JPH05262658A - Immunostimulant - Google Patents

Abstract

PURPOSE:To provide the immunostimulant containing the hot water or warm water extract of glycyrrhiza after the extraction of glycyrrhizin as an active ingredient. CONSTITUTION:Glycyrrhiza after the extraction of glycyrrhizin with water or an alkali aqueous solution is extracted with warm water or hot water at 40-120 deg.C for 0.5-24hr, preferably at 50-95 deg.C for 5-16hr. After the extraction, the supernatant is separated from the residue and then concentrated or dried. Or, a polymeric substance fraction is separated from the extraction solution or concentration solution by a membrane separation method, by an alcohol precipitation method, etc. The immunostimulant contains the extraction solution, concentration solution or polymeric substance fraction as an active ingredient. The immunostimulant is employed as an immunological function-activating agent and also for quasi-medicines, cosmetics, foods, specific healthy foods, beverages. feeds, etc.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to the use of a hot water or hot water extract of licorice after extraction with glycyrrhizin as an immunopotentiator.

[0002]

2. Description of the Related Art We have found a strong immunopotentiating effect on a fraction previously extracted from licorice under acidic acetic acid, particularly a fraction obtained by further extracting a hot water extract of licorice from Afghanistan with acetic acid ( Further, it was found that these fractions have an antitumor activity which is considered to be due to an immunopotentiating action, and they were found to be used as an antitumor agent (Japanese Patent Laid-Open No. 3-255032). Publication).

Conventionally, in the case of producing this substance, the hot water extract of untreated licorice root was adjusted to be acidic, the supernatant excluding the formed precipitate was adjusted to neutral, and then the formed precipitate was formed. Equal amount of alcohol is added to the depleted supernatant and the resulting precipitate is collected. Further, an equal amount of alcohol is added to the 1N acetic acid extract of this precipitate to remove salts from the precipitate formed.

[0004]

However, this method is not suitable for mass production due to the complicated steps, and it is difficult to obtain other useful ingredients in licorice such as glycyrrhizin, which makes it difficult to use for water and livestock applications. Is not very economically favorable.

Further, feeds for water and livestock containing licorice extract and feed additives (JP-A-3-173827, JP-A-3-175934, JP-A-2-250832) have already been used. However, its effectiveness is mainly due to detoxification and antibacterial effects of glycyrrhizin and the like, and those containing an immunopotentiating substance in licorice other than glycyrrhizin as an active ingredient have not been known until now.

The object of the present invention is to provide a method for producing a novel immunopotentiator, which overcomes the drawbacks of the conventional methods.

[0007]

Means for Solving the Problems As a result of intensive studies to achieve the above object, the present inventors have found that hot or hot water extract of licorice after extracting glycyrrhizin with water or an aqueous alkali solution. The present invention was completed by recognizing a strong immunopotentiating activity in and its high molecular substance fraction. On the other hand, the hot water extract of licorice that had not been subjected to the extraction operation with water or an aqueous alkaline solution had a weak immunopotentiating activity. This is considered to be due to the immunosuppressive action of low molecular weight substances such as flavonoids extracted together with glycyrrhizin. Therefore, this extraction operation is essential to obtain the substance of the present invention. The immunopotentiating activity was specifically shown in Examples.

That is, the immunopotentiator of the present invention is effective in extracting hot water or hot water of licorice after extracting glycyrrhizin with water or an aqueous alkaline solution, a concentrated solution thereof, a dried product thereof, or a high molecular substance fraction thereof. It is characterized in that it is contained as a component.

The licorice to be used in the present invention is not limited in origin and species, but it is particularly produced in Afghanistan (Glycyrrhiza).
It is preferable to use licorice from glabla) or Shinji (Glycyrrhiza glandulifera). Glycyrrhizin is extracted with water or an alkaline aqueous solution, preferably 0.1 to 0.
It is desirable to use 5% ammonia water and to perform the extraction at a temperature of 5 to 40 ° C. and an extraction time of 1 to 24 hours.

Further, hot water or hot water extraction conditions of licorice after glycyrrhizin extraction are 40 to 120 ° C. and 0.5 to 24.
The time is preferably 50 to 95 ° C. and the time is in the range of 5 to 16 hours. The pressure during extraction is usually slightly increased, but normal pressure may be used, and reduced pressure may be used if desired.

After the extraction is completed, the supernatant and the residue are separated, and the supernatant is concentrated or dried. Although it shows sufficient immunopotentiating activity even when used in this state, it has a higher effect by concentrating or drying the polymer substance fraction separated by the membrane separation method or the alcohol precipitation method. Can be obtained. These immunopotentiating activities were measured using the mitogenic activity against mouse lymphocytes and the antibody production promoting activity as indicators.

The immunopotentiator of the present invention can be used for various purposes. The first use is as an immune function activator or animal animal immune function activator that makes full use of its immune enhancing effect. The second use is as a quasi drug, a cosmetic, a food, a food for specified health use, a drink, a feed, etc., which is mixed in the expectation of its immunopotentiating effect.

[0013]

EXAMPLES The present invention will now be described in detail with reference to examples, but the present invention is not limited thereto.

(Example 1) Acquisition of immunopotentiator: 100 g of licorice root powder was added to 100 ml of 0.3% ammonia water, and the mixture was extracted for 16 hours. Then, it was separated into a supernatant and a residue by filtration, and the residue was diluted with 50 ml of distilled water. Wash and filter to obtain a residue. Add 100 ml of distilled water to this residue and add 5
After extraction at 0 ° C. for 12 hours and at 95 ° C. for 1 hour, the supernatant was freeze-dried to obtain a brown powder in a yield of 3 to 5%.
These are designated as samples A and B, respectively. Also, 95 ℃
Methanol was added to the supernatant of the extraction for 1 hour in an equal amount, and the produced precipitate was freeze-dried to obtain a brownish gray powder in a yield of 1 to 2%. This is sample C. In addition, the above ammonia-extracted supernatant was neutralized to pH 7 and lyophilized to be sample D.

Separately from the above, 100 ml of licorice root powder was added with 100 ml of distilled water and extracted for 16 hours, and the mixture was separated into a supernatant and a residue by a filtration operation, and 100 ml of distilled water was added to the residue for a further 16 minutes.
After extracting for a period of time, the residue is obtained by filtration. To this residue was added 100 ml of distilled water, extraction was carried out at 50 ° C for 12 hours and at 95 ° C for 1 hour, and then the supernatant was freeze-dried to give a brown powder of 5-7.
Obtained in% yield. These are designated as samples E and F, respectively. Also, methanol was added in an equal amount to the supernatant of extraction at 95 ° C. for 1 hour, and the resulting precipitate was lyophilized to give 1 to brown powder.
Obtained in a yield of 3%. This is sample G. Further, a sample H is obtained by freeze-drying the above water-extracted supernatant.

(Example 2) Measurement of mitogenic activity: Mouse spleen cells and a sample were added to a 96-well plate and cultured for 48 hours. Then, live cells were stained with propidium iodide (PI) and the fluorescence intensity was measured. The amount of cell proliferation was quantified. The results are shown in Figure 1.

From the results shown in FIG. 1, samples A, B,
It was found that all of C, E, F and G have strong mitogenic activity. Samples D and H were inactive and tended to be suppressed.

(Example 3) Antibody production promoting activity test: Mouse spleen cells and a sample are added to a 96-well plate and cultured for 48 hours, and then the culture supernatant is collected. And the antibody (IgM) in the culture supernatant
Was measured by the fluorescent antibody method. The results are shown in Figure 2.

From the results of FIG. 2, samples A, B, C,
It was found that all of E, F and G have an action of promoting antibody production. On the other hand, samples D and H tended to be suppressed.

[0020]

As shown in the above examples, it was confirmed that the substance of the present invention has a strong immunopotentiating activity and can be produced inexpensively by a simple operation without damaging useful components in licorice such as glycyrrhizin. ..

[Brief description of drawings]

FIG. 1 is an actual measurement diagram showing the mitogenic activity of the immunopotentiator of the present invention.

FIG. 2 is an actual measurement diagram showing the antibody production promoting activity of the immunopotentiator of the present invention.

Claims (6)

[Claims] 1. An immunopotentiator, characterized by using licorice after glycyrrhizin extraction as a raw material. 2. An immune enhancer characterized by using licorice as a raw material after extracting glycyrrhizin with water or an aqueous alkaline solution. 3. An immunopotentiator, which comprises extracting licorice after extraction with glycyrrhizin with warm water or hot water. 4. An immunopotentiator comprising, as an active ingredient, a hot or hot water extract of licorice after glycyrrhizin extraction, a concentrated solution thereof, or a dried product thereof. 5. An immunopotentiator, which comprises, as an active ingredient, a high molecular substance fraction in a hot water or hot water extract of licorice after glycyrrhizin extraction or a concentrate thereof. 6. A method for producing an immunopotentiator using licorice after extraction with glycyrrhizin as a raw material.