JP3152402B2 - Antitumor agent and method for producing the same - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an antitumor agent containing, as an active ingredient, a fruiting body of a soft basidiomycete Hydanashitake Coralum genus Yamabushitake or a hot water extract of a liquid culture mycelium, and a method for producing the same.

[0002]

2. Description of the Related Art Yamabushitake itself is an edible mushroom, which is also present in Japan, but is called monkey head hair (yuyukinhe) in China and is used as a health food. It is said that its effects include physical strength enhancement, blood pressure lowering, and immune recovery.

[0003] There have been a great many studies on the antitumor activity of polysaccharides and polysaccharide protein complexes isolated from fruiting bodies of basidiomycetes and cultured mycelium, and some of them have been practically used as antitumor agents. Some have been converted. But,
Many of them do not have very strong antitumor activity, and their effectiveness is questioned, and the drug efficacy has recently been limited by review. Also, the antitumor activity of soft basidiomycetes was not known at all.

[0004]

Means for Solving the Problems The present inventors have widely searched for more effective mushrooms, including those from overseas.

[0005] As a result, the present inventors have found that the fruit body of the soft basidiomycete, the Japanese name Yamabushitake, and a cultured mycelium of a soft mycelia fungus called Yuyukinhe, which is used as a health food in China, is contained in the hot water extract. The present inventors have found that the component has an extremely remarkable antitumor effect, and as a result of intensive studies, completed the present invention.

That is, the antitumor agent of the present invention is obtained by adding 0.5 to 5 parts by weight of alcohol to 1 part by weight of a hot water extract of a fruit body or a liquid culture hypha of Yamabushitake or a concentrated solution thereof.
The alcohol precipitate fraction produced is characterized as an active ingredient.

[0007]

The yamabushitake used for this purpose may be any of those belonging to Hericium erina- ceus . The antitumor effect was specifically shown in the following examples.

Although the antitumor effect of Yamabushitake has been traditionally described, it has not been confirmed at the research level, and the active substance is not clear. Further, since the fruiting body has a low growth rate, industrially obtaining these active substances has a problem in economical efficiency. Moreover, the antitumor effect of the cultured mycelium was not known at all.

The culture medium for liquid culture is not particularly limited, and any of the commonly used mushroom medium compositions may be used. Particularly, good results can be obtained by using yeast extract and glucose. On the other hand, the presence of air (oxygen) is essential to increase the culture speed. If the amount is too small, the growth of mycelium is slow, and if it is too large, autolysis occurs and the yield of mycelium decreases. Generally, about 0.1 vvm to 1 vvm is suitable (vvm represents air (oxygen) supply per minute per medium volume). An alcohol such as methanol, ethanol, or isopropyl alcohol is added to the hot water extract of the mycelium and the fruiting body thus obtained in an amount of 0.5 to 5 parts by weight per 1 part by weight of the hot water extract or a concentrate thereof to produce. Alcow
By drying the precipitate fraction, the antitumor component of the present invention can be obtained.

[0010] The antitumor effect of Yamabushitake was determined from the tumor cell growth inhibitory effect of macrophages or changes in the diameter of solid tumors implanted in mice as shown in the Examples.

[0011]

【Example】

(Example 1) Acquisition of antitumor component in fruiting body: Yamabushitake fruiting body was homogenized as wet cells, added with 10 times the amount of wet cell weight of water, extracted with hot water for 1 hour, and extracted. Concentrate. Methanol was added to the concentrate, and the resulting precipitate was centrifuged.
Next, the precipitate is dissolved in water, and the solution is freeze-dried.
A gray-brown powder was obtained. This is designated as Sample A.

(Example 2) Acquisition of antitumor component in liquid culture mycelium: Inoculation of Yamabushitake inoculum in a 2.6 liter jar containing 1.5 liter of 1% by weight of yeast extract, 3% by weight of glucose, and pH 5 medium 25
After culturing for 7 days at 0.3 ° C. in air at 0.3 ° C., mycelium was separated by centrifugation or filtration. The mycelium is homogenized as wet cells, water is added in an amount 10 times the weight of the wet cells, hot water is extracted for 1 hour, and the extract is concentrated. Methanol was added to the concentrate, and the resulting precipitate was centrifuged.
The precipitate is then dissolved in water, the lysate is lyophilized,
A brown powder was obtained. This is designated as Sample B.

Example 3 Macrophage Tumor Cell Growth Inhibition Test: To 96-well plate, mouse peritoneal exudate cells and sample induced with thioglycolate medium, and mouse mast cell tumor culture cell P815 as target cells were added. After culturing for 46 hours, tritium thymidine was added, and the cells were further cultured for 2 hours. After completion of the culture, the P815 cells were collected on a filter paper, and the tritium incorporated in the P815 cells was measured with a liquid scintillation counter to measure the P815 cell proliferation inhibitory effect of macrophages. The results are shown in Table 1.

[0014]

[Table 1]

As shown above, the hot-water-extraction-alcohol-precipitating components of the fruit body of Yamabushitake and the cultured mycelium activated the cell growth inhibitory action of macrophages more strongly than the commercially available mushroom carcinogens Lentinan and Krestin.

(Example 4) Effect on sarcoma 180 solid tumor: Ascites tumor cells of sarcoma 180 grown intraperitoneally in ICR mice were treated with IC
1 × 10 ⁶ cells / animal were implanted subcutaneously in the axilla of R mice, and the antitumor agent of the present invention was intraperitoneally administered 24 hours after the implantation. The administration frequency was once a day for 10 days. Four weeks after transplantation, the tumor diameter (short diameter and long diameter) was measured, the product was calculated, and the product was compared with the control group to calculate the tumor suppression rate. Six weeks later, the percentage of complete tumor regression was examined. Table 2 shows the results.

[0017]

[Table 2]

The administered antitumor agent was used in the form of a suspension in physiological saline, and the control group received only physiological saline.

[0019]

As shown in the above Examples, the present invention has been completed by recognizing that the hot water extraction-alcohol precipitate of the fruit body of Yamabushitake and cultured mycelium can be used as a powerful antitumor agent.

For reference, an infrared chart of hot water extraction of alcoholic fruit bodies of Yamabushitake and cultured mycelia (samples A and B) is attached.

[Brief description of the drawings]

FIG. 1 is an infrared chart of Sample A obtained in Example 1.

FIG. 2 is an infrared chart of Sample B obtained in Example 2.

──────────────────────────────────────────────────続 き Continued on the front page (58) Fields surveyed (Int. Cl. ⁷ , DB name) A61K 35/84 A61P 35/00 BIOSIS (DIALOG) CA (STN) MEDLINE (STN)

Claims (5)

(57) [Claims] 1. An antitumor comprising adding 0.5 to 5 parts by weight of alcohol to 1 part by weight of a hot water extract of a fruiting body of Yamabushitake or a concentrated liquid thereof, and containing an alcohol precipitated fraction as an active ingredient. Agent. 2. An antitumor comprising adding 0.5 to 5 parts by weight of alcohol to 1 part by weight of a hot water extract of a fruit body of Yamabushitake or a concentrated solution thereof, and containing an alcohol precipitated fraction as an active ingredient. Formulation of the agent. 3. An antibacterial method comprising adding 0.5 to 5 parts by weight of alcohol to 1 part by weight of a hot water extract of a liquid culture mycelium of Yamabushitake or a concentrated liquid thereof, and containing an alcohol precipitated fraction as an active ingredient. Tumor agents. 4. An antibacterial method comprising adding 0.5 to 5 parts by weight of alcohol to 1 part by weight of a hot water extract of a liquid culture mycelium of Yamabushitake or a concentrated liquid thereof and containing an alcohol precipitated fraction as an active ingredient. Formulation of tumor agents. 5. The method according to claim 1, wherein the alcohol is methanol, ethanol or isopropyl alcohol.
5. The method for producing an antitumor agent or an antitumor agent according to any one of claims 1 to 4.