JP2006028070A - Antineoplastic composition - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an antineoplastic composition having high safety and antineoplastic activity. <P>SOLUTION: The antineoplastic composition comprises a hot water extract of Agaricus blazei Murrill and a bovine globulin concentrate. Specifically, the hot water extract is obtained from either one of the following: the carpophore, its cell wall broken product, cultured product, its cell wall broken product of Agaricus blazei Murrill and dried products thereof. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention has an excellent antitumor effect comprising an extract of "A scientific name" Agaricus blazei Murrill "official Japanese name" Himematsutake (one name, Kawariharatake) and bovine globulin concentrate, which are mushrooms of the genus Agaricus (Agaricus). Relates to antitumor compositions.

  Antitumor drugs are drugs that inhibit the growth of cancer cells, and are indirectly inhibited through the immune mechanism and cytotoxic drugs that exert their effects by directly inhibiting the synthesis of nucleic acids and proteins required for growth. Biological response modifiers (BRMs).

As for the Japanese matsutake mushrooms classified as BRM, one of the inventors of the present invention, Hitoshi Ito, was reported at the 1980 general meeting of the Japanese Cancer Association, and more than 60 publications have already been published in public societies and academic journals. (For example, see Patent Documents 1 to 3).
However, there is no cytotoxic drug having no adverse effect, and a multi-drug combination therapy is carried out to increase the effectiveness while reducing the adverse effect by combining a plurality of antitumor drugs having different action mechanisms one by one.

Therefore, generally used are Chinese medicines such as mushrooms, seaweeds, bear moths, royal jelly, shark cartilage, carrots, Juzentaihoto, etc. belonging to the so-called BRM, and combined use of cytotoxic drugs and BRM. Done. However, the actual situation is that satisfactory safety and antitumor effects have not yet been obtained.
JP-A 64-67194 JP-A 64-67195 JP-A-2-78630

  This invention is made | formed in view of such the conventional actual condition, and makes it a subject to provide the composition which has the outstanding safety | security and antitumor action.

  This invention solves the said subject with the anti-tumor composition characterized by including the hot-water extract of Japanese matsutake and a cow globulin concentrate.

  Since the composition of the present invention has no toxicity, is excellent in safety, and has a high antitumor action, it can effectively suppress and treat tumors by oral administration.

  In the present invention, a hot water extract of Himematsutake is obtained from any one or more of Himematsutake fruiting bodies, Himematsutake cultures, cell wall destruction products of Himematsutake fruiting bodies, cell wall destruction products of Himematsutake cultures, and dried products thereof. However, it is preferable to extract from the dried product in terms of storage stability, handleability and extraction efficiency.

  In the present invention, prior to the hot water extraction, the fruit bodies of Japanese matsutake, himematsutake culture, or their cell wall destruction products and their dried products are extracted with an organic solvent or a water-containing organic solvent, and odor components peculiar to these It is desirable to remove the pigment component. The organic solvent used here includes methanol, ethanol, ethyl acetate, ether, and the like, and the water-containing organic solvent generally includes water-containing methanol, water-containing ethanol, etc. in a range of 30% or less. About 80% ethanol is preferred in terms of organic solvent.

  The hot water extract of the present invention may be any one of a hot water extract, a vacuum concentrated solution thereof, or a lyophilized product thereof. The hot water extract is concentrated under reduced pressure, and then subjected to a precipitation treatment with ethanol. Then, it is desirable to perform fractionation by liquid chromatography, followed by dialysis and lyophilization.

  In addition, the cell wall-disrupted dried material of the fruit body of Himematsutake is extracted with double the amount of hot water for 2 hours, and the hot water extract is further concentrated under reduced pressure, fractionated by liquid chromatography, dialysis and freeze-dried. For example, the extract has the following chemical composition. Crude ash content 5.36% (wt%, the same applies hereinafter), crude protein 41.20%, crude lipid 3.54%, crude fiber 6.20%, carbohydrate 43.57%, ergosterol 0.13%. When these crude ash, crude protein, crude lipid and carbohydrate are further analyzed, they have the compositions of Table 1, Table 2, Table 3 and Table 4, respectively.

  An extract having the above composition does not exhibit a certain decomposition point and melting point, and is carbonized by ignition, but is extremely stable. It is stable at room temperature for at least 3 years, and no decrease in activity is observed even after sterilization at 120 ° C. for 20 minutes.

  Bovine globulin concentrate is a 100% natural product purified from the serum of edible cattle at a facility for high-purity products that meets the GMP standards specified by the US Department of Agriculture, and is highly stable and stable. The ingredients of the bovine globulin concentrate are shown in Table 5. This figure is data from Proliant USA.

  In the composition of the present invention, the hot water extract of Japanese matsutake and the beef globulin concentrate as described above are blended, and the blending ratio of both is 0.5 to 20 parts by mass of the latter with respect to 1 part by mass of the former. It is preferable to do this.

Moreover, in the present invention, examples of the method used as a food or drink include the following methods.
1) A method in which the above-described composition is used as a sprinkle or filled in a tea pack or capsule.
2) A method of preparing and using saccharides, acids, salts, fragrances, etc. in the composition as described above.
3) Baking products, fermented products, kneaded products, dairy products, fat products, seasonings, foods such as confectionery, or coffee, cocoa, tea, fruit juice, vegetable juice, fermented beverages, soft drinks, etc. A method of adding and using in the manufacturing process of beverages.

  The following examples further illustrate the present invention.

Example 1
An extract was obtained from a Japanese matsutake mushroom culture (cultured mycelium and culture filtrate) as follows.
First, the mycelium of Himematsutake was cultured with shaking at 30 ° C. for 30 days in a liquid medium (pH 5.5) consisting of 20 g of glucose, 5 g of yeast extract, and 1 L of water. Seven times the amount of water was added to the centrifuged mycelium, and the mixture was extracted by heating at 95 ° C. for 2 hours. Ethanol was added to the extracted filtrate to precipitate the polysaccharide. The polysaccharide was collected by centrifugation, washed with acetone and then ether to obtain 21 mg of hot water extract per gram of mycelium. The complex component was a glucomannan protein complex with a molecular weight of 10 ⁵ to 10 ⁷ (gel filtration method), [α] _D + 57 ° (c = 2.0, water), a protein content of 5%, and a small amount of galactose and ribose. It was.
Next, the filtrate obtained by culturing the above-mentioned mycelium was concentrated under reduced pressure to 1/6 volume, and an equal volume of ethanol was added thereto and left at 4 ° C. overnight. The resulting precipitate was centrifuged, washed with acetone and ether, and vacuum-dried (room temperature) to obtain 575 mg of dried product per liter of culture filtrate.
The dried product was a mannan protein complex containing a molecular weight of 10 ⁵ to 10 ⁷ (gel filtration method), [α] _D + 63 ° (c = 2.0, water), trace amounts of glucose, galactose and ribose.
Thus, all the components isolated from the Japanese matsutake mushroom culture were glucomannan protein complexes.

  The hot water extract obtained from the above-mentioned Japanese matsutake mushroom culture does not exhibit a certain decomposition point and melting point, and is carbonized by ignition, but is extremely stable. At room temperature, it was stable for at least 3 years, and no decrease in activity was observed even after sterilization at 120 ° C. for 20 minutes.

  Next, 50 mg of the hot water extract (including the dried product) obtained above and 240 mg of bovine globulin concentrate were uniformly mixed to obtain the composition of the present invention.

Example 2
Purified water (1000 mL) was added to 100 g of dried cell wall-disrupted fruit bodies of Himematsutake fruiting bodies, and hot water extraction was performed for 2 hours on a water bath with gentle stirring. The same operation was repeated twice, and the two hot water extracts were combined, and then concentrated under reduced pressure to 200 mL. Ethanol was added to the concentrated solution under reduced pressure until the final ethanol concentration reached 70%, followed by centrifugation to separate 13.3 g of ethanol precipitate. The ethanol precipitate was used as a stationary phase and subjected to column chromatography packed with DEAE-Toyopearl Gel (trade name, manufactured by Toyo Soda Kogyo Co., Ltd.). . The eluted fraction was dialyzed and lyophilized to obtain 5.6 g of a hot water extract. The obtained hot water extract had the composition and physicochemical properties as shown in Tables 1 to 4 above.

  Next, 100 mg of the hot water extract obtained above and 240 mg of bovine globulin concentrate were uniformly mixed to obtain the composition of the present invention.

Test example 1
Each of the compositions of the present invention obtained in Examples 1 and 2, the extracts obtained in Examples 1 and 2 and the bovine globulin concentrate were confirmed by oral administration for confirming antitumor activity as follows.

Anti-tumor test Ten ICR / Slc female mice aged 5 weeks were used as one group. To this, Sarcoma 180 tumor cells (5 × 10 ⁶ cells) collected from mice 7 days after transplantation were subcutaneously transplanted, and a fixed amount of each specimen was obtained twice daily in the morning and evening for 20 days, 24 hours later. Oral administration was forcibly administered with a gastric sonde. On the 21st day of transplantation, the tumor volume (cm ³ ), (4 / 3πa ² b / 2, where a: minor axis mm, b: major axis mm) was measured, and tumor suppression rate (% ) = (1-T / C) × 100. However, T: Volume of the tumor in the sample administration group, C: Volume of the tumor in the control group. Further, the number of mice in which the tumor completely disappeared by the 28th day after transplantation and the number of mice that survived until the 28th day after the transplantation were confirmed. The results are shown in Table 6.

  In an oral administration experiment on mice, each dose was adjusted with physiological saline so that the prescribed oral dose was 0.2 mL per 10 g of the body weight of the mouse. Daily, twice in the morning and evening, every day for 20 days. Oral administration was forcibly performed with a sonde.

  As is apparent from Table 6, in the test groups (1) and (3) which are the compositions of the present invention, the complete disappearance of the tumor, as compared with the groups (2) and (4) which were administered with the hot water extract alone. A significant antitumor enhancing effect was observed.

  From the results of animal experiments, when drugs are administered to the human body, it is necessary to consider the drug sensitivity between humans and animals (mouses). Therefore, in this experiment, the sensitivity difference between mice and humans (Emil J. Freireich et al .: Cancer chemotherapy reports, 50 (4)) , 219, 1966), it is preferable to add 50-1000 mg / 60 kg (human) of bovine globulin concentrate to 50-100 mg / 60 kg (human) of the hot water extract of Himematsutake.

Test example 2
The test group (1) [combination group] and the test group (2) [single administration group] described in Table 6 were selected, and the mechanism for the antitumor enhancing effect of Japanese matsutake by adding bovine globulin concentrate was examined.

  The immunological mechanism was examined by observing the effects of the control group, (1) and (2) on the various immunocompetent cells under the same experimental conditions listed in Table 6. 20 days after oral administration, the mice were exsanguinated 24 hours after the final administration, and L3T4-positive cells and asialo GM1-positive cells present in the spleen were determined by the method of Hitoshi Ito (Anti-Cancer Drug Design, Vol. 8, 193- 202, 1993), measured by flow cytometry. The results are shown in Table 7.

  The results shown in Table 7 show that the hot water extract used in the present invention has an activation effect on the suppression of production of L3T4-positive cells and asialo GM1-positive cells even when administered alone. A remarkable activation effect was observed, and it was confirmed that the antitumor effect shown in Table 6 correlated well.

Test example 3
The composition of the present invention obtained in Examples 1 and 2 was subjected to an acute toxicity test by oral administration. The LD50 for mice was more than 3000 mg / kg, and the LD50 for rats was more than 3500 mg / kg. Further, from the results of subacute toxicity tests on rats and the results of general pharmacology tests on rabbits, the composition of the present invention did not have any problems related to toxicity.

Example 3
After obtaining a hot water extract in the same manner as in Example 1, 5 kg of beef globulin concentrate, 100 g of sugar, 15 g of honey, 5 g of caramel, 0.75 g of ascorbic acid, and 0.3 g of citric acid were added to 1 kg of the hot water extract. And 0.2g of lemon-based fragrances were added and mixed to obtain the health drink of the present invention.

Example 4
After obtaining an ethanol precipitate in the same manner as in Example 1, 50 g of beef globulin concentrate, 0.5 g of ascorbic acid and 2 kg of apple juice were added to and mixed with 10 g of the freeze-dried product of the ethanol precipitate. I got apple juice.

Example 5
After obtaining a freeze-dried extract in the same manner as in Example 1, 5 g of the extract and 25 g of bovine globulin concentrate were collected together with some calcium chloride and sodium triphosphate and dehydrated by water exposure. Thereafter, it was added to 2 kg of pre-cooled fish meat in the grinding step and frozen to produce the frozen savory taste of the present invention.

Claims (4)

  An antitumor composition comprising a hot water extract of Japanese matsutake and a bovine globulin concentrate.   A hot water extract of Himematsutake was obtained from any one or more of the fruit bodies of Himematsutake, the cell wall destruction of the fruit body of Himematsutake, the culture of Himematsutake, the cell wall destruction of Himematsutake culture, and their dried products. The antitumor composition according to claim 1, wherein   The hot water extract is characterized in that the hot water extract of Himematsutake is concentrated under reduced pressure, then precipitated with ethanol, then fractionated by liquid chromatography, then dialyzed and lyophilized. The antitumor composition according to claim 1 or 2.   The antitumor composition according to any one of claims 1 to 3, which is a medicine or a health food.