WO2005004892A1

WO2005004892A1 - Plant worms mycelium extracat fraction and composition for oral intake

Info

Publication number: WO2005004892A1
Application number: PCT/JP2004/009550
Authority: WO
Inventors: Yoshihiro Yamaguchi; Eiji Kunitomi; Kenji Uchida
Original assignee: Kobayashi Pharmaceutical Co., Ltd.
Priority date: 2003-07-14
Filing date: 2004-07-06
Publication date: 2005-01-20
Also published as: CN1822847A; JP4746260B2; CN100589812C; HK1088559A1; JP2005035928A

Abstract

[PROBLEMS] To provide a fraction containing the active ingredient contained in plant worms. To provide a means efficacious in immunopotentiation, an effective means of preventing or treating cancer and a composition useful in producing health foods. [MEANS FOR SOLVING PROBLEMS] It is intended to provide a precipitation fraction obtained by extracting plant worms mycelium with hot water and adding ethanol to the thus obtained extract. It is also intended to provide a composition for oral intake which contains the above precipitation fraction. It is further intended to provide foods, drinks, immunopotentiators, anticancer agents and so on containing the above composition for oral intake which are characterized by having an anticancer effect, an effect of preventing cancer or an immunopotentiation effect. It is furthermore intended to provide a method of effectively producing the above-described composition for oral intake.

Description

Specification

Fractionated Cordyceps mycelium extract and composition for oral consumption

Technical field

[0001] The present application relates to a fraction derived from Cordyceps sinensis having a pharmacological effect, and a method for producing the fraction. Furthermore, the present invention relates to an orally ingested composition, a pharmaceutical composition, a food and drink composition containing the fraction, or a pharmaceutical, a quasi-drug, a food or drink having an immunostimulatory effect and / or used for the prevention or treatment of cancer. It relates to things.

^ Scenic technology

[0002] Cordyceps is a collective term for mushrooms arising from insects and the like, and is positioned as a genus of the Aspergillus fungus' Aperidomycete '' Aperidomyceae. Among them, Cordyceps s inensis (Berkely) Saccardo is a mushroom that parasitizes lepidopterous bat larvae and is distributed in the alpine belts of China, Tibet, Nepal, the Himalayas, 3000-4000 m above sea level. is there. In China, it has long been prized and handed down as a traditional Chinese medicine for immortality and tonic. Recent research has shown that the components contained in Cordyceps sinensis have various physiological activities, and there have been reports on immunostimulatory effect, antitumor activity, hypoglycemic effect, hypotensive effect and vasodilatory effect. (See Non-Patent Document 1-6). Attempts have been made to efficiently ingest active ingredients by paying attention to such physiological activities (see Patent Document 1), but the content of the active ingredients in the extract and the active aspect are sufficient. It ’s hard to say.

[0003] Further, due to recent advances in culture technology, it is possible to produce cordyceps mycelium cultures that are almost equivalent to natural products. Therefore, the mycelium of Cordyceps sinensis is more suitable as a raw material for industrial production than the part of the fruit body conventionally used. Therefore, there is a demand for a method for efficiently removing a fraction containing the active ingredient of Cordyceps sinensis using the mycelium culture.

Patent Document 1: Japanese Patent Laid-Open No. 11-228440

Non-Patent Document 1: Biol. Pharm. Bull., No. 22 (No. 9), 966-970, 1990 Non-Patent Document 2: Jpn. J. Pharmacol., 79, 505-508 1999 Non-Patent Document 3: J. Kanazawa Med. Univ., Vol. 16, pp. 46-54, 1991 Non-Patent Document 4: Biol. Pharm. Bull., Vol. 16 (No. 12), 1291-1293 Page, 1993

Non-Patent Document 5: Phytochemistry, 51st, 891-898, 1999

Non-Patent Document 6: Life Science, Vol. 66 (No. 14), pp. 1369-1376, 2000 Disclosure of the Invention

Means for solving the problem

[0004] As a result of diligent research to solve the problem, the present inventor has found a method for preparing a fraction of Cordyceps mycelium having specific physiological activity.

An object of the present invention is to provide a fraction derived from Cordyceps mycorium having an immunostimulatory effect and an effect of preventing or treating Z or cancer, a composition for oral consumption containing the fraction, and the composition for oral intake. It is to provide foods and drinks, immunostimulants, anticancer agents and cancer preventives.

[0005] That is, according to one aspect of the present invention, there is provided a precipitate fraction obtained by adding ethanol to an extract obtained by hot water extraction of Cordyceps mycelium. The amount of ethanol added here may be, for example, 0.5 to 2.0 times the volume ratio of the extract.

[0006] According to another aspect of the present invention, 0.5 to 2.0 times as much ethanol as the volume ratio of the extract is added to the extract, and the resulting precipitate fraction is obtained as a supernatant. The precipitate fraction obtained by further adding ethanol in such an amount that the extract contains 70% -90% of ethanol in a volume ratio is provided.

[0007] According to another aspect of the present invention, the precipitate fraction is obtained by extracting a Cordyceps mycorrhizal mycelium with a hot water extract and / or adding a certain amount of ethanol to the extract. The removed supernatant is concentrated before adding ethanol, and then ethanol is added to provide a precipitate fraction. According to another aspect of the present invention, a residue obtained by heat extraction with ethanol as a pretreatment is used as the Cordyceps mycorrhizal mycelium to obtain ethanol by adding ethanol to an extract obtained by hot water extraction. Precipitate fraction is provided. Furthermore, according to another aspect of the present invention, the precipitate fraction has a molecular weight of 100,000 or more. The precipitated fraction is characterized in that it comprises a polysaccharide. Furthermore, according to another aspect of the present invention, there is provided the precipitate fraction characterized by having an activity of enhancing production of interferon γ (IFN γ) or tumor necrosis factor _α (TNF-ct) in mammals. Is done.

[0008] According to another aspect of the present invention, there is provided a fraction having a molecular weight of 100,000 or more obtained by further subjecting the precipitate fraction to molecular weight fractionation by dialysis.

According to another aspect of the present invention, there is provided a composition for ingestion comprising the precipitate fraction or the fraction having a molecular weight of 100,000 or more. According to another aspect of the present invention, a food or drink containing the composition for oral consumption is provided. Furthermore, according to another aspect of the present invention, an immunostimulant comprising the composition for oral consumption is provided. Furthermore, according to another aspect of the present invention, an anticancer agent or cancer preventive agent comprising the composition for oral intake is provided.

[0009] According to still another aspect of the present invention, there is provided a method for producing the fraction,

a) a process in which Cordyceps mycelium is heated and extracted with water at 85-100 ° C to obtain an extract; and

b) The above production method is provided, which comprises a step of collecting 0.5 to 2.0 times the amount of ethanol in the extract with respect to the extract and collecting the generated precipitate. Furthermore, according to another aspect of the present invention, there is provided a method for producing the fraction,

a) A step of heat extraction of Cordyceps mycelium with water at 85-100 ° C to obtain an extract; b) 0.5-2. 0 times as much ethanol as the extract to the extract. Calorie, removing the generated precipitate and recovering the supernatant; and

c) recovering the precipitate obtained by further adding ethanol in an amount such that the supernatant contains 70% -90% ethanol in a volume ratio to the supernatant. Is also provided.

[0010] Hereinafter, the present invention will be described more specifically.

The cordyceps in the present invention means a genus of the mycomycetes 'Aperidomyceae', which is, for example, Cordyceps sinensis (Berkely) Saccardo.

[0011] The Cordyceps mycelium in the present invention is natural even if it is produced by culture. For example, a dry powder of a culture can be used.

The hot water extraction performed in the present invention is performed, for example, at 70-120 ° C. at the liquid temperature. The preferred temperature is 85-100 ° C, more preferably 90-95 ° C. The extraction time is, for example, 1 hour to 24 hours, preferably 2 to 12 hours, and more preferably 37 hours. The hot water extraction may be performed a plurality of times, for example, 1 to 4 times, preferably 2 to 3 times. The pH value of the water used for extraction is not particularly limited, but for example, distilled water can be used.

[0012] The extract obtained by the hot water extraction can be used as it is, but the extract is concentrated and used in order to reduce the amount of ethanol used and obtain a fraction efficiently. You can also. The concentration ratio is not particularly limited, but may be, for example, 80 to 20% by volume, preferably 60% to 25%, and more preferably 50% to 30%. The concentration can be performed under normal pressure or reduced pressure, but is preferably performed under reduced pressure.

[0013] The amount of ethanol added to the extract is not particularly limited. For example, the volume ratio of the extract to the extract after concentration may be 0 · 5–2 · 0 times, preferably 0 · 8–1 · This is the same even when it is not concentrated. The obtained precipitate fraction can be purified by a method well known in the art, for example, by dissolving once again in water and then adding ethanol to obtain a precipitate fraction one or more times. It can also be purified.

[0014] Further, a precipitate fraction can be further obtained by adding ethanol to the extract obtained as a supernatant after recovering a precipitate fraction produced by adding a certain amount of ethanol to the extract. . The extract obtained as the supernatant can be used as it is, but can also be used after being concentrated. The concentration rate is not particularly limited. For example, the volume ratio is preferably 80 to 20%, preferably 60% to 25%, and more preferably 50% to 30%. The concentration can be performed under normal pressure or reduced pressure, but is preferably performed under reduced pressure. The amount of ethanol added first is not particularly limited, but may be, for example, 0.5 to 2.0 times the volume ratio with respect to the concentrated extract, preferably 0.8. 2 times. The amount of ethanol contained in the extract after the second ethanol is added is not particularly limited, but may be, for example, 70% to 90% by volume. More preferably, it is 75% -85%. For example, by adding ethanol in a volume ratio of 0 · 5-2.0 times as much as the volume of the extract, the resulting precipitate fraction is recovered, and then concentrated to, for example, vacuum to remove the ethanol, and then the volume ratio is 3.0. — 5. Precipitate fraction can be obtained by adding 0 times ethanol.

[0015] The composition for oral consumption in the present invention includes, for example, a food and drink composition, a pharmaceutical composition, and the like.

In order to obtain a fraction with a high content of the active ingredient, fractions of cordyceps mycelium subjected to hot water extraction in the present invention may be pretreated. The pretreatment is not particularly limited, and examples thereof include heat extraction using an alcohol having 11 to 14 carbon atoms or the aqueous alcohol solution, and ethanol is preferably used. The temperature at the time of heating is, for example, 70 to 120 ° C., preferably <is 80 to 100 ° C., more preferably <is 90 to 95 ° C. The extraction time is, for example, 1 to 1 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. For example, after extraction with 99.5% ethanol, the residue is extracted with 60-80% ethanol aqueous solution by volume ratio, and the residue is extracted. Can be used as raw material for hot water extraction.

[0016] The precipitate fraction obtained in the present invention may contain, for example, polysaccharides as its components. The polysaccharide is not particularly limited, and is composed of naturally occurring monosaccharides. At one or more sites in the molecule, modifications such as asylation and phosphorylation, or proteins and nucleic acids are bound. You may receive modification by doing. The molecular weight of the polysaccharide is not particularly limited, but may be, for example, 100,000 or more and 2 million or less. The sugar content of the fraction of the present invention can be measured by a method well known to those skilled in the art, for example, phenol-sulfuric acid method. The sugar content of the present invention is not particularly limited, but it is, for example, 40 to 90% by weight, preferably 50 to 80% by weight, more preferably 60 to 80% by weight in terms of D-gnolecose.

[0017] The precipitate fraction can be fractionated into, for example, a fraction having a molecular weight of less than 50,000, a fraction having a molecular weight of 50,000 or more and less than 100,000, and a fraction having a molecular weight of 100,000 or more by means such as dialysis. Since the immunostimulant of the present invention has an action of enhancing production of interferon-γ and / or tumor necrosis factor- _α in mammals, it is used as a prophylactic and / or therapeutic agent for subjects who require immunostimulation. can do. Further, since the anticancer agent of the present invention exhibits a significant antitumor effect, it can be an effective prophylactic and therapeutic agent 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 commonly used ingredients, such as one or more pharmaceutically acceptable excipients, diluents, wetting agents, emulsifiers, dispersants, adjuvants, May contain preservatives, buffers, binders, stabilizers, etc.

[0018] The dose of the immunostimulant and anticancer agent of the present invention can be appropriately selected depending on the patient's body shape, age, physical condition, degree of disease, elapsed time after onset, etc. / Day / used at adult dose.

[0019] Further, the ingested composition of the present invention can be used as a functional food as it is, and can also be used as a component for quasi drugs, foods and drinks, food additives and the like. The use enables daily and continuous ingestion of the oral ingestion composition having the immunostimulatory effect and antitumor effect of the present invention, and improves the constitution by effective immunostimulation, treatment of cancer and onset of cancer. Prevention is possible. Examples of the use of the oral ingestion composition of the present invention as a food material include functional foods, health foods, and general foods (juices, confectionery, processed foods) that have an immunostimulatory effect, a cancer therapeutic effect, or a preventive effect. Etc.) and dietary supplements (such as energy drinks).

The invention's effect

[0020] Since the orally ingested composition of the present invention has an immunostimulatory action and Z or antitumor action, the present invention provides an effective means for immunostimulation and an effective preventive or therapeutic means for cancer. And a composition useful in the manufacture of health foods.

BEST MODE FOR CARRYING OUT THE INVENTION

[0021] Preferred embodiments of the present invention will be described in more detail below. However, the present invention is not limited to these examples.

[Example 1] Preparation of fraction of Cordycebus sinensis mycelium

In practicing the present invention, the powder of Cordycebus sinensis mycelium as a raw material is, for example, “fermented worm fungus powder” commercially available in China (product name: CORBRIN CAPSULE, manufacturer: Hangzhou Zhonghua East Pharmaceutical Co., Ltd.) Company) can be used.

[0022] The raw cordyceps mycelium powder was extracted with 99.5% ethanol at 90-95 ° C for 6 hours.

The amount of ethanol was 2 mL per lg mycelium. The residue was separated by filtration, and the residue was extracted again by this method and separated into an extract and residue by filtration. The extracts obtained by the second extraction were combined and concentrated under reduced pressure to obtain 15.0% (weight ratio) of a viscous dark brown liquid ethanol extract (Et Ext) as a raw material. Next, the residue was extracted with a 70% aqueous ethanol solution (volume ratio) at 90-95 ° C for 6 hours. The amount of 70% ethanol was 2 mL per lg of residue. The residue was separated by filtration, and the residue was extracted again by this method, and separated into an extract and residue by filtration. The extracts obtained by the two extractions were combined and concentrated under reduced pressure until the volume reached about 40%, and freeze-dried to obtain a dark brown powder of 70% ethanol extract (70% Et Ext) as a raw material. % (Weight ratio) was obtained. Next, the residue was extracted with distilled water at 90-95 ° C for 6 hours. The amount of distilled water was 4 mL per lg residue. The residue was separated by filtration, and the residue was extracted again by this method and separated into an extract and residue by filtration. The extracts obtained by the two extractions were combined and concentrated under reduced pressure until the volume reached about 40%, and an equal volume of 99.5% ethanol was added in a volume ratio, and the mixture was allowed to stand in a single cold place. . The precipitate is recovered by centrifugation, dissolved in distilled water, frozen, and freeze-dried to obtain a highly hygroscopic 50% ethanol precipitate fraction (50% EP) of 19.9% (weight) Ratio). The supernatant was concentrated under reduced pressure until the volume reached about 40%, 4 volumes of 99.5% ethanol was added by volume ratio, and the mixture was allowed to stand in a cold place. The precipitate was collected by centrifugation, dissolved in distilled water, frozen, and freeze-dried to obtain a brown powder 80% ethanol precipitate fraction (80% EP) in a raw material equivalent of 6.0% (weight ratio).

[0023] Dissolve the 50% ethanol precipitate in distilled water and dialyze against distilled water using SpectraZPor (registered trademark) CE Membrane MWC 0: 100,000 manufactured by Spectrum. The amount of distilled water used to dissolve the 50% ethanol precipitate fraction was 50 mL per lg fraction. Freeze-dried the dialysis solution to make a fraction (Ml) with a molecular weight of 100,000 or more (weight ratio) in terms of raw material (Ml) Got. The dialyzed external solution was concentrated under reduced pressure and dialyzed against distilled water using Spectra / Por (registered trademark) 6 Membrane MWCO: 50,000 manufactured by Spectrum. By lyophilizing the dialysis internal solution, a fraction (M2) having a molecular weight of 50,000 to less than 100,000, which is 3.94% (weight ratio) in terms of raw material, was obtained. The dialyzed external solution was concentrated under reduced pressure and lyophilized to obtain a fraction (M3) having a molecular weight of less than 50,000 of 3.45% (weight ratio) in terms of raw material.

[0024] As a result of quantification of the sugar content of the Ml fraction by the phenol-sulfuric acid method, the sugar content of the Ml fraction was 69.9% in terms of D (+)-gnolecose. The phenol-sulfuric acid method was performed by a general method (for example, “New Experimental Chemistry Course 20 Biochemistry [II]”, page 1085, edited by The Chemical Society of Japan, October 20, 1978, Maruzen Co., Ltd.) checking ...

) o

[0025] [Production Example 1] Preparation example of hot water extract

The hot water extract used as a comparative control in the examples was prepared by the following method. Cordyceps mycelium powder was extracted with distilled water at 90-95 ° C for 4 hours with stirring. The amount of distilled water was 20 mL per lg mycelium. The residue was separated by filtration, and the filtrate was concentrated under reduced pressure and freeze-dried to obtain a hot water extract (HWE) which was a hygroscopic brown powder.

[0026] [Example 2] Measurement of immunostimulatory effect using human peripheral blood

Peripheral blood 500 / i L collected from human force and RPMI1640 medium in an amount equal to that of peripheral blood were added to a 48-well cell culture plate. The test substance was supplemented to a final concentration of 200 μg / mL and cultured under conditions of 37 ° C and 5% CO for 24 hours. Collect the culture supernatant and use interferon

2

-y (IFN-y) and tumor necrosis factor-a (TNF-a) were each quantified using a measurement kit (manufactured by Biosource) by enzyme immunoassay (ELI SA). The method of using the kit and the method of analyzing the quantification results were in accordance with the procedure attached to the kit.

[0027] 50% ethanol precipitation fraction (50% EP), 80% ethanol precipitation fraction (80% EP), ethanol extract (Et Ext), 70% ethanol extract (70% Et Ext) prepared in Example 1 Figure 1 and Figure 2 show the results of measuring IFN-production and TNF-production for the hot water extract (HWE) prepared in Production Example 1. From the measurement results, in both IFN-γ and TNF-chicken, 50% EP and 80% EP showed a significant increase in production compared to HWE, Et Ext, and 70% Et Ext. Thus, the effect of the fraction of the present invention in a test system using human-derived peripheral blood was confirmed. It was also revealed that 50% EP had a higher enhancement effect than 80% EP.

[0028] 50% ethanol precipitation fraction (50% EP) molecular weight fraction prepared in Example 1 Ml (fraction with a molecular weight of 100,000 or more), M2 (molecular weight of 50,000 or more and less than 100,000) Figure 3 and Figure 4 show the results of measuring the production of IFN-γ and TNF-spider for M3 (fraction of less than 50,000) and M3 (fraction of molecular weight less than 50,000), respectively. From the measurement results, it was found that both IFN-γ production and TNF-spider have a production enhancement effect with Ml exceeding 50% 50. In addition, the production-enhancing activity is concentrated only in Ml (fraction with a molecular weight of 100,000 or more), and the IFN-γ and TNF-spider production-promoting activity at 50% EP is a compound with a molecular weight of 100,000 or more. Was found to be deeply involved.

[0029] [Example 3] Measurement of anti-tumor effect using tumor-bearing mice

6-week-old crossing group purchased from Japan SLC Co., Ltd. Sic: 32 BDF1 mice 32 mice were divided into 4 groups, and 4 mice were acclimated for 1 week. A 7-week-old mouse was inoculated with the mouse tumor Sarcomal80 under the right underarm skin at 1 × 10 ⁶ cells / mouse. Prior to the inoculation, the hair of the inoculation part was shaved so that the tumor colonization and growth were observed more accurately. Tumor growth was determined by measuring the short axis and long axis of the tumor with a digital caliper, and calculating the size of the tumor (major axis x minor axis length 2 mm ³ ). Group 4 is a control group (control group), hot water extract group (HWE group), 50% ethanol precipitation fraction group (50% EP group) and 80% ethanol precipitation fraction group (80% EP group). The group was orally administered the test substance with a sonde every day at 10 am from the day of tumor inoculation. In the control group, physiological saline was administered at a dose of 0.1 mL per 10 g of body weight, and in the HWE group, HWE was administered in physiological saline at 250 mg / kg. 50% EP group is 50% EP strength S44. Dissolved in physiological saline to 8 mg / kg, 80% EP group is 80% EP strength S15.0 and dissolved in physiological saline to be 0 mg / kg. Administered.

[0030] FIG. 5 shows the measurement results of tumor volume on day 7 and day 10. From the measurement results, the antitumor activity of the fraction of the present invention was confirmed in an in vivo test system. In addition, the dose should be 50% EP of HWE 1Z5 or less. At the same time, this result supported the result in Example 2.

[0031] [Example 4] A431 cell growth inhibition test

A431 cells (human squamous cell carcinoma cell line; obtained from Dainippon Pharmaceutical Co., Ltd.) are suspended in Dulbecco's modified Eagle's medium (manufactured by Sigma) containing 10% FBS at a concentration of ⁴ × 10 ⁴ cells / mL. 50 μL (2 × 10 ³ pcs / tool) was added to each well of the hole mic plate (Sumitomo Bakelite). After incubation for 6 hours (37 ° C, 5% C ○), HWE, 70% Et Ext,

2

50 μL of medium containing 50% EP and 80% EP each containing 2000 × g / ml and 10% FBS was added to each well (final concentration 1000 μg / ml). A medium containing 200 μΜ Genistein with 50 a L added was used as a positive control. After 4 days of culture, 5 μl of Cell counting kit-8 (Wako Pure Chemical Industries, Ltd.) was added to each well, and the absorbance (wavelength 450 nm) after 60 minutes was measured. The inhibition rate was calculated by comparison with a system not containing a test substance.

[0032] The test results are shown in Table 1 below. In the test results, 80% EP was remarkably high, especially when all the test substances showed the effect of suppressing cell proliferation.

[0033] [Table 1]

Depreciation control rate

Genistein 100 μΜ 619%

HWE lOOOM W A5 &%

70% EtE t 361%

50% EP ιοοθ © ω 49.6%

80% EP 835%

[Example 5] A431 cytotoxicity test

A431 cells (human squamous cell carcinoma cell line; obtained from Dainippon Pharmaceutical Co., Ltd.) are suspended in Dulbecco's modified Eagle's medium (manufactured by Sigma) containing 10% FBS so that the concentration becomes ⁵ × 10 ⁵ cells / mL. 100 μL (5 × 10 ⁴ pieces / well) was added to each well of the holed micro mouth plate (manufactured by Sumitomo Bakelite). After 24 hours of culture (37 ° C, 5% CO), HWE, 70% Et Ext, 50% EP,

2

The medium was replaced with serum-free medium containing 1000% / ig / mL of 80% EP. Cultured for 48 hours Then, 5 μL of Cell counting kit_8 (Wako Pure Chemical Industries) was added to each well, and the absorbance (wavelength 450 nm) after 30 minutes was measured. The survival rate was calculated by comparison with a system that did not contain the test substance.

[0035] The test results are shown in Table 2 below. In the test results, HWE and 70% Et Ext showed some cytotoxicity, whereas the fractions of the present invention, 50% EP and 80% EP, had no effect on the viability of A431 cells. There was also a power not shown. This confirmed that the cell growth inhibitory effect exhibited by 50% EP and 80% EP in Example 4 was not due to cytotoxicity.

[0036] [Table 2] Table 2

Final enemy survival rate

HWE 1000 μ§ω 665%

70% EtExt 1000 μ§ω 74.4%

50% EP looo μ @ ω 115%

80% EP _ lOOOOugtnl 116% Brief description of drawings

[0037] [Fig. 1] Fig. 1 shows hot water extraction fraction (HWE), 50% ethanol precipitation fraction (50% EP), 80% ethanol precipitation fraction (80% EP), ethanol extract (Et Ext ) And 70% ethanol extract (70Et Ext) show the results of measuring the IFN-γ production enhancing activity by in vitro tests using human peripheral blood.

[Figure 2] Figure 2 shows the hot water extraction fraction (HWE), 50% ethanol precipitation fraction (50% EP), 80% ethanol precipitation fraction (80% EP), ethanol extract (Et Ext), 70 This shows the results of measuring in vitro test using human peripheral blood the TNF production enhancement activity of% ethanol extract (70Et Ext).

[Figure 3] Figure 3 shows the fractions obtained by dialysis molecular weight fractionation (Ml) with a molecular weight of 100,000 or more, fractions with a molecular weight of 50,000 or more but less than 100,000 (M2), and fractions with a molecular weight of less than 50,000. (M3), hot water extraction fraction (HWE) and 50% ethanol precipitation fraction (50% EP) IFN- y production It shows the results of measuring the enhancement activity by in vitro test using human peripheral blood

[Figure 4] Figure 4 shows the fraction obtained by dialysis molecular weight fractionation (Ml) with a molecular weight of 100,000 or more, the fraction with a molecular weight of 50,000 to less than 100,000 (M2), and the fraction with a molecular weight of less than 50,000. (M3), hot water extract fraction (HWE) and 50% ethanol precipitation fraction (50% EP) show the results of measuring the TNF-a production enhancing activity by in vitro tests using human peripheral blood Is a thing

[FIG. 5] FIG. 5 shows tumors produced by administering a hot water extraction fraction (HWE), a 50% ethanol precipitation fraction (50% EP), and an 80% ethanol precipitation fraction (80% EP) to mice. It shows the change of volume.

Claims

The scope of the claims

[I] By adding 0.5 to 2.5 times the volume of ethanol to the extract obtained by hot water extraction of the mycelium of Cordyceps sinensis (Berkely) Saccardo The resulting precipitate fraction.

[2] The precipitate fraction according to claim 1, wherein the resulting supernatant is removed by adding 0.5 to 2.0 times the amount of ethanol to the extract in a volume ratio to the extract. The precipitate fraction obtained by further adding an amount of ethanol in which the extract contains 70% -90% of ethanol in a volume ratio to the obtained extract.

[3] The precipitating fraction according to any one of claims 1 and 2, wherein the extract is obtained by concentrating before the ethanol is obtained. Precipitation fraction as described.

[4] The precipitate fraction according to any one of claims 1 to 3, wherein the mycelium is a residue after heat extraction with ethanol as a pretreatment.

[5] The precipitate fraction according to any one of claims 1 to 4, which comprises a polysaccharide having a molecular weight of 100,000 or more.

[6] The precipitated fraction according to any one of [4] to [5], which has an activity of enhancing production of interferon γ or tumor necrosis factor α in a mammal.

[7] A fraction having a molecular weight of 100,000 or more obtained by further subjecting the precipitate fraction according to claim 1 to molecular weight fractionation by dialysis.

8. The fraction according to claim 7, wherein the sugar content is 60 80% by weight in terms of D-glucose.

[9] A composition for oral consumption comprising the fraction according to any one of claims 1 to 8.

[10] A food or drink comprising the composition for oral consumption according to claim 9.

[I I] The food or drink according to claim 10, which has an anticancer effect, a cancer preventive effect, or an immunostimulatory effect.

[12] An immunostimulant comprising the composition for oral consumption according to claim 11.

[13] An anticancer agent comprising the composition for oral consumption according to claim 11.

[14] The method for producing a fraction according to any one of claims 1 to 8,

a) Process for obtaining extract by heating extraction of Cordyceps mycelium at 85-100 ° C with water; And

b) The above-mentioned production method comprising the steps of: adding 0.5 to 2.0-fold amount of ethanol to the extract by volume ratio with respect to the extract and recovering the generated precipitate.

[15] The method for producing a fraction according to any one of claims 2 to 8,

a) A step of heating extract of Cordyceps mycelium with water at 100 ° C to obtain an extract; b) 0.5-5. 0 times the amount of ethanol in the extract Removing the generated precipitate and recovering the supernatant; and

c) recovering the precipitate obtained by further adding ethanol in an amount such that the supernatant contains 70% -90% ethanol in a volume ratio to the supernatant. .