KR101568823B1 - Composition containing an extract of vegetable worms containing germanium - Google Patents

Abstract

The present invention relates to a pharmaceutical composition, a food composition, a cosmetic composition and a manufacturing method for preventing or treating cancer, which comprises extracting a germanium-containing Cordyceps sinensis extract and selectively killing cancer cells without killing the mesenchymal stem cells . The present invention also relates to a medium composition for promoting mesenchymal stem cell proliferation comprising a germanium-containing Cordyceps extract.

Description

[0001] The present invention relates to a composition containing a germanium-containing cordyceps extract,

The present invention relates to a pharmaceutical composition, a food composition, a cosmetic composition and a manufacturing method for preventing or treating cancer, which comprises extracting a germanium-containing Cordyceps sinensis extract and selectively killing cancer cells without killing the mesenchymal stem cells . The present invention also relates to a medium composition for promoting proliferation of mesenchymal stem cells comprising a germanium-containing Cordyceps mellitus extract.

Cordyceps is a kind of mushroom which is a kind of mushroom that various fungi enter into insects and form a fruit body by host or form spore phloem. It contains fungi which are parasitic on some fungi in addition to insects, . Among the typical Cordyceps genus, there is a complete generation of ovarian reproductive organs, including the genus Ascycycetes, Cordyceps belonging to Clavicipitaceae, Paecilomyces genus of incomplete fungi, Torrubiella genus And Podonectria. It is known that about 300 species of caterpillar fungus belonging to fruiting bodies are distributed worldwide.

Due to the physiological activity of several active ingredients such as polysaccharides, cordycepin (3'-deoxyadenosine), ergosterol, mannitol, etc., they are now used for various medical applications . In relation to cancer, the extracts of Millitarian Cordyceps sinensis, which was administered to cancer cells transplanted with cancer cells, demonstrated anti-cancer effects that inhibited the growth of solid tumors. These anti-cancer effects were observed in natural killer (NK) cells T cells, and increased IL-2 production in splenocytes (Lee H, Lee Y, Park T. 2004. J Korean Soc Food Sci Nutr 33: 59-65). In addition, it is known that chordysepin, an indicator component of Cordyceps militaris, is inserted into cancer cell DNA and RNA instead of adenine to have an anticancer effect (Cunningham KG. 1951. J Chem Soc 1: 2299-3200) .

However, Cordyceps is toxic to kill not only cancer cells but also normal stem cells. In the present invention, it was confirmed that mesenchymal stem cells, which are normal human adult stem cells, are killed in a concentration-dependent manner. Therefore, if it is used as a medicine or a health functional food that can affect the body without improving the toxicity of Cordyceps, it may cause a dysfunction due to stem cell death as well as a pure function due to the anticancer effect. Until now, studies on Cordyceps sinensis have focused only on the pure function, and the problem of the toxicity of Cordyceps sinensis has not been clarified yet. So far, there has been little research on the relationship between the potency and toxicity of Cordyceps.

On the other hand, the medical effect of germanium was first discovered in 1930 when the spring water of Lourdes, the border province of France and Spain, was reported to have a great effect on various diseases, Is known to be very high. Thereafter, studies on organic germanium capable of pharmacological action without remaining in the body proceed actively, and a relatively large amount of organic germanium is contained in herbal medicine such as ginseng, garlic, ganoderma, And research is continuing to use organic germanium for the treatment of refractory adult diseases.

Under these circumstances, the inventors of the present invention confirmed that the extract of Cordyceps sinensis containing germanium has a similar anticancer effect without toxicity to normal cells, unlike the extract of Cordyceps sinensis, which does not contain germanium. Further, the extract of Cordyceps sinensis containing germanium Promotes the proliferation of normal mesenchymal stem cells. Thus, the present invention has been completed.

An object of the present invention is to provide a pharmaceutical composition for prevention or treatment of cancer, which comprises less than 100 ug / ml of germanium-containing Cordyceps mushroom extract and selectively kills cancer cells without killing mesenchymal stem cells will be.

Another object of the present invention is to provide a food composition for cancer prevention or improvement, which comprises less than 100 ug / ml of germanium-containing Cordyceps Extract and which selectively kills cancer cells without killing mesenchymal stem cells .

Another object of the present invention is to provide a cosmetic composition comprising Cordyceps mellifera extract containing 190 to 220 ppm of germanium. At this time, the extract of Cordyceps mellifera may be prepared by cultivating the insect-stricken bacterium strain in a culture medium containing 200 to 1000 ppm of germanium for 14 to 28 days after inoculation and culturing the mycelium of the genus Cordyceps; And a method for producing the same.

It is another object of the present invention to provide a medium composition for promoting mesenchymal stem cell proliferation comprising a germanium-containing Cordyceps mellitus extract.

It is still another object of the present invention to provide a method for producing a bacterium of the present invention, which comprises culturing the bacterium of the present invention in a medium containing 200 to 1000 ppm of germanium for 14 to 28 days after inoculation, Which method comprises the step of obtaining a Cordyceps mellifera extract containing 190 to 220 ppm of mesenchymal stem cells, and selectively killing the cancer cells without killing the mesenchymal stem cells, thereby providing a method for producing germanium-containing Cordyceps Extract .

In order to achieve the above object, the present invention provides a method for treating cancer, which comprises the step of extracting germanium-containing Cordyceps sinensis extract in an amount of more than 100 g / ml and selectively killing cancer cells without killing mesenchymal stem cells And to a pharmaceutical composition for preventing or treating such diseases.

In another embodiment, the present invention provides a preventive or remedial food for cancer characterized by containing less than 100 ug / ml of germanium-containing Cordyceps sinensis extract and selectively killing cancer cells without killing mesenchymal stem cells ≪ / RTI >

Preferably, the Cordyceps sinensis may be Cordyceps militaris.

Also, preferably, the germanium-containing Cordyceps extract may contain 190 to 220 ppm of germanium.

Also preferably, the cancer may be lung cancer, liver cancer, neuroblastoma or lymphoma.

Preferably, the germanium-containing Cordyceps mushroom extract is produced by inoculating and culturing the Cordyceps mushroom strain in a medium containing germanium to produce a mushroom mycelium, and extracting the culture solution containing the mushroom mycelium May be produced by the method of the present invention.

More preferably, the medium containing germanium may contain 200 to 1000 ppm of germanium.

More preferably, the step of culturing the organisms in the step of inoculating and culturing the organisms in the medium containing germanium may be 14 to 28 days.

More preferably, the step of extracting the culture medium containing the mycelia of Cordyceps mellifera may include a step of subjecting the culture medium containing the mycelia of Cordyceps mellus to hot water extraction.

In another aspect, the present invention relates to a cosmetic composition comprising Cordyceps mellifera extract containing 190 to 220 ppm of germanium. Preferably, the extract of Cordyceps mellifera may be prepared by culturing the mycelia of Cordyceps sinensis after culturing for 14 to 28 days inoculum with a medium containing 200 to 1000 ppm of germanium, and extracting the culture solution containing the mycelium of Cordyceps mellifera And a method for producing the same.

In another aspect, the present invention relates to a medium composition for promoting mesenchymal stem cell proliferation comprising a germanium-containing Cordyceps mucus extract.

Preferably, the Cordyceps sinensis may be Cordyceps militaris.

Also, preferably, the germanium-containing Cordyceps extract may contain 190 to 220 ppm of germanium.

Preferably, the germanium-containing Cordyceps mushroom extract is produced by inoculating and culturing the Cordyceps mushroom strain in a medium containing germanium to produce a mushroom mycelium, and extracting the culture solution containing the mushroom mycelium May be produced by the method of the present invention.

More preferably, the medium containing germanium may contain 200 to 1000 ppm of germanium.

More preferably, the period of incubation in the step of inoculating and culturing the organisms with the germanium-containing medium is 14 to 28 days.

More preferably, the step of extracting the culture medium containing the mycelia of Cordyceps mellifera may include a step of subjecting the culture medium containing the mycelia of Cordyceps mellus to hot water extraction.

According to another aspect of the present invention, there is provided a method for producing a microorganism of the present invention, comprising the steps of: culturing a microorganism of the genus Cordyceps; culturing the microorganism for 14 to 28 days after inoculation of the microorganism strain in a culture medium containing 200 to 1000 ppm of germanium; And a step of obtaining a Cordyceps sinensis extract containing 190 to 220 ppm of germanium, the method comprising the step of selectively killing cancer cells without killing the mesenchymal stem cells will be.

Hereinafter, the present invention will be described in more detail.

The present invention provides a pharmaceutical composition, a food composition, a cosmetic composition and a manufacturing method for preventing or treating cancer based on the fact that the germanium-containing Cordyceps extract can selectively kill cancer cells without killing the mesenchymal stem cells do. The present invention also provides a medium composition for promoting mesenchymal stem cell proliferation comprising germanium-containing Cordyceps mushroom extract by confirming that germanium-containing Cordyceps sinensis extract can promote proliferation of mesenchymal stem cells.

In a specific embodiment of the present invention, in order to maximize the germanium content in the extract of Cordyceps sinensis containing germanium and to develop an efficient culture and extraction method therefor, various concentrations of germanium and culture conditions are varied in a medium containing germanium And extracted with germanium by homogenization and sonication extraction, hot water extraction, and methanol extraction (Experimental Examples 1 and 2).

As a result, it was confirmed that it was the most efficient method by increasing the content of germanium in the extract of Cordyceps sinensis containing germanium by culturing the cordyceps in a medium containing 200 to 1000 ppm of germanium for about 3 weeks and extracting it by the hot water extraction method 2 and experimental results 2).

In another embodiment of the present invention, in order to compare the toxicity of the extract of Cordyceps mellifera and the effect of the extract of the present invention, the extract of Cordyceps mellifera and the extract of the present invention is administered to mesenchymal stem cells and cancer cells (B cell lymphoma, T cell lymphoma, Cells, liver cancer cells) by concentration (Experimental Example 3).

As a result, when the Cordyceps sinensis - free extract was treated with cancer cells, the proliferation of human normal mesenchymal stem cells was inhibited as well as the cancer cell proliferation was dose - dependently treated. On the other hand, it was confirmed that the extract of Cordyceps sinensis containing germanium not only inhibited the proliferation of cancer cells in a concentration-dependent manner but also promoted the proliferation of normal mesenchymal stem cells. Through these effects, the extract of the present invention can selectively kill only cancer cells without being toxic to mesenchymal stem cells (experimental results 3-1 to 3-5).

In addition, in the present invention, cancer cells were identified by subcutaneous injection of cancer cells into mice, and the effect of the extract of Cordyceps sinensis and the extract of Cordyceps sinensis containing germanium was administered to the mice at various concentrations (Experimental Example 4).

As a result, it was confirmed that the extract of Cordyceps sinensis-free Cordyceps sinensis was effective against the death of nude mice but the effect of germanium-containing Cordyceps sinensis extract was more effective against the death of nude mice. Furthermore, Since the effect was reduced when the extract was administered at a dose of 66 μl as compared with when the extract was administered at a dose of 33 μl, the appropriate concentration of the germanium-containing Cordyceps Extract was proved to be less than 100 μg / ml, which corresponds to the concentration of the 66 μl Results 4-5).

In another embodiment of the present invention, in order to compare the toxicity of the extract of Cordyceps mellifera and the effect of the extract of the present invention in vivo, The extracts were administered by concentration and after one week clinical chemistry measurements were compared (Experimental Examples 4-6).

As a result, the abnormal test value was confirmed in the Cordyceps sinensis extract - treated group, and the value close to the abnormal level was confirmed at the value confirmed to be normal. On the other hand, no abnormality was detected in the group treated with 33 μl of germanium-containing Cordyceps extracts. However, the germanium-containing cordyceps hydrothermal extract was found to be abnormal when 66 ㎕ was administered. Thus, it was confirmed again that the germanium-containing Cordyceps coca herbal extracts had the effect of killing only the cancerous mass without toxicity, and that the most effective concentration to kill the cancerous mass without causing toxicity was less than 100 ug / ml -6).

Therefore, it has been proved that the Cordyceps sinensis extract containing germanium of the present invention can selectively kill cancer cells without killing mesenchymal stem cells, and can promote the proliferation of mesenchymal stem cells.

The composition of the present invention comprises less than 100 ug / ml of germanium-containing Cordyceps Extract, preferably less than 100 ug / ml, 10 to 90 ug / ml, 20 to 80 ug / ml, 30 to 70 ug / ml, 40 To 60 ug / ml or 50 ug / ml. If the concentration of the germanium-containing Cordyceps sinensis extract is more than 100 ug / ml, the killing effect on the cancer mass may be decreased, and the toxicity may increase in vivo. The concentration corresponding to 33 μl of the experimental example of the present invention is 50 ug / ml, and the concentration corresponding to 66 μl is 100 ug / ml.

The term "germanium" in the present application is one of the metal elements belonging to group 4B of the periodic table, and the element symbol is Ge. It is a by-product of mainly zinc and copper and is produced in the form of oxides. It is also contained in coal. The germanium contained in the extract of the present invention means not only highly toxic germanium contained in natural minerals and soil but also organic germanium contained in plants, mushrooms and the like. In addition, the organic germanium may be germanium extracted from plant, natural biosynthetic germanium, germanium produced using genetic recombination technology, or chemically synthesized organic germanium. Preferably, the term " germanium " in the present invention refers to germanium naturally present in the Chinese cabbage by culturing the Chinese cabbage on a medium containing germanium.

In the present invention, the term "vegetable worms" is a type of mushroom which is a type of mushroom that various fungi enter into insects to form a fruit body as a host or form spore veins on the body. It also includes fruiting bodies that have been parasitized on some fungi. In an embodiment of the present invention, Cordyceps militaris was used. However, the present invention is not limited thereto, and the present invention is not limited thereto. Examples include Cordyceps sphecocephala, Paecilomyces japonica, Cordyceps scarabaeicola, Paecilomyces japonica, Paecilomyces tenuipes, Cordyceps sinensis), but it is not limited to a germanium-containing cordyceps, but is included in the scope of the present invention.

The term " germanium-containing caterpillar fungus "refers to caterpillar fungus containing germanium in the interior, which may be a caterpillar fungus in which germanium is present. The Chinese caterpillar fungus containing germanium may be a biosynthesizing germanium in the cordyceps using a recombinant technology and may be the administration of the organic germanium synthesized through chemical synthesis into the cordyceps. Preferably, the germanium-containing cordyceps of the present invention can be obtained by cultivating cordyceps in a culture medium containing germanium.

In one embodiment, the medium for culturing Cordyceps sinensis can contain 200-1,000 ppm of germanium, and preferably 500-1000 ppm, in order to allow germanium to naturally exist within the cordyceps. Cordyceps can be most efficiently cultured in a medium containing such a germanium concentration. The Cordyceps can also be cultured for 14 to 28 days, preferably for 17 to 25 days.

For example, the germanium-containing Cordyceps sinensis extract may be prepared by a method comprising the step of preparing a mycelia of Cordyceps mellifera by inoculating and culturing the Cordycephalosporium sp. Strain in a medium containing germanium, and extracting a culture containing the mycelia of Cordyceps mellifera .

Extraction of the culture solution containing the mycelia of Cordyceps mellus is included in the scope of the present invention, as long as it contains germanium, regardless of the extraction method and the extraction solvent. For example, the culture medium containing the mycelia of Cordyceps mellifera can be extracted with a solvent selected from the group consisting of water, C3 to C5 ketones, C1 to C4 linear or branched alcohols, and mixed solvents thereof. Furthermore, the extract can be fractionated and separated into at least one selected from the group consisting of water, C5 to C7 alkanes, C1 to C4 linear or branched alcohols, methylene chloride, and ethyl acetate. As another example, it can be extracted by sonication after homogenization by dissolving the mycelia of caterpillar fungus cultivated in germanium-containing medium in sterilized distilled water and centrifuging.

In addition, in the present invention, the extraction of germanium-containing Cordyceps horticulture can be carried out according to a conventional method for producing a plant or mushroom extract, and can be performed by hot water extraction, pressure extraction, reflux extraction, warm- However, the present invention is not limited thereto. In addition, the extract prepared as described above can be concentrated or removed by concentration filtration and / or lyophilization in a subsequent vacuum filtration step. Therefore, the germanium-containing Cordyceps mushroom extract of the present invention is used to mean a dried liquid, a concentrated liquid extract from which the extraction solvent is removed, or a solid extract. The extract of germanium-containing Cordyceps sinensis extracted in this manner may contain 190 to 220 ppm of germanium. Preferably 200 to 220 ppm of germanium.

Specifically, the germanium-containing cordyceps mushroom extract of the present invention is produced by inoculating and culturing a mushroom strain to a medium containing 200 to 1000 ppm of germanium to produce a mushroom mycelium, extracting the mushroom-containing mushroom-containing culture medium with hot water, And a step of obtaining a cordyceps extract containing 190 to 220 ppm of germanium.

Unlike the germanium-free Cordyceps sinensis extract, the extract of Cordyceps sinensis containing germanium of the present invention is not toxic and has an activity of selectively killing cancer cells without killing mesenchymal stem cells, May be used as an effective component of a pharmaceutical composition.

The cancer may refer to a tumor and includes all diseases caused by a condition where abnormally proliferating abnormal cells to be normally killed are caused. The cancer includes solid cancer and blood cancer. The present invention relates to the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of cancer such as cerebrospinal fluid, head and neck cancer, lung cancer, breast cancer, thymoma, mesothelioma, esophageal cancer, gastric cancer, colon cancer, liver cancer, pancreatic cancer, Endometrial cancer, lymphoma, acute leukemia, chronic leukemia, multiple myeloma, sarcoma, malignant melanoma, skin cancer, and the like. In an embodiment of the present invention, the cancer may be lung cancer, liver cancer, neuroblastoma or lymphoma.

The pharmaceutical compositions of the present invention may be administered to mammals, including humans, by a variety of routes. The mode of administration may be any mode conventionally used and may be administered, for example, orally, rectally or by intravenous, intramuscular, subcutaneous, or injection. In addition, they may be formulated into parenteral formulations in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, percutaneous preparations, suppositories and sterilized injection solutions, .

In one embodiment, the pharmaceutical composition of the present invention may additionally contain adjuvants such as physiologically acceptable carriers, excipients, diluents, etc. in addition to germanium-containing Cordyceps extracts. Examples of carriers, excipients and diluents that can be included in the composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil and the like. When the composition of the present invention is formulated, diluents such as fillers, extenders, binders, wetting agents, disintegrants, surfactants and excipients which are commonly used may be used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like. These solid preparations may contain at least one excipient such as starch, calcium carbonate, sucrose ), Lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate talc may also be used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, syrups and the like, and water, liquid paraffin, and / or various excipients such as wetting agents, sweeteners, . ≪ / RTI > Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, transdermal preparations and the like. Examples of non-aqueous solvents or suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The dosage of the composition of the present invention may be varied depending on the patient's age, weight, sex, dosage form, health condition and disease severity, and may be administered once to several times a day You may. Dosages may be higher or lower depending on individual differences.

The term " patient " refers to a mammal including a human, for example, a human requiring treatment for cancer.

In addition, the food composition of the present invention may comprise an acceptable food-aid additive and may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of functional foods.

In the present invention, the term " food " means a natural product or a processed product containing one or more nutrients, preferably a state of being able to be directly eaten through a certain processing step, , Health functional foods, beverages, food additives and beverage additives. Examples of the food include various foods, beverages, gums, tea, vitamin complex, and functional foods. In addition, in the present invention, the food may contain special nutritional foods (e.g., crude oil, spirits, infant food, etc.), meat products, fish products, tofu, jelly, noodles (Such as soy sauce, soybean paste, hot pepper paste, mixed sauce), sauces, confectionery (eg snacks), dairy products (eg fermented milk, cheese), other processed foods, kimchi, pickled foods But are not limited to, natural flavors (eg, ramen soup, etc.), vitamin complexes, alcoholic beverages, alcoholic beverages and other health supplement foods. The health functional food, beverage, food additive or beverage additive can be produced by a usual production method.

In addition, in the food, the amount of the germanium-containing cordyceps extract may be 0.00001% to 50% by weight of the whole food. When the food is a beverage, 0.001 g To 50 g, and preferably from 0.01 g to 10 g.

For example, when the extract of the present invention is used as a cosmetic composition, it can be used as a cosmetic composition in which all kinds of ingredients usable in ordinary cosmetics such as fragrances, coloring matters, bactericides, antioxidants, preservatives, moisturizers, thickeners, A synthetic polymer material, and the like, and the content thereof can be appropriately adjusted depending on the purpose of the final product and the intended use.

As another example, the extract of the present invention can be used as a medium composition for promoting mesenchymal stem cell proliferation. In addition to the germanium-containing Cordyceps mellifera extract, the culture medium composition may further include a component for enhancing the proliferation and / or viability of the stem cells at a rate higher than the minimum rate. It may also include essential and nonessential amino acids, vitamins, energy sources, lipids and trace elements necessary for minimal growth and / or survival of nutrients and cells that nourish the cells generally contained in the cell culture medium. As a specific example, the culture medium composition of the present invention may include but is not limited to more than 0 to 100 ug / ml of germanium-containing cordyceps extract.

The medium prepared from the medium composition of the present invention not only promotes the proliferation of mesenchymal stem cells but also can promote the proliferation of mesenchymal stem cells while selectively killing cancer cells when cultured together with cancer cells.

The present invention provides a composition comprising a germanium-containing Cordyceps mushroom extract that selectively kills cancer cells without killing mesenchymal stem cells. Using the germanium-containing Cordyceps mushroom extract of the present invention, Prevent, treat and / or ameliorate mesenchymal stem cell proliferation and promote mesenchymal stem cell proliferation.

FIG. 1A shows the content of germanium in the mycelia of Chinese caterpillar fungus, according to the concentration of germanium in the medium. "ppm" represents the concentration of germanium in the medium.
FIG. 1B shows the recovery of germanium in the mycelium of Chinese cabbage organs according to the concentration of germanium in the medium. "ppm" represents the concentration of germanium in the medium.
1C is a graph showing the germanium content of the extract of Cordyceps mellus mycelia according to the extraction method measured by absorbance at 525 nm.
In Figs. 2A, 3A, 4A, 5A and 6A, the horizontal axis represents the amount (l) of the treated hot-water extract and the vertical axis represents the survival rate (%).
"C" is a group treated with 33 .mu.l of a hot water extract of Cordyceps, "D" is a group treated with Cordyceps waxy water, and ""E" represents the treated group of 66.7 μl of the Cordyceps melliflora extract, and "F" represents the treated group of the Cordyceps militaris hot water extract of 133 μl.
In Fig. 2C, 3C, 4C, 5C and 6C, "A" is a control (control), "B" is a treated group of 6.6 μl of a cordyceps hot water extract containing germanium and "C" is a treated group of 33 μl of a germanium- Group "D" was treated with 46.7 μl of cordierite hydrothermal extract containing germanium, "E" was treated with 66.7 μl of cordierite hydrothermal extract containing germanium and "F" was treated with 133 μl of germanium- .
FIG. 2A is a graph comparing the cytotoxic effect of B cell lymphoma cancer cells on the concentration of hot-water extract of Cordyceps mellifera and germanium-containing Cordyceps.
FIG. 2B shows the B cell lymphoma cell death killing effect according to the concentration of the Cordyceps;
FIG. 2c shows B cell lymphoma cell death kinetics depending on the concentration of germanium-containing Cordyceps waxy hot-water extract.
FIG. 3A is a graph comparing the cytotoxic effects of T cell lymphoma cancer cells on the concentration of hot water extract of Cordyceps mellifera and germanium-containing Cordyceps mellitus.
FIG. 3B shows the cytotoxic effect of T cell lymphoma cancer cells on the concentration of Cordyceps militaris hot water extract.
FIG. 3c shows the T cell lymphoma cell killing effect depending on the concentration of the organophosphate-containing hot-water extract containing the organic germanium.
4A is a comparison of stem cell death effects according to the concentration of the hot water extract of Cordyceps mellonus containing the hot water extract of Cordyceps mellifera and the organic germanium.
FIG. 4B shows the stem cell death effect according to the concentration of the Cordyceps militaris hot water extract.
FIG. 4c shows the stem cell killing effect according to the concentration of the organophosphate-containing hot water extract containing germanium.
FIG. 5a is a graph comparing lung cancer cell killing effect according to the concentration of the hot water extract of Cordyceps mellonus containing the hot water extract of Cordyceps mellifera and the organic germanium.
FIG. 5B shows lung cancer cell killing effect depending on the concentration of the extract of Cordyceps mellifera.
FIG. 5c shows lung cancer cell killing effect according to the concentration of the organophosphate-containing hot water extract containing germanium.
FIG. 6a is a graph comparing the killing effect of hepatocyte apoptosis with the concentration of hot water extract of Cordyceps mellonus containing the hot water extract of Cordyceps mellifera and the organic germanium.
6B shows the effect of hepatic cancer cell death on the concentration of the Cordyceps mizugi hot water extract.
FIG. 6c shows the effect of hepatic cancer cell death according to the concentration of the organophosphate-containing hot water extract containing germanium.
In FIGS. 7A to 10B, "CTL" is a control group, "CM" is a group treated with 33 μl of the Cordyceps hot water extract, "CMGe" is a group treated with 33 μl of a cordyceps hot water extract containing germanium, "CMGe 2" And 66 쨉 l of the extract.
7A is a graph comparing changes in the volume of a cancer mass in a nude mouse with a hot water extract of Cordyceps mellifera and a germanium-containing Cordyceps.
FIG. 7B shows the average amount of change in the volume of the cancer mass in nude mice of the Cordyceps mellitus hot water extract containing the caterpillar fungus extract and germanium after one week of treatment.
FIG. 8a shows the weight change of the liver by the treatment with the hot water extract of Cordyceps mellifera containing the hot water extract of Cordyceps mellifera and germanium.
8B shows the change in weight of the kidney by the treatment with the hot water extract of Cordyceps mellifera containing the hot water extract of Cordyceps mellifera and germanium.
FIG. 9a shows the effect of the germanium-containing Cordyceps mothera hot-water extract on spleen by MTT assay. "CTL" represents a control group in which only water is administered, "CM" represents a group administered orally to Cordyceps sinensis extract, and "CMGe" represents a group administered orally to Cordyceps sinensis extract containing germanium.
FIG. 9B shows the effect of the organophosphate-containing hot water extract on the active oxygen in the splenocytes. "CM + H2O2" is a group that is administered orally after the oxidative stress induction with hydrogen peroxide, "CM + + H2O2" is a group that is administered only by water, "CTL + H2O2" H2O2 "is a group of oral administration of germanium-containing Cordyceps sinensis extract after oxidative stress induced by hydrogen peroxide, and" ROS "represents reactive oxygen species.
FIG. 10A shows the mass of the cancer taken from the nude mouse of the untreated group.
Fig. 10B shows the mass of the cancer taken from the nude mouse in the group treated with the Cordyceps mizugi extract.
Fig. 10C shows the mass of the cancer taken from a nude mouse in a group treated with 33 μl of germanium-containing Cordyceps militaris hot-water extract.
Fig. 10D shows the mass of cancer taken from a nude mouse in a group treated with 66 ㎕ of germanium-containing cordyceps hot-water extract.
Fig. 11 shows the result of comparing the in vivo toxicity of the hot water extract of Cordyceps mellifera containing the hot water extract of Cordyceps mellifera and the organic germanium. "CM1-5" germanium-containing cordyceps hot water extract 33, "CM1-5" germanium-containing caterpillar fungus hydrothermal extract 33, "blue" was lower than the normal value and red was higher than the normal value. ≪ / RTI > treated group, "CMGe21-5" germanium-containing cordyceps hot water extract.
12A shows an open field test container for activity measurement.
FIG. 12B is a comparison of the activity of the nude mouse treated with the hot water extract of Cordyceps mellifera and the germanium-containing Cordyceps mellitus with the open field test.

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited by the following examples.

Experimental Example  1. Strain and Culture

Cordyceps militaris used in this experiment was purchased from Korea Microorganism Preservation Center (60303, KCCM). The culture medium used for the culture was PDB (Aceta, USA). In order to develop optimum organismic organism culture, mycelial culture was grown in PDB medium without germanium at 2% concentration in 6 different PDB media (germanium (Sigma Aldrich, USA)) at room temperature After culturing at 165 rpm, the weight was measured.

Next, the concentration of organic germanium in the mycelium was induced, and the concentration of organic germanium was measured every week.

Experimental Example  2. Preparation of germanium-containing Cordyceps Extract

2-1. Ultrasonic Extraction Homogenizing  & sonication extraction )

The mycelia of Cordyceps militaris and germanium were grown in 30 ml of sterile distilled water and homogenized for 30 minutes at 30,000 rpm and sonicated for 20 minutes at 20 KHz. The supernatant was collected by centrifugation at 8000 rpm for 10 minutes.

2-2. Hot water extraction ( Water extraction )

When the final amount of the mycelium of Cordyceps mushroom grown in the medium containing germanium mushroom mycelium and germanium was adjusted to 24 ml with sterilized distilled water and the final amount was reduced to 50 ml, the supernatant was collected by centrifugation at 8000 rpm for 10 minutes.

2-3. Methanol extraction ( MeOH extraction )

5 ml of 80% methanol (MeOH) was added to 5 g of the remaining pellet after hot water extraction and the mixture was stirred at 25 ° C for 24 hours. The next day, 5 ml of 80% methanol was further added to the pellet. And the supernatant was collected.

2-4. Germanium content measurement

1ml phenylfluorone solution and 1ml cyclohexanol were added to the germanium extracts of Cordyceps mellifera and germanium in order to measure the germanium content of the mycelium extracts. The extracts of Cordyceps mushroom mycelium grown in the medium containing germanium were diluted 100 times and 1 ml was added thereto. After incubation at 25 ° C for 20 minutes, absorbance was measured at 525 nm.

Experimental Example  3. MTT assay On by Intermediate lobe  Measuring effects on stem cells and cancer cells

3-1. Intermediate lobe  Extract treatment of stem cells and cancer cells

2 × 10 ⁴ mesenchymal stem cells (MSC) (Lonza, USA), lung cancer cells (H157, A549, H460) (KCLB: Korean cell line bank), liver cancer cells (HepG2, Huh7) 1% L-glutamine, 1% penicillin-streptomycin, and 1% L-glutamylcysteine (KCLB) And cultured in DMEM (Dulbecco's Modified Eagle's Medium) or RPMI-1640 containing L-glutamine for 2 days at 37 ° C in a 5% CO ₂ environment. The extracts of Cordyceps mellifera mycelia were treated with 6.6 μl, 33 μl, 46.7 μl, 66.7 μl, and 133 μl per 1 ml of the medium, respectively. At this time, the concentration corresponding to 33 μl is 50 ug / ml.

3-2. MTT assay  Cell proliferation assay

Cell proliferation assay kit (promega, USA)) was used for 24 hours after transferring 100 μl of the Cordyceps sinensis extract and germanium-containing Cordyceps sinensis extract to cancer cells and stem cells for 2 days. . Treatment of 15 ㎕ of the tetrazolium component of yellow produces a purple formazan product, MMT (3- (4,5-dimethylthiazol-2-yl) -2,5 -diphenyl-tetrazolium bromide). After culturing the cells at 37 ° C for 2 hours and 30 minutes, the cells were treated with 100 μl of Solubilization Solution / Stop Mix for 1 hour to solubilize the formazan product and measure the absorbance at 570 nm.

Experimental Example  4. Mouse-assisted Organic Germanium  Identification of the effect of the insecticide extract

4-1. Cancer cell culture

2 × 10 ⁴ cancer cells (U87MG) (KCLB) were cultured in DMEM or RPMI-1640 containing 10% FBS, 1% penicillin-streptomycin, 1% L-glutamine for 2 days in a 37 ° C. 5% CO 2 environment. When the cells were confluent, they were washed twice with HBSS (Hank's balanced salt solution), trypsinized, and the cells were removed and washed twice with fresh culture medium.

4-2. Cancer cell xenotransplantation in nude mice ( Xenograft )

Balb / c nude mice (male, 9-11 weeks, n = 21) (Koatech, Korea), approximately 21-25 g, were adapted in a bacterial environment. The animal room conditions were maintained at temperature (22 ± 2 ° C), light (12 hour light / dark cycle) and humidity (50 ± 10%). All experimental food and bedding were sterilized. A total of 2 × 10 ⁶ cancer cells (U87MG) were dissolved in 0.1 mL HBSS and subcutaneously injected into the neck of each nude mouse using a 26-gauge needle. After 2 weeks, the amount of cancer mass was compared between the administration group of caterpillar fungus extracts and the group treated with germanium-containing caterpillar fungus hydrothermal extract and the control group for 1 week after the confirmation of cancerous mass, and the clinical chemical test values of each group Were measured using a drychem.

4-3. MTT assay On by Organic Germanium  Caterpillars Of hot-water extract  Measuring the effect on the spleen

The MTT assay was performed to determine the proliferative activity of spleen cells containing various immune cells after oral administration of hot water extract of Cordyceps mellifera and hot water extract of Cordyceps mellifera containing organism germanium at a concentration of 150 mg per kg body weight per day for 4 months. The MTT assay was performed in the same manner as described above.

4-4. Organic Germanium  Caterpillars Of hot-water extract Splenocyte  Measuring the Effect on My Active Oxygen

In order to measure the level of active oxygen in the cells of spleen cells, oxidative stress was induced with 5 mM hydrogen peroxide, followed by washing with PBS three times, and treated with 0.025 mM DCFH-DA (dichlorofluorescin diacetate) for 30 minutes. After washing three times with PBS, excitation wavelength was measured at 485 nm and emission wavelength was measured at 530 nm.

4-5. Cordyceps Hot water extract and Organic Germanium  Caterpillars Of hot-water extract  Nude mouse In  Change in weight of a cancer mass

The water extracts of Cordyceps mellifera extract containing hot water extract of Cordyceps melliflorus and organic germanium were treated for one week in nude mice that had been grown for 2 weeks and then weighed out from each nude mouse.

4-6. Cordyceps Hot water extract and Organic Germanium  Caterpillars Of hot-water extract  in vivo  Toxicity comparison

In order to compare the in vivo toxicity of Cordyceps melliflora extract containing hot - water extract and organic germanium, we performed a clinical chemistry test after nude mice were fed a weekly hot water extract of Cordyceps melliflora containing hot water extract and organic germanium.

4-7. Cordyceps Hot water extract and Organic Germanium  Caterpillars Hot water extract  Measuring the Effect on Nude Mouse Activity

An open field test (Fig. 12a) was performed to measure the activity of nude mice after feeding nude mice for 1 week with the Cordyceps militaris hot water extract containing the hot water extract of Cordyceps mellifera and the organic germanium.

Experiment result

1. Measurement of germanium content in cells by various extraction methods

After measuring the weight after culture (Table 1), it was confirmed that 500 ppm of germanium was added for 3 weeks in order to maximize the germanium content in the mycelia of the Chinese cabbage.

0 ppm 100 ppm 200 ppm 500 ppm 700 ppm 1000 ppm 7 days 1.60 + 0.08 2.10 ± 0.16 2.20 ± 0.29 1.53 ± 0.29 1.93 + - 0.33 1.93 + 0.19 14 days 2.03 ± 0.19 1.67 ± 0.59 2.20 ± 0.22 2.53 ± 0.19 2.60 ± 0.57 2.63 ± 0.45 21st 3.30 ± 0.29 3.27 ± 0.33 3.70 ± 0.14 3.90 + - 0.33 4.00 + - 0.57 3.10 ± 0.14

(Unit: g / 50 ml)

As a result of measuring the content of organic germanium in the mycelia of the Chinese cabbage cultivated in the PDB medium supplemented with germanium, the content of organic germanium in the mycelium was highest as the days of culture were increased in the medium containing 1000 ppm of germanium (FIG. . However, the recovery rate of organic germanium per germanium body weight after culturing in germanium - containing medium was 500 ppm, which is the most effective for recovering organic germanium, and the recovery rate was significantly higher as the culture days were increased. On the contrary, when the concentration of germanium in the medium was 1000 ppm, the recovery was not even half (FIG.

Based on these results, 500ppm of germanium was added to the culture medium, and the mycelia of the caterpillar fungus were cultured in the PDB liquid medium at 25 ° C and 180rpm for 3 weeks. As a result, 52.71g of caterpillar fungus was obtained per liter.

2. Cordyceps by various extraction methods Within the mycelium  Germanium content measurement

In the case of homogenization and sonication extraction, the contents of germanium were 2.158ppm and 189ppm of germanium in the extracts of Cordyceps sinensis extract and germanium-containing cordyceps, respectively.

The amount of germanium increased by 88 times in the mycelial extract of Cordyceps mellifera cultured in media containing germanium. When the extract of Cordyceps mellifera was obtained by hot water extraction method, it contained 2.77ppm and 219ppm of germanium, respectively and showed an increase of germanium content about 79 times.

In the methanol extraction method, the extracts contained 1.93 ppm and 195 ppm of germanium in the extracts and the extracts of germanium, respectively, and the germanium content was increased by about 101 times.

Methanol extraction method showed the highest increase in the amount of germanium, but the highest germanium content was obtained when the extract of Cordyceps mellifera was obtained by hot water extraction method.

3. MTT assay On by Intermediate lobe  Effects on stem cells and cancer cells

3-1. MTT assay B by cell  Lymphoma Ramos ) Deadly effect

When human b - cell lymphoma (Ramos) was treated with germanium - free Cordyceps waxy hydrothermal extract, cancer cell proliferation was inhibited in a concentration - dependent manner. Although the effect of the organophosphate-containing extract of organism germanium was inhibited by the concentration-dependent inhibition of cancer cell proliferation, the effect was slightly lower than that of the germanium-free organophosphate extract (Figs. 2a to 2c). When the cell viability of the untreated control group was taken as 100%, 6.7 당, 33,, 46.7,, 66.7 당, and 133 농도 concentration per 1 ml of culture medium were added to the human B cell lymphoma (Ramos) Respectively, survival rates were 15.6%, 1.3%, 0.9%, 0.6% and 0.2%, respectively. On the other hand, 84.5%, 37.8%, 32.3%, 18.9%, and 8.7% of viability was obtained when the organophosphate-containing hot water extract containing germanium was treated (FIG.

This result shows that the germanium-free Cordyceps waxy hydrothermal extract has a strong effect on cancer cell death, but it is more toxic than the germanium-containing Cordyceps waxy hydrothermal extract.

3-2. MTT assay T by cell  Lymphoma CEM ) Measurement of killing effect

In the same manner as in B-cell lymphoma, the treatment of human T cell lymphoma (CEM) with the germanium-free Cordyceps wax extract showed the effect of inhibiting cancer cell proliferation in a concentration-dependent manner (FIGS. 3A to 3C). Similar to B cell lymphoma, when treated with the organism germanium-containing Cordyceps, the effect of inhibiting cancer cell proliferation was dose-dependently lower than that of germanium-free Cordyceps waxy hydrothermal extract. When the cell viability of the untreated control group was taken as 100%, 6.6 당, 33,, 46.7,, 66.7,, and 133 농도 concentration per 1 ml of culture medium were added to the human T cell lymphoma (CEM) Respectively, the survival rate was 35.9%, 10.6%, 6.5%, 2.5% and 3.8%, respectively. On the other hand, the survival rate was 75.2%, 30.5%, 24.9%, 15.0%, and 6.8%, respectively, when the organelle germanium containing hot water extract was treated.

This study also shows that the germanium-free Cordyceps waxy hydrothermal extract has a strong effect on blood cancer cell death, but it is more toxic than the germanium-containing Cordyceps waxy hydrothermal extract. In order to prove this effect, the effect of Cordyceps sinensis extract containing Cordyceps waxy hot water extract and organic germanium on human mesenchymal stem cells was examined.

3-3. MTT assay Human by Intermediate lobe  Measuring effects on stem cell death and proliferation

In order to prove that the germanium-free caterpillar fungus hydrothermal extract has a stronger effect on the cancer cell death, it is more toxic than the germanium-containing caterpillar fungus hydrothermal extract. In order to prove that the human mesenchymal stem cells contain the caterpillar fungus hydrothermal extract and the organic germanium The effect of the extracts of Cordyceps sinensis was confirmed.

When mesenchymal stem cells (MSC) treated with the germanium-free Cordyceps wax extract were treated with MSC, the proliferation of stem cells was inhibited in a concentration-dependent manner as in the case of cancer cells (FIGS. 4A to 4B). However, when the organelle germanium - containing Cordyceps extracts were treated, the proliferation of stem cells was similar to that of the untreated control group or rather increased. When the cell viability of the untreated control group was assumed to be 100%, when treated with 6.6 μl, 33 μl, 46.7 μl, and 66.7 μl per 1 ml of the culture medium, the extract of Cordyceps mellitus without germanium was 128% 27.9%, 17.1%, and 10.0%, respectively (Fig. 4A). On the other hand, the survival rate of 110%, 178.1%, 176.3%, and 172.2% of the organelle germinated caterpillar fungus hydrothermal extract were respectively.

Experimental results show that the germanium-free caterpillar fungus hydrothermal extract has a strong effect on cancer cell death, but it also proves that it is more toxic than the germanium-containing caterpillar fungus hydrothermal extract because it kills normal stem cells necessary for human survival Respectively. Therefore, the hot water extract of Cordyceps mugwort containing organogermanium has a slightly weaker effect than the germanium-free Cordyceps waxy hydrothermal extract for cancer cell death, but it is more stable in human body and more killing effect on cancer cells, considering toxicity to stem cells And it was confirmed that there was an excellent effect.

3-4. MTT assay Of lung cancer cells (A549)

In order to measure the killing effect on more cancer cells, the killing effect of the germanium-free Cordyceps wax extract was measured on human lung cancer cells (A549). The treatment with human lung cancer cells (A549) inhibited cancer cell proliferation in a concentration-dependent manner (Figs. 5A and 5B). Although the effect of the organophosphate extract containing germanium was inhibited in a concentration-dependent manner, the effect was less than that of the germanium-free Cordyceps waxy hydrothermal extract (Figs. 5A and 5C). When the cell viability of the untreated control group was taken as 100%, 6.6 μl, 33 μl, 46.7 μl, 66.7 μl, and 133 μl per 1 ml of the medium were added to the human lung cancer cell line (A549) containing no germanium The survival rates were 33.6%, 17.5%, 13.5%, 11.0%, and 9.3%, respectively. On the other hand, when treated with the germanium - containing hot water extract of Cordyceps, the survival rate was 127%, 42.2%, 43.8%, 43.9% and 29.3%, respectively.

It is also shown that the extract of Cordyceps sinensis not containing germanium is more effective than the extract of Cordyceps sinensis, which contains organ germanium, for lung cancer cell death, but it is more toxic than the germanium-containing cordyceps hydrothermal extract .

3-5. MTT assay On by Liver cancer cell line  ( HepG2 ) Measurement of killing effect

Finally, to determine the killing effect of hepatocellular carcinoma cells, keratinocyte - free hot - water extracts of germanium were treated with human hepatoma cell line (HepG2) to determine the killing effect. When hepatocyte hepatoma cell line (HepG2) was treated with the germanium-free hot-water extract of Cordyceps mellifera, cancer cell proliferation was inhibited in a concentration-dependent manner (Figs. 6A to 6C). The effect of germanium - containing Cordyceps sinensis extract was inhibited in a concentration - dependent manner, but the effect was less than that of germanium - free Cordyceps waxy hydrothermal extract. When the cell viability of the untreated control group was taken as 100%, 6.6 당, 33,, 46.7,, 66.7 ㎕, and 133 ㎕ per 1 ml of medium were added to the human liver cancer cell line (HepG2) The survival rates were 16.1%, 0.7%, 0.8%, 0.8%, and 0.9%, respectively. On the other hand, the survival rate was 72.9%, 30.7%, 21.8%, 15.9%, and 9.7%, respectively, when the organelle germanium containing hot water extract was treated.

The results of this study also show that the germanium-free cadmium-free hot-water extract is more effective than the germanium-containing cadmium-hydrothermal extract for liver cancer cell death, but it is more toxic than the germanium-containing cadmium- .

4. Mouse-assisted Organic Germanium  Identification of the effect of the insecticide extract

4-1. Cordyceps Hot water extract and  Caterpillar fungus containing germanium Of hot-water extract  Comparison of mortality of nude mice

The volume of cancer mass after administration of germanium free hot water extract of Cordyceps mellifera to nude mice showed that the volume of cancer mass was reduced after 1 week compared to the control group. The volume of the cancer mass after the administration of the organelle germanium-containing Cordyceps sinensis extract to nude mice also effectively reduced the volume of the cancer mass (Fig. 7A). After 1 week, when the volume of cancer mass in the untreated control group was taken as 100%, it was decreased to 88.7% when 33 μl of the germanium-free cadmium-free hot water extract was added, and 33 μl of germanium- And 48.8% when it was fed. When 66 쨉 l of 33 ㎕ of germanium-containing cordyceps hot-water extract was fed, it decreased to 74.6% (Fig. 7B). The results showed that the germanium - free Cordyceps waxy hydrothermal extract showed an effect on the cancer death of the nude mice, but the germanium - containing cordyceps waxy hydrothermal extract showed the better effect on the cancer death of nude mice. Especially, when 66 ㎕ of 33 ㎕ of hot water extract containing germanium was added to the extract, the effect was reduced. The concentration of the composition corresponding to 33 추출 of the extract is 50 ug / ml.

4-2. Organic Germanium  Caterpillars Hot water extract  Weight change of each organ when treated with nude mouse

Changes in weight of liver, kidney and spleen were measured in nude mice after treatment for 1 week in nude mice in which a lump of carcass was grown for 2 weeks with hot water extract of Cordyceps mellifera containing caterpillar fungus extract and organic germanium 8b). In the liver and kidney, no significant changes were observed in the group treated with the hot water extract of Cordyceps melliflorus and the group treated with the Cordyceps mellitus containing the organic germanium.

4-3. MTT assay On by Organic Germanium  Caterpillars Of hot-water extract  Effect on the spleen

When the survival rate of spleen cells in the control group was 100%, the spleen cells of the group administered orally with the extract of the Chinese cabbage extract showed a 15% proliferative effect, and the spleen extract containing the organic germanium was orally administered to the spleen Cells increased about 16% (Fig. 9A). The results showed that the extracts of Cordyceps sinensis extract and organo - germanium - containing Cordyceps sinensis effectively inhibited the growth of spleen cells containing the greatest number of immune cells.

4-4. Organic Germanium  Caterpillars Of hot-water extract Splenocyte  Effect on My Active Oxygen

When oxidative stress was induced by 5 mM hydrogen peroxide, active oxygen production increased by 2.38-fold (238%) when the degree of reactive oxygen species production of splenocytes isolated from mice in the control group was only 100%. However, in the spleen cells of rats receiving daily doses of Cordyceps sinensis extract, only 34% of the active oxygen was not elevated. In spite of oral germanium-containing Cordyceps sinensis extract, Did not do it. Thus, it was confirmed that the extracts of Cordyceps mugwort or Cordyceps mugwort containing organic germanium had an effect of reducing the generation of active oxygen by about 5 to 7 times (FIG. 9b).

4-5. Cordyceps Hot water extract and Organic Germanium  Caterpillars Of hot-water extract  Nude mouse In  Change in weight of a cancer mass

It was confirmed that the weight of the cancer mass was effectively reduced after 1 week when the organism germanium-containing Cordyceps sinensis extract was treated with nude mice (FIGS. 10A to 10D). After 1 week, when the weight of the cancerous mass of the control group was taken as 100%, it was increased to 128.8% when 33 μl of the germanium-free Cordyceps waxy hydrothermal extract was fed, and 33 μl of the germanium- And 63.9% when it was fed. When 66 .mu.l of germanium-containing cordyceps hot-water extract was doubled as 33 .mu.l, it was increased to 106.6%. This confirms that the extract of Cordyceps mellifera containing organic germanium has a stronger effect on the cancer death of nude mice.

4-6. Cordyceps Hot water extract and Organic Germanium  Caterpillars Of hot-water extract  in vivo  Toxicity comparison

In the untreated group and in the group treated with the Cordyceps melliflora extracts, abnormal test values were found to be high, and even the values confirmed to be normal were almost close to abnormal levels (FIG. 11). However, no abnormality was detected in the group treated with 33 μl of the germanium-containing cordyceps extract. Therefore, it was confirmed that administration of germanium-containing Cordyceps waxy hot-water extract exerts an effect of killing only the cancerous mass and does not exert toxicity. However, when the concentration of the germanium-containing Cordyceps waxy hot-water extract was doubled (66 μl), abnormalities were found. Therefore, it was confirmed that 33 μl of the extract of Cordyceps mellifera containing organogermanium is the most effective concentration capable of killing the cancer mass without causing toxicity. The concentration of the composition corresponding to 33 추출 of the extract is 50 ug / ml.

4-7. Cordyceps Hot water extract and Organic Germanium  Caterpillars Hot water extract  Measuring the Effect on Nude Mouse Activity

When the activity of the untreated control group was taken as 100%, the activity of the group fed with the organophosphate-containing hot water extract containing the organic germanium was the best. When 33 μl of the germanium-free cordyceps hydrothermal extract was fed, the activity decreased to 91.3%, and when 33 μl of the germanium-containing cordyceps hydrothermal extract was fed, the activity was effectively increased to 111.3% (FIG. ).

Claims (25)

delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete Germanium < / RTI >
A medium composition for promoting mesenchymal stem cell proliferation.
19. The method of claim 18, wherein the germanium-
Inoculating and culturing the Cordyceps sinensis strain in a medium containing germanium to produce a mycelia of Cordyceps;
And extracting the culture containing the mycelia of Cordyceps mellifera.
20. The composition of claim 19, wherein the germanium-containing medium comprises 200 to 1000 ppm of germanium.
The composition according to claim 19, wherein the culture time is 14 to 28 days in the step of inoculating and culturing the organisms in a culture medium containing germanium.
20. The method according to claim 19, wherein the step of extracting the culture medium containing the mycelium of Cordyceps mellifera is performed by hot water extraction of the culture solution containing the mycelia of Cordyceps mellifera.
19. The composition of claim 18, wherein the caterpillar fungus is Cordyceps militaris.
19. The composition of claim 18, wherein the germanium-containing Cordyceps extract comprises between 190 and 220 ppm of germanium.
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