JPH0678366B2 - Manufacturing method of chrysanthemum polysaccharides - Google Patents

Description

Detailed Description of the Invention [Industrial applications]

TECHNICAL FIELD The present invention relates to a method for producing a chrysalis polysaccharide having an action of activating the human immune system such as enhancing interferon producing ability and enhancing natural killer activity.

[Prior art]

Interferon is a natural product produced by cells and a kind of hormone-like substance that acts on cells.Since its discovery, it has been regarded as an effective substance for viruses, immunosuppression, some cancers, etc. It is still controversial because it is not absorbed by administration. There are many studies on interferon, NH
Actions mediated by cells (natural killer; natural cancer destroying action), anti-cancer action due to cell differentiation being promoted or induced by interferon, viral disease of NH-IFN system in animals and humans, and onset of smallpox Studies on how important the relationship between extension, metastasis, and prognosis have been promoted and announced. In any case, if interferon is deficient or significantly lowered, it is considered that it is easy to be infected with a viral disease even if it does not involve macrophages and NH cells, and it is likely to cause serious injury, and cooperation with NH, interleukin and other lymphokines, Furthermore, it is presumed that the relationship between macrophages, polynuclear leukocytes, and immunocompetent cells may be impaired, resulting in easy carcinogenesis.

[Problems to be Solved by the Invention]

In order to prevent the lack or reduction of interferon from the aspect of problem solving by administering interferon, conversely, increasing interferon in the body can suppress the production of infectious virus as described above and prevent the onset of disease. From a viewpoint, substances with excellent interferon-producing ability,
The development of NH active activators is awaited.

[Means for solving problems]

The present inventors have increased interferon production ability and
By conducting a search for substances that also enhance the activity, it was found that among many substances, the salmonella jellyfish exhibits excellent potency for this purpose, the manufacturing method thereof was established, and the present invention was completed here. That is, the fungus is extracted with hot water, the extract is ultrafiltered, and a lower alcohol is added to the obtained ultrafiltration inner solution,
This is a method for producing a fungus, a jellyfish polysaccharide, which is characterized in that the formed precipitate is collected. Asteraceae is a plant of the family Asteraceae, Auricula
The fruiting body of ria auricula (L.ex Hook) Underw is produced in Sichuan, Fujian and Jiangsu provinces of China. It is said that this drug has medicinal properties, and according to the Chu Andai Dictionary, the medicinal properties are mainly due to bleeding suppressive action and are used as a material of Chinese medicinal herbs. In Japan, about 400 tons are imported and used annually. The Pleurotus cornucopiae polysaccharide according to the present invention can be produced by purifying an extract of Pleurotus cornucopiae with hot water by a specific method. The outline of the manufacturing method is described below. First, the dried fungus is thoroughly washed with cold water, then the soluble fraction is extracted with hot water, the solid content is removed, and then clarified by centrifugation to obtain a fungus extract. The obtained clear liquid is subjected to ultrafiltration as it is without being concentrated. Ultrafiltration is carried out under pressure according to a conventional method using an ultrafiltration membrane having a molecular weight cut off of about 10,000 to 200,000. The filtration membrane may be one having the above-mentioned molecular weight cut-off,
There is no particular limitation on the material. Examples of filtration membranes used here include Toyo Roshi Kaisha Ltd. products UK-10 (fraction molecular weight, 10,000), UP-20 (fraction molecular weight, 20,000), UK-50 (fraction molecular weight, 50,000), UK -20
0 (molecular weight cutoff, 200,000), NT of Nitto Denko Corporation product
U-2000 series (molecular weight cutoff, 20,000 to 100,000), NT
U3000 series (molecular weight cutoff, 8,000 to 100,000), NTU-
The 4000 series (molecular weight cutoff, 6,000 to 200,000) are listed. Of these, the UK-200 and NTU-2000 series are particularly preferable. Here, the pressure applied during filtration is suitably 0.5 to 2.0 kg / cm ² . A lower alcohol such as methanol, ethanol or isopropyl alcohol is added to the liquid obtained by ultrafiltration to cause precipitation, which is filtered off. The purified product precipitated is dried by a conventional method to give a product.
From the viewpoint of quality, the freeze-drying method is preferable. The lyophilized product of the obtained Kikulag polysaccharide is a yellowish brown powder, which is soluble in water, and contains methanol, ethanol,
It is insoluble in organic solvents such as acetone, ether, chloroform, ethyl acetate, benzene and hexane. The color reaction is positive for phenol-sulfuric acid reaction and anthrone-sulfuric acid reaction. The ultraviolet absorption spectrum shows no maximum absorption value when measured in an aqueous solution, and shows only terminal absorption. This is shown in FIG. The infrared absorption spectrum is shown in FIG.

【Example】

The present invention will be described in detail below with reference to examples and application examples, but it goes without saying that the present invention is not limited to these examples. [Example-1] After thoroughly washing 50 g of the fungus Jellyfish with cold water, 3000 g of water was added, extraction was carried out at 100 ° C for 1 hour, and solid matter was removed using a case to obtain 1000 g of an extract. 1000 g of water was added to the above solid content again, extraction was carried out at 100 ° C. for 1 hour, and the solid content was removed using a case to obtain 1000 g of an extract. After combining the extract with the extract obtained first,
To 2000 g (Bx0.2) of the extract, 10 g of Celite 545 as a filter aid was added and filtered to obtain a transparent filtrate. This filtration was performed at room temperature under a pressure of 2.0 kg / cm ² using an ultrafilter UHP-90 (manufactured by Toyo Roshi Kaisha, Ltd.) and a filtration membrane UK-200 (molecular weight cutoff of 200,000; manufactured by Toyo Roshi Kaisha, Ltd.). When 400 g of filtrate was obtained, 400 g of water was added to the residual filtrate, and ultrafiltration was continued, and the same operation was repeated twice to obtain a residual filtrate of about 200 g (Bx2 .0), 800 ml of ethanol was added thereto, and the resulting precipitate was separated by filtration.The purified product of the precipitate was lyophilized to obtain 3.6 g of a jellyfish polysaccharide. 900 kg was added and extraction was carried out for 1 hour at 100 ° C. Then, the solid content was removed by applying a pulper finisher to obtain about 300 kg of extract liquid. Then, post-treatment is carried out in the same manner as in [Example-1] to obtain about 30
0 kg of extract was obtained. This extract is combined with the extract obtained first to give about 600 k
After centrifuging the extract of g, 2.0 kg of Celite 545 was added as a filter aid, and the mixture was stirred and filtered using a spakura filter to obtain 560 kg of a transparent filtrate (Bx0.2). This filtrate was used as an ultrafilter RW-1 type (manufactured by Nitto Denko Corporation), a filtration membrane NTU-2000-P.18-B (fraction molecular weight 100,0).
00; manufactured by Nitto Denko Co., Ltd.). When 420 kg of the filtrate was obtained, 140 kg of water was added and ultrafiltration was continued, and the same operation was repeated twice to obtain about 140 kg of a polysaccharide fraction as a residue. This product was concentrated under reduced pressure to about 60 kg (Bx2.0), and 240 kg of ethanol was added to this, and the resulting precipitate was filtered off, followed by freeze-drying to obtain 1.0 kg of the jellyfish polysaccharide (hereinafter referred to as " Goods). [Application example-1] 2.1g of this product per day will be served to 5 men and women (ages 20s to 50s) every day
It was administered 30 minutes before for 14 consecutive days. The day after the start of administration (the second day of administration), the eighth day,
On the 15th and 22nd days, the effect on the α-type interferon (IFN-α) -producing ability was measured. The measurement method was a competitive method using ¹²⁵ I · IFN-α. The IFN-α production ability was revealed to be significantly higher after administration than before administration of this product, demonstrating that this product is effective against interferon production ability. The measurement results are shown in [Table-1]. [Application Example-2] 2.1 g of this product was administered to 5 males and 5 females (aged 20s to 50s) daily for 14 days 30 minutes before lunch. The day after the start of administration (the second day of administration), the eighth day,
The natural killer activity (NH activity) was measured on the 15th and 22nd days. The measurement method was labeled with ⁵¹ Cr as target cells.
Cytotoxic assay using HeLa cells was used. NH activity shows a clear increase by administration of this product,
The effectiveness of this product for activation of NH activity was proved. The measurement results are shown in [Table-2].

【The invention's effect】

The jellyfish polysaccharide according to the present invention is a drug, food, or nutrition for diseases that can be expected to have preventive or therapeutic effects on cancer, colds, hepatitis, etc. by activating the human immune system such as IFN production ability and NH activity. It can be used and applied to supplement foods.

[Brief description of drawings]

FIG. 1 is a diagram showing an ultraviolet absorption spectrum of the jellyfish polysaccharide of the present invention, and FIG. 2 is a diagram showing the infrared absorption spectrum of the same.

Claims (2)

[Claims] 1. A fungus fungus, characterized in that the fungus is extracted with hot water, the extract is subjected to ultrafiltration, a lower alcohol is added to the obtained ultrafiltration inner solution, and the resulting precipitate is collected. Polysaccharide manufacturing method. 2. An ultrafiltration membrane having a molecular weight cutoff of 10,000 to 200,
The method for producing the chrysalis polysaccharides according to claim 1, which is 000.