JPS635077B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a method for producing a substance with antitumor activity. Specifically, the present invention relates to a method for producing a substance having an antitumor effect by extraction from Basidiomycetes. In recent years, it has come to be understood that the causes of many diseases are due to immune function regulation in vivo. These diseases include not only the so-called autoimmune diseases, but also gastric ulcers, cancer, diabetes, and even the aging phenomenon, where abnormalities in biological regulatory functions are thought to be caused by immunity. .
Recently, not only basic research has been conducted on how such immune function is regulated and what causes abnormalities, but also drugs and drugs that can be involved in immune phenomena to maintain normal homeostasis in the body. Research on drugs for the treatment of diseases caused by abnormal immune function has become widespread. For example, lentinan and PS-K, which are basidiomycete polysaccharides extracted from shiitake mushrooms and versicolor versicolor, are known to have antitumor effects through immune function regulation. From this point of view, the present inventor conducted intensive research to develop a new drug with fewer side effects and toxicity and a high immunomodulatory effect, and discovered that an extract of a certain basidiomycete can meet the purpose. Reached. That is, the gist of the present invention resides in a method for producing a substance having an antitumor effect, which is characterized by extracting the substance having an antitumor effect from a fungus belonging to the genus Morinokarebataceae of the family Shimejifamily, using a solvent. The present invention will be explained in detail below. The basidiomycetes used as raw materials in the method of the present invention are
It is a fungus that belongs to the family Shimejitake and the genus Morinokarebatake. Examples of such fungi include Tricholomataceae Collybia acervata.
(Fr. Kummer), Tricholomataceae Collybia butyraceae and Tricholomataceae Collybia
maculata). As a raw material for the method of the present invention, whole fungi belonging to the genus Morinocera can be used, such as their fruiting bodies. The fungi used as raw materials in the method of the present invention are usually washed with water, physiological saline, various buffer solutions, organic solvents, etc., and then used for solvent extraction. Examples of such solvents include water, lower alkanols such as methanol and ethanol, lower ketones such as acetone, mixed solvents of water-miscible organic solvents and water such as ethers such as dioxane, and lower alkanols. A mixed solvent of lower alkanol and water is used. During extraction, it is preferable to pulverize the bacterial cells from the viewpoint of extraction efficiency. The amount of extraction solvent is usually 3 to 3
15 times the amount (weight), preferably 5 to 10 times the amount (weight)
That's about it. If the amount of the solvent is too small, the extraction will be incomplete, and if the amount of the solvent is too large, no particular improvement in the accompanying effect will be observed, so both are unfavorable. The temperature during extraction is usually room temperature (15 to 25°C), but heating may be used. Extraction time varies depending on various conditions, but usually
It lasts from several hours to several weeks. The extraction operation may be repeated. After extraction, the bacterial cells and the extract are separated by a well-known method such as filtration or centrifugation, and the extract is concentrated to obtain a substance having the desired immune function regulating effect. The substance having antitumor action obtained by the method of the present invention is useful as an antitumor agent when administered in a conventional manner. The present invention will be explained in more detail by giving examples below, but the present invention will not exceed the gist thereof.
The following examples are not intended to limit the invention. Example 1 The fruiting body of Eseolymiki (fresh weight 126 g) was pulverized in a mixer after removing mud and leaf debris. At this time, 1000 ml of 70% ethanol-water mixture is used as a solvent, and the pulverized liquid is placed in a glass container and left at room temperature for 2 hours. During this time, the pulverized solution is occasionally stirred with a glass rod to increase extraction efficiency. After 2 weeks, the extract is suction filtered using a Bützfner funnel and concentrated under reduced pressure at below 40°C. Repeat the same operation 2 to 3 times (in this case,
Leave the grinding solution for about one week. ) Each extract was combined to obtain about 3 g of a brown oily substance. The infrared spectrum of this product is shown in FIG. Example 2 Mud and leaf debris were removed from the fruiting bodies of Turnip chinensis (fresh weight 150 g), and they were ground in a mixer. At this time, 1000ml of 70% ethanol-water mixture was used as the solvent, and the crushed liquid was placed in a glass container and
Leave at room temperature for a week. During this time, occasionally stir the pulverized solution with a glass rod to increase extraction efficiency. After 2 weeks, the extraction was filtered with suction in a Bützfner funnel, and the
Concentrate under reduced pressure below ℃. Repeat the same operation 2 to 3 times (in this case,
(The pulverized solution may be left to stand for about one week.) Each extract was combined to obtain about 3.1 g of a brown oily substance. Test Example 1 The antitumor effect of the extract of the present invention was confirmed by the following screening test.
Sarcoma-180 was used as a typical tumor type, and a subcutaneous nodular tumor was used. The extract-administered group of the present invention used 4 mice per group, and the control group used 10 mice per group. Test method Experimental animals were 5-6 week old ddN male mice (body weight
22-25g) was used. The cancer type was implanted into the abdominal cavity of a mouse, and on the 7th day after sufficient proliferation, the cells were collected, and 4×10 ⁶ cells were implanted subcutaneously in the groin region of an experimental mouse to form a solid tumor. From 24 hours after transplantation, the extract of the present invention was dissolved in physiological saline and administered intraperitoneally. The administration volume was adjusted to 0.2 ml per animal, and administration was performed once a day for 10 consecutive days. The control group received only physiological saline. After transplant
Tumors were excised on the 30th day, and the average tumor weight (T) of the extract-administered group of the present invention and the average tumor weight (C) of the control group were
was measured and T/C (%) was calculated. The results are shown in Table 1.

[Table] T/C at Sarcoma-180
(%) 25 or less is considered to be a significant response.

[Brief explanation of the drawing]

FIG. 1 is a graph showing the infrared spectrum of the product of Example 1.

Claims (1)

[Scope of Claims] 1. A method for producing a substance having antitumor activity, which comprises extracting a substance having the following physicochemical properties and antitumor activity from a fungus belonging to the genus Morinokarebataceae of the family Shimejifamily, using a solvent. Physical and chemical properties: Brown oily substance. Infrared absorption is 1550-1680cm, 1400cm, 1150cm,
Located around 1080cm, 1050cm, and 990cm. 2. In the method described in claim 1,
A method characterized in that the solvent is water. 3. In the method described in claim 1,
A method characterized in that the solvent is a mixed solvent of water and an organic solvent miscible with water. 4. In the method described in claim 1,
A method characterized in that the solvent is a lower alkanol. 5. The method according to any one of claims 1 to 4, characterized in that the fungus is Versaceae. 6. The method according to any one of claims 1 to 4, wherein the fungus is Aeseolymiki.