HK1088559B - Plant worms mycelium extracat fraction and composition for oral intake - Google Patents

Description

Extract of Cordyceps sinensis mycelia and composition for oral intake

Technical Field

The present invention relates to a Cordyceps sinensis-derived isolate having pharmacological effects and a method for producing the isolate. The present invention further relates to an orally ingestible composition, a pharmaceutical composition, a food or beverage composition, or a pharmaceutical, pharmaceutical external product, food or beverage having an immunostimulating effect and/or for preventing or treating cancer, containing the isolate.

Background

Cordyceps sinensis is a generic name of fungi produced by insects and the like, and belongs to the genus Ascomycetes, Clavicipitales, Clavicipitaceae. Wherein the Cordyceps sinensis (Berkely) Sacchardo) is parasitized on larva of Hepialus hepialid Chen et Shen, and is a strain distributed in mountain area of China, Tibet, Nipol, Himalayan, and barren grass with elevation of 3000-4000 m. Since ancient times in China, the Chinese medicinal composition is regarded as a Chinese medicament which is not old and can strengthen essence and bones, and is born by the curiosity and inheritance. In recent years, it has been known through studies that components contained in cordyceps sinensis have various physiological activities, and immune activation effects, antitumor activities, blood glucose level lowering effects, blood pressure lowering effects, and vasodilatation effects have been reported (see non-patent documents 1 to 6). Taking such physiological activities into consideration, attempts have been made to efficiently ingest active ingredients (see patent document 1), but it is still difficult to say that the content and activity of active ingredients in the extract are sufficiently high.

In addition, with the progress of culture techniques in recent years, a mycelium culture of Cordyceps sinensis, which is almost the same as that of a natural product, can be produced. Therefore, the mycelia of Cordyceps sinensis are more suitable as a raw material for industrial production than the fruiting body part used in the past. Therefore, a method for efficiently extracting a fraction containing an effective component of Cordyceps sinensis by using the mycelium culture has been sought.

Patent document 1: japanese unexamined patent publication No. 11-228440

Non-patent document 1: pharm Bull, vol.22 (9), p.966-970, 1990

Non-patent document 2: jpn.j.pharmacol, volume 79, page 505-

Non-patent document 3: kanazawa Med. Univ., Vol.16, pp.46-54, 1991

Non-patent document 4: pharm Bull, vol.16 (12), page 1291-1293, 1993

Non-patent document 5: phytochemistry, Vol.51, pp.891-898, 1999

Non-patent document 6: life Science, Vol.66 (No. 14), p.1369-

Disclosure of Invention

The present inventors have made intensive studies to solve the above problems and found a method for preparing a specific and physiologically active isolated product of cordyceps sinensis mycelia.

The purpose of the present invention is to provide a isolate derived from cordyceps sinensis mycelia, which has an immunostimulating effect and/or an effect of preventing or treating cancer, a composition for oral ingestion containing the isolate, and a food or drink, an immunostimulating agent, an anticancer agent and a cancer preventive agent containing the composition for oral ingestion.

That is, according to one aspect of the present invention, there is provided a precipitated fraction obtained by adding ethanol to an extract obtained by extracting cordyceps sinensis mycelia with hot water. The amount of ethanol added here may be, for example, 0.5 to 2.0 times the volume ratio of the aforementioned extract.

In another aspect of the present invention, ethanol is added to the extraction solution in an amount of 0.5 to 2.0 times by volume to the extraction solution, the precipitate formed is removed, the supernatant is used as the obtained extraction solution, and ethanol is further added thereto in an amount such that the ethanol content in the extraction solution is 70 to 90% by volume, thereby providing the precipitate obtained.

The precipitation part is provided through other side of the present invention, and is a precipitate obtained by adding a certain amount of ethanol to a hot water extract of cordyceps sinensis mycelia and/or the extract to obtain a supernatant from which a generated precipitation part is removed, concentrating the supernatant before adding ethanol, and then adding ethanol thereto. According to another aspect of the present invention, there is provided a method for pretreating a precipitate of cordyceps sinensis mycelia, which comprises adding ethanol to an extract obtained by hot water extraction of cordyceps sinensis mycelia to a residue obtained by heat extraction with ethanol. Further, the present invention provides the aforementioned precipitate containing polysaccharides having a molecular weight of 10 ten thousand or more, according to another aspect of the present invention. Further, according to another aspect of the present invention, there is provided the above-mentioned precipitate fraction having an activity of promoting the production of interferon- γ (IFN-. gamma.) or tumor necrosis factor- α (TNF-. alpha.) in mammals.

In another aspect of the present invention, there is provided a fraction having a molecular weight of 10 ten thousand or more obtained by further subjecting the precipitate fraction to molecular weight separation by dialysis.

According to another aspect of the present invention, there is provided a composition for oral ingestion, which contains the above-mentioned precipitated fraction or fraction having a molecular weight of 10 ten thousand or more. In another aspect of the present invention, there is provided a food or beverage containing the composition for oral ingestion. Further, according to another aspect of the present invention, there is provided an immunostimulant comprising the composition for oral ingestion. Further, according to another aspect of the present invention, there is provided an anticancer agent or a cancer preventive agent containing the composition for oral ingestion.

According to still another aspect of the present invention, there is provided a method for manufacturing the above-mentioned part, comprising the steps of,

a) heating and extracting cordyceps sinensis mycelia with water at 85-100 ℃ to obtain an extraction liquid;

b) and a step of adding ethanol to the extract in an amount of 0.5 to 2.0 times by volume of the extract, and recovering the precipitate.

Further, according to another aspect of the present invention, there is provided a method for manufacturing the above-mentioned part, comprising the steps of,

a) heating and extracting cordyceps sinensis mycelia with water at 85-100 ℃ to obtain an extraction liquid;

b) adding ethanol in an amount of 0.5 to 2.0 times by volume of the extraction solution to the extraction solution, removing the generated precipitate, and recovering the supernatant;

c) and a step of recovering the obtained precipitate by further adding ethanol to the supernatant in an amount such that the ethanol content in the supernatant is 70% to 90% by volume.

The present invention will be described in further detail below

The Cordyceps sinensis in the present invention refers to a genus of Ascomycetes, Clavicipitales, Clavicipitaceae, such as Cordyceps sinensis (Berkely) Sacchardo).

The cordyceps sinensis mycelia of the present invention may be produced by culturing or may be obtained naturally, and for example, dried powder of the culture may be used.

The hot water extraction in the present invention is carried out, for example, at a liquid temperature of 70 to 120 ℃. The preferable temperature is 85 to 100 ℃, and the more preferable temperature is 90 to 95 ℃. The extraction time is, for example, 1 to 24 hours, preferably 2 to 12 hours, and more preferably 3 to 7 hours. The hot water extraction may be carried out a plurality of times, for example, 1 to 4 times, preferably 2 to 3 times. The pH of the water used for extraction is not particularly limited, and may be, for example, distilled water.

The extract obtained by the hot water extraction can be used as it is, but the extract can be used after concentrating the extract in order to reduce the amount of ethanol used and to efficiently obtain a separated product. The concentration ratio is not particularly limited, and may be, for example, 80 to 20%, preferably 60 to 25%, and more preferably 50 to 30% by volume. The concentration may be carried out under either atmospheric pressure or reduced pressure, but is preferably carried out under reduced pressure.

The amount of ethanol added to the extract is not particularly limited, and may be, for example, 0.5 to 2.0 times, preferably 0.8 to 1.2 times, the volume ratio to the concentrated extract, or the like, when the extract is not concentrated. The obtained precipitate can be purified by a method known in the art, for example, by dissolving the precipitate in water and adding ethanol to obtain a precipitate, and further purifying the precipitate one or more times.

Further, a certain amount of ethanol is added to the extract, and after the generated precipitate is collected, the precipitate can be obtained by further adding ethanol to the extract obtained as the supernatant. The extract obtained as the supernatant may be used as it is or may be concentrated for use. The concentration ratio is not particularly limited, and may be, for example, 80 to 20%, preferably 60 to 25%, and more preferably 50 to 30% by volume. The concentration may be carried out under either atmospheric pressure or reduced pressure, but is preferably carried out under reduced pressure. The amount of ethanol to be initially added is not particularly limited, and may be, for example, 0.5 to 2.0 times, preferably 0.8 to 1.2 times, the volume ratio to the concentrated extract. The amount of ethanol contained in the extract after the second addition of ethanol is not particularly limited, and may be, for example, 70 to 90% by volume, preferably 75 to 85% by volume. For example, the precipitated fraction can be obtained by adding 0.5 to 2.0 times the volume ratio of ethanol to the extract, recovering the precipitated fraction, removing the ethanol by, for example, concentration under reduced pressure, and then adding 3.0 to 5.0 times the volume ratio of ethanol to the supernatant.

The orally ingestible composition in the present invention includes, for example, a food or beverage composition, a pharmaceutical composition, and the like.

In the present invention, the cordyceps sinensis mycelia extracted with hot water may be pretreated in order to obtain a fraction having a high content of active ingredients. The pretreatment is not particularly limited, and examples thereof include heat extraction using an alcohol having 1 to 4 carbon atoms or an aqueous alcohol solution thereof, and ethanol is preferably used. The temperature during heating is, for example, 70 to 120 ℃ of the liquid temperature, preferably 80 to 100 ℃, and more preferably 90 to 95 ℃. The extraction time is, for example, 1 to 24 hours, preferably 2 to 12 hours, and more preferably 3 to 7 hours. The ethanol extraction may be performed a plurality of times, for example, 1 to 4 times, preferably 2 to 3 times. The extraction may be performed a plurality of times using different solvents, and for example, the extraction may be performed using 95 to 99.5% ethanol, and then the residue may be extracted with 60 to 80% ethanol aqueous solution by volume ratio, and the residue may be used as a raw material for hot water extraction.

The precipitated fraction obtained in the present invention may contain, for example, polysaccharides as a component thereof. The polysaccharide is not particularly limited, and is a naturally occurring monosaccharide, and 1 or 2 or more sites in the molecule thereof may be subjected to modification such as acylation or phosphorylation, or modification by protein or nucleic acid binding. The molecular weight of the polysaccharide is not particularly limited, and may be, for example, 10 to 200 ten thousand. The sugar content of the precipitated fraction of the present invention can be measured by a method known to those skilled in the art, for example, phenol-sulfuric acid method. The sugar content in the present invention is not particularly limited, and is, for example, 40 to 90% by weight, preferably 50 to 80% by weight, and more preferably 60 to 80% by weight in terms of D-glucose.

The above-mentioned precipitate fraction can be separated into, for example, a fraction having a molecular weight of less than 5 ten thousand, a fraction having a molecular weight of 5 ten thousand or more and less than 10 ten thousand, and a fraction having a molecular weight of 10 ten thousand or more by a method such as dialysis.

The immunoactivator of the present invention has an action of promoting the production of interferon-gamma and/or tumor necrosis factor-alpha in mammals, and therefore can be used as a prophylactic and/or therapeutic agent for a subject in need of immune activation. In addition, the anticancer agent of the present invention exhibits a significant antitumor effect, and therefore can be used as a prophylactic or therapeutic agent effective for cancer. Therefore, the immunostimulant and anticancer agent of the present invention can be used as an active ingredient of a pharmaceutical composition. The pharmaceutical composition may contain various components generally used, and for example, may contain 1 or more kinds of pharmaceutically acceptable excipients, diluents, wetting agents, emulsifiers, dispersants, auxiliary agents, preservatives, buffers, binders, stabilizers and the like.

The dose of the immunostimulant and anticancer agent of the present invention can be suitably selected depending on the body form, age, physical condition, disease state, and time elapsed after onset of disease of a patient, and is, for example, 1 to 5000 mg/day per adult.

The orally ingestible composition of the present invention can be used as it is as a functional food, and can also be used as a pharmaceutical external product, a component of a food or drink, a food additive, or the like. By using such a composition, the orally ingestible composition having an immunostimulating effect and an antitumor effect of the present invention can be ingested daily and continuously, and can improve the body condition, treat cancer and prevent the occurrence of cancer by effective immunostimulating. Examples of the use of the orally ingestible composition of the present invention as a food material include functional foods, health foods, general foods (fruit juices, snacks, processed foods, and the like), and nutritional supplements (nutritional drinks and the like) having an immune activation effect, a cancer treatment effect, or a cancer prevention effect.

The oral composition of the present invention has an immune activation effect and/or an antitumor effect, and therefore the present invention provides an effective means for immune activation, an effective means for preventing or treating cancer, and a composition useful for producing a health food.

Drawings

FIG. 1 is a graph showing the results of measuring IFN-. gamma.production-promoting activity of a hot water-extracted fraction (HWE), a 50% ethanol-precipitated fraction (50% EP), an 80% ethanol-precipitated fraction (80% EP), an ethanol extract (Et Ext) and a 70% ethanol extract (70Et Ext) by an in vitro assay using human peripheral blood.

FIG. 2 is a graph showing the results of measuring the TNF-. alpha.production promoting activity of a hot water-extracted fraction (HWE), a 50% ethanol-precipitated fraction (50% EP), an 80% ethanol-precipitated fraction (80% EP), an ethanol extract (Et Ext) and a 70% ethanol extract (70Et Ext) by an in vitro assay using human peripheral blood.

FIG. 3 is a graph showing the results of measuring IFN-. gamma.production-promoting activity of a fraction having a molecular weight of 10 ten thousand or more (M1), a fraction having a molecular weight of 5 ten thousand or more and less than 10 ten thousand (M2), a fraction having a molecular weight of less than 5 ten thousand (M3), a hot water-extracted fraction (HWE) and a 50% ethanol-precipitated fraction (50% EP) by an in vitro test using human peripheral blood.

FIG. 4 is a graph showing the results of measuring the TNF-. alpha.production amount-promoting activity of a fraction having a molecular weight of 10 ten thousand or more (M1), a fraction having a molecular weight of 5 ten thousand or more and less than 10 ten thousand (M2), a fraction having a molecular weight of less than 5 ten thousand (M3), a hot water extraction fraction (HWE) and a 50% ethanol precipitation fraction (50% EP) by an in vitro test using human peripheral blood.

FIG. 5 is a graph showing changes in tumor volume by administration of a hot water extraction fraction (HWE), a 50% ethanol precipitation fraction (50% EP), and an 80% ethanol precipitation fraction (80% EP) to mice.

Detailed Description

Hereinafter, preferred examples of the present invention will be described in further detail, but the present invention is not limited to these examples.

EXAMPLE 1 preparation of an isolated product of Chinese Caterpillar fungus mycelia

In the practice of the present invention, as the powder of the cultured Chinese caterpillar fungus mycelia to be used as the raw material, for example, "fermented Cordyceps fungus powder" (product name: CORBRIN CAPSULE, manufacturing company: China eastern Mey, Ltd.) commercially available in China can be used.

Extracting the powder of the cordyceps sinensis mycelia with 99.5% ethanol at 90-95 ℃ for 6 hours. The amount of ethanol was 2mL per 1g of mycelia. The residue is filtered, the residue is extracted again in this way, and the extract and the residue are separated by filtration. The extracts obtained by the 2 extractions were combined and concentrated under reduced pressure to obtain a viscous, concentrated brown liquid ethanol extract (Et Ext) in terms of raw materials (15.0% by weight). The residue is then extracted with 70% ethanol aqueous solution (by volume) at 90-95 ℃ for 6 hours. The amount of 70% ethanol was 2mL per 1g of residue. The residue is filtered, the residue is extracted again in this way, and the extract and the residue are separated by filtration. The extracts obtained by the 2 extractions were combined and concentrated under reduced pressure until the volume became about 40%, and the mixture was lyophilized to obtain a 70% ethanol extract (70% Et Ext) as a concentrated brown powder (45.8% by weight in terms of the starting material). The residue is then extracted with distilled water at 90-95 ℃ for 6 hours. The amount of distilled water was 4mL per 1g of the residue. The residue is filtered, the residue is extracted again in this way, and the extract and the residue are separated by filtration. The extracts obtained by the 2 extractions were combined and concentrated under reduced pressure until the volume was about 40%, and 99.5% ethanol was added thereto in an equal volume ratio, and the mixture was allowed to stand overnight in a cold and dark place. The precipitate was recovered by centrifugation, dissolved in distilled water and frozen, and freeze-dried to obtain a 50% ethanol precipitated fraction (50% EP) of a highly hygroscopic dark brown powder of 17.9% (by weight) in terms of the starting material. After the supernatant was concentrated under reduced pressure to about 40% in volume, 99.5% ethanol was added in an amount of 4 times the volume, and the mixture was allowed to stand overnight in a cold and dark place. The precipitate was recovered by centrifugation, dissolved in distilled water and frozen, and freeze-dried to obtain a brown powder 80% ethanol precipitate (80% EP) in an amount of 6.0% by weight in terms of the starting material.

The 50% ethanol precipitate was dissolved in distilled water, and the mixture was purified by using a Spectrum/Por (registered trademark) CE Membrane MWCO manufactured by Spectrum: 100000 dialyzed against distilled water. The amount of distilled water to dissolve the 50% ethanol precipitate portion was 50mL per 1g of the precipitate portion. A fraction (M1) having a molecular weight of 10 ten thousand or more was obtained by freeze-drying the dialyzed internal solution in terms of 10.0% by weight of the starting material. The dialyzed external solution was concentrated under reduced pressure, and the resultant was purified by using a Spectrum/Por (registered trademark) 6Membrane MWCO manufactured by Spectrum corporation: 50000 dialyzed against distilled water. A fraction (M2) having a molecular weight of 5 ten thousand or more and less than 10 ten thousand, in terms of 3.94% by weight of the starting material, was obtained by freeze-drying the dialyzed internal solution. The dialyzed external solution was concentrated under reduced pressure and lyophilized to obtain a fraction (M3) having a molecular weight of less than 5 ten thousand in terms of 3.45% (by weight) of the starting material.

The sugar content of the M1 fraction was quantified by the phenol-sulfuric acid method, and the sugar content of the M1 fraction was 69.9% in terms of D (+) -glucose. Here, the phenol-sulfuric acid method is carried out by a general method (for example, see "New Experimental chemistry lecture 20 biochemistry [ II ]", page 1085, edited by Japan chemical society, 10.20.1978, Bolus Kabushiki Kaisha)

Production example 1 preparation example of Hot Water extract

The hot water extracts used as comparative control in the examples were prepared in the following manner. Extracting the cordyceps sinensis mycelium powder with distilled water at 90-95 ℃ for 4 hours while stirring. The amount of distilled water was 20mL per 1g of mycelia. The residue was filtered, and the filtrate was concentrated under reduced pressure and lyophilized to obtain a hygroscopic dark brown powder Hot Water Extract (HWE).

Example 2 measurement of immune activation Effect using human peripheral blood

To a 48-well cell culture plate, 500. mu.L of peripheral blood collected from human and an equal amount of RPMI1640 medium to the peripheral blood were added. Adding the substance to be detected to a final concentration of 200. mu.g/mL, and adding 5% CO at 37 deg.C₂Cultured under the conditions of (1) for 24 hours. Recovering culture supernatant by enzyme linked immunosorbent assayThe assay kit (manufactured by BIOSOURCE corporation) for the method (ELISA) quantitatively determined interferon- γ (IFN-. gamma.) and tumor necrosis factor- α (TNF-. alpha.) respectively. The method of using the kit and the method of analyzing the quantitative result were carried out in accordance with the instructions attached to the kit.

IFN-. gamma.production and TNF-. alpha.production were measured for the 50% ethanol-precipitated fraction (50% EP), the 80% ethanol-precipitated fraction (80% EP), the ethanol extract (Et Ext), the 70% ethanol extract (70% Et Ext) prepared in example 1 and the Hot Water Extract (HWE) prepared in production example 1, and the results are shown in FIGS. 1 and 2, respectively. From the results of the measurement, it was found that 50% EP and 80% EP have a significant effect of promoting the production of IFN-. gamma.or TNF-. alpha.as compared with HWE, Et Ext and 70% Et Ext, and the effect of the isolate of the present invention in a test system using human-derived peripheral blood was confirmed. In addition, it was also found that 50% EP has a higher accelerating effect than 80% EP.

For the molecular weight isolates M1 (fraction having a molecular weight of 10 ten thousand or more), M2 (fraction having a molecular weight of 5 ten thousand or more and less than 10 ten thousand) and M3 (fraction having a molecular weight of less than 5 ten thousand) of the 50% ethanol precipitation fraction (50% EP) prepared in example 1, IFN-. gamma.and TNF-. alpha.production were measured, and the results are shown in FIG. 3 and FIG. 4, respectively. From the results of this measurement, it was found that M1 had an effect of promoting the production of EP by more than 50% in both IFN-. gamma.production and TNF-. alpha.production. In addition, it was confirmed that the activity of promoting the production of IFN-. gamma.and TNF-. alpha.was concentrated only on M1 (fraction having a molecular weight of 10 ten thousand or more), and that the activity of promoting the production of IFN-. gamma.and TNF-. alpha.of 50% EP was deeply related to the compound having a molecular weight of 10 ten thousand or more.

EXAMPLE 3 measurement of antitumor Effect Using cancer-afflicted mice

6-week-old crossbreed S1c purchased from SLC corporation of Japan 32 male mice BDF1 were divided into 4 groups of 8 mice, and the 4 mice were kept in one cage for 1 week. 7 weeks old mice were inoculated subcutaneously under the right axilla with 1X 10⁶Tumor Sarcoma180 of cell/mouse. In addition, the inoculation sites were shaved off prior to inoculationThe body hair of (2) enables observation of adhesion and growth of a tumor to be more accurately performed. The growth of tumor is measured by digital vernier caliper to obtain the short and long diameters of tumor, and the size of tumor (long diameter × short diameter) is calculated²/2mm³). The 4 groups were a control group (control group), a hot water extract group (HWE group), a 50% ethanol precipitation fraction group (50% EP group), and an 80% ethanol precipitation fraction group (80% EP group), and each group was orally administered with a probe at 10 am every day from the day of tumor inoculation to the test substance. The dose was 0.1mL per 10g body weight in the control group, 250mg/kg of HWE dissolved in physiological saline was administered in the HWE group, 44.8mg/kg of 50% EP dissolved in physiological saline was administered in the 50% EP group, and 15.0mg/kg of 80% EP dissolved in physiological saline was administered in the 80% EP group.

The measurement results of the tumor volumes on day 7 and day 10 are shown in fig. 5. From the results of the measurement, it was confirmed that the isolate of the present invention had antitumor activity in the in vivo test system. Furthermore, 50% EP administered in an amount of 1/5 or less of HWE clearly showed an antitumor effect exceeding that of HWE, and this result also supports the result of example 2.

EXAMPLE 4A 431 cell proliferation inhibition assay

A431 cells (human squamous cell carcinoma cell line; prepared from Japan) were suspended in Dulbecco's modified Eagle's medium (SIGMA corporation) containing 10% FBS so as to be 4X 10⁴Each at 50. mu.L (2X 10)³One/well) was added to each well of a 96-well microplate (manufactured by sumitomo BAKELITE). After 6 hours of culture (37 ℃ C., 5% CO)₂) To each well, a medium containing HWE, 70% Et Ext, 50% EP, 80% EP and 10% FBS as a test substance at 2000. mu.g/ml was added at 50. mu.L (final concentration: 1000. mu.g/ml). As a positive control, 50. mu.L of a medium containing 200. mu.M Genistein was added. After 4 days of culture, 5. mu.L of Cell counting kit-8 (Wako pure chemical industries, Ltd.) was added to each well, and the absorbance (wavelength: 450nm) was measured after 60 minutes. The inhibition ratio was calculated from the comparison with a system containing no test substance.

The test results are shown in table 1 below. In the test results, all the substances to be tested showed the effect of inhibiting cell proliferation, and among them, the inhibition rate of 80% EP was remarkably high.

TABLE 1

EXAMPLE 5A 431 cell Damage test

A431 cells (human squamous cell carcinoma cell line; prepared from Japan) were suspended in Dulbecco's modified Eagle's medium (SIGMA corporation) containing 10% FBS so as to be 5X 10⁵Each at a concentration of 100. mu.L (5X 10)⁴One/well) was added to each well of a 96-well microplate (manufactured by sumitomo BAKELITE). After 24 hours of culture (37 ℃ C., 5% CO)₂) The medium was exchanged with serum-free medium containing 1000. mu.g/mL HWE, 70% Et Ext, 50% EP, 80% EP, respectively. After 48 hours of culture, 5. mu.L of Cell counting kit-8 (Wako pure chemical industries, Ltd.) was added to each well, and the absorbance (wavelength 450nm) was measured after 30 minutes. The survival rate was calculated from the comparison with a system containing no test substance.

The test results are shown in table 2 below. In this assay result, HWE and 70% Et Ext showed some cytotoxicity, while the isolates of the invention, 50% EP and 80% EP, did not show any effect on the survival of a431 cells. From this, it was confirmed that the cell growth inhibitory effects exhibited by 50% EP and 80% EP in example 4 were not due to cell damage.

TABLE 2

Claims (15)

1. A mycelium extract of Cordyceps sinensis is characterized in that a precipitate is obtained by adding ethanol in an amount of 0.5-2.0 times by volume of an extract obtained by extracting the mycelium of Cordyceps sinensis with hot water.

2. An extract of mycelia of Cordyceps sinensis, which is characterized by being obtained by removing the precipitate of claim 1 from the supernatant as an extract and further adding ethanol to the extract in an amount such that the ethanol content in the extract is 70 to 90% by volume.

3. The extract of mycelia of chinese caterpillar fungus according to claim 1 or 2, wherein the extract is a substance obtained by concentration before adding ethanol.

4. The extract of the mycelium of chinese caterpillar fungus according to claim 1 or 2, wherein the mycelium is a residue after heat extraction with ethanol as a pretreatment.

5. The extract of mycelia of chinese caterpillar fungus according to claim 1 or 2, which contains polysaccharides having a molecular weight of 10 ten thousand or more.

6. The extract of the mycelium of chinese caterpillar fungus according to claim 1 or 2, which has an activity of promoting the production of interferon- γ or tumor necrosis factor- α in mammals.

7. A mycelium extract of Cordyceps sinensis, characterized in that it is a substance having a molecular weight of 10 ten thousand or more obtained by further subjecting the precipitate fraction of claim 1 to molecular weight separation by dialysis.

8. The extract of mycelia of Cordyceps sinensis as claimed in claim 7, wherein the content of sugar is 60 to 80% by weight in terms of D-glucose.

9. A composition for oral ingestion, comprising the extract of mycelia of Cordyceps sinensis according to any one of claims 1 to 8.

10. A food or drink characterized by containing the composition for oral intake according to claim 9.

11. The food or drink according to claim 10, which has an anticancer effect, a cancer-preventing effect, or an immune-activating effect.

12. An immunostimulant comprising the oral composition according to claim 9.

13. An anticancer agent comprising the composition for oral ingestion according to claim 9.

14. The method for producing the extract of mycelia of Cordyceps sinensis as claimed in any one of claims 1 to 8, comprising the steps of,

a) heating and extracting cordyceps sinensis mycelia with water at 85-100 ℃ to obtain an extraction liquid;

b) and a step of adding ethanol to the extract in an amount of 0.5 to 2.0 times by volume of the extract, and recovering the precipitate.

15. The method for producing the extract of mycelia of Cordyceps sinensis as claimed in any one of claims 2 to 8, comprising the steps of,

a) heating and extracting cordyceps sinensis mycelia with water at 85-100 ℃ to obtain an extraction liquid;

b) adding ethanol in an amount of 0.5 to 2.0 times by volume of the extraction solution to the extraction solution, removing the generated precipitate, and recovering the supernatant;

c) and a step of adding ethanol to the supernatant in an amount such that the ethanol content in the supernatant is 70 to 90% by volume, and recovering the precipitate thus obtained.