CN1822847A - Plant worms mycelium extracat fraction and composition for oral intake - Google Patents

Abstract

The invention provides a fraction containing the active ingredient contained in plant worms. The invention also provides a means efficacious in immunopotentiation, an effective means of preventing or treating cancer and a composition useful in producing health foods. 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

The separator of Cordyceps mycelium extract and oral uptake compositions

Technical field

The present invention relates to have the separator that is derived from Cordyceps of pharmacological effect and the manufacture method of this separator.The invention further relates to the oral uptake compositions, medical composition, the diet product compositions that contain this separator, or have the pharmaceuticals immune activation effect and/or that be used to prevent or treat cancer, the outside product of medicine, diet product etc.

Background technology

Cordyceps is the general name by the bacterium of generations such as insecticide, is in the genus of ascomycetes Ergota Zoopagales Clavicipitaceae.Wherein Chinese Cordyceps (Cordyceps sinensis (Berkely) Saccardo) parasitizes on the larva of lepidoptera Hepialus armorieanus Oberthur, is the bacterium on the waste grassland of 3000～4000 meters of alpine belt, height above sea level being distributed in China, Tibet, Nepal, Himalaya.In China since ancient times just and under extremely valuing, passing on as the Chinese medicine of alive for evermore, strong able-bodied bone.In recent years, known that by research contained composition has various physiologically actives in the Cordyceps, carried out reporting (with reference to non-patent literature 1～6) for immune activation effect, anti-tumor activity, the effect of blood sugar lowering value, hypotensive activity and vasorelaxation action.Pay close attention to such physiologically active, also made trial (with reference to patent documentation 1) in order to absorb effective ingredient effectively, but also very abundant hardly for content of effective in the extract and active aspect.

In addition, along with the progress of culture technique in recent years, can produce mycelium culture with natural goods Cordyceps much at one.Thereby the part of the sporophore that utilized with the past compares, and the mycelium of Cordyceps is more suitable for as the raw material on commercial production.Therefore, utilize this mycelium culture, be used for the method for part that efficient extracting contains the effective ingredient of Cordyceps and just sought.

Patent documentation 1: the spy opens flat 11-228440 communique

Non-patent literature 1:Biol.Pharm.Bull., the 22nd volume (No. 9), 966-970 page or leaf, nineteen ninety

Non-patent literature 2:Jpn.J.Pharmacol., the 79th volume, 505-508 page or leaf, non-patent literature 3:J.Kanazawa Med.Univ. in 1999, the 16th volume, 46-54 page or leaf, 1991 years

Non-patent literature 4:Biol.Pharm.Bull., the 16th volume (No. 12), 1291-1293 page or leaf, 1993 years

Non-patent literature 5:Phytochemistry, the 51st volume, 891-898 page or leaf, 1999 years

Non-patent literature 6:Life Science, the 66th volume (No. 14), 1369-1376 page or leaf, 2000 years

Summary of the invention

The inventor has carried out meticulous research for solving described problem, found that the modulator approach of the separator of the Cordyceps mycelium that has physiologically active specifically.

The separator that is derived from Cordyceps mycelium that the purpose of this invention is to provide effect with immune activation effect and/or prevention or treatment cancer, and contain the oral uptake compositions of this separator and the diet product, immunoactivator, anticarcinogen and the cancer prevention agent that contain this oral uptake compositions.

Promptly by a side of the present invention, can provide the precipitation part, this precipitation part is to obtain by adding ethanol in the extract that gets at the hot-water extraction Cordyceps mycelium.In the alcoholic acid amount of this adding, can be for example to be 0.5～2.0 times amount with respect to the volumetric ratio of above-mentioned extract.

By other sides of the present invention, the volumetric ratio that adds with respect to described extract in above-mentioned extract is the ethanol of 0.5～2.0 times of amount, remove the precipitation part that generates, with supernatant as the extract that obtains, count the ethanol of 70%～90% amount by volumetric ratio to wherein further adding the amount of alcohol that makes this extract contain, thereby resulting precipitation part is provided.

Provide the precipitation part by other sides of the present invention, described precipitation part is to add a certain amount of ethanol in the hot-water extraction liquid of Cordyceps mycelium and/or this extract, obtained removing the supernatant of the precipitation part that generates, before adding ethanol, it is concentrated, and then to wherein adding ethanol, thereby the precipitation that obtains.Provide by other sides of the present invention, as pretreatment will with ethanol heat extracting and residue as above-mentioned Cordyceps mycelium, by the precipitation part that obtains of adding ethanol in the extract that get through hot-water extraction.And then, by other sides of the present invention provide with contain have 100,000 or the polysaccharide of higher molecular weight be the above-mentioned precipitation part of feature.And then by other sides of the present invention, it is the above-mentioned precipitation part of feature that the activity to have the generation of promotion interferon-(IFN-γ) or tumor necrosis factor-alpha (TNF-α) for mammals is provided.

By other sides of the present invention, providing by above-mentioned precipitation part is further carried out molecular weight that the molecular weight separation obtains with dialysis is 100,000 or higher part.

By other sides of the present invention, provide that to contain above-mentioned precipitation part or molecular weight be 100,000 or the oral uptake compositions of higher part.By other sides of the present invention, also provide and contain the diet product of above-mentioned oral uptake with compositions.And then, by other sides of the present invention, provide and contain the immunoactivator of above-mentioned oral uptake with compositions.Further, by other sides of the present invention, provide and contain of anticarcinogen or the cancer prevention agent of above-mentioned oral uptake with compositions.

By further other side of the present invention, the manufacture method of above-mentioned part also is provided, this method contains following operation,

A) Cordyceps mycelium is heated extracting at 85～100 ℃ of following waters, obtain the operation of extract;

B) volumetric ratio that adds with respect to described extract in extract is the ethanol of 0.5～2.0 times of amount, reclaims the sedimentary operation that produces.

Further by other side of the present invention, also provide the manufacture method of above-mentioned part, this method contains following operation,

A) Cordyceps mycelium is heated extracting at 85～100 ℃ of following waters, obtain the operation of extract;

B) volumetric ratio that adds with respect to described extract in extract is the ethanol of 0.5～2.0 times of amount, the precipitation that produces is removed, reclaimed the operation of supernatant;

C) count the ethanol of 70%～90% amount by volumetric ratio by further in described supernatant, adding the amount of alcohol make supernatant contain, thereby reclaim the sedimentary operation that obtains.

Below further specify the present invention

Cordyceps among the present invention is meant the genus of ascomycetes Ergota Zoopagales Clavicipitaceae, for example Chinese Cordyceps (Cordyceps sinensis (Berkely) Saccardo).

Cordyceps mycelium among the present invention can be the material of producing by cultivating, and also can be by the material that obtains naturally, for example can use the dried powder of culture etc.

The hot-water extraction of carrying out among the present invention is for example carried out under 70～120 ℃ of temperature of liquid.Preferred temperature is 85～100 ℃, more preferably 90～95 ℃.The extracting time for example is 1 hour～24 hours, preferred 2～12 hours, and more preferably 3～7 hours.Hot-water extraction can repeatedly be carried out, and for example 1～4 time, preferably carries out 2～3 times.There is no particular limitation to be used for the pH value of extractive water, for example can use distilled water to carry out.

By above-mentioned hot-water extraction extract, can be used as it is, but for the alcoholic acid amount that reduces use and obtain separator efficiently, use after can concentrating this extract.There is no particular limitation for enrichment factor, for example can be that volumetric ratio is 80～20%, and preferred 60%～25%, further preferred 50%～30%.In addition, this concentrates and can carry out under normal pressure or under the arbitrary condition under the decompression, but preferably under reduced pressure carries out.

There is no particular limitation to add alcoholic acid amount in extract, for example can be that volumetric ratio with respect to the extract after concentrating is 0.5～2.0 times amount, preferred 0.8～1.2 times, as the same when not concentrating.The precipitation that obtains part can be made with extra care with the known method of this technical field, for example can add operation that ethanol obtains precipitating part by back that it is dissolved in the water and carry out one or many again and make with extra care.

In addition, in above-mentioned extract, add a certain amount of ethanol, with the precipitation that generates partially recycled after, in the extract that obtains as supernatant, by further adding ethanol and then can obtain precipitating part.As this supernatant and extract, can be used as it is, also can concentrate the back and use.There is no particular limitation for enrichment factor, for example can be that volumetric ratio is 80～20%, and preferred 60%～25%, further preferred 50%～30%.In addition, this concentrates and can carry out under normal pressure or under the arbitrary condition under the decompression, but preferably under reduced pressure carries out.In addition, there is no particular limitation for the initial alcoholic acid amount that adds, and for example can be that volumetric ratio with respect to the extract after concentrating is 0.5～2.0 times a amount, preferred 0.8～1.2 times.There is no particular limitation to add for the second time behind the ethanol in the extract contained alcoholic acid amount, for example can be that volumetric ratio is 70%～90%, and preferred 75%～85%.For example add volumetric ratio with respect to extract and be 0.5～2.0 times ethanol, in the supernatant after reclaiming the precipitation part that generates, for example carry out after concentrating under reduced pressure removes ethanol, add volumetric ratio and be 3.0～5.0 times ethanol, also can obtain precipitating part.

Oral uptake compositions among the present invention for example comprises diet product compositions and medical composition etc.

For obtaining the high separator of active constituent content, for the Cordyceps mycelium that carries out hot-water extraction, can carry out pretreatment in the present invention.There is no particular limitation in this pretreatment, can exemplify out the use carbon number and be 1～4 the alcohol or the heating extracting of this alcohol-water solution etc., preferably uses ethanol.Temperature during heating is 70～120 ℃ of temperature of liquid for example, preferred 80～100 ℃, more preferably 90～95 ℃.The extracting time is for example 1 hour～24 hours, preferred 2～12 hours, and more preferably 3～7 hours.The ethanol extracting can be carried out repeatedly, and for example 1～4 time, preferred 2～3 times.Also can use different solvents repeatedly to carry out, for example can use 95～99.5% ethanol to carry out extracting after, be that 60～80% ethanol water carries out extracting with its residue volumetric ratio, its residue is used as the raw material that carries out hot-water extraction.

The precipitation part that obtains among the present invention can contain for example polysaccharide etc. as its composition.There is no particular limitation for this polysaccharide, is the material that is made of naturally occurring monosaccharide, and the modification of acidylate, phosphorylation etc. can be accepted in its molecule 1 or 2 or more a plurality of position, or by protein, the bonded modification of nucleic acid.There is no particular limitation for the molecular weight of this polysaccharide, for example can be 100,000～2,000,000.The sugared content of precipitation part of the present invention can be measured with the known method of those skilled in the art, for example can use phenol sulfuric acid process etc.There is no particular limitation for sugar content of the present invention, for example is scaled 40～90 weight % with the D-glucose, preferred 50～80 weight %, and then preferred 60～80 weight %.

Above-mentioned precipitation part, for example can be separated into molecular weight for example by methods such as dialysis is 50,000 or higher and be 100,000 or higher part less than 100,000 part, molecular weight less than 50,000 part, molecular weight.

Immunoactivator of the present invention owing to have the effect that promotes that interferon-in the mammals and/or tumor necrosis factor-alpha produce, thereby for the testee who needs immune activation, can be used as preventive drug and/or curative and uses.In addition, anticarcinogen of the present invention is because demonstrate significant antitumous effect, so can become effective preventive drug of cancer and curative.Therefore, immunoactivator of the present invention and anticarcinogen can be used as the effective ingredient of medical composition and use.This medical composition can contain general employed various compositions, for example can contain a kind or more kinds of at pharmaceutically permissible excipient, diluent, wetting agent, emulsifying agent, dispersant, adjuvant, antiseptic, buffer agent, binding agent, stabilizing agent etc.

The dosage of immunoactivator of the present invention and anticarcinogen can suitably be selected according to elapsed time after patient's the bodily form, age, health, severity extent, the morbidity etc., and for example, general use amount is 1～5000mg/ day/adult.

In addition, oral uptake compositions of the present invention can former state be used as functional food, can also use as the composition of the outside product of medicine, diet product etc., food additives etc.By this use, make the oral uptake compositions with immune activation effect and antitumous effect of the present invention reach routinely picked-up in routine ground, can improve the generation of body constitution, treatment cancer and prophylaxis of cancer by effective immune activation.The example that oral uptake compositions of the present invention is used as food material can be enumerated functional food, health food, normal food (fruit juice, dessert, processed food etc.), dietary supplement (nutritious drink etc.) with immune activation effect or treatment cancer effect or prophylaxis of cancer effect.

Oral uptake compositions of the present invention is owing to have immune activation effect and/or antitumor action, thereby the invention provides the effective means that is used for immune activation and, also be provided at compositions useful in the manufacturing of health food for the effective preventive means or the treatment means of cancer.

Description of drawings

Fig. 1 promotes active result's figure for showing generation that in vitro by personnel selection tip blood tests the IFN-γ that measures hot-water extraction part (HWE), 50% ethanol precipitation part (50%EP), 80% ethanol precipitation part (80%EP), ethanolic extract (Et Ext), 70% ethanolic extract (70Et Ext).

Fig. 2 promotes active result's figure for showing generation that in vitro by personnel selection tip blood tests the TNF-α that measures hot-water extraction part (HWE), 50% ethanol precipitation part (50%EP), 80% ethanol precipitation part (80%EP), ethanolic extract (Et Ext), 70% ethanolic extract (70Et Ext).

Fig. 3 for show in vitro by personnel selection tip blood test the molecular weight of measuring with dialysis separate the molecular weight that obtains be 100,000 or higher part (M1), molecular weight be 50,000 or higher less than 100,000 part (M2) and molecular weight less than 50,000 part (M3), hot-water extraction part (HWE) and 50% ethanol precipitation partly the generation of the IFN-γ of (50%EP) promote active result's figure.

Fig. 4 for show in vitro by personnel selection tip blood test the molecular weight of measuring with dialysis separate the molecular weight that obtains be 100,000 or higher part (M1), molecular weight be 50,000 or higher less than 100,000 part (M2) and molecular weight less than 50,000 part (M3), hot-water extraction part (HWE) and 50% ethanol precipitation partly the generation of the TNF-α of (50%EP) promote active result's figure.

Fig. 5 is for by giving the figure that gross tumor volume that Mus hot-water extraction part (HWE), 50% ethanol precipitation part (50%EP), 80% ethanol precipitation part (80%EP) demonstrate changes.

The specific embodiment

Below, be described in further detail with regard to preferred embodiment of the present invention, but the present invention is not limited to these embodiment.

The modulation of the separator of [embodiment 1] Chinese Cordyceps mycelium

In enforcement of the present invention, the powder that becomes the Chinese Cordyceps mycelium culture of raw material can use for example Chinese commercially available " fermented Cordyceps powder " (trade name: CORBRIN CAPSULE, manufacturing company: Zhongmei Huadong Pharmaceutical Co., Ltd. Hangzhou).

With raw material Cordyceps mycelium powder end with 99.5% ethanol 90～95 ℃ of following extractings 6 hours.Amount of alcohol is every 1g mycelium 2mL.Filter residue, once more with residue with this method extracting, separate extract and residue with filtering.2 resulting extracts of extracting are carried out concentrating under reduced pressure altogether, obtain being scaled the heavy-gravity persimmon liquid ethanol extract (Et Ext) of 15.0% (weight ratio) with raw material.Then with residue with 70% ethanol water (volume ratio) 90～95 ℃ of following extractings 6 hours.70% alcoholic acid amount is every 1g residue 2mL.Filter residue, use the method extracting residue once more, with filtering separately extract and residue.The extract of 2 extracting gained is carried out concentrating under reduced pressure altogether, become approximately 40% until volume, obtain being scaled persimmon powder 70% ethanolic extract (70%Et Ext) of 45.8% (weight ratio) by lyophilization with raw material.Then use distilled water 90～95 ℃ of following extractings 6 hours residue.The distillation water yield is every 1g residue 4mL.Filter residue, use the method extracting residue once more, with filtering separately extract and residue.With 2 extractings and extract carry out concentrating under reduced pressure altogether, be about 40% until volume, add volume ratio and be 99.5% ethanol of equivalent, be statically placed in one evening of cold and dark place.Reclaim precipitate by centrifugalize, it is also freezing that it is dissolved in the distilled water, obtains being scaled with raw material the bibulous Umber powder 50% ethanol precipitation part (50%EP) of 17.9% (weight ratio) through lyophilization.Supernatant is carried out concentrating under reduced pressure after volume is about 40%, and adding volume ratio is 99.5% ethanol of 4 times of amounts, is statically placed in one evening of cold and dark place.Reclaim precipitate by centrifugalize, it is also freezing that it is dissolved in the distilled water, obtains being scaled with raw material the dark brown powder 80% ethanol precipitation part (80%EP) of 6.0% (weight ratio) through lyophilization.

50% ethanol precipitation is partially dissolved in the distilled water, distilled water is dialysed with Spectrum corporate system Spectra/Por (registered trade mark) CE Membrane MWCO:100000.The amount of dissolving the distilled water of 50% ethanol precipitation part is every 1g precipitation part 50mL.Obtaining with the molecular weight that raw material is scaled 10.0% (weight ratio) by the lyophilization dialyzed solution is 100,000 or higher part (M1).With the extracellular fluid dialysis concentrating under reduced pressure, distilled water is dialysed with Spectrum corporate system Spectra/Por (registered trade mark) 6Membrane MWCO:50000.By the lyophilization dialyzed solution obtain with raw material be scaled 3.94% (weight ratio), molecular weight is 50,000 or higher and less than 100,000 part (M2).With the extracellular fluid dialysis concentrating under reduced pressure, by lyophilization obtain with raw material be scaled 3.45% (weight ratio), molecular weight is less than 50,000 part (M3).

For the M1 part, with the phenol sulfuric acid process sugar content is done quantitatively, the sugar content of M1 part is scaled 69.9% with D (+)-glucose as a result.At this, the phenol sulfuric acid process is carried out (for example with reference to " new experimental chemistry lecture 20 biochemistrys [II] ", 1085 pages, Japanization association compiles, and on October 20th, 1978 issued, and ball is apt to Co., Ltd.) with general method

The modulation example of [Production Example 1] hot-water extraction thing

The hot-water extraction thing that contrasts usefulness as a comparison among the embodiment is modulated with following method.With Cordyceps mycelium powder end with distilled water extracting while stirring 4 hours under 90～95 ℃.The distillation water yield is every 1g mycelium 20mL.Filter residue, obtained hygroscopic dark brown powder hot-water extraction thing (HWE) by concentrating under reduced pressure filtrate postlyophilization.

[embodiment 2] end user's tip hematometry immune activation effect

On 48 porocytes are cultivated with plate, add the tip blood 500 μ L that gather from the people and with the RPMI1640 culture medium of tip blood equivalent.Adding detected material, to make its ultimate density be 200 μ g/mL, at 37 ℃, 5%CO ₂Condition under cultivated 24 hours.Reclaim culture supernatant, use and utilize the mensuration test kit (BIOSOURCE corporate system) of enzymoimmunoassay (ELISA) respectively interferon-(IFN-γ) and tumor necrosis factor-alpha (TNF-α) to be done quantitative assay.The using method of test kit and the analytic method of quantitative result are gone according to the appended operating instruction of test kit.

For synthetic hot-water extraction thing (HWE) in synthetic 50% ethanol precipitation part (50%EP) in embodiment 1,80% ethanol precipitation part (80%EP), ethanolic extract (Et Ext), 70% ethanolic extract (70%Et Ext) and the Production Example 1, measure IFN-γ generation and TNF-α generation, the result is shown in respectively among Fig. 1 and Fig. 2.By this measurement result as can be known, compare with HWE, Et Ext and 70%Et Ext, 50%EP and 80%EP have the effect of significant promotion generation for IFN-γ or for TNF-α, have confirmed to be derived from use the effect of separator of the present invention in people's the test system of tip blood.In addition, also clear and definite 50%EP compares with 80%EP has higher facilitation effect.

For the molecular weight separator M1 of the part of synthetic 50% ethanol precipitation in embodiment 1 (50%EP) (molecular weight be 100,000 or higher part), M2 (molecular weight be 50,000 or higher and less than 100,000 part) and M3 (molecular weight less than 50,000 part), measure IFN-γ and TNF-α generation, the result is shown in respectively among Fig. 3 and Fig. 4.By this measurement result as can be known, no matter be that M1 has the effect above the promotion generation of 50%EP for IFN-γ generation or for TNF-α generation.In addition, the activity of having confirmed this promotion generation only concentrates on M1 (molecular weight be 100,000 or higher part), the promotion IFN-γ of 50%EP and the generation activity of TNF-α, with molecular weight be 100,000 or higher chemical compound very dark relation is arranged.

[embodiment 3] are used and are suffered from cancer Mus mensuration antitumous effect

Buying 32 of the male Mus of hybridization group Slc:BDF1 in 6 ages in week from Japanese SLC Co., Ltd., is that a component becomes 4 groups with 8, puts into a cage with 4 and raises and train for 1 week.To 7 the week ages Mus in the subcutaneous vaccination 1 * 10 of right oxter ⁶The tumor Sarcoma180 of the Mus of cell/mouse.In addition, before inoculation, shave off the chaeta of inoculation position earlier, make and more correctly carry out for the observation of adhering to and growing up of tumor.The growth of tumor is with the minor axis of digital vernier caliper measurement tumor and major diameter, calculates the size (major diameter * minor axis of tumor ²/ 2mm ³).4 groups be respectively matched group (control group), hot-water extraction thing group (HWE group), 50% ethanol precipitation partly organizes (50%EP group) and 80% ethanol precipitation is partly organized (80%EP group), each group gives detected material from beginning 10 of every mornings inoculated tumour day with the probe per os.Administered dose is, control organizes every 10g body weight and gives normal saline 0.1mL, HWE group is dissolved in HWE to become 250mg/kg in the normal saline and gives, 50%EP group is dissolved in 50%EP to become 44.8mg/kg in the normal saline and gives, and the 80%EP group is dissolved in 80%EP to become 15.0mg/kg in the normal saline and gives.

The measurement result of the gross tumor volume of the 7th day and the 10th day is shown among Fig. 5.Can confirm that by this measurement result separator of the present invention has anti-tumor activity in the test system of in vivo.And then administered dose is that 1/5 or the lower 50%EP of HWE expresses the antitumous effect above HWE, and this result has also supported the result of embodiment 2 simultaneously.

The test of [embodiment 4] A431 cell inhibitory effect

With A431 cell (people's squamous epithelium JEG-3; Get from big Japanese pharmacy) outstanding turbid in the Eagle culture medium (SIGMA corporate system) that the Dulbecco ' s that contains 10%FBS changes, make it become 4 * 10 ⁴Individual/mL, respectively with 50 μ L (2 * 10 ³Individual/hole) add in each hole of the little culture plate in 96 holes (Sumitomo BAKELITE system).Cultivate (37 ℃, 5%CO after 6 hours ₂), with contain as detected material, respectively the do for oneself culture medium of HWE, 70%Et Ext, 50%EP, 80%EP and 10%FBS of 2000 μ g/ml is added (final concentration is 1000 μ g/ml) in each hole to 50 μ L respectively.To be added with the material of culture medium of the Genistein that contains 200 μ M of 50 μ L as positive control.Cultivate after 4 days, in each hole, add Cell counting kit-8 (with the pure medicine of light) 5 μ L respectively, measure absorbance (wavelength 450nm) after 60 minutes.According to the system that does not contain detected material relatively calculate suppression ratio.

Result of the test is shown in the following table 1.In this result of the test, all detected materials all demonstrate the effect that suppresses cell proliferation, and wherein particularly the suppression ratio of 80%EP is high significantly.

Table 1

	Final concentration	Suppression ratio
			Genistein HWE
70％Et Ext 50％EP 80％EP	100μM 1000μg/ml 1000μg/ml 1000μg/ml 1000μg/ml	67.9％ 45.8％ 36.1％ 49.6％ 83.5％

[embodiment 5] A431 primary cellular defect test

With A431 cell (people's squamous epithelium JEG-3; Get from big Japanese pharmacy) outstanding turbid in the Eagle culture medium (SIGMA corporate system) that the Dulbecco ' s that contains 10%FBS changes, make it become 5 * 10 ⁵Individual/mL, respectively with 100 μ L (5 * 10 ⁴Individual/hole) add in each hole of the little culture plate in 96 holes (Sumitomo BAKELITE system).Cultivate (37 ℃, 5%CO after 24 hours ₂), exchange with the serum-free medium that contains 1000 μ g/mL HWE, 70%Et Ext, 50%EP, 80%EP respectively.Cultivate after 48 hours, in each hole, add the Cell counting kit-8 (with the pure medicine of light) of 5 μ L respectively, measure the absorbance (wavelength 450nm) after 30 minutes.According to the system that does not contain detected material relatively calculate survival rate.

Result of the test is shown in the following table 2.In this result of the test, HWE and 70%Et Ext demonstrate some cytotoxicities, and be relative therewith, and separator 50%EP of the present invention and 80%EP do not demonstrate any influence to the survival rate of A431 cell.Thus, confirmed that the shown cell inhibitory effect effect of in embodiment 4 50%EP and 80%EP is not because of due to the primary cellular defect.

Table 2

	Final concentration	Survival rate
			HWE
70％Et Ext 50％EP 80％EP	1000μg/ml 1000μg/ml 1000μg/ml 1000μg/ml	66.8％ 74.4％ 11.5％ 11.6％

Claims (15)

1. the precipitation part is characterized by, with the mycelium of Chinese Cordyceps through hot-water extraction and extract in, be that the ethanol of 0.5～2.0 times of amount obtains by adding volumetric ratio with respect to extract.

2. precipitation part, it is characterized by, to have removed by add volumetric ratio with respect to extract in above-mentioned extract is that supernatant after that the ethanol of 0.5～2.0 times of amount generates, the described precipitation part of claim 1 is as resulting extract, by counting by volumetric ratio to wherein further adding the amount of alcohol that makes this extract contain that the ethanol of 70%～90% amount obtains.

3. claim 1 or 2 described precipitation parts is characterized by, and described extract is by concentrating the material that obtains before adding ethanol.

4. each described precipitation part in the claim 1～3, wherein, described mycelium is as pre-treatment and heat residue after the extracting with ethanol.

5. each described precipitation part in the claim 1～4 is characterized by, and contains to have 100,000 or the polysaccharide of higher molecular weight.

6. each described precipitation part in the claim 1～5 is characterized by, and for mammal, has the activity that promotes that interferon-or tumor necrosis factor-alpha produce.

7. to separate the molecular weight that obtains be 100,000 or higher part by the described precipitation part of claim 1 is further carried out molecular weight by dialysis.

8. the described part of claim 7 is characterized by, and sugar content is scaled 60～80 weight % with the D-glucose.

9. the oral uptake compositions is characterized by, and contains each described part in the claim 1～8.

10. the diet product is characterized by, and contain the described oral uptake compositions of claim 9.

11. the described diet product of claim 10 is characterized by, and have antitumaous effect, prophylaxis of cancer effect or immune activation effect.

12. immunoactivator is characterized by, and contains the described oral uptake compositions of claim 11.

13. anticarcinogen is characterized by, and contains the described oral uptake compositions of claim 11.

14. the manufacture method of each described part is characterized by in the claim 1～8, contains following operation,

A) Cordyceps mycelium is heated extracting at 85～100 ℃ of following waters, obtain the operation of extract;

B) volumetric ratio that adds with respect to described extract in extract is the ethanol of 0.5～2.0 times of amount, reclaims the sedimentary operation that produces.

15. the manufacture method of each described part is characterized by in the claim 2～8, contains following operation,

A) Cordyceps mycelium is heated extracting at 85～100 ℃ of following waters, obtain the operation of extract;

B) volumetric ratio that adds with respect to described extract in extract is the ethanol of 0.5～2.0 times of amount, the precipitation that produces is removed, reclaimed the operation of supernatant;

C) in described supernatant, further add the amount of alcohol that makes described supernatant contain is counted 70%～90% amount by volumetric ratio ethanol, reclaim the sedimentary operation that obtains thus.

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