JP2001010970A - Antineoplastic fraction from agaricus blazei murill - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a novel medicine or health food which exhibits excellent pharmacological activity and can easily be absorbed by digestive tracts. SOLUTION: This antineoplastic agent or health food includes a chromatography main elution fraction of molecular weight 100-2,000 obtained by the process for extracting fruit bodies of Agaricus blazei Murill with hot water, the process for dialyzing the extract and the process for treating the dialyzed outer solution by chromatography as an active ingredient. The antineoplastic agent or health food is obtained by including a dialyzed outer solution, which is obtained by the process for extracting the fruit bodies with hot water, the process for mixing the extract with ethanol and centrifuging the mixture to divide into a precipitate and a supernatant, the process for mixing the supernatant with ethanol and centrifuging the mixture to divide into a precipitate and a supernatant and then the process for dissolving the precipitate into distilled water to dialyze, as an active ingredient.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an antitumor agent or health food containing a substance having a relatively low molecular weight, which is obtained by subjecting fruiting bodies of Kawariharatake to hot water extraction.

[0002]

2. Description of the Related Art Kawariharatake is known to have antitumor activity, and its powder or various extracts are used orally as health foods. In particular, many polymer substances such as protein polysaccharides derived from Kawariharatake have been reported.

[0003]

Generally, high-molecular substances such as protein polysaccharides cannot always be said to have good absorption from the digestive tract. Substances having lower molecular weight and excellent pharmacological activity, especially substances which can be orally administered, are always required as pharmaceuticals and health foods.

[0004]

Means for Solving the Problems As part of the search for an antitumor active ingredient, an activity test based on oral administration to mice has been studied. As a result, Agaricus blaz
Since the antitumor activity was recognized in the relatively low-molecular-weight hot-water-extracted fraction of ei Murill (Kawariharatake, Kyowa agaricus mushroom), the present invention was carried out by fractionating the active ingredient.

[0005] The present invention is effective for a main elution fraction of chromatography having a molecular weight of 100 to 2,000 obtained by a step of extracting a fruit body of Kawariharatake with hot water, a step of dialysis of the extract, and a step of chromatography of an outer dialysate. It relates to an antitumor agent or health food as a component.

[0006] The present invention also provides a step of extracting the fruit body of Kawariharatake with hot water, a step of adding ethanol to the extract and mixing the mixture, and centrifuging the mixture to separate a precipitate and a supernatant.
Ethanol is added to the supernatant and mixed, and the mixture is centrifuged to separate into a precipitate and a supernatant, and the precipitate is dissolved in distilled water and the dialysis process is performed, and the outer dialysis solution obtained is used as an active ingredient. Related to antitumor agents or health foods.

[0007]

DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, a method for producing an antitumor active ingredient of the present invention will be described. The fruit body of Kawariharatake used in the present invention may be used in natural products or artificial cultures,
Commercially available dried products can also be conveniently used. It is carried out by adding 5 to 10 times water to the dried fruit body of Kawariharatake and heating and refluxing for 1 to 3 hours. Hot water extraction is performed by repeating heating and refluxing of the hot water extraction residue. The hot water extract thus obtained is freeze-dried to obtain a dried product (hereinafter referred to as dried product A). 5 to 20
Add water twice, place in dialysis tubing, and add 10 times with distilled water.
A dried product having an antitumor activity (hereinafter referred to as a dried product C) can be obtained by dialysis for up to 15 hours and freeze-drying the outer dialysate.

Next, the inner dialysis solution is further dialyzed for 20 to 40 hours in running water, dialyzed twice with distilled water for several hours each, and then lyophilized to obtain a dried product having antitumor activity. (Hereinafter referred to as dried product B). The obtained dried product C was dissolved in about 10 times the volume of distilled water, and chromatography was carried out using distilled water as an eluent.
Separate by mL to obtain many fractions. The main eluted fraction in the middle of the obtained fraction, having a molecular weight of 100 to
2000 fractions are the fractions showing high tumor inhibitory activity of the present invention. Analysis of these fractions by reverse phase chromatography using ODS, ion exchange chromatography using DEAE-TOYOPEARL650, etc. confirmed that they contained several components in addition to arginine, lysine and mannitol. Have been.

[0009] Further, an equal amount of ethanol is added to the hot water extract obtained by the above method, mixed and centrifuged to separate a precipitate and a supernatant. The obtained supernatant is further added with 1 to 3
A double volume of ethanol is added and mixed, the precipitate obtained by centrifugation is dissolved in distilled water, and the resulting dialysis solution obtained by dialysis of the solution is a low-molecular fraction, which is a high tumor-inhibiting activity of the present invention. Is a fraction showing The fraction having high tumor-inhibiting activity thus obtained can be used as a pharmaceutical preparation as it is or together with various excipients used in the production of oral pharmaceutical preparations. The fraction can be used as it is or as a health food or drink with other foods.

[0010]

EXAMPLES Example 1 2 L of distilled water was added to 300 g of Agaricus blazei Murill (Kyowa agaricus mushroom), and the mixture was heated under reflux for 2 hours. This was filtered and separated into a filtrate (hot water extract) and a residue. To the residue, 2 L of distilled water was added again, and the mixture was heated under reflux for 2 hours to perform hot water extraction. After obtaining a filtrate, the residue was subjected to another similar hot water extraction, and the filtrates were combined. The combined filtrate (hot water extract) was freeze-dried to obtain 153 g (51%) of dried product A. Add 50 mL of distilled water to 50 g of dried product A and add dialysis tube (Spectra
/ Por Membrane 50x31,8mm id x30 m, FE 0526-65)
Put in. This was dialyzed against 3 L of distilled water for 12 hours. This outer dialysis solution is freeze-dried and dried matter C27g (53%)
I got The inner dialysis solution was further dialyzed in running water for 30 hours, then dialyzed twice with distilled water (4 hours each, 8 hours in total), and the inner dialysis solution was freeze-dried to obtain 11 g (22%) of dried product B (22%). Obtained. Subsequently, 3 g of the dried product C was dissolved in 30 mL of distilled water and TO
Chromatography was performed using YOPEARL HW40C (40 mm id x 420 mm). All effluent solvents used distilled water. Fractions 1 to 30 were obtained by fractionating 20 mL of each fraction. These fractions were divided into the following five groups with reference to thin layer chromatography. The weight was as follows. Fractions 1 to 11 (75 mg, 2.5%),
Fractions 12-15 (920 mg, 30.7%), Fractions 16-17 (1570 mg,
52.3%), fractions 18-19 (270 mg, 9%), fractions 20-28 (97 m
g, 3,2%). The infrared absorption spectrum (IR) data is as follows. Fraction 16: IR (KBr) 3390, 3325, 3285, 2940, 2920, 164
1, 1634, 1622, 1615, 1600,1595, 1405, 1394, 1084,
1020. Molecular weight (gel filtration method) 100-2000

The antitumor test of the Agaricus hot water extract component and the dialyzed fraction was carried out as follows. 1) Administration sample 1 to 10 mg / mL of the following (A) to (D)
A concentrated aqueous solution was used. (A) Agaricus hot water extract (dry matter A, freeze-dried hot water extract) (B) High molecular fraction (dry matter B) (C) Low molecular fraction (dry matter C) (D ) Low molecular fraction (chromatographic fraction 16) 2) ICR / JCL mice (five-week-old female, 6 /
Group)-6 animals x 3 groups x 6 species were used. 3) Tumor cell line: Sarcoma 180 cells. The cells were subcultured at 37 ° C. (Method) In vivo assay using ICR mouse (oral administration) 1) 2-5X10,00 Sarcoma 180 cells subcutaneously under one foot of mouse
The transplant was performed at a rate of 0,000 cells / mouse. 2) An aqueous solution of the drug was orally administered for 1 to 10 days. The drug concentration was 10 mg / mL for each drug. 3) The tumor volume was observed every half week for 5 weeks, and the tumor mass was excised and weighed on the last day. The tumor volume was calculated from the major axis and minor axis of the tumor mass. (Results) The tumor inhibition rates of the administered samples were as follows. (A) 68%, (B) 65%, (C) 71%, (D) 8
3%

Example 2 The same hot water extraction as in Example 1 was performed to obtain 6 L of a combined filtrate (hot water extract). The filtrate is concentrated under reduced pressure to 1 L
Then, 1 L of ethanol was added thereto, mixed, and centrifuged to obtain a precipitate and a supernatant. 3 L of ethanol was further added to the supernatant, mixed, and the precipitate obtained by centrifugation was dissolved in distilled water and dialyzed. The tumor growth inhibition rate of the external dialysate was determined in the same manner as in Example 1 and was found to be 86%.

[0013]

Industrial Applicability According to the present invention, an antitumor agent and a health food containing an antitumor fraction obtained by extracting Kawariharatake with hot water are provided.

 ──────────────────────────────────────────────────続 き Continuation of the front page (72) Inventor Tomohisa Ota 2-28-60 Heiwacho, Kanazawa-shi, Ishikawa (72) Inventor Takuma Sasaki 4-12-5-401 Izunocho, Kanazawa-shi, Ishikawa F-term (reference) 4B018 LE03 MD82 ME08 MF01 MF04 MF06 4C088 AA07 AC16 AC17 BA05 BA23 BA24 CA02 CA14 CA16 MA52 NA11 NA14 ZB26

Claims (2)

[Claims] 1. A step of extracting the fruit body of Kawariharatake with hot water, a step of dialysis of the extract, a step of chromatography of the obtained outer dialysis solution, and a molecular weight of 10
An antitumor agent or health food comprising a chromatographic main elution fraction of 0 to 2000 as an active ingredient. 2. A step of extracting the fruit body of Kawariharatake with hot water, a step of adding and mixing ethanol to the extract, a step of separating the mixture by centrifugation into a precipitate and a supernatant, and adding and mixing ethanol to the supernatant. And separating the mixture into a precipitate and a supernatant by centrifugation, dissolving the precipitate in distilled water for dialysis, and an antitumor agent or health food containing the obtained dialysate as an active ingredient.