, research on antiviral efficacy of herbal extracts and plant extracts is underway. However, since the current situation is inadequate, it is necessary to develop a composition using an herbal extract capable of overcoming the disadvantages of existing antiviral preparations and exhibiting excellent innate immunity promoting effect and antiviral activity with little toxicity and side effects. .

On the other hand, Schizonepetae Spica is a year old with Lamiaceae. Temper is known to be nontoxic and tasteless. The main ingredients are d-menthone, d1-menthone and small amount of d-limonene, as well as heparidine and ursolic acid. ), Luteolin, daucesterol and the like have been reported (Zhang YH, et al., Zhongguo Zhong Yao Za Zhi., 31 (15), pp 1247-57, 2006). It has been reported that the extracts have antihistamine and antipruritic activity (Tohda C, et al., Biol Pharm Bull., 23 (5), pp599-601, 2000), and essential fats, which are the main components, have anti-worm and antifungal action. Has been reported (Park IK. Et al., J Econ Entomol., 99 (5), pp1717-21, 2006).

In addition, Korean Patent Publication No. 2013-0058790 discloses a pharmaceutical composition for promoting growth growth or osteoporosis prevention and treatment containing an active ingredient extract hyeonghyeong, and Korean Patent Publication No. 2013-0058791 to prevent and improve osteoporosis A health functional food containing an effective mold extract is disclosed. However, there have been no reports on innate immunity enhancing effect and antiviral activity using the extract of Hyeop.

The present invention was derived by the above-mentioned demands, and the present invention completed the present invention by confirming that the mold extract can activate the macrophages, which are the main cells of innate immunity, to inhibit the proliferation of various viruses. In addition, the present invention is effective in preventing the mold extract, which can be effectively used in pharmaceutical compositions, health functional foods or livestock feed compositions for the prevention or treatment of infectious diseases caused by various viruses or bacteria while ensuring long-term safety with little toxicity and side effects. An object of the present invention is to provide an innate immunity and antiviral composition, an innate immunity enhancement and an antiviral activity enhancement method of an animal.

In order to achieve the above object, the present invention provides a pharmaceutical composition for innate immunity enhancement and antiviral containing the extract of hyunggwa as an active ingredient.

The present invention also provides a dietary supplement for innate immunity and anti-viral health food containing the extract as an active ingredient.

In another aspect, the present invention provides a feed composition for innate immunity enhancement and antiviral containing the extract of the hyungpang as an active ingredient.

The present invention also provides a method for enhancing innate immunity and antiviral activity of an animal, comprising administering a mold extract to an animal in need of enhancement of innate immunity except for humans.

According to the present invention, by using a mold-opening extract exhibiting excellent innate immunity promoting effect can be effectively applied to the prevention and treatment of various viral and bacterial infectious diseases, it can contribute to the enhancement of immunity. In addition, the innate immunity enhancement and antiviral composition containing the mold extract according to the present invention can be used with confidence even for long-term use for prophylactic purposes because it rarely causes toxicity or side effects. Therefore, the composition according to the present invention can be applied for the prevention and treatment of infectious diseases caused by bacteria and viruses, pharmaceutical compositions or health functional foods for improving and regulating the immunity of patients with reduced or suppressed immunity, and of animals It can be widely used as feed composition for immuno-stimulation for the purpose of strengthening anti-disease. In addition, according to the present invention can be effectively and safely enhance the immunity of the animal by administering to the animal necessary for the enhancement of innate immunity.

1 is a result of antiviral activity analysis of influenza virus (PR8-GFP virus) of the mold extract according to an embodiment of the present invention. Medium is a group of cells treated with nothing as a negative control; PR8-GFP is a virus infection group; IFN-β / PR8-GFP is a positive control group, including PR8-GFP virus infection and 1,000 Units / ml IFN-β treated group; Mold opening / PR8-GFP is a treatment group for PR8-GFP virus infection and mold opening extract.

Figure 2 shows the results of the antiviral activity analysis of bullous stomatitis virus (VSV-GFP virus) of the extract according to an embodiment of the present invention. Medium is a group of cells treated with nothing as a negative control; VSV-GFP is a virus infection group; IFN-β / VSV-GFP is a positive control group with VSV-GFP virus infection and 1,000 Units / ml IFN-β treated group; Mold opening / VSV-GFP is a VSV-GFP virus infection and mold opening extract treatment group.

Figure 3 is an antiviral activity analysis results for herpes simplex virus (HSV-GFP virus) of the extract according to an embodiment of the present invention. Medium is a group of cells treated with nothing as a negative control; HSV-GFP is a virus infection group; IFN-β / HSV-GFP is a positive control group with HSV-GFP virus infection and 1,000 Units / ml IFN-β treated group; Mold opening / HSV-GFP is the HSV-GFP virus infection and mold opening extract treatment group.

Figure 4 shows the results of antiviral activity analysis of Newcastle disease virus (NDV-GFP virus) of the extract according to an embodiment of the present invention. Medium is a group of cells treated with nothing as a negative control; NDV-GFP is a virus infection group; IFN-β / NDV-GFP is a positive control group with NDV-GFP virus infection and 1,000 Units / ml IFN-β treated group; Mold opening / NDV-GFP are NDV-GFP virus infection and mold opening extract treatment groups.

Figure 5 is an antiviral activity analysis results for the enterovirus (EV-71 virus) of the mold extract according to an embodiment of the present invention. Medium is a group of cells treated with nothing as a negative control; EV-71 is a viral infection group; IFN-β / EV-71 is a positive control group with EV-71 virus infection and 1,000 Units / ml IFN-β treated group; Mold opening / EV-71 is an EV-71 virus infection and mold opening extract treatment group.

Figure 6 shows the results of the pre-inflammatory cytokine induction assay by the mold extract according to an embodiment of the present invention.

7 is a result confirming the suppression of infection of influenza viruses (H1N1 and H5N2) by the mold extract according to an embodiment of the present invention. H1N1 and H5N2 are the influenza virus-treated (1.0 MOI) group, Medium is the negative control group, which is the MDCK cell group that has not been treated with anything. ) Is processed simultaneously.

The present invention relates to a pharmaceutical composition for innate immunity enhancement and antiviral containing the extract of hyunggwa as an active ingredient.

The mold extract is preferably extracted with one or more solvents selected from water and alcohols having 1 to 4 carbon atoms, more preferably water, methanol, ethanol, or butanol, and even more preferably. Hydrothermal extraction using water as a solvent. The hot water extraction is 1) adding 5 to 30 times the distilled water on the basis of the mold opening weight; 2) extracting hot water at a temperature of 100-130 ° C. for 2-5 hours; And 3) filtering the hot water extract; but is not limited thereto.

In addition, the said mold-opening extract shall contain any of the extract obtained by an extraction process, the dilution or concentrate of an extract, the dry matter obtained by drying an extract, or a crude product or a purified product.

One example of a virus having an antiviral activity of the mold extract of the present invention is Orthomixoviridae, Rhabdoviridae, Paramixoviridae, Herpesviridae and Piconaviridae. (Picornaviridae) is one or more viruses, preferably influenza virus, Newcastle disease virus, bullous stomatitis virus (Vesicular stomatitis virus), Cossackie virus, enterovirus type 71 (Enterovirus-71), herpes simplex virus (Herpes Simplex Virus), rhinovirus, respiratory syncytial virus (RSV), foot and mouth disease virus, Colorado true tick fever virus, leo virus, human immune deficiency virus, type B Hepatitis Virus, Hepatitis C Virus, Swine Fever Virus, Bovine Viral Diarrhea Virus, Porcine reproductive and respiratory syndrome virus, Swine Ozeski disease virus, Rotavirus, Parvovirus, Porcine epidemic diarrhea virus, and more preferred examples of influenza virus, Newcastle disease virus , Vesicular stomatitis virus (Vesicular stomatitis virus), herpes simplex virus, enterovirus type 71, rhinovirus (Rhinovirus), respiratory syncytial virus (RSV), but is not limited thereto.

The mold extract of the present invention exhibits a strong ability to induce immune factors, increases the innate immunity of the individual, and has the effect of inhibiting the infection and proliferation of the virus. Characterized in that it can. When infecting pathogens, one of the defense mechanisms of innate immunity is to secrete immune factors.The secretion of these immune factors (TNF-α, IL-6 and IFN-β) protects the pathogens, thus inducing an appropriate level of cytokine response. Can be a prophylactic and treatment method for various infectious disease pathogens. In particular, it can enhance the immunity to the virus.

Pharmaceutical composition for innate immunity enhancement and antiviral of the present invention may further comprise a pharmaceutically acceptable carrier, excipient or diluent. Pharmaceutically acceptable carriers, excipients or diluents which may be used in the present invention are not particularly limited so long as they do not impair the effects of the present invention, and include, for example, fillers, extenders, binders, wetting agents, disintegrants, surfactants, lubricants, Sweeteners, fragrances, preservatives and the like. Representative examples of pharmaceutically acceptable carriers, excipients or diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, maltitol, starch, gelatin, glycerin, acacia rubber, alginate, calcium phosphate, calcium carbonate, calcium Silicates, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, propylene glycol, polyethylene glycol, vegetable oil, injectable Ester, utopsol, macrogol, tween 61, cacao butter, lauridge, etc. are mentioned. Pharmaceutical composition for innate immunity enhancement and antiviral of the present invention may be in the form selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories and injections. The method of formulating the pharmaceutical composition may be carried out according to conventional methods known in the art, and is not particularly limited.

The pharmaceutical composition for congenital immunity enhancement and antiviral of the present invention may be administered orally or parenterally, and the dosage is depending on the age, sex, weight, condition, degree of disease, type of drug, route of administration and duration of administration. Although appropriately selected, generally about 5 to 500 mg / kg, preferably about 100 to 250 mg / kg may be administered once or three times a day.

It will be apparent to those skilled in the art that the method, dosage, route of administration, components, etc., of the innate immunity enhancing and antiviral pharmaceutical compositions of the present invention may be appropriately selected from conventional techniques known in the art.

The pharmaceutical composition for innate immunity and antiviral of the present invention can be used for the prevention and treatment of bacterial or viral infectious diseases. The pharmaceutical composition for innate immunity enhancement and antiviral of the present invention may be used in combination with a pharmaceutical composition including other pharmaceutically active ingredients, or other active ingredients, in addition to the mold extract as an active ingredient.

In addition, the present invention relates to a dietary supplement for innate immunity and anti-viral health food containing the extract of the hyunggwa. The dietary supplement for innate immunity and antiviral of the present invention may further comprise a food supplement acceptable food supplement. Food-acceptable food supplement additives which can be used in the present invention include sugars such as glucose, fructose, maltose, sucrose, dextrin, cyclodextrin and natural carbohydrates such as sugar alcohols such as xylitol, sorbitol, erythritol, taumate , Natural flavors such as stevia extract, synthetic flavors such as saccharin, aspartamic acid, colorants, pectic acid or salts thereof, alginic acid or salts thereof, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, glycerin, Alcohols, carbonating agents, and the like, but are not limited thereto. Health functional foods for innate immunity enhancement and antiviral of the present invention may be in the form selected from the group consisting of powders, granules, tablets, capsules, candy, chewing gum, jelly and beverages. The content of the mold extract in the dietary supplement for innate immunity and antiviral foods may be appropriately selected in consideration of the form, flavor, and taste of the food, for example, 0.01 to 30% by weight based on the total weight of the dietary supplement. It can be a range. It will be apparent to those skilled in the art that the form, composition, and manufacturing method of innate immunity enhancing and antiviral dietary supplements of the present invention may be appropriately selected from conventional techniques known in the art.

The present invention also relates to an innate immunity enhancing and antiviral feed composition containing the extract of the hyunghwa as an active ingredient. The content of the mold extract in the feed composition for innate immunity and antiviral may be appropriately selected according to the species, age, weight, and breeding conditions of the feed livestock, and 0.01 to 95% by weight, preferably based on the total weight of the feed composition. It may be a ratio of 0.1 to 80% by weight. The feed composition for innate immunity and antiviral of the present invention may be prepared according to a feed production method known in the art, for example, after mixing various feed ingredients or blended feed with the mold extract of the present invention, It may be produced by further performing a processing process, for example, molding into pellets or cutting into granules and the like. It will be apparent to those skilled in the art that the components, compositions, methods of preparation, and methods of feeding innate immunity enhancing and antiviral feed compositions of the present invention may be appropriately selected from conventional techniques known in the art.

In addition, the present invention relates to a method for enhancing innate immunity and antiviral activity of an animal, characterized in that the administration of a mold extract to an animal in need of enhancement of innate immunity except for humans. In the enhancement of innate immunity and antiviral activity of the animal of the present invention, it is apparent to those skilled in the art that the dosage, route of administration, timing of administration, etc. of the extract may be appropriately selected from conventional techniques known in the art. .

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for explaining the present invention in more detail, it is obvious to those skilled in the art that the scope of the present invention is not limited by them.

Example 1 Preparation of Extract

The mold extract was added 20 times of distilled water to the mold and extracted hot water at 115 ° C. for 180 minutes, filtered first at 0.45 μm, and filtered at 0.22 μm to remove the precipitate. The pH was then adjusted to 7.0, 1 ml aliquoted into 1.5 ml Ep-tubes and stored at -20 ° C for use in all assays.

Example 2. Antiviral Activity Assay of Mold Extract

Influenza virus (PR8), Vesicular stomatitis virus, Herpes simplex virus (HSV), Newcastle disease virus and Enterovirus 71 (Enterovirus-71; EV-71 The antiviral activity analysis of the mold extract was carried out.

(1) Antiviral Activity Assay

Influenza virus (PR8), vesicular stomatitis virus (VSV), herpes simplex virus (HSV-GFP), and Newcastle disease virus (Raw 264.7 cells, 8 × 10 ⁵⁾ cell / well), and enterovirus 71 (Enterovirus-71; EV-71) was analyzed by infecting Hela cells.

After culturing the cells in a 12-well TC plate, 1 μg / ml of open mold extract (pH adjustment) prepared in Example 1 was treated in DMEM to which 1% FBS was added. The negative control was treated only with DMEM with 1% FBS, and the positive control was treated with mouse IFN-β (500 units / ml). After 12 hours of treatment with the mold extract, PR8-GFP (MOI: 1.0), VSV-GFP (MOI: 1.0), HSV-GFP (MOI: 3.0), NDV-GFP (MOI: 3.0) and EV-71 (MOI: 1.0) was inoculated respectively. After 2 hours of inoculation, the inoculum was removed, washed three times with PBS, and after 12 and 24 hours, the extent of virus infection was checked.

(2) Antiviral Activity Assay

1) Analysis result of influenza virus (PR8-GFP virus)

Antiviral activity assay results for influenza virus are shown in FIG. Figure 1 (a) shows a green fluorescent protein (GFP) fluorescent image after 12 hours of infection, Figure 1 (b) shows a green fluorescent protein (GFP) fluorescent image after 24 hours of infection, Figure 1 (c) shows infection 12 Cell viability after hours and 24 hours is shown, and FIG. 1 (d) shows the number of viruses present in the cells. As a result of treatment with the mold extract, the infection rate of influenza virus was significantly decreased, and the cell survival rate was also increased compared to the virus infection group (Fig. 1).

2) Analysis result of bullous stomatitis virus (VSV-GFP virus)

The results of the antiviral activity assay for bullous stomatitis virus are shown in FIG. 2. FIG. 2 (a) shows a green fluorescent protein (GFP) fluorescence image 12 hours after infection, FIG. 2 (b) shows a green fluorescent protein (GFP) fluorescence image 24 hours after infection, and FIG. 2 (c) shows a infection 12 Cell viability after hours and 24 hours is shown, and FIG. 2 (d) shows the number of viruses present in the cells. As can be seen from Figure 2, as a result of treatment with a mold extract, the infection rate of bullous stomatitis virus was significantly reduced, the virus titer was also reduced, the cell survival rate was also increased compared to the virus infection group.

3) Analysis result of herpes simplex virus (HSV-GFP virus)

The results of the antiviral activity assay for herpes simplex virus are shown in FIG. 3. Figure 3 (a) shows a GFP fluorescence image after 12 hours of infection, Figure 3 (b) shows a GFP fluorescence image after 24 hours of infection, Figure 3 (c) shows the cell viability after 12 hours and 24 hours of infection, Figure 3 (d) shows the number of viruses present in the cell. As can be seen from Figure 3, as a result of treatment with a mold extract, the infection rate of herpes simplex virus significantly decreased, the cell survival rate also increased compared to the virus infected group.

4 ) Analysis result of NDV-GFP virus

The results of the antiviral activity assay for Newcastle disease virus are shown in FIG. 4. Figure 4 (a) shows the GFP fluorescence image 24 hours after infection, Figure 4 (b) shows the relative amount of GFP fluorescence. As shown in FIG. 4, when the Newcastle disease virus-infected cells were treated with a mold extract, it was confirmed that the Newcastle disease virus was significantly reduced.

5 ) Analysis result of enterovirus (EV-71 virus) type 71

The results of the antiviral activity assay for enterovirus 71 are shown in FIG. 5. 5 (a) shows an optical image 12 hours after infection, and FIG. 5 (b) shows an optical image 24 hours after infection. As shown in FIG. 5, it was confirmed that enterovirus 71 was significantly reduced as a result of treating the mold extract with cells infected with enterovirus 71.

Example 3 Analysis of Pro-inflammatory Cytokine Induction in Mouse Penis Using Macrophage Cell Line

In order to confirm the immune factor induction effect of the extract hyunggae extract, a pre-inflammatory cytokine induction assay was performed.

(1) Inflammatory cytokine induction assay

Raw 264.7, a mouse macrophage line, was grown and used for analysis. After culturing the cells in a 6-well TC plate, it was treated by adding the above-described type of extract (pH adjustment) to DMEM to which 1% FBS was added. The negative control group treated only DMEM with 1% FBS, and the positive control group was a cytokine secreted in response to foreign substances such as viruses and cancer cells. Interferon, a substance that induces an anti-cancer and antiviral action and induces an immune response -β (IFN-β) was treated at 1000 Units / ml. The samples were treated with 1 μg / ml of open mold extract and the amount of TNF-α, IL-6 and IFN-β (pg / ml) was measured by ELISA on cells after 12 hours and after 24 hours.

(2) Results of whole inflammatory cytokine induction assay

The results of the proinflammatory cytokine induction assay by the mold extracts are shown in FIG. 6. Tumor necrosis factor-α (TNF-α) is mainly secreted by activated macrophages, the most important role is the regulation of immune cells. It is also known to have the ability to inhibit viral replication. Interleukin 6 (IL-6) is an important cytokine that activates B-cells to increase antibody production and promote antigen-specific immune responses Interferon (IFN) is a cell regulator that functions very well. For example, it protects cells from viruses, inhibits cell division in tissue culture or bone marrow, regulates T cell function, induces the functioning of natural immune cells (NK cells), increases phagocytosis, and special cancer cells. It is known to suppress cleavage of.

As shown in FIG. 6, it can be seen that TNF-α, IL-6, and IFN-β, which are proinflammatory cytokines, are induced by a mold extract in Raw 264.7 cells.

Example 4 Analysis of Inhibition of H1N1 and H5N2 Virus Infection by Extraction Extract

(1) Analysis method

After 24 hours treatment with 1 μg / ml open mold extract and 1.0 multiplicity of infection virus in MDCK cell line, 10 μl of Ez-Cytox reagent was treated and incubated for 12 hours to measure cytotoxicity. Absorbance was measured.

(2) Analysis result

Analysis of cell viability caused by H1N1 and H5N2 virus infection by H. N. and H5N2 virus infection showed that cell survival rate was reduced to about 20% by H1N1 and H5N2 virus infection. In one group, the cell survival rate was found to be 50-80%, which prevented a decrease in cell viability due to viral infection (FIG. 7).

Preparation Example 1 Injection

Hungyeong extract prepared in Example 1: 100 mg

Sodium metabisulfite: 3.0 mg

Methylparaben: 0.8 mg

Propylparaben: 0.1 mg

Sterile Distilled Water for Injection: Appropriate

The ingredients were mixed and prepared in a conventional manner to a final volume of 2 ml, filled in ampoules and sterilized to prepare an injection.

Preparation Example 2 Tablet

Hyojeung extract prepared in Example 1: 200 mg

Potato starch: 100 mg

Lactose: 100 mg

Colloidal silicic acid: 16 mg

Magnesium Stearate: Appropriate

The above ingredients were mixed, compressed into tablets and prepared according to a conventional tablet preparation method.

Preparation Example 3 Capsule

Hungyeong extract prepared in Example 1: 100 mg

Lactose: 50 mg

Starch: 50mg

Talc: 2 mg

Magnesium Stearate: Appropriate

The capsules were prepared by mixing the ingredients and filling the gelatin capsules according to a conventional capsule production method.

Preparation Example 4 Pill

Hungyeong extract prepared in Example 1: 120 mg

Corn starch: 100 mg

Sterilized Distilled Water: Appropriate

The above ingredients were mixed and refilled into spheres having the appropriate size according to a conventional pill preparation method to prepare pills.

Preparation Example 5 Health Functional Foods

1) health drink

Oligosaccharide (2%), liquid fructose (0.5%), sugar (2%), salt (0.5%), water (75%) and the like in a suitable amount of the mold extract extract prepared in Example 1 was mixed and sterilized Thereby preparing a beverage.

2) functional food

The functional food containing the mold extract was prepared by appropriately mixing the mold extract extracted in Example 1 with various vitamins and mineral-containing functional food.

Preparation Example 6 Feed Composition

An appropriate amount of the mold extract extracted in Example 1 was mixed with the animal feed, to prepare a feed composition, and then pelletized and granulated.

Claims (13)

1. Congenital immunity enhancement and antiviral pharmaceutical composition, characterized in that it contains an extract of hyeonggae as an active ingredient.
2. The pharmaceutical composition of claim 1, wherein the extract is extracted with water or one or more solvents selected from alcohols having 1 to 4 carbon atoms.
3. The method of claim 1, wherein the virus is one selected from orthomixoviridae, Rhabdoviridae, Paramixoviridae, Herpesviridae, and Piconaviridae. Congenital immunity enhancement and antiviral pharmaceutical composition, characterized in that the above virus.
4. According to claim 3, Orthomixoviridae is Influenza virus (Influenza virus), Rhabdoviridae (Rhabdoviridae) is Vesicular stomatitis virus (Vesicular stomatitis virus), Paramixoviridae (Paramixoviridae) is Newcastle disease virus, Herpesviridae is a herpes simplex virus, and Piconaviridae is Entero virus 71 for innate immunity enhancement and antiviral Pharmaceutical composition.
5. The pharmaceutical composition of claim 1, further comprising a pharmaceutically acceptable carrier, excipient or diluent.
6. The method of claim 1, wherein the pharmaceutical composition is in tablets, pills, powders, granules, capsules, suspension emulsions, syrups, aerosols, external preparations, suppositories, and injections, characterized in that it has a formulation selected from the group consisting of Pharmaceutical composition for antiviral.
7. Congenital immunity enhancement and anti-viral health functional food containing the extract as an active ingredient.
8. 8. The dietary supplement for innate immunity and antiviral of claim 7, wherein the dietary supplement further comprises a food supplement acceptable food supplement.
9. 8. The dietary supplement for innate immunity and antiviral of claim 7, wherein the dietary supplement has a formulation selected from the group consisting of powders, granules, tablets, capsules, candy, chewing gum, jelly and beverages.
10. The method of claim 7, wherein the virus is one selected from Orthomixoviridae, Rhabdoviridae, Paramixoviridae, Herpesviridae and Piconaviridae. Congenital immunity enhancement and antiviral health functional foods, characterized in that the above-mentioned virus for the innate immunity enhancement and antiviral.
11. The method of claim 10, wherein the Orthomixoviridae is an Influenza virus, the Rhabdoviridae is a Vesicular stomatitis virus, and the Paramixoviridae is 11. Newcastle disease virus, Herpesviridae is a herpes simplex virus, and Piconaviridae is Entero virus 71 for innate immunity enhancement and antiviral Health functional food.
12. Congenital immunity enhancement and antiviral feed composition containing a mold extract as an active ingredient.
13. A method for enhancing innate immunity and antiviral activity of an animal, characterized in that the extract is administered to an animal in need of enhancement of innate immunity except for humans.

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