KR20050050618A - Functional health food product containing the extract of african phellinus mushroom for protecting and improving viral diseases - Google Patents

Abstract

The present invention relates to a dietary supplement containing African extract Phellinus baumi. Pilat showing antiviral activity.

African situation mushroom extract according to the present invention exhibits excellent antiviral activity, it can be used as an effective and safe health food for the prevention and improvement of viral diseases.

Description

Functional health food product containing the extract of African phellinus mushroom for protecting and improving viral diseases}

The present invention relates to a composition containing an extract of Phellinus baumi. Pilat, which exhibits antiviral activity, and more particularly, an African situation mushroom exhibiting excellent antiviral activity against enterovirus and bullous stomatitis virus. It relates to a dietary supplement containing the extract.

As modern medicine advances, bacterial diseases are markedly suppressed by the development of antibiotics and preventive vaccines, but viral diseases are not yet conquered, but rather are reported to increase through air pollution and environmental pollution. However, only a few viral diseases in which prophylactic vaccines are developed, and fundamental treatments for many diseases, including viral flu, hepatitis, aseptic meningitis, and the like, have not yet been developed.

Enteroviruses are single-stranded (+) RNA viruses belonging to the family Picornaviridae . It is a representative human pathogen that causes aseptic meningitis, epidemic keratoconjunctivitis (Apollo's eye disease), and hand, foot and mouth disease in children and infants. In Japan, 5000-6000 cases (5804 cases in 1998) are isolated from enteroviral diseases every year, and sporadic cases of enteroviral cases have been reported in Korea since the early 1990s.

However, isolation and identification of viruses through cell culture for diagnosis in the event of disease (Kim et al. 1994. Journal of Korea Society of Virology 24: 77-86), or genetic diagnosis using RT-PCR methods are only used. Zoll et al. 1992. Journal of Clinical Microbiology 30: 160-165. Vaccine preparations aimed at defense against enteric viral diseases have not yet been developed.

Vesicular Stomatitis Virus (hereinafter referred to as VSV virus) belongs to Rhabdoviridae , Vesiculovirus , and the virus particles exhibit a shell type of 170 × 70 nm. Single-stranded RNA virus with encapsulation and high sensitivity to lipid solvents and fungicides. In particular, bullous disease caused by bullous stomatitis virus in cattle and pigs has been of constant concern due to the difficulty in differential diagnosis between foot and mouth disease.

Sichuan mushrooms are different from China since ancient times, and they grow on the heartwood of hardwoods such as mulberry, elm, aspen and alder, and are mud mushrooms of Hymenochaetaceae A white fungus belonging to the genus Phellinus Ouel.Em.lmaz . In Korea, the situation mushroom refers to the woody mud mushroom ( Phellinus Linteus ). Situation mushrooms have been used for the treatment of gynecological diseases such as uterine bleeding and menstrual irregularities. Pharmacological and medical researches on the pharmacological action of situational mushrooms have been known to activate immune function following chemotherapy following liver cancer surgery, including gastric cancer, esophageal cancer, duodenal cancer, colon cancer, and rectal cancer, which are cancers of the digestive system. . Since the efficacy of the situation mushroom was found to be caused by polysaccharide beta glucan, research on polysaccharides has been actively conducted. In general, mushrooms contain potassium, calcium, magnesium, vitamins B2, B3 C, and fibrous amino acids. In addition to these ingredients, mushrooms contain polysaccharide β-glucan. Beta-glucan reinforces the body's immunity, repels bacteria and foreign substances that have invaded the body, and acts as a suppressor of infection. As a conventional invention related to a situation mushroom, Korean Patent Application No. 2001-0003264 discloses a Pelinus linteus strain and an anticancer immunoactive polysaccharide isolated and purified therefrom, and Korean Patent Application No. 1998-0015617 discloses a genus of Pelinus. Disclosed is a separation and purification of a novel polysaccharide material exhibiting a large amount of mycelium or fruiting body from the strain and showing immuno-enhancing activity therefrom. Korean Patent Registration No. 10-0174433 discloses an anticancer active polysaccharide isolated from Mycelium of Felinus linteus strain, a preparation method thereof, and a pharmaceutical composition comprising the same, and Korean Patent Application No. 2000-0054319 describes The use of the mushroom extract as an anticancer agent is disclosed, and Korean Patent Registration No. 10-0348115 discloses a seedling composition containing a natural situation mushroom extract and a method of preparing the same. In addition, a lot of research on domestic and international medicines and health foods using the situation mushroom and its extracts are being made.

However, not all mushrooms belonging to mud mushrooms have this effect, and to date, it has been found to be wood mud mushrooms, horse mud mushrooms and fir mud mushrooms.

African Phellinus of the present invention (Phellinus baumi. Pilat) is pine scales belonging to mushrooms and (Hymenochaetaceae) mud mushrooms in (Phellinus), growing the parasitic like hardwood belonging to Moraceae, which grows in the wild in alpine give way zone of Kenya It is a mushroom. The ecological feature of this area is the altitude of 5000m above sea level, although it is a tropical region near the equator, it has the optimal environment for mushroom growth. Perennial, the surface is grayish brown or dark gray, and has a hard shell and smooth. The underside is hollow, light brown, and porous. Spores are small and globose. Each year there is an increase in layers, and the increased layers are clearly distinguished and can be easily counted so that age can be accurately measured. Fruiting bodies are perennial hardwoods. The lampshade is generally horseshoe-shaped, 11cm high, 20cm wide and 8cm thick. When grown, the upper part is blackened, dark gray, and the surface is rough. When fully grown, the lampshade is tanned and flattened. African situation mushrooms decay live or dead wood, producing white decay in white matter and heartwood.

The medical efficacy of treating diseases has been well known in oriental medicine for a long time. It has anti-cancer effects, and the extract has also been proved by clinical trials as having an excellent therapeutic and preventive effect against liver cancer, lung cancer and many other cancers. However, the antiviral activity of the African situation mushroom has not been studied.

Kenya is known to have more than 50% of residents living with viral diseases. The inventor of the present invention focuses on the fact that the mortality rate of the inhabited area of African situation mushrooms is significantly lower than that of other regions, resulting in viral mortality, and thus, the antiviral activity of the mushrooms used as a traditional treatment of the inhabitants. As a result of the study, it was confirmed that the African situation mushroom extract exhibited excellent antiviral activity against antiviral activity, in particular, enteric virus and VSV virus, and completed the present invention.

The present invention is to provide a dietary supplement for the prevention and improvement of viral diseases containing African situation mushroom extract showing excellent antiviral activity.

In order to achieve the above object, the present invention provides a dietary supplement for the prevention and improvement of viral diseases, including African situation mushroom extract and food acceptable food supplement additives.

The virus includes enterovirus or VSV virus.

In addition, the health functional food is characterized in that the powder, granules, tablets, capsules or beverage form.

Hereinafter, the present invention will be described in detail.

African situation mushroom extract of the present invention, after cutting and grinding the dried African situation mushroom, about 5 to 25 times the weight (kg), preferably about 10 to 15 times the water, 1 to 4 carbon atoms Lower alcohols or their mixed solvents having a mixing ratio (kg / l) of about 1: 0.1 to 1:10, preferably 1: 0.2 to 1: 5, are extracted at 50 to 100 ° C., preferably 70 to 80 ° C. Soluble extract according to water, lower alcohol, or a mixed solvent thereof is extracted, concentrated under reduced pressure and freeze-dried by extraction methods such as solvent extraction and hot water extraction for about 1 hour to 1 day, preferably 2 hours to 4 hours at a temperature. It can be obtained by.

The present invention provides a health functional food comprising an extract of African situation mushrooms and a food supplement acceptable food additive which exhibits the effect of preventing and improving the viral diseases described above.

Compositions comprising the extracts of the present invention can be used in a variety of food and beverages for the prevention and improvement of viral diseases. Examples of the foods to which the extract of the African situation mushrooms can be added include various foods, for example, candy, chocolate, beverages, gum, tea, vitamin complexes, health functional foods, and the like, powders, granules, tablets and capsules. Or in the form of a beverage.

At this time, the amount of the extract in the food or beverage, in the case of the health functional food composition of the present invention can generally be added to 0.01 to 50% by weight, preferably 0.1 to 20% by weight of the total functional food composition, In the case of 0.02 to 10 g, preferably 0.3 to 1 g, based on 100 ml.

The health functional beverage composition of the present invention is not particularly limited in the liquid component except for containing the extract as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates, etc. as additional ingredients, as in general beverages. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; Conventional sugars such as polysaccharides such as dextrin, cyclodextrin and the like and sugar alcohols such as xylitol, sorbitol, erythritol and the like. The proportion of such natural carbohydrates is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.

Other flavors may be advantageously used natural flavors (tautin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.). The composition of the present invention is a variety of nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese, chocolate), pectic acid and salts thereof, alginic acid and salts thereof, Organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, etc. Other compositions of the present invention may be used in natural fruit juices and fruit juice beverages and vegetable beverages. The components may be used independently or in combination The ratio of such additives is not critical but is zero per 100 parts by weight of the composition of the present invention. It is generally selected from the range of about 20 parts by weight.

 African situation mushroom extract itself of the present invention has almost no toxicity and side effects, so can be used with confidence even for prolonged administration.

The present invention is described in more detail based on the following examples and examples, but the present invention is not limited to the examples or experimental examples.

Example 1. Preparation of African Mushroom Extract

2.1 kg of dried African mushrooms (collected from Phellinus baumi. Pilat, Kenya, Africa) were chopped and powdered, and 5 L of methanol was added thereto and then deposited at 70 ° C. for 2 hours. The methanol solution extracted from the sample was filtered through a filter paper (filter No. 1, Whatama, USA), concentrated under reduced pressure using a vacuum condenser (EYELA, Japan), and lyophilized to extract 85 g of African situation mushrooms. Got.

Example 2. Preparation of African Mushroom Extract

50 g of dried African mushrooms were hydrothermally extracted from a 1000 ml of distilled water using an electronic shaker (Heating mentle, manufactured by M-tops) for at least 3 hours. The extract was filtered, concentrated under reduced pressure using a vacuum condenser, and lyophilized to obtain an African mushroom extract having a dry weight of 20.8 g.

Experimental Example 1. Intestinal virus inhibitory activity of the situation mushroom extract

Each of the intestinal viruses of the African situation mushroom extract obtained in Example 1 was tested using a cell viability test (MTT Assay). The MTT test is a colorimetric measurement of the activity of mitochondrial dehydrogenase, which is mainly used to determine mitochondrial redox potential or cell viability (Mosmann et al. 1983).

For this experiment, RD-A cell line (Rhabdomysarcoma, National Institute of Health pediatric machine virus, October 18, 2002) was used, and enterovirus as coxsackie virus (Coxsackie B Virus 100 TCID50 (10 ^-5 / ml), Hereinafter, CB2 virus) was used (National Institutes of Pediatric Virology, October 18, 2002).

RD-A cells were cultured in Dulbecco's Modified Eagle Medium (DMEM) with 10% FBS (Fetal Bovine Serum). When the cells grew properly, the cells were washed three times with PBS (Phosphate Buffered Saline) and then PBS was removed. Trysin was treated to allow cells to fall out of the culture plate (culture plate) to be separated by one colony, followed by centrifugation (1000 rpm / 10 minutes) to form a pallet. A certain amount of DMEM was added, mixed well, and then transferred to a culture plate. African Dense Mushroom Extract solution of Example 1 in which RD-A cells (3 × 10 ⁵ cells / well) were serially diluted four-fold from the same concentration of CB2 virus (10 ⁻⁵ ) and a reference concentration of 30 ug / ml Was added to a microtiter plate with 96 wells and incubated at 37 ° C. in a 5% CO ₂ incubator for 3-4 days. At this time, both the cell control group (hereinafter referred to as CC) and the virus control group (hereinafter referred to as VC) were added with PBS solution instead of the extract of the present invention, and the cell control group was not infected with the virus. MTT reagent (Sigma, USA); 3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyl tetrazolium bromide (3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyl tetrazolium bromide) After dissolving in PBS (phosphate buffered saline) at a concentration of mg / ㎖ was used by sterile filtration with a 0.22um membrane filter (membrane filter). 50 μl of MTT (0.5 mg / ml) was added to each well, incubated at 37 ° C. for 3 hours, and then 10 μl of 0.1M HCl was added to stop MTT-formazan formation. Live cells with active mitochondria break down the tetrazolium ring to form a dark blue formazan, so to dissolve it, add 100 μl of 10% sodium dodecylsulfate (SDS) overnight. Was allowed to fully dissolve MTT-formazan and then absorbance (OD) was measured at a wavelength of 570 nm.

When the virus penetrates the cells, the cells are killed and the OD value is significantly decreased during MTT staining. However, cells containing drugs will maintain a constant OD value at a certain concentration by antiviral activity.

No. Titer Treatment Average Absorbance (Ave.OD) One Cell control CC 1.178 2 Virus control group (10 ^-5 ) VC 0.264 3 Extract of Reference Concentration 30 µg / ml 0.477 4 4 ^(-1) of the reference concentration 7.5 µg / ml 0.642 5 4 ^(-2) of the reference concentration 1.8 µg / ml 0.699 6 4 ^(-3) of the reference concentration 450ng / ml 1.315 7 4 ^(-4) of the reference concentration 110ng / ml 1.362 8 4 ^(-5) of the reference concentration 20ng / ml 1.534 9 4 ^(-6) of the reference concentration 5ng / ml 1.791 10 4 ^(-7) of the reference concentration 1.25ng / ml 1.604 11 4 ^(-8) of the reference concentration 312 pg / ml 1.645 12 4 ^(-9) of the reference concentration 78 pg / ml 0.115

CC: Cell control without added virus and extract.

VC: Virus control group with only virus but no extract.

Experimental results, as shown in Table 1 and Figure 1, in the case of cells without addition of the extract of the present invention, the OD value is significantly decreased during MTT staining, the cells added with the extract of the present invention is the term of the African situation mushroom extract OD values were kept constant at constant concentrations by viral activity.

The results of three replicates showed that the extract of the African situation mushroom (Example 1) showed antiviral activity at a concentration of 1.8 ug / ml-0.4 pg / ml against the CB2 virus.

Experimental Example 2. VSV virus inhibitory activity of African situation mushroom extract

In order to measure the inhibitory ability of the extract of the present invention against VSV virus, MDCK (Marbin & Darby Canine Kidney, National Institute of Health respiratory tract virus, October 18, 2002) cell line was used, and VSV virus (Vesicular Stomatitis Virus 100 TCID50). (10 ^-5 / mL), National Institute of Health Gastroenterology, October 18, 2002).

The experiment was conducted in the same manner as in Experimental Example 1.

No. Titer Treatment Average Absorbance (Ave.OD) One Cell control CC 1.178 2 Virus control group (10 ^-5 ) VC 0.297 3 Extract of Reference Concentration 30 µg / ml 0.078 4 4 ^(-1) of the reference concentration 7.5 µg / ml 0.416 5 4 ^(-2) of the reference concentration 1.8 µg / ml 0.945 6 4 ^(-3) of the reference concentration 450ng / ml 0.988 7 4 ^(-4) of the reference concentration 110ng / ml 1.815 8 4 ^(-5) of the reference concentration 20ng / ml 1.164 9 4 ^(-6) of the reference concentration 5ng / ml 1.964 10 4 ^(-7) of the reference concentration 1.25ng / ml 1.378 11 4 ^(-8) of the reference concentration 312 pg / ml 1.000 12 4 ^(-9) of the reference concentration 78 pg / ml 0.262

CC: Cell control without added virus and extract.

VC: Virus control group with only virus but no extract.

As a result of three repeated experiments, as shown in Table 2 and FIG. 2, antiviral activity was exhibited at a concentration of 0.5 µg / ml to 7.3 pg / ml for the VSV virus.

Experimental Example 3. Toxicity Test

In order to test the toxicity of the African situation mushroom extract of the present invention, animal experiments were performed.

The African situation of the present invention by dividing 25 ± 5g of ICR mice (Central experimental animals) and 235 ± 10g of SPF Sprague Dawley (Biogenomics) rats into three groups of 3 mice each Mushroom extract (Example 1) Toxicity was observed for 24 hours after intraperitoneal administration at a dose of 20 mg / kg, 10 mg / kg, 1 mg / kg, respectively.

As a result, no deaths were observed in all three groups, and no significant symptoms were found in weight gain and feed intake. Therefore, it could be confirmed that the African situation mushroom extract of the present invention is a safe drug.

African situation mushroom extract of the present invention can be administered in the following formulations, the following formulation examples are merely to illustrate the invention, whereby the content of the present invention is not limited.

Formulation Example 1 Preparation of Healthy Food

1000 mg of extract of Example 1

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B ₁ 0.13 mg

Vitamin B ₂ 0.15 mg

Vitamin B ₆ 0.5 mg

0.2 μg of vitamin B ₁₂

Vitamin C 10 mg

10 μg biotin

Nicotinic Acid 1.7 mg

Folate 50 ㎍

Calcium Pantothenate 0.5mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. The granules may be prepared and used for preparing a health food composition according to a conventional method.

Formulation Example 2 Preparation of Healthy Drinks

1000 mg of extract of Example 1

Citric acid 1000 mg

100 g oligosaccharides

Plum concentrate 2 g

1 g of taurine

Add 900 ml of purified water

After mixing the above components in accordance with a conventional healthy beverage production method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed sterilization and then refrigerated and stored in the present invention For the preparation of healthy beverage compositions.

African situation mushroom extract according to the present invention exhibits antiviral activity can be used as a health functional food useful for the prevention and improvement of various viral diseases.

1 is a diagram showing the antiviral activity of the African situation mushroom extract against Koksagi virus,

Figure 2 is a diagram showing the antiviral activity of the African situation mushroom extract against VSV virus.

Claims (3)

 A dietary supplement for the prevention and improvement of viral diseases comprising African situation mushroom extract and food acceptable food supplement additives. The health functional food according to claim 1, wherein the virus is an enteric virus or Vesicular Stomatitis Virus.  The dietary supplement of claim 1 in the form of a powder, granule, tablet, capsule or beverage.