KR20060116280A - Composition and extracts of clavicorona pyxidata for curing alzheimer's disesase - Google Patents

Abstract

A composition for treating Alzheimer's disease comprising the extract of Clavicorona pyxidata is provided to inhibit activity of acetylcholine esterase(AChE), propyl endopeptidase(PEP) and beta-secretase which cause Alzheimer's disease without side effects. The composition for treating Alzheimer's disease comprises the extract of Clavicorona pyxidata, wherein the Clavicorona pyxidata extract is prepared by extracting fruit bodies and mycelium of Clavicorona pyxidata with ethyl alcohol and water, filtering the Clavicorona pyxidata extract, concentrating and freeze-drying the filtered Clavicorona pyxidata extract; and the composition is formulated as powder, granule, tablet, capsule, liquid and injection.

Description

Composition and extracts of Clavicorona pyxidata for curing Alzheimer's disesase for treating Alzheimer's disease

Figure 1 is a schematic diagram of the preparation method of the extract for each part of the wormwood mushroom.

Figure 2 is a schematic drawing of the preparation method of the extract for each organic solvent of the wormwood mushroom.

The present invention relates to the preparation of the extract of the bark mushrooms having the efficacy of preventing and treating Alzheimer's disease, a pharmaceutical composition and a method for producing a dietary supplement. The creeper mushroom is a edible mushroom belonging to the species of Hymenomycetidae, Aphyllophorales, and Clavariaeae, which is 15 cm in height and butterfly, and the shape of the mushroom is different from that of ordinary mushrooms. Unlike the whole tree branches or corals. Branches are gradually branched upward from the thick white stem with a diameter of 3-5cm, and many small branches are concentrated at the tip. The wormwood mushroom is commercially used as an edible mushroom with excellent meat quality and has been reported to be inhibitory effect of reverse transcriptiase of AMV virus, but it has been reported to be related to the physiological, physiological and pharmacological characteristics. The information is very poor.

To date, the causes of Alzheimer's disease have been shown to be a sign of significant decline in memory and cognition, suggesting evidence that it is closely related to the cholinergic nervous system. In the brain of AD patients, β, γ-secretase Due to activity Abnormal toxic proteins called β-amyloid are deposited, causing neural plaques and nerve knots to cause impaired cognition. Current trends in the treatment of Alzheimer's disease are focused on the development of drugs that can improve the reduced cognitive function by supplementing the deteriorated nervous system, even if it is not a causal treatment method. Representative acetylcholinesterase (AchE) inhibitors, Cognex, Aricept, and Galantamine, are available on the market with FDA approval. These drugs are synthetic, mono-drug drugs that are temporary and weak in effect, and selectively in the brain. Because it does not work, it is difficult to administer the drug for a long time due to various side effects. In addition, the inhibitory activity of β-secretase related to β-amyloid deposition, which is expected as the next generation Alzheimer's treatment, has not been developed yet. Prolyl endopeptidase (PEP) inhibitors reported to date are mostly synthetic inhibitors or microbial origin, but there are few reports on mushroom-derived PEP inhibitors. Synthetic agents that are currently used as a treatment for Alzheimer's disease have problems such as temporary efficacy and serious side effects, which require urgent improvement.

In the present invention, in order to solve the problems of the current side effects of synthetic therapeutic drugs, extracts of each part and culture filtrate, mycelium, and fruiting bodies of edible green mushrooms, which proved to be safe for treating Alzheimer's disease, and for improving brain function It is characterized by a pharmaceutical composition and a method of preparing a dietary supplement.

The present invention will be described in detail the extract of each part and fraction, extract and brain composition for improving brain function and a method for producing health functional foods having the effect of preventing and treating Alzheimer's disease. In order to achieve the above object, the present invention comprises the steps of preparing a mycelium and fruiting body of the wormwood mushroom, the culture filtrate of the prepared wormwood mushroom, the step of preparing the extract for each part of the mycelium and fruiting body, cultivation of the mushroom It provides a manufacturing method comprising the step of obtaining the fractional extract of the filtrate, mycelium and fruiting body, the pharmaceutical composition for improving brain function and the manufacturing of health functional food.

Aspen mushrooms used in the present invention were used to compare the strains distributed from the Korea Institute of Biotechnology Gene Bank (KCTC) and the wild botanical mushrooms fruiting body and the mycelium was purely cultured from the fruiting body in a conventional manner. The method for preparing mycelium mycelium mycelium extract having the effect of preventing and treating Alzheimer's disease was incubated at 120 rpm for 20 days at 120 rpm after inoculating 5% (w / v) in YMG liquid medium at pH 4-9. . In the method of preparing fruiting bodies, seedlings and seedlings were inoculated with the seedlings of the barley mushroom in a conventional manner, and the fruiting bodies produced after incubating the mycelia under 15 to 28 ° C. and 95% humidity for 15 to 30 days were separated.

Hereinafter, the method of preparing the extract of Zapsiaceae mushrooms having an effect of preventing and treating Alzheimer's disease according to the present invention will be described in detail. However, these examples are merely illustrative of the present invention, the present invention is not limited thereto.

Example 1.

Preparation of extracts of different parts of hawthorn mushrooms (see Fig. 1)

1) Korea Biotechnology genes to receive pre-sale a little from the bank (KCTC) Prepare the wood Hagi mushrooms (Clavicorona pyxidata KCTC16042) mycelium, pH 4 ~ 9, YMG 1 ~ 5% in liquid medium (w / v) after vaccination, 20 - 120 rpm was incubated at 28 degreeC for 15 to 30 days.

2) The manufacturing process of the extract of cultivated mushrooms of Pleurotus eryngii is as follows. The prepared liquid culture was filtered with filter paper to separate the supernatant and mycelium. The supernatant obtained after the filtration was frozen and then freeze-dried at minus 70 ℃, 0.01 atm and used as a culture filtrate extract sample.

3) The manufacturing process of mycelium mycelium mycelium extract is as follows. After filtration of the culture filtrate, 4 times of ethyl alcohol was added to the isolated mycelium, and extraction was performed at 40 to 60 ° C. for 4 hours. After extraction, the filtrate was separated, the supernatant was separated, and concentrated under reduced pressure at 65 to 80 ° C. to prepare an ethyl alcohol extract of the mycelium mycelia. Water extract of the mycelium was added to the mycelium and 4 times the water together, hot water extraction for 4 hours at 100 ℃, and the supernatant was separated through a filtration process. Thereafter, the supernatant was frozen, freeze-dried at -70 ° C and 0.01 atm, and used as mycelium hydrothermal extract.

4) The manufacturing process of the extract of fruiting body of the bark mushrooms is as follows. Fruiting body extract was put into fruiting body with 4 times the number of ethyl alcohol, and then pulverized at 1000 rpm using a crusher and extracted for 4 hours at 40 ~ 60 ℃. Filtration was performed after extraction to separate the supernatant, and concentrated under reduced pressure at 70 ° C. to prepare an ethyl alcohol extract of the fruiting body of Pleurotus eryngii. The fruit extract of the fruiting body was put into the fruiting body and 4 times the number of water, after the grinding process, hot water extraction at 100 ℃ for 4 hours, and the supernatant was separated through a filtration process. Thereafter, the supernatant was frozen, freeze-dried at -70 ° C and 0.01 atm, and used as mycelium hydrothermal extract.

Example 2.

A process for preparing extracts from fractions of cultured filtrate, mycelium, and fruiting body of Pleurotus eryngii (see Figure 2)

1) Preparation of the extract of each fraction of the mushroom mushroom culture filtrate, mycelium, fruiting body was carried out by the following method. To each sample of the part of the bark mushrooms in 4 times of ethyl alcohol and extracted for 4 hours at 60 ℃, filtered using a filter paper to obtain a supernatant. The obtained supernatant was concentrated under reduced pressure at 70 ° C., to prepare an ethyl alcohol extract of Shiratake mushroom.

2) The method of preparing the hexane fraction is as follows. The ethyl alcohol extract was dissolved in 4 times distilled water, and then dissolved in a separatory funnel by adding 1: 1 volume of hexane, followed by standing at room temperature for 24 hours. After 24 hours, the hexane layer and the water layer were separated, and the separated hexane layer was hot-air dried in an oven at 70 ° C. to prepare a hexane fraction.

3) The method for preparing the chloroform fraction is as follows. The water layer separated in step 2 was added by mixing 1: 1 volume of chloroform in a separatory funnel, and then allowed to stand at room temperature for 24 hours. After 24 hours, the chloroform layer and the water layer were separated, and the separated chloroform layer was hot-air dried in an oven at 70 ° C. to prepare a chloroform fraction.

4) The preparation method of ethyl acetate fraction is as follows. The water layer separated in step 3) was added to the separatory funnel by adding 1: 1 volume of ethyl acetate, and then allowed to stand at room temperature for 24 hours. After 24 hours, the ethyl acetate layer and the water layer were separated, and the separated ethyl acetate layer was hot-air dried in an oven at 70 ° C. to prepare an ethyl acetate fraction.

5) The manufacturing method of the water layer is as follows. The water layer separated in step 4) was frozen in a freezer at -20 ° C and then freeze-dried at -70 ° C 0.01 atm to prepare a water fraction.

Experimental Example 1.

Determination of Acetylcholinesterase (AChE) Inhibitory Activity of Extracts from Different Part of Mushrooms.

The inhibition activity of acetylcholinesterase (AChE) was measured using the method of Ellman et al. As a positive control, tacrine, which is currently approved by the FDA and used as a treatment for Alzheimer's disease, was used.

In each well of a 96-well plate, 95 µl of buffer solution (50 mM phosphate buffer, pH 8.0), 15 µl of extract of Botanical mushrooms and 20 µl of acetylcholinesterase ( AChE) was added and reacted at 37 ° C. for 15 minutes. Then, 100 μl of substrate (Ellmans reaction solution) was added to each well and reacted at 37 ° C. for 10 minutes. Then, the absorbance was measured at 412 nm of an ELISA reader and the change was used. AChE enzyme inhibitory activity was measured.

Table 1. Determination of Acetylcholinesterase (AChE) Inhibitory Effect of Extracts from Different Part of Mushrooms

Sample (10 ug / ml) Acetylcholinesterase inhibition rate (%) Mycelium Extract Culture filtrate extract Fruiting body extraction Ethanol extraction Water extraction Ethanol extraction Water extraction Shiitake mushroom 78.3 ± 1.7 80.1 ± 2.1 88.0 ± 5.2 65 ± 2.1 70 ± 1.3 Taclean 87.4 ± 6.5

※ The measurement was repeated three times, and the result is expressed as the average value ± standard deviation.

※ Tacrine, a positive control, is sold with FDA approval and reported side effects such as hepatotoxicity.

As shown in Table 1, it can be seen that the extract of each part of the wormwood mushroom has an inhibitory effect on acetylcholinesterase (AChE) comparable to tacrine, which is a synthetic monopharmaceutical drug, and especially in the case of a culture filtrate extract, a positive control Inhibition of acetylcholinesterase was superior to that of intacrine. Considering that the positive control tacrine is a high-purity purified drug, it can be seen that the acetylcholinesterase (AChE) inhibitory activity of the unrefined bastard mushrooms is much better than tacrine, a positive control.

Experimental Example 2.

Determination of the prolyl endopeptidase inhibitory activity of the extracts of different parts of the mushrooms.

Inhibition activity of prolyl endopeptidase (PEP) was measured by Toda method, and Z-Pro-prolinal was used as a positive control. 800 μl buffer solution (100 mM phosphate buffer pH 7.0), 40 μl extract of mushroom mushrooms and 80 μl prolyl endopeptidase (PEP) were added to the cuvette for absorbance measurement. I was. Subsequently, 80 μl of substrate, Z-Gly-Pro-pNA, was added and reacted at 37 ° C. for 15 minutes, and the change in absorbance at 410 nm was measured. Enzyme inhibitory activity was measured.

Table 2. Determination of Prolyl Endopeptidase (PEP) Enzyme Inhibitory Effect of Extracts from Various Parts of Prickly Pear Mushroom

Sample (10 ug / ml) Prolyl endopeptidase inhibition rate (%) Mycelium Extract Culture filtrate extract Fruiting body extraction Ethanol extraction Water extraction Ethanol extraction Water extraction Shiitake mushroom 86.2 ± 4.2 81.8 ± 2.1 91.3 ± 3.4 82 ± 1.6 84 ± 2.4 G-Pro-Final 90.4 ± 2.3

※ The measurement was repeated three times, and the result is expressed as the average value ± standard deviation.

※ G-Pro-prorinal, a positive control, is a synthetic drug and not a drug.

As shown in Table 2, the extract of each part of the wormwood mushroom showed the best prolyl endopeptidase (PEP) inhibitory activity in the culture filtrate extract, and showed an excellent inhibitory effect on the extract of each part. The G-pro-prorinal used as a positive control showed an inhibition rate of about 90.4%, but it is a synthetic agent to test the inhibition of enzyme activity and cannot be used as a medicine. To date, studies on the inhibition of enzymatic activity of prolyl endopeptidase (PEP) for the development of a therapeutic agent for Alzheimer's disease have not been conducted with edible mushrooms.

Experimental Example 3.

Determination of Acetylcholinesterase (AChE) Inhibitory Activity of Extracts from Culture Filtrate, Mycelium and Fruiting Body Fractions of Pleurotus eryngii Mushroom.

The inhibition activity of acetylcholinesterase (AChE) was measured using the method of Ellman et al. As a positive control, tacrine, which is currently approved by the FDA and used as a treatment for Alzheimer's disease, was used. Each well of a 96-well plate contains 95 μl of buffer solution (50 mM phosphate buffer, pH 8.0), 15 μl of mushroom mushroom culture filtrate, mycelium, fruiting body fraction extract and 20 μl Acetylcholinesterase (AChE) was added and reacted at 37 ° C. for 15 minutes. Then, 100 μl of substrate (Ellman's reaction solution) was added to each well and reacted at 37 ° C. for 10 minutes. Then, the absorbance was measured at 412 nm of ELISA reader and the change value was used. AChE enzyme inhibitory activity was measured.

Table 3. Inhibitory Effects of Extracts from Fractions of Cultured Ficus, Mycelium and Fruiting Botanical Mushrooms on Acetylcholinesterase (AChE) Activity

Sample (10 ug / ml) Acetylcholinesterase inhibition rate (%) extract Fraction ethanol Hexane Chloroform Ethyl acetate water Cultivation of mushrooms 85.2 ± 1.1 85.4 ± 1.6 83.3 ± 3.5 85.2 ± 3.1 74.5 ± 3.8 Myrtle mycelium 74.2 ± 6.1 87.4 ± 5.6 83.3 ± 7.5 81.2 ± 2.1 87.5 ± 3.8 Bodhi tree fruiting body 86 ± 1.7 85.3 ± 3.4 84 ± 1.4 81.3 ± 2.0 84.1 ± 4.1 Tacrine 82.3 ± 4.5

※ The measurement was repeated three times, and the result is expressed as the average value ± standard deviation.

※ Tacrine, a positive control, is sold with FDA approval and reported side effects such as hepatotoxicity.

As shown in Table 3, ethanol and mycelium mycelium mycelium showed excellent acetylcholinesterase inhibitory effect in the extract of each fraction of culture filtrate, mycelium and fruiting body. In the tendency of each fraction, both the polar and non-polar fractions showed similar inhibition of enzyme activity, and the extracts of different parts of the mushrooms showed similar inhibition rate as the positive control. Tacrine, a positive control, was purified to a high purity, and considering that the present invention was not purified, it was found to be superior in acetylcholinesterase inhibitory effect than tacrine.

Experimental Example 4.

Determination of Prolyl Endopeptidase (PEP) Inhibitory Activity of Extracts from Culture Filtrate, Mycelia and Fruiting Body

Inhibition activity of prolyl endopeptidase (PEP) was measured by Toda method, and Z-Pro-prolinal was used as a positive control. Into the absorbance measuring cuvette, put 800 µl buffer solution (100 mM phosphate buffer pH 7.0), 40 µl basin mushroom culture filtrate, mycelium and fruiting body fraction extract and 80 µl prolyl endopeptidase (PEP) at 37 ℃. The reaction was carried out for 10 minutes. Subsequently, 80 µl of the substrate, Z-Gly-Pro-pNA, was added and reacted at 37 ° C. for 15 minutes, and then the amount of change in absorbance was measured at 410 nm. Inhibition of PEP enzyme activity was measured.

Table 4. Inhibitory Effects of Extracts of Profil Endopeptidase (PEP) on Fractions of Cultured Filtrate, Mycelium and Fruiting Body

Sample (10 ug / ml) Prolyl endopeptidase inhibitory activity (%) extract Fraction ethanol Hexane Chloroform Ethyl acetate water Cultivation of mushrooms 65.3 ± 2.4 83 ± 2.7 76 ± 1.9 80 ± 1.3 80 ± 2.7 Myrtle mycelium 75.2 ± 3.1 83.5 ± 1.6 81.3 ± 3.5 89.2 ± 2.4 81.4 ± 1.1 Bodhi tree fruiting body 76.4 ± 1.2 83.2 ± 0.2 82.1 ± 1.3 84.2 ± 2.6 81.3 ± 0.1 Z-Pro-prolinal 91.3 ± 2.5

※ The measurement was repeated three times, and the result is expressed as the average value ± standard deviation.

※ G-Pro-prorinal, a positive control, is a single-agent compound and not a drug.

As shown in Table 4, the extracts of mycelium ethanol and culture filtrate, mycelium, and fruiting body fractions of P. eryngii mushroom showed overall excellent prolyl endopeptidase (PEP) inhibitory effect, and the extract of each part was also excellent prolyl endopeptide. Daype (PEP) inhibitory effect was shown. G-pro-prolinal, an artificial synthetic material used as a positive control, shows an effect of about 91% of prolyl endopeptidase (PEP) but cannot be used as a medicine, so prolyl endopeptide for treatment of Alzheimer's disease It can be seen that the agent (PEP) inhibitors are excellent extracts of the fractions of the mushrooms.

Experimental Example 5.

Measurement of β-secretase Inhibitory Activity of Ethyl Alcohol Extract of Pleurotus eryngii Mycelia

Beta-secretase inhibitory activity was measured using a β-secretase Activity Assay Kit from Biovision, and a β-secretase inhibitor was used as a negative control. Each well of a 96-well plate contains 50 μl of buffer, 1.25 μl of Ethyl alcohol extract of mycelium mycelium mycelium, 50 μl of 2X reaction buffer and 1.25 μl of substrate. The solution (ethanesil and Davylsil solution) was added and reacted for 15 minutes in a 37 ℃ incubator blocked light. Thereafter, 1.25 μl of beta secretase was added to each well, and reacted for 120 minutes in a 37 ° C. incubator in which light was blocked. Thereafter, absorbance was measured at 340 nm and 500 nm of ELISA reader, and beta secretase enzyme inhibitory activity was measured using the change value.

Table 5. Inhibitory Effects of Ethyl Alcohol Extracts on the Beta-Secretase of Myrtle Mycelia

Sample (10 ug / ml) Beta-secretase inhibition rate (%) Myrtle Myrtle Extract 100 Beta secretase inhibitors 84.7 ± 3.4

※ The measurement was repeated three times, and the result is expressed as the average value ± standard deviation.

※ The sample is ethyl alcohol extract of the mycelium mycelia.

※ Beta-secretase inhibitor, a negative control, is an enzyme synthesized by molecular biological techniques.

As shown in Table 5, the ethyl alcohol extract of the mycelium mycelium mushroom mycelium showed higher beta secretase (β-secretase) inhibitory effect than the negative control synthesized by molecular biological techniques.

Example 3.

Pharmaceutical Formulation and Functional Health Food Preparation of Pleurotus eryngii Extract (See Formulation Example)

The extract of the present invention may be mixed with a pharmaceutically acceptable carrier as an active ingredient to prepare a composition for preventing and treating Alzheimer's disease. The pharmaceutical composition may further include conventionally used excipients, sweeteners, lubricants, flavoring agents, etc., tablets, capsules, powders, granules, suspensions, emulsifiers, syrups, solutions or It may be formulated in unit dosage forms, such as for parenteral administration, or in a pharmaceutical formulation for multiple administration. The pharmaceutical composition containing the extract of the present invention may be administered parenterally or orally according to the desired method.

In addition, the health functional food is a food prepared by adding the extract (or fractions) to the general food, capsules, powdered, suspensions, etc., means that bringing a specific health effect when ingested do.

The 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 invention is not limited.

Formulation Example 1 Preparation of Injection

100 mg of extract of Examples 1 and 2

Sodium Metabisulfite 3.0 mg

Methylparaben 0.8 mg

Propylparaben 0.1 mg

Appropriate sterile distilled water for injection

The above ingredients are mixed and prepared in a conventional manner to have a final volume of 2 ml, followed by filling into a 2 ml ampoule and sterilizing to prepare an injection.

Formulation Example 2 Preparation of Tablet

200 mg of extract of Examples 1 and 2

Lactose 100 mg

Starch 100 mg

Stearic Acid Magnesium Proper

A tablet is prepared by mixing and tableting the above components according to a conventional tablet manufacturing method.

Formulation Example 3 Preparation of Capsule

Example 1 and Example 2 100 mg extract

Lactose 50 mg

Starch 50 mg

Talc 2 mg

Magnesium stearate proper amount

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Formulation Example 4 Preparation of Health Functional Food

100 mg of extract of Examples 1 and 2

Vitamin mixture proper amount

70 μl of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

0.2 μl of vitamin B12

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

Potassium Diphosphate 55mg

Potassium Citrate 90 mg

100 mg potassium carbonate

Magnesium chloride 24.8 mg

The composition ratio of the above-mentioned vitamin and mineral mixtures is a composition that is relatively suitable for health foods as a preferred embodiment, but may be arbitrarily modified, and the above ingredients may be mixed according to a general health functional food manufacturing method. The granules may be prepared and used for preparing the nutraceutical composition according to a conventional method.

Formulation Example 5 Preparation of Health Beverage

1000 mg of extract of Examples 1 and 2

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.

Although the composition ratio is mixed with a relatively suitable component for a preferred beverage in a preferred embodiment, the compounding ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and usage.

The present invention, according to the extraction method of each part and extract of the mycelium fractions of the mycelia of the mycelia, Alzheimer's disease-inducing enzymes acetylcholinesterase (AChE), prolyl endopeptidase (PEP), beta secretase (β The method of preparing extracts with inhibitory effects of -secretase), pharmaceutical compositions for improving brain function, and methods of preparing health functional foods. Since the existing treatment for Alzheimer's disease is limited to acetylcholinesterase (AChE) inhibitory effects, it does not show the inhibitory effects of prolyl endopeptidase (PEP) and beta secretase (β-secretase). In addition, the current therapeutic agent has a disadvantage that contains a lot of various side effects as a single-agent synthetic drug. However, the extract and the brain composition for improving the brain function and health functional foods provided by the present invention is a acetylcholinesterase (AChE), prolyl endopeptidase (PEP), known as Alzheimer's disease-causing enzyme Beta-secretase (β-secretase) all have the effect of inhibiting the enzyme activity, and also by using an edible mushroom and its extract as an excellent formulation with little side effects is a characteristic object of the present invention.

Claims (5)

Food additives, dietary supplements and pharmaceutical compositions for the purpose of preventing, treating and improving cognition of Alzheimer's disease using fruiting bodies, cultured mycelium and culture filtrate of Clavicorona pyxidata . According to claim 1, Food additives, health functional foods and the like comprising a sample obtained by extracting 2-4 times the number of ethyl alcohol and water to the culture mycelia and culture mycelium with a solvent, the fraction obtained by filtration and lyophilized Pharmaceutical compositions. According to claim 1, after extracting the fruiting body and cultivated mycelium of the bark mushrooms with ethyl alcohol, and concentrated to the residue obtained by adding an organic solvent, such as organic solvent hexane, chloroform, ethyl acetate, and concentrated the freeze-dried sample Food additives, nutraceuticals and pharmaceutical compositions comprising. The food additive according to claim 1, wherein the fraction obtained by adding the organic culture supernatant and supernatant liquid of the mycelium mycelium mycelium liquid to the supernatant and the organic solvent such as hexane, chloroform, and ethyl acetate are concentrated and lyophilized. And pharmaceutical compositions. The pharmaceutical composition according to claim 2, wherein the form of the pharmaceutical preparation is any one selected from powders, granules, tablets, capsules, liquids and injections.

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