KR101837448B1 - Composition for enhancing innate immunity and antivirus comprising Piperis Longi Fructus extract as effective component - Google Patents

Abstract

The present invention relates to a concomitant immunization enhancing and antiviral pharmaceutical composition containing a petroleum extract as an active ingredient, a congenital immunization containing an active ingredient, a congenital immunity enhancing and antiviral health functional food, The present invention relates to a method for promoting innate immunity and antiviral activity of an animal, which comprises administering a feed composition for enhancing and antivirus and a petal extract to an animal in need of innate immune enhancement other than humans. The composition for congenital immunity enhancement and antiviral according to the present invention exhibits antiviral activity and innate immune enhancement efficacy and can be applied for prevention and treatment of bacterial and viral infectious diseases and can be applied to patients suffering from immunosuppression or immune suppression Medicines for improving and controlling immunity, functional foods for health, and congenital immunity enhancement and antiviral feeds for enhancing anti-disease of animals. In addition, since the composition for congenital immunity enhancement and antiviral according to the present invention hardly causes toxicity or side effects, it can be safely used for prolonged use even for prophylactic purposes.

Description

[0001] The present invention relates to a composition for enhancing innate immunity and antivirus comprising an extract of Pepal as an active ingredient,

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

Immunity can be divided into innate immunity, which is born from birth, and acquired immunity, which is obtained by adapting to life. Congenital immunity is also called "natural immunity" and is characterized by nonspecific responses to antigens and no special memory effect.

Recently, interferon-derived congenital immunity has been attracting attention in the in vivo innate defense immune system because activation of interferon-mediated immunity can be a fundamental preventive measure against various infectious pathogens. Therefore, studies on the interferon activation mechanism and development of immunoregulatory agents capable of inducing interferon have been actively conducted.

On the other hand, immune factors such as cytokines are secreted as a defense mechanism against congenital immune due to infection of pathogens. Immune response occurs and defense against pathogens occurs. Therefore, it can be a preventive and therapeutic method for various infectious disease pathogens by inducing innate immune response, and it is necessary to study the enhancing agent for congenital immune which can induce it.

Virus is a Latin word for toxic substances, a group of infectious pathogenic particles that pass through bacterial filter paper (0.22 μm). Viruses can be classified into bacteriophages, plant viruses, and animal viruses according to the type of host cell. DNA viruses and RNA viruses can be classified according to the type of nucleic acid. Recently, various virus diseases such as H1N1, AI, and foot-and-mouth disease have caused a great social problem, and the concern about effective measures for viral diseases has raised a great interest in society.

Currently, vaccination is the best way to prevent viral diseases. However, in case of viral diseases, vaccine efficiency due to the generation of many viral serotypes (subtypes) is important. The development and dissemination of antiviral inhibitors that can overcome the problems of these vaccines is an important issue. For this purpose, a preventive agent that enhances immunity of an individual animal by stimulating the in vivo innate immune system, Can be an important method of drug development.

Amantadine and rimantadine are two typical antiviral agents that inhibit the proliferation of influenza virus. However, these two antiviral agents are effective only for the serotype A influenza virus and the serotype B It has been confirmed that it is not effective against influenza virus. In addition, amantadine and rimantadine have been found to have a disadvantage in that the mutant virus, which does not affect the ion channel function of the influenza virus M2 protein, appears very easily when used. To overcome this drawback, zanamivir and oseltamivir have been developed as antiviral agents effective against all 16 serotype A influenza viruses and serotype B influenza viruses. However, there is a disadvantage that Zanamivir should be administered by inhalation and intravenous injection. Ocelaminivir can be administered orally, but it is pointed out as a disadvantage due to recent reports of the emergence of resistant virus and side effects such as vomiting and dizziness when administered orally.

In addition, as the main control method of vesicular stomatitis virus is the inability to completely cure diseases, prevention and blocking prevention as well as foot-and-mouth disease and eradication of susceptible domestic livestock are the best methods.

In addition, vaccines against Newcastle Disease virus are classified into virulence vaccine and Sadox vaccine. It is known that the most widely used Newcastle disease virulence vaccine, B1 strain and La Sota strain (including Clone strain) The multivitamins combined vaccine oil vaccine, a Newcastle Disease vaccine that is used globally, can prevent three or more diseases at the same time by a single vaccination. However, in the case of laying hens farms, cases of Newcastle disease caused by immunity reduction are increasing.

There are also relatively small viruses among RNA viruses. These are called 'pico', which means small, and 'RNA', which are called picornaviruses. These viruses are called picona viridae. Enteroviruses belonging to the family of picornaviruses contain about 70 serotypes that cause various clinical symptoms such as aseptic meningitis, hand-foot disease, herpetic myelitis, acute hemorrhagic conjunctivitis and poliovirus. , Cossackie virus and echo virus, and other enteroviruses. The diameter of the enterovirus is about 20 to 30 nm, and it has a single strand RNA as a gene. Most are infected to the respiratory organs and central nervous system as well as the digestive organs of the vertebrates, but often do not show any obvious symptoms. Coxsackie virus (CXV) is a human enterovirus belonging to the picornaviridae, and is largely divided into A type and B type (Pallansch MA and Roos RP, Fields Virology, 4th edi, pp723-775 , 2001).

In recent years, high-risk enteroviruses and mutant viruses (enterovirus type 71 (EV-71) and coxsackievirus A24 mutant strains) have been newly discovered and become popular all over the world. Is required.

Enteroviruses including Coxsackie virus are infected to respiratory organs and central nervous system including vertebrate animal digestive organs and cause various clinical symptoms. Therefore, it is urgently required to prepare countermeasures at the national level. However, the types and serotypes of viruses are very various Thus, effective commercialized vaccines and therapeutic agents have not been developed.

Herpes Simplex Virus (hereinafter referred to as HSV) is a relatively large virus having a size of about 175 nm as a DNA virus belonging to the genus Herpesvirus, and is an infectious agent widely spread to humans. Herpes simplex infection is an infectious disease of HSV type 1 (hereinafter referred to as 'HSV-1') and HSV type 2 (hereinafter referred to as 'HSV-2'), It is a common infectious disease in people with HSV-1 mainly forms around the mouth and eye, and HSV-2 forms blisters around the penis. In addition, it has been reported that HSV infection is a serious cause of cervical cancer in immunocompromised patients (Melnick, JL, Adam, E. and Rawls, W., Concer, 1355-1385, 1974). In addition, it is known that newborns and fetuses can not produce HSV antibodies themselves, and that antibodies to maternal antibodies can not be passed on to the fetus. Currently, about 500,000 cases of HSV-1-associated ocular disease occur in the United States alone in a year, of which more than 1,000 are known to undergo eye transplant surgery. Approximately 95% of HSV-2 infections are known to be transmitted through sexual contact with opponents with active lesions. About 20% to 30% of US adults are infected with HSV-2, of which 20 to 50% (Johnson RE et al., N. Engl. J. Med., 321, 7, 1989).

After HSV infection, HSV begins to replicate in skin or mucosal epidermal cells and travels along the nervous system. After HSV infection, HSV is latent in the spinal ganglia throughout the entire life span, and reactivates when the immune function deteriorates. ≪ / RTI > Acyclovir, a nucleoside derivative, is known to be highly effective for early infections (Bryson, YJ et al., N. Engl. J. Med. 308, 916, 1983). In order to maintain the efficacy of this drug, it is necessary to continuously administer the drug, and when the administration is discontinued, the infectious disease caused by HSV recurs (Mindel A. et al., Lancet i: Straus SE et al., N. Engl. J. Med., 310, 1545 (1984)). It has also been reported that recurrence after treatment for the first infection of HSV has a high mortality (Nahmias, A.J. and Coleman, R.M., Immunobiology of Herpes Simplex Virus Infection CRC Press, Boca Raton, 92-102,1984).

Although a vaccine has been developed to prevent the infection of HSV, live attenuated vaccine, which weakens the virus itself, has been shown to be directly involved in the oncogenesis of the HSV genome. (Cappel, R., Sprecher, S., De Cuyper, F. and De Braekeleer, J., J. Med. Virol., 16, 137-145, 1985).

Under these circumstances, many efforts have recently been made to overcome the disadvantages of existing antiviral agents both at home and abroad. One of them is research on the antiviral efficacy of herbal medicine extracts and plant extracts in Korea , It is necessary to develop a composition containing the herbal medicine extract as an active ingredient which can overcome the disadvantages of the existing antiviral agent and exhibit the innate immune enhancing effect and the antiviral activity with almost no toxicity and side effects .

Meanwhile, Piperis Longi Fructus has been used for thousands of years in India to heal leprosy, horns, horns, chills, chills, bruises, and blood clots. These are mainly cultivated in India, imported from Korea and used as medicinal herbs have. Significant research has been carried out on the activity of the petioles, and medicinal anti-allergic effects, anti-inflammatory effects, and liver protective effects have been reported. Pepal is a dried fruit of a hinoki, and is used to warm the inside and calm the excitement with a spicy, hot medicine. It is the less ripe fruit of the piper longum Linne of the pepper and contains palmitic acid, piperidine, and piperine. Abdominal pain, vomiting, loss of appetite, diarrhea, dysentery, and toothache. It has been reported that the pharmacological action is insecticidal action, anticonvulsant action, skin vasodilating action, antioxidant action, hypothalamic and antiepileptic action, and hypolipidemic action.

Korean Patent Laid-Open Publication No. 2013-0103882 discloses a composition for suppressing obesity that contains a petroleum extract. Korean Patent Laid-Open Publication No. 2013-0023176 discloses a composition for treating pancreatic cancer and a health functional food Lt; / RTI > However, there has been no report on the effect of the extract on the development of the innate immunity and the antiviral activity.

Accordingly, the present invention has been accomplished on the basis of this finding that the present invention can inhibit the proliferation of various viruses by activating macrophages, the main cells of innate immunity. In addition, the present invention provides a method for the prevention and treatment of infectious diseases caused by various viruses or germs, while ensuring safety for a long period of time with little toxicity and side effects, and is capable of effectively using a petal extract that can be effectively used for a health food or a livestock feed composition And a method for promoting innate immunity and antiviral activity of an animal.

In order to accomplish the above object, the present invention provides a concomitant immunization enhancing and antiviral pharmaceutical composition comprising a petroleum extract as an active ingredient.

In addition, the present invention provides a congenital immunity enhancing and antiviral health functional food containing an extract as an active ingredient.

In addition, the present invention provides a congenital immunity enhancing and antiviral feed composition containing a petroleum extract as an active ingredient.

In addition, the present invention provides a method for enhancing innate immunity and antiviral activity of an animal, which comprises administering a petal extract to an animal that requires congenital immunity enhancement except for a human.

According to the present invention, it is possible to effectively apply to the prevention and treatment of various viruses and bacterial infectious diseases by using a petal extract that exerts excellent innate immune enhancing effect, and can contribute to enhancement of immunity.

In addition, the composition for congenital immunity enhancement and antivirus comprising the extract of the present invention hardly causes toxicity or side effects, so that it can be safely used for prolonged use for preventive purposes. Accordingly, the composition according to the present invention can be applied for the prevention and treatment of infectious diseases caused by bacteria and viruses, and can be applied to pharmaceutical compositions and health functional foods for immunity enhancement and control of immunocompromised patients, And an immunity enhancing feed composition aimed at enhancing anti-disease.

In addition, according to the present invention, immunization of an animal can be effectively and safely promoted by administering a petal extract to an animal necessary for congenital immunity enhancement.

1 is a graph showing the results of analysis of antiviral activity against influenza virus (PR8-GFP virus) of a petroleum extract according to an embodiment of the present invention. Medium is a negative control group of cells that have not been treated; PR8-GFP is a group of virus infections; IFN-? / PR8-GFP as a positive control, PR8-GFP virus infection and 1,000 units / ml IFN-? Treatment group; Lilipop / PR8-GFP is a group treated with PR8-GFP virus infection and petal extract.
FIG. 2 is a graph showing the results of antiviral activity analysis of the virus extract of VSV-GFP virus according to an embodiment of the present invention. Medium is a negative control group of cells that have not been treated; VSV-GFP is a virus-infected group; IFN-β / VSV-GFP was a positive control group, with VSV-GFP virus infection and 1,000 units / ml IFN-β treatment group; Lilium / VSV-GFP is a group treated with VSV-GFP virus infection and Liliaceae extract.
FIG. 3 is a graph showing the results of antiviral activity analysis of HSV-GFP virus of a filial extract according to an embodiment of the present invention. Medium is a negative control group of cells that have not been treated; HSV-GFP is a virus-infected group; IFN-β / HSV-GFP as a positive control, HSV-GFP virus infection and 1,000 units / ml IFN-β treatment group; Pigmented / HSV-GFP is a group treated with HSV-GFP virus infection and Liliaceae extract.
FIG. 4 is a graph showing the results of antiviral activity analysis of a Newcastle disease virus (NDV-GFP virus) of a filial extract according to an embodiment of the present invention. Medium is a negative control group of cells that have not been treated; NDV-GFP is a virus-infected group; IFN-? / NDV-GFP is a positive control group, NDV-GFP virus infection and 1,000 units / ml IFN-? Treatment group; Lilium / NDV-GFP is a group treated with NDV-GFP virus infection and petal extract.
FIG. 5 is a graph showing the results of antiviral activity analysis of Enterobacteriaceae extract (EV-71 virus) according to an embodiment of the present invention. Medium is a negative control group of cells that have not been treated; EV-71 is a virus-infected group; IFN-beta / EV-71 was a positive control group, with EV-71 viral infection and 1,000 Units / ml IFN-? Treatment group; Pigment / EV-71 is a group treated with EV-71 virus infection and petal extract.
FIG. 6 shows the results of the proinflammatory cytokine induction assay by the extract of the present invention according to the present invention.
FIG. 7 shows the results of confirming the inhibition of infection of influenza viruses (H1N1 and H5N2) by the petroleum extract according to the embodiment of the present invention. H1N1 and H5N2 were treated with influenza virus alone (1.0 MOI). Medium was negative control MDCK cell line. Lilium / H1N1 and pemphigus / H5N2 were treated with 1 ㎍ / ) At the same time.

The present invention relates to a concomitant immunity enhancing and antiviral pharmaceutical composition containing a petroleum extract as an active ingredient.

The wilt extract is preferably extracted with at least one solvent selected from water and an alcohol having 1 to 4 carbon atoms, more preferably extracted with a solvent of water, methanol, ethanol, or butanol, And extracted with hot water using water as a solvent. The hot water extraction comprises: 1) adding 5 to 30 times of distilled water based on the weight of the waste; 2) hot water extraction at a temperature of 100 to 130 ° C for 2 to 5 hours; And 3) filtering the hot-water extract. However, the present invention is not limited thereto.

The wilted extract preferably includes any one of an extract obtained by the extraction treatment, a diluted or concentrated liquid of the extract, a dried product obtained by drying the extract, or a controlled preparation or a purified product.

Examples of viruses having the antiviral activity of the present extract of the present invention include Orthomixoviridae, Rhabdoviridae, Paramixoviridae, Herpesviridae and Piconaviridae, (Picornaviridae), and preferably one or more viruses selected from the group consisting of influenza virus, Newcastle disease virus, Vesicular stomatitis virus, Cossackie virus, Enterovirus-71, Herpes simplex virus, rhinovirus, respiratory syncytial virus (RSV), foot and mouth disease virus, Colorado tick fever virus, reovirus, human immunodeficiency virus, B type Hepatitis virus, hepatitis C virus, swine fever virus, bovine viral diarrhea virus, Porcine reproductive and respiratory syndrome virus, porcine Ozeki disease virus, rotavirus, parvovirus, porcine epidemic diarrhea virus, and the like. Among the more preferable viruses are influenza virus, Newcastle disease virus , Vesicular stomatitis virus, herpes simplex virus, enterovirus type 71, rhinovirus, respiratory syncytial virus (RSV).

The present invention extract exhibits a strong immunological factor-inducing ability and increases the innate immunity of an individual to inhibit the infection and proliferation of viruses. Moreover, since it shows little cytotoxicity or side effects, it can be safely used . The secretion of these immune factors (TNF-α, IL-6, and IFN-β) protects the pathogen, so that the appropriate level of cytokine induction Can be a preventive and therapeutic method for a variety of epidemic pathogens. In particular, it can strengthen the immunity against viruses.

The pharmaceutical composition for congenital immunity enhancement and antiviral of the present invention may further comprise a pharmaceutically acceptable carrier, excipient or diluent. The pharmaceutically acceptable carrier, excipient or diluent which can be used in the present invention is not particularly limited so long as the effect of the present invention is not impaired. For example, a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, Flavoring agents, 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 Methylcellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, propylene glycol, polyethylene glycol, vegetable oil, injectable Ester, witepsol, macrogol, tween 61, cacao paper, and laurie paper. The pharmaceutical composition for congenital immunity enhancement and antiviral of the present invention may be in the form of a tablet, a pill, a powder, a granule, a capsule, a suspension, an emulsion, a syrup, an aerosol, a external preparation, a suppository and an injection. The method of preparing the pharmaceutical composition may be performed according to a conventional method known in the art, and is not particularly limited.

The pharmaceutical composition for congenital immunity enhancement and antiviral use according to the present invention may be administered orally or parenterally. The dosage may be adjusted according to the age, sex, weight, condition, degree of disease, drug form, May be appropriately selected, but generally about 5 to 500 mg / kg, preferably about 100 to 250 mg / kg, can be administered one to three times a day.

It will be apparent to those skilled in the art that the method of concomitant immunization enhancement and formulation of pharmaceutical compositions for antiviral use of the present invention, dosage, route of administration, constituents, etc., can be appropriately selected from conventional techniques known in the art. The secretion of these immune factors (TNF-α, IL-6, and IFN-β) protects the pathogen, so that the appropriate level of cytokine induction Can be a preventive and therapeutic method for a variety of epidemic pathogens. In particular, it can strengthen the immunity against viruses.

The congenital immunity enhancing and antiviral pharmaceutical compositions of the present invention can be used for the prevention and treatment of bacterial infectious diseases or viral infectious diseases. The pharmaceutical composition for congenital immunity enhancement and antiviral use according to the present invention may be used as an active ingredient in combination with a pharmaceutical composition other than a wilt extract or a pharmaceutical composition containing other active ingredients.

In addition, the present invention relates to a health functional food for congenital immunity enhancement and antiviral including a petroleum extract. The congenital immunity enhancing and antiviral health functional food of the present invention may further include a food acceptable food supplementary additive. Food-acceptable food supplementary additives that may be used in the present invention include sugars such as glucose, fructose, maltose, sucrose, dextrin, cyclodextrins, natural carbohydrates such as sugar alcohols such as xylitol, sorbitol and erythritol, , Natural flavor such as stevia extract, synthetic flavor such as saccharin and aspartame, colorant, pectic acid or its salt, alginic acid or its salt, organic acid, protective colloid thickener, pH adjusting agent, Alcohols, carbonates, and the like. The congenital immunity enhancing and antiviral health functional food of the present invention may be in a form selected from the group consisting of powder, granule, tablet, capsule, candy, chewing gum, jelly and beverage. The content of the volumetric extract in the health functional food for congenital immunity enhancement and antiviral treatment can be appropriately selected in consideration of the form, flavor, taste, etc. of the food, and for example, 0.01 to 30% Lt; / RTI > It is apparent to those of ordinary skill in the art that the form, composition and manufacturing method of the health functional food for congenital immunity enhancement and antiviral of the present invention can be appropriately selected from conventional techniques known in the art.

In addition, the present invention relates to a congenital immunity enhancing and antiviral feed composition comprising a wilt extract. The content of the petal extract in the feed composition for congenital immunity enhancement and antiviral treatment may be appropriately selected according to species, weight, weight and feeding conditions of the feed livestock, and is preferably 0.01 to 95% by weight, 0.1 to 80% by weight. The congenital immunity enhancing and antiviral feed composition of the present invention can be prepared according to a feed production method known in the art. For example, after mixing various feed ingredients or compound feed with the present extract of the present invention, For example, a step in the form of a pellet or a step in the form of granules or the like. It is apparent to those skilled in the art that the composition, composition, manufacturing method, feeding method, and the like of the congenital immunity enhancing and antiviral feed composition of the present invention can be appropriately selected from conventional techniques known in the art.

The present invention also relates to a method for enhancing innate immunity and antiviral activity of an animal, which comprises administering a petal extract to an animal in need of congenital immunity enhancement other than human. It is apparent to those of ordinary skill in the art that the dose, administration route, administration time, and the like of the pemphigus extract in the promotion of innate immunity and antiviral activity of the animal of the present invention can be appropriately selected from conventional techniques known in the art .

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited thereto.

Example  1. Preparation of Lilium extract

Twenty times as much distilled water was added to the volumetric flour based on the weight of the volumetric flour, and the mixture was subjected to hot water extraction at 115 ° C for 180 minutes, followed by primary filtration at 0.45 μm and secondary filtration at 0.22 μm to remove the precipitate. Subsequently, the pH was adjusted to 7.0, and 1 ml of each was dispensed into a 1.5 ml Ep-tube and stored at -20 캜 for use in all analyzes.

Example  2. Antiviral activity analysis of Lilium extract

Influenza virus (PR8), Vesicular stomatitis virus, Herpes simplex virus (HSV), Newcastle disease virus and enterovirus type 71 (Enterovirus-71; EV-71 ) Were analyzed for antiviral activity.

(1) Antiviral activity assay method

Influenza virus (Influenza virus; PR8), vesicular stomatitis virus (Vesicular stomatitis virus; VSV), a Raw 264.7 cells (8 × 10 ⁵ to herpes simplex virus (HSV-GFP) and Newcastle disease virus (Newcastle disease virus) mouse macrophage cell line (Enterovirus-71; EV-71) was infected with Hela cells and analyzed.

Cells were cultured on a 12-well TC plate, and then treated with 1 / / ml of petal extract (pH adjusted) prepared in Example 1, respectively, in DMEM supplemented with 1% FBS. Negative controls were treated with DMEM supplemented with 1% FBS, and positive controls were treated with mouse IFN-β (500 units / ml). GFP (MOI: 1.0), HSV-GFP (MOI: 3.0), NDV-GFP (MOI: 3.0) and EV-71 (MOI: 1.0), respectively. After 2 hours of inoculation, the inoculum was removed, washed three times with PBS, and after 12 and 24 hours, the degree of virus infection was confirmed.

(2) Antiviral activity assay results

1) Influenza virus ( PR8 - GFP virus ) Analysis result

The results of analysis of antiviral activity against influenza virus are shown in Fig. Fig. 1 (a) shows GFP (green fluorescent protein) fluorescence image after 12 hours of infection, and Fig. 1 (b) shows GFP (green fluorescent protein) fluorescence image after 24 hours of infection. As a result, it was confirmed that infection rate of influenza virus was remarkably decreased (Fig. 1).

2) Of vesicular stomatitis virus (VSV-GFP virus)  Analysis

The results of antiviral activity assay for vesicular stomatitis virus are shown in Fig. FIG. 2 (a) shows GFP (green fluorescent protein) fluorescence image after 12 hours of infection, and FIG. 2 (b) shows GFP (green fluorescent protein) fluorescence image after 24 hours of infection. As can be seen from FIG. 2, the treatment with a petal extract showed that the infection rate of vesicular stomatitis virus was significantly lowered.

3) Herpes simplex virus ( HSV - GFP virus ) Analysis result

The results of analysis of antiviral activity against herpes simplex virus are shown in Fig. Fig. 3 (a) shows GFP fluorescence image after 12 hours of infection, and Fig. 3 (b) shows GFP fluorescence image after 24 hours of infection. As can be seen from FIG. 3, treatment with the extract of L. elata showed that the infection rate of the herpes simplex virus was remarkably decreased.

4 ) Analysis of Newcastle disease virus (NDV-GFP virus)

The results of antiviral activity assay for Newcastle disease virus are shown in FIG. Fig. 4 (a) shows GFP fluorescence image after 24 hours of infection, and Fig. 4 (b) shows the relative GFP type light amount. As shown in FIG. 4, when the cells infected with Newcastle Disease Virus were treated with L. extract, it was confirmed that Newcastle Disease Virus was remarkably decreased.

5 ) Results of analysis of enterovirus ( EV- 71 virus ) type 71

The results of antiviral activity assay for enterovirus type 71 are shown in Fig. Fig. 5 (a) shows an optical image after 12 hours of infection, and Fig. 5 (b) shows an optical image after 24 hours of infection.

As shown in FIG. 5, when the cells infected with the enterovirus type 71 were treated with the extract of petroleum extract, it was confirmed that the type 71 of enterovirus was significantly decreased.

Example  3. Using mouse macrophages Petal  Proinflammatory cytokine induction ( pro - inflammatory Cytokine Induction ) analysis

The proinflammatory cytokine induction assay was performed to confirm the induction of the immune factor induction of Lilium extract.

(1) proinflammatory cytokine induction assay method

Mouse macrophage line Raw 264.7 was used for the analysis. Cells were cultured on a 6-well TC plate and then treated with DMEM supplemented with 1% FBS by adding the above-prepared extract (pH adjusted). The negative control group was treated with DMEM supplemented with 1% FBS, and the positive control group was a cytokine secreted in response to external substances such as viruses and cancer cells, and the interferon, which is an immune response inducing substance, -β (IFN-β) was treated at 1000 units / ml. The amount of TNF-α, IL-6, and IFN-β produced (pg / ml) was measured by ELISA on the cells treated with 1 μg / ml of the extract and 12 hours and 24 hours later.

(2) Results of proinflammatory cytokine induction assay

The results of the induction of proinflammatory cytokine by the petal extract are shown in Fig. Tumor necrosis factor-α (TNF-α) is mainly secreted by activated macrophages, the most important role being 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 stimulates B-cell activation and stimulates antigen-specific immune responses by increasing antibody production. Interferon (IFN) For example, cell protection from viruses, inhibition of cell division in bone marrow, suppression of T cell function, and hyperactivity of naturally-occurring immune cells (NK cells), thereby increasing phagocytosis, Which is known to inhibit the cleavage.

As shown in FIG. 6, it can be confirmed that the proinflammatory cytokines TNF-.alpha., IL-6 and IFN-.beta. Are strongly induced by the extract of Rhodiola from Raw 264.7 cells.

Example  4. By the extract H1N1  And H5N2  Virus Infection Inhibition Analysis

(1) Analysis method

MDCK cell line was treated with 10 μl of Ez-Cytox reagent at the same time and treated with 1 μg / ml of petal extract and 1.0 MOI (multiplicity of infection) virus for 24 hours and then cultured for 12 hours at 450 nm Absorbance was measured.

(2) Analysis results

Analysis of cell viability by H1N1 and H5N2 virus infections by Lilium extract showed that cell viability was reduced to about 20% by H1N1 and H5N2 virus infection and 1μg / ml of Lilium extract and 1 MOI virus were treated simultaneously In one group, the cell viability was 50 to 60%, which was confirmed to prevent the decrease of cell viability due to viral infection (Fig. 7).

Manufacturing example  1. Injection

The petroleum extract prepared in Example 1: 100 mg

Sodium metabisulfite: 3.0 mg

Methylparaben: 0.8 mg

Propyl paraben: 0.1 mg

Sterile sterilized distilled water for injection:

The above ingredients were mixed and made into a final volume of 2 ml by a conventional method, filled in an ampoule and sterilized to prepare an injection.

Manufacturing example  2. Refining

The petroleum extract prepared in Example 1: 200 mg

Potato starch: 100 mg

Lactose: 100 mg

Colloidal silicic acid: 16 mg

Magnesium stearate:

The ingredients were mixed and tableted according to a conventional tablet preparation method to prepare tablets.

Manufacturing example  3. Capsule

The petroleum extract prepared in Example 1: 100 mg

Lactose: 50 mg

Starch: 50 mg

Talc: 2 mg

Magnesium stearate:

The above components were mixed according to a conventional capsule manufacturing method and filled in gelatin capsules to prepare capsules.

Manufacturing example  4. Pill

The petroleum extract prepared in Example 1: 120 mg

Corn starch: 100 mg

Sterilized distilled water: suitable amount

The above ingredients were mixed and pelletized to spheres of appropriate size according to conventional pellet manufacturing methods to produce pellets.

Manufacturing example  5. Health functional foods

1) Health drinks

The desired amount of the extract prepared in Example 1 was mixed with the beverage ingredients such as oligosaccharide (2%), liquid fructose (0.5%), sugar (2%), salt (0.5%) and water (75% To prepare a beverage.

2) Functional food

A functional food containing a palatable extract was prepared by mixing the petal extract prepared in Example 1 with various vitamins and mineral-containing functional foods in an appropriate amount.

Manufacturing example  6. Feed composition

The petri dish extract prepared in Example 1 was mixed with an appropriate amount of animal feed to prepare a feed composition, which was then pelletized and granulated.

Claims (13)

The present invention relates to a vaccine composition for the prevention and treatment of congenital immune enhancement and influenza virus, Vesicular stomatitis virus, Newcastle disease virus, Herpes Simplex Virus, Enterovirus 71. The pharmaceutical composition according to any one of claims 1 to 5, 2. The antiviral pharmaceutical composition according to claim 1, wherein the wilt extract is extracted with at least one solvent selected from the group consisting of water and an alcohol having 1 to 4 carbon atoms. delete delete The antiviral pharmaceutical composition according to claim 1, further comprising a pharmaceutically acceptable carrier, excipient or diluent. The method of claim 1, wherein the pharmaceutical composition has a formulation selected from the group consisting of tablet, pill, powder, granule, capsule, suspension emulsion, syrup, aerosol, external preparation, A pharmaceutical composition for antiviral therapy for viral infection. Influenza virus, Vesicular stomatitis virus, Newcastle disease virus, Herpes Simplex virus and enterovirus type 71, which contain an extract of Pepavar as an active ingredient, (Entero virus 71). ≪ / RTI > 8. The method of claim 7, wherein the health functional food further comprises a food-acceptable food-aid additive. 9. The method of claim 7, wherein the health functional food further comprises a pharmaceutically acceptable food supplementary additive. An antiviral health functional food for a virus selected from the group consisting of Newcastle disease virus, Herpes Simplex virus, and enterovirus 71 (Entero virus 71). 8. The method of claim 7, wherein the health functional food has a formulation selected from the group consisting of powder, granule, tablet, capsule, candy, chewing gum, jelly, and beverage. An antiviral health function for any one virus selected from the group consisting of Vesicular stomatitis virus, Newcastle disease virus, Herpes Simplex virus and Enterovirus 71 food. delete delete Influenza virus, Vesicular stomatitis virus, Newcastle disease virus, Herpes Simplex virus and enterovirus type 71, which contain an extract of Pepavar as an active ingredient, (Entero virus 71). ≪ / RTI > delete

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