KR101930778B1 - Composition for antivirus comprising Ulmi Cortex extract as effective component - Google Patents
Composition for antivirus comprising Ulmi Cortex extract as effective component Download PDFInfo
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- KR101930778B1 KR101930778B1 KR1020150042939A KR20150042939A KR101930778B1 KR 101930778 B1 KR101930778 B1 KR 101930778B1 KR 1020150042939 A KR1020150042939 A KR 1020150042939A KR 20150042939 A KR20150042939 A KR 20150042939A KR 101930778 B1 KR101930778 B1 KR 101930778B1
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Abstract
The present invention relates to an antiviral pharmaceutical composition containing an extract of a milky lotion as an active ingredient, an antiviral health food containing an extract of a milky lotion as an active ingredient, an antiviral feed composition containing an extract of a milky lotus as an effective ingredient, The present invention relates to a method for promoting antiviral activity of an animal, which comprises administering a bodily extract to an animal in need of innate immune enhancement other than human. The antihypertensive astrocyte composition according to the present invention exhibits antiviral activity and innate immune enhancing effect and can be applied for the prevention and treatment of bacterial and viral infectious diseases and can be used for immunity enhancement and immunosuppression Medicines for the control, health functional foods, and antiviral feeds for enhancing the anti-disease of animals. Further, since the antiviral composition according to the present invention hardly causes toxicity or side effects, it can be safely used for prolonged use even for prophylactic purposes.
Description
The present invention relates to an antiviral composition containing an extract of a milky lotion as an active ingredient, and more particularly to an antiviral composition containing an extract of a milky lotion which can be used as a pharmaceutical, a health functional food or a feed composition as an active ingredient will be.
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).
In this circumstance, recently efforts have been made to overcome the disadvantages of existing antiviral agents both at home and abroad, one of which is research on antiviral efficacy against 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 .
On the other hand, Ulmi Cortex is a medicinal herb that has been used as a folk medicine for long time in Korea and Northeast Asia as a bark of the root of the elm (Ulmus macrocarpa Hance) which belongs to the elm family and the bark of the bark cork . In the Donghak Dictionary, there is a tendency to develop swelling, urination, diarrhea, constipation, and swelling of the blood vessels. It is also known to be applied to relieving, treating, solitary, mastitis. Studies on the components of the milky blood have so far been carried out with the use of sterols (β-sitosterol, phytosterol, stigmasterol), hydrolyzable tannins, resins, fatty oils, catechins and catechin complexes Procyanidins, and the like.
It has been reported that catechin polymer called procyanidin oligomer isolated from the extract of Eurasia japonica inhibits bacterial collagenase and inhibits periodontal disease (J Periodontal Res. 2003 Jun; 38 (3): 282-289 ). Korean Patent Laid-Open Publication No. 2006-0095062 discloses a composition for promoting hair growth through hair-promoting action of milky blood. Korean Patent Publication No. 2006-0094704 discloses a composition for inhibiting collagen A < RTI ID = ≪ / RTI > However, the antiviral activity using the extract of Milkweed has not been reported yet.
Accordingly, the present invention has been accomplished on the basis of the above findings. It is an object of the present invention to provide a method for inhibiting the proliferation of various viruses by activating macrophages, the major cells of innate immunity. The present invention also relates to a pharmaceutical composition for preventing or treating infectious diseases caused by various viruses or germs, a milky oil extract which can be effectively used for a health functional food or a livestock feed composition, while ensuring safety for a long period of time with little toxicity and side effects An antiviral composition containing the compound as an active ingredient, and a method for promoting the antiviral activity of an animal.
In order to achieve the above object, the present invention provides an antiviral pharmaceutical composition comprising an extract of a milky lotion as an active ingredient.
Further, the present invention provides an antiviral health functional food containing an extract of a milky lotion as an active ingredient.
In addition, the present invention provides an antiviral feed composition containing an extract of a milky lotus as an active ingredient.
In addition, the present invention provides a method for promoting antiviral activity of an animal, which comprises administering an astringent bark extract to an animal requiring innate immune enhancement except for a human.
According to the present invention, by using the extract of Eucalyptus japonica, which exerts an excellent innate immunity-enhancing effect, it can be effectively applied to the prevention and treatment of various viruses and bacterial infectious diseases and can contribute to the enhancement of immunity.
In addition, the antiviral composition comprising the extract of the present invention can be safely used for prophylactic purposes for a prolonged period of time since it does not cause toxicity or side effects. 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, the immunization of an animal can be effectively and safely promoted by administering the extract of the present invention 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 whey protein 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; Milkweed / PR8-GFP is a group treated with PR8-GFP virus infections and whey protein extract.
FIG. 2 is a graph showing the results of antiviral activity assay for the vesicular stomatitis virus (VSV-GFP virus) of the white-and-white extract 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; Milky white / VSV-GFP is a group treated with VSV-GFP viral infection and milky blood extract.
FIG. 3 is a graph showing the results of antiviral activity analysis of HSV-GFP virus 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; HSV-GFP is a group treated with HSV-GFP viral infection and whey protein extract.
FIG. 4 is a graph showing the results of antiviral activity assay of NDV-GFP virus of a Milkweed 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; Milkweed / NDV-GFP is a group treated with NDV-GFP viral infection and milky blood extract.
FIG. 5 is a graph showing the results of antiviral activity analysis of enterobacteriaceae (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; Milky white / EV-71 is a group treated with EV-71 virus infection and milky blood extract.
FIG. 6 shows the results of the proinflammatory cytokine induction assay by the extract of a milky white blood cell according to an embodiment of the present invention.
FIG. 7 shows the results of confirming the inhibition of infection of influenza viruses (H1N1 and H5N2) by the extract of a milky white blood cell according to an embodiment of the present invention. H1N1 and H5N2 were treated with influenza virus only (1.0 MOI), Medium was a negative control MDCK cell group, and milky blood / H1N1 and milky blood / H5N2 were obtained from 1 μg / ml of milky blood extract and influenza virus (1.0 MOI) at the same time.
The present invention relates to an antiviral pharmaceutical composition containing an extract of a milky lotion as an active ingredient.
Preferably, the above-mentioned extract is extracted with at least one solvent selected from the group consisting of water and an alcohol having 1 to 4 carbon atoms, more preferably extraction with a solvent of water, methanol, ethanol or butanol, Is a hot-water extraction 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 milky blood; 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.
In addition, the above-mentioned milky skin extract preferably includes any one of the extract obtained by the extraction treatment, the diluted or concentrated liquid of the extract, the dried product obtained by drying the extract, or the adjusted product or the purified product.
An example of a virus having an antiviral activity of the present invention of the present invention is Orthomixoviridae, Rhabdoviridae, Paramixoviridae, Herpesviridae and Piconazole (Vesicular stomatitis virus, Cossackie virus, Enterovirus-71, Enterococcus faecalis, Enterococcus faecalis, Enterovirus-71, Viruses such as Herpes Simplex Virus, Rhinovirus, respiratory syncytial virus (RSV), foot and mouth disease virus, Colorado tick fever virus, reovirus, human immunodeficiency virus, B Hepatitis virus, hepatitis C virus, swine fever virus, bovine viral diarrhea virus, Porcine reproductive and respiratory syndrome virus, porcine Ozeki disease virus, rotavirus, parvovirus and porcine epidemic diarrhea virus. Examples of the more preferable virus include influenza virus, Newcastle disease virus , Vesicular stomatitis virus, herpes simplex virus,
The astaxanthin extract of the present invention strongly shows the immunity-inducing ability and increases the innate immunity of the individual to inhibit the infection and proliferation of the virus. In addition, since it shows little cytotoxicity or side effects, Can be 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 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 antiviral pharmaceutical composition of the present invention may be in a form selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories and injections. 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 antiviral pharmaceutical composition of the present invention may be administered orally or parenterally. The dosage may be appropriately selected depending on the age, sex, weight, condition, degree of disease, drug form, 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 formulation, dose, route of administration, constituents, etc. of the pharmaceutical composition for antiviral of the present invention can be suitably selected from conventional techniques known in the art.
The pharmaceutical composition for antiviral of the present invention can be used for the prevention and treatment of bacterial infectious diseases or viral infectious diseases. The antiviral pharmaceutical composition of the present invention may be used as an active ingredient in combination with a pharmaceutically active ingredient in addition to the extract of a milky lotion, or in combination with a pharmaceutical composition containing other effective ingredient.
The present invention also relates to an antiviral health functional food comprising a milk extract. The anti-viral health functional food of the present invention may further comprise a pharmaceutically acceptable food-aid 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, , A natural flavor such as stevia extract, a synthetic flavor such as saccharin and aspartame, a coloring agent, a pectic acid or a salt thereof, an alginic acid or a salt thereof, an organic acid, a protective colloid thickener, a pH adjusting agent, a stabilizer, Alcohols, carbonates, and the like. The 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 eucaryotic extract in the anti-viral health functional food may be appropriately selected in consideration of the form, flavor, taste, etc. of the food, and may be, for example, in the range of 0.01 to 30% have. It is apparent to those of ordinary skill in the art that the form, composition and manufacturing method of the antiviral health functional food of the present invention can be suitably selected from conventional techniques known in the art.
In addition, the present invention relates to an antiviral feed composition comprising a milky lotion extract. The content of the dandruff extract in the antiviral feed composition may be appropriately selected according to the kind, age, body weight, rearing condition, etc. of the feed livestock, and is 0.01 to 95% by weight, preferably 0.1 to 80% % ≪ / RTI > by weight. The antiviral feed composition of the present invention may be prepared according to a method of manufacturing feeds known in the art. For example, after mixing various feed materials or compound feeds with the whey protein extract of the present invention, For example, it can be produced by pelletizing or by further performing a cutting 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 antiviral feed composition of the present invention can be suitably selected from conventional techniques known in the art.
The present invention also relates to a method for promoting the antiviral activity of an animal, which comprises administering an astringent bark extract to an animal requiring congenital immunity enhancement except for a human. It is obvious to those of ordinary skill in the art that the dose, route of administration, time of administration, etc. of the dermatophyte extract may be suitably selected from conventional techniques known in the art in the promotion of the antiviral activity of the animal of the present invention.
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 a milky skin extract
The extract was prepared by adding 20-fold distilled water to the whitened blood, extracting it at 115 ° C for 180 minutes, firstly filtering it to 0.45 μm and then filtering it to 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 the extracts
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 DMEM supplemented with 1% FBS was treated with 1 쨉 g / ml of milky white extract (pH adjusted) prepared in Example 1, respectively. 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 : 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. 1 (a) shows GFP (green fluorescent protein) fluorescence image after 12 hours of infection, FIG. 1 (b) shows green fluorescent protein (GFP) fluorescence image after 24 hours of infection, And cell survival rate after 24 hours, and Fig. 1 (d) shows the number of viruses present in the cells. As a result of the treatment with the extract of Eucalyptus japonicus, the infection rate of influenza virus was remarkably decreased and the cell survival rate was also higher than that of the virus infection group (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, FIG. 2 (b) shows green fluorescent protein (GFP) fluorescence image after 24 hours of infection, And cell survival rate after 24 hours, and Fig. 2 (d) shows the number of viruses present in the cells. As can be seen from FIG. 2, as a result of the treatment with the extract of Milkweed, the infection rate of the vesicular stomatitis virus was remarkably decreased, the virus level was decreased, and the cell survival rate was also higher than that of the virus infection group.
3) Herpes simplex virus ( HSV - GFP virus ) Analysis result
The results of analysis of antiviral activity against herpes simplex virus are shown in Fig. 3 (a) shows GFP fluorescence image after 12 hours of infection, FIG. 3 (b) shows GFP fluorescence image after 24 hours of infection, FIG. 3 (c) shows cell viability after 12 hours and 24 hours of infection, Figure 3 (d) shows the number of viruses present in the cells. As can be seen from FIG. 3, the infection rate of herpes simplex virus was significantly lowered and the cell survival rate was also higher than that of virus infection group.
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 the Newcastle disease virus were treated with the extract of the Eustachian tube, it was confirmed that the Newcastle disease virus was significantly reduced.
5 ) Results of analysis of enterovirus ( EV- 71 virus )
The results of antiviral activity assay for
Example 3. Using mouse macrophages Milky blood Proinflammatory cytokine induction ( pro - inflammatory Cytokine Induction ) analysis
The proinflammatory cytokine induction assay was carried out to confirm the immune factor induction effect of the extract of the whitish blood.
(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 treated with DMEM supplemented with 1% FBS by adding the above-prepared milky white 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 dandruff extract and after 12 hours and 24 hours.
(2) Results of proinflammatory cytokine induction assay
The results of the proinflammatory cytokine induction assay by the dermatophyte 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-a, IL-6 and IFN-? Are strongly induced by the extract of Ehrlichia cylindrica in Raw 264.7 cells.
Example 4. by the extract of milky blood H1N1 And H5N2 Virus Infection Inhibition Analysis
(1) Analysis method
MDCK cell line was treated with 1 쨉 g / ml of dermatophyte extract and 1.0 MOI (multiplicity of infection) at the same time, and after 24 hours, 10 쨉 l of Ez-Cytox reagent was treated for 12 hours to measure cytotoxicity, The absorbance was measured.
(2) Analysis results
The cell viability by H1N1 and H5N2 virus infection by HLA - B and H5N2 virus infection was decreased to 20% by H1N1 and H5N2 virus infection and 1 μg / In the group treated at the same time, the cell survival rate was 80 to 90%, and it was confirmed that the decrease in cell viability due to virus infection was prevented (FIG. 7).
Manufacturing example 1. Injection
The whitish blood 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
200 mg of the milky skin extract prepared in Example 1
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 whitish blood 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 dandruff 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 milk material of Example 1 was mixed with an appropriate amount of beverage ingredients such as oligosaccharide (2%), liquid fructose (0.5%), sugar (2%), salt (0.5% The beverage was prepared by sterilization.
2) Functional food
The functional food containing the extract of Eucalyptus japonica was prepared by mixing the Eucalyptus extract prepared in Example 1 with various vitamins and mineral-containing functional foods in an appropriate amount.
Manufacturing example 6. Feed composition
The animal feed composition was mixed with the milky white extract prepared in Example 1 in an appropriate amount to prepare a feed composition, which was then pelletized and granulated.
Claims (13)
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| KR1020150042939A KR101930778B1 (en) | 2015-03-27 | 2015-03-27 | Composition for antivirus comprising Ulmi Cortex extract as effective component |
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| KR1020150042939A KR101930778B1 (en) | 2015-03-27 | 2015-03-27 | Composition for antivirus comprising Ulmi Cortex extract as effective component |
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| KR101930778B1 true KR101930778B1 (en) | 2018-12-19 |
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Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| KR101947271B1 (en) | 2017-07-03 | 2019-02-12 | 동국대학교 경주캠퍼스 산학협력단 | Composition comprising ulmi cortex extracts for inhibiting ovotoxicity |
| KR102345933B1 (en) * | 2021-04-23 | 2022-01-03 | 강원대학교산학협력단 | Antiviral composition containing extract of high content of catechin glycoside derived from Ulmus species as an active ingredient |
| KR102558581B1 (en) * | 2021-06-17 | 2023-07-24 | 주식회사 첨단환경 | Composition for preventing or treating PEDV comprising extract of Ulmus davidiana var. japonica supercritical extract residue and Chamaecyparis Obtusa emulsion complex |
| CN114652731B (en) * | 2022-03-16 | 2023-12-12 | 华中农业大学 | Application of tannic acid in preparation of medicine for preventing and treating African swine fever |
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| JPS62142181A (en) * | 1986-12-08 | 1987-06-25 | Nippon Shinyaku Co Ltd | Novel tannin |
| JP2738908B2 (en) * | 1994-01-20 | 1998-04-08 | 保芦 将人 | Antiviral powder material and antiviral extract |
| KR100348870B1 (en) * | 2000-01-07 | 2002-08-17 | 삼성전자 주식회사 | Peptido-glyco Compounds from Chinese elm and Process for Preparing the Same |
| KR100508401B1 (en) * | 2003-01-24 | 2005-08-19 | 오수진 | An Extract having anticancer activity |
| JP2006265260A (en) * | 2006-05-15 | 2006-10-05 | Hoashi Masahito | Food and drink |
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Non-Patent Citations (2)
| Title |
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| Acta Chinese Medicine and Pharmacolog, 2006, 34(6) pp. 27-30* |
| The Korean Journal of Pesticide Science, 2010, 14(3), pp. 280-283* |
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