JP2000086526A - Immunity activator and beverage containing the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an immunity activator, which is made of a highly safe natural substance, easily produced at low cost, showing no side-effect, good in flavor, and added to a food or the like to be ingested every day. SOLUTION: This activator contains, as the effective ingredient, an extract of cacao beans or cacao husk as the cacao bean skin, and is a high-molecular- weight fraction having a molecular weight of 12,000 or more, where the stock is preferably extracted with water and/or water-soluble organic solvent.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to cocoa beans (Theobr
The present invention relates to an immunostimulant containing, as an active ingredient, an extract of cacao husk, which is the hull of oma cacao seed) and / or cacao beans, and a food and drink containing the immunostimulant and having an immunostimulatory action.

[0002]

2. Description of the Related Art The immune system is particularly important among many elements constituting a defense mechanism of a living body, and is susceptible to invasion of foreign viruses and microorganisms, and to foreign self-materials such as cancer cells. On the other hand, it functions to maintain the homeostasis of the living body.
The immunity function, such as aging, stress, fatigue, environmental factors,
It is thought that various diseases and infectious diseases are caused when the decrease is caused by some factor. For this reason, it is considered that enhancing immunity, which is a defense function of a living body, is important for maintaining homeostasis of the living body.

[0003] In recent years, attempts have been made to use natural extracts such as food ingredients, Chinese herbal prescriptions, and crude drugs to activate the defense mechanism of the living body, which results in lymphocyte blastogenesis (mitogenic activity) and enhanced antibody production. Action, macrophage activation, cytokine production, tumor necrosis facto
r: TNF) has been evaluated from various viewpoints such as production.

[0004] In particular, the blastogenesis of lymphocytes is a prerequisite for the expression of immune functions, and is therefore considered to be one index for evaluating the immunostimulatory action of a living body. Regarding the lymphocyte blastogenesis, the cell components of Bifidobacterium longum (JP-A-3-120222), herbal medicine materials (JP-A-56-97233), and peptides in milk (JP-A-Heisei 56-97233) JP-A-8-73499), the hot water extract of Knotweed rhizome (JP-A-6-56685) and the licorice extract (JP-A-5-262658) are reported. These are patent applications as immunopotentiators.

[0005] By the way, there has been no report on the immunostimulatory effect of cacao beans as a raw material of chocolate or cocoa or cacao husk as its outer skin. That is, cacao beans or cacao husks have been used in the past to prevent diabetic complications (JP-A-9-234).
018), a carcinogenesis preventive action (JP-A-9-2246)
No. 06), an anti-stress effect (JP-A-9-20602)
No. 6), a gastric ulcer prevention effect (Japanese Patent Application Laid-Open No. 7-274894).
Japanese Patent Application Laid-Open No. 7-238028)
JP, JP-A-7-213251), fat absorption inhibitory action (JP-A-6-321796), cholesterol elevation inhibitory action (JP-A-6-987).
No. 18, JP-A-3-181494), lactic acid bacteria growth promoting action (JP-A-8-196268),
A blood pressure rise inhibitory action (JP-A-3-181495);
Cholesterol gallstone formation inhibitory action (JP-A-3-1814)
No. 96) is merely reported.

[0006]

DISCLOSURE OF THE INVENTION The present inventors have found that natural products having high safety can be used as raw materials, production can be easily performed at low cost,
Various food materials and plant extracts for the purpose of providing immunostimulants and foods and drinks having immunostimulatory effects that have no side effects, have a good flavor, can be added to foods, etc., and can be taken daily on a daily basis. The product was screened using the spleen cells of mice for lymphocyte blastogenesis (mitogenic activity) as an indicator. As a result, the extract obtained from cacao beans and / or cacao husk, which is the hull of cacao beans,
The inventors have found a strong effect of transforming lymphocytes into immature cells, and have completed the present invention.

[0007]

That is, the immunostimulant of the present invention contains cocoa beans and / or an extract obtained from cacao husk, which is the hull of cocoa beans, as an active ingredient. Comprises such an immunostimulant.

The above extract is an extract obtained by extraction with water and / or an organic solvent compatible with water, and the extract has a molecular weight of 12,000.
It is preferable that it is the above-mentioned high molecular fraction.

The cocoa beans used in the present invention are seeds in the fruits of evergreen tree of the family Aguiridae, and are generally used as raw materials for chocolate and cocoa. The cocoa beans consist of cocoa nibs and cocoa husks overlying them and can be used separately or in whole, and can be used raw or heat-treated. You may use it. The extract of cacao husk, which is the hull of cacao beans, has a particularly high mitogenic activity.
The use of cacao husks is recognized to be effective because chocolate and cocoa are treated as unneeded materials that are removed from the cocoa beans and production costs are reduced.

[0010] The method for obtaining the extract from the cacao beans or cacao husk of the present invention is not particularly limited. That is, for example, chloroform, ethyl acetate, methanol, ethanol, butanol, acetone, propylene glycol,
It can be obtained by extraction using a solvent such as glycerin or water, or a mixture thereof, but it is preferable to use water, an organic solvent compatible with water, or a mixture thereof.

As a solvent compatible with water, only one of methanol, ethanol, acetone, propylene glycol, glycerin and the like may be used, or a mixture of two or more of them may be used. In this case, use of water or a mixed solution of water and ethanol is most preferable in terms of easy extraction and concentration, safety for living bodies, and the like.

The extraction temperature may be any of high temperature, room temperature and low temperature, but it is preferable to extract at 50-100 ° C. for about 1-5 hours. The extract may be filtered, and after distilling off the extractant, the filtrate may be concentrated under reduced pressure or may be freeze-dried.

As the immunostimulant of the present invention, the extract obtained by the above method may be used directly as it is, but it is generally dissolved or dispersed in an appropriate liquid carrier, or Mixed with a carrier or adsorbed thereto, and in some cases, further emulsifiers,
A dispersing agent, a suspending agent, a spreading agent, a penetrating agent, a wetting agent, a stabilizer and the like are added and used as a formulation such as an emulsion, an oil, a wettable powder, a powder, a tablet, a capsule, and a liquid.

[0014] The amount of the extract to be added when used as a preparation varies depending on the form of the preparation.
% Or more is preferable, and there is no problem in safety.

When the above-mentioned immunostimulant is added to food and drink to obtain the food and drink of the present invention having an immunostimulating action, the method of adding the immunostimulant to food and drink is generally performed. Although a method may be used, it is preferable to add it in the final step so as to add a dye, a fragrance and the like.

The foods and drinks include confectionery such as chewing gum, candy, tablet confectionery, gummy jelly, chocolate, biscuit or snack, ice confectionery such as ice cream, sherbet or ice confection, beverage, pudding, jam, dairy product, etc. Daily intake of these foods and drinks can enhance immunity. The addition amount of the immunostimulant of the present invention to these foods and drinks depends on the form of the food and drink, but is preferably 0.001% by weight or more, and from the viewpoint of palatability, 5% by weight or less is preferable. .

[0017]

BEST MODE FOR CARRYING OUT THE INVENTION First, Examples 1 to 5 and test examples of the immunostimulant of the present invention will be specifically described below. However, the present invention is not limited to these.

Example 1 1 kg of cocoa beans was ground and extracted twice with 5 liters of hexane to remove fat. After air-drying the defatted cocoa beans, reflux extraction was performed with 5 L of water for 2 hours. The extract was centrifuged and the supernatant was filtered. The filtrate was concentrated under reduced pressure and freeze-dried to obtain 105 g of a brown powder (the immunostimulator of the present invention).

Example 2 Cacao husks, which are hulls, were separated from cocoa beans. 1 kg of the cacao husk was pulverized and subjected to reflux extraction with 5 liters of water for 2 hours. The extract was centrifuged and the supernatant was filtered. After concentrating the filtrate under reduced pressure,
By freeze-drying, 115 g of a brown powder (the immunostimulant of the present invention) was obtained.

Example 3 A cocoa husk, an outer hull, was separated from cocoa beans. 1 kg of the cacao husk was ground and extracted twice with 5 liters of hexane to remove fat. After air-drying the defatted cacao husk, it was subjected to reflux extraction with 5 liters of water for 2 hours. The extract was centrifuged and the supernatant was filtered. The filtrate was concentrated under reduced pressure and freeze-dried to obtain 120 g of a brownish powder (the immunostimulant of the present invention).

Example 4 The defatted cacao husk prepared by the method shown in Example 3 was subjected to reflux extraction with 5 liters of 30% ethanol for 2 hours. Centrifuge the extract,
The supernatant was filtered. The filtrate was concentrated under reduced pressure and freeze-dried to give a brown powder (the immunostimulant of the present invention) 125 g.
I got

Example 5 2.0 g of a cocoa husk water extract obtained by the method described in Example 2 was mixed with 150 ml of purified water.
And filled in a dialysis tube having a molecular weight cut off of 12,000, and dialyzed once in 3 liters of purified water for 12 hours for 3 times. Thereafter, the content of the dialysis tube is freeze-dried, and the high molecular fraction of the cacao husk water extract (the immunostimulant of the present invention)
1.04 g was obtained.

[Mitogen activity test] The immunostimulants of the present invention according to Examples 1 to 5 were used as samples to examine mitogen activity. Mitogen activity was measured in ICR mice (5-8
(Week-old, male), and using spleen cells, the dehydrogenase activity of the cells was measured using WST-1 reagent (Dojindo Laboratories) as an index. The spleen removed from the mouse was replaced with RPMI 1
The suspension was gently ground using a wire mesh in 640 medium to obtain a single cell suspension. After washing the cells three times with the same medium, the cells are washed with the same medium containing 10% fetal bovine serum (FBS) for 3.5 times.
× 10 diluted to ⁶ cells and placed in a 96-well microplate at 190
Dispensed in μl.

Further, two samples for measuring mitogen activity were prepared.
A solution prepared to a specified concentration with a 0% ethanol aqueous solution (blank is a 20% ethanol aqueous solution) was added to each well in an amount of 10 μl.
(Final concentration: 200, 20, 2 μg / m
l). After culturing for 72 hours in an environment of 5% CO ₂ and 37 ° C., 20 WST-1 reagent (final concentration 5 mM) was added to each well.
μl was added, and the mixture was further cultured after stirring for 4 hours. After the culture, the absorbance was measured at a measurement wavelength of 415 nm and a reference wavelength of 630 nm using a microplate reader to compare mitogenic activities.

The test results are shown in the graph of FIG. From the test results, mitogenic activity was detected in all of the immunostimulants of the present invention of Examples 1 to 5. Further, it was confirmed that the cocoa husk extract shown in Examples 2, 3, and 4 had higher activity than the cocoa bean extract of Example 1. Furthermore, since the high molecular fraction of the cocoa husk water extract of Example 5 having a molecular weight of 12,000 or more had an increased mitogenic activity, the active component was considered to be a high molecular component.

Next, the food and drink of the present invention formulated according to Examples 6 to 11 will be described. However, the present invention is not limited to these.

[Example 6-Formulation of powder] Lactose 70.0% Potato starch 20.0 Cocoa husk water extract (Example 2) 10.0 100.0%

Example 7-Formulation of tablets D-mannitol 40.0% Lactose 35.0 Microcrystalline cellulose 10.0 Hydroxypropylcellulose 5.0 Cocoa husk water extract (Example 3) 10.0 100.0%

Example 8 Formulation of Chewing Gum Gum Base 20.0% Sugar 54.3 Glucose 15.0 Syrup Cane 9.3 Peppermint Oil 0.4 Cacao Husk 30% Ethanol Extract (Example 4) 1.0 100.0%

Example 9-Candy Formula Sugar 50.0% starch syrup 34.0 citric acid 1.0 peppermint oil 0.2 cacao husk 30% ethanol extract (Example 4) 1.0 water 13.8 100.0%

Example 10-Formulation of Tablet Confectionery Sugar 75.6% Glucose 19.0 Sucrose Fatty Acid Ester 0.2 Cacao Husk Water Extract High Molecular Fraction (Example 5) 2.0 Water 3.2 100.0%

Example 11-Beverage Formula Orange Fruit Juice 30.0% Isomerized Sugar 15.0 Citric Acid 0.1 Vitamin C 0.1 Fragrance 0.1 Cocoa Husk Water Extract High Molecular Fraction (Example 5) 1.0 Water 53.7 100.0%

[0033]

As is apparent from the above description,
According to the present invention, the following effects can be obtained.

The immunostimulant of the present invention has a strong mitogenic activity and has an effect of enhancing the immunity of a living body. In particular, since it is a natural product with high safety, it is an extract using cocoa beans and / or cacao husk, which is the hull of cocoa beans, as a raw material. Moreover, since the raw material itself is inexpensive, it can be easily produced at low cost.

The food and drink of the present invention have a good flavor, and by eating the food and drink, it is possible to ingest an immunostimulatory component on a daily basis, and to reduce immunity caused by aging, stress, fatigue, environmental factors and the like. It can recover and prevent various diseases and infectious diseases.

[Brief description of the drawings]

FIG. 1 shows the results of a mitogen activity test of the immunostimulant of the present invention.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A23L 2/38 A61K 9/20 B A61K 9/20 A23L 2/00 FF term (Reference) 4B014 GB06 GB08 GB13 GG06 GK06 4B017 LC03 LG04 LG14 4B018 MD57 ME14 MF01 4C076 AA12 AA13 AA30 BB01 CC07 DD38A DD67A EE31A EE32A EE38A EE58A FF01 FF11 FF12 GG41 GG44 4C088 AB12 AC04 BA08 BA09 MA10 BA26 BA27 MA27

Claims (5)

[Claims] 1. An immunostimulant comprising an extract obtained from cocoa beans as an active ingredient. 2. An immunostimulant comprising, as an active ingredient, an extract obtained from cacao husk, which is the hull of cacao beans. 3. The immunostimulant according to claim 1, wherein the extract is an extract obtained by extraction with water and / or an organic solvent compatible with water. 4. A cocoa bean and / or a cocoa husk, which is the hull of cocoa beans, obtained by extracting with water and / or an organic solvent compatible with water and having a molecular weight of preferably 1
An immunostimulator comprising an extract of a polymer fraction of 2,000 or more as an active ingredient. 5. A food or drink comprising the immunostimulating agent according to claim 1 and having an immunostimulating action.