JP2000239175A - Antiallergic agent - Google Patents

Abstract

(57) [Problem] To provide an antiallergic agent which is derived from natural products, has no side effects, has high safety, and can be used in foods and drinks such as pharmaceuticals, foods for specified health use, health foods and the like. The antiallergic agent is characterized by containing sour milk whey, and is effective for immediate allergy or delayed allergy, and is orally and / or transdermally administered.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION The present invention relates to a medicine, which is effective against inflammation such as allergic rhinitis and atopic dermatitis.
The present invention relates to an antiallergic agent that can be used as a food for specified health use, a health food, and the like.

[0002]

2. Description of the Related Art In recent years, the number of patients suffering from allergic diseases has been increasing due to changes in the living environment due to an increase in the dietary habits and the opportunity to come into contact with chemical substances, etc. Above all, the increase in hay fever and atopic dermatitis is remarkable, and has become a social problem.

[0003] Allergic reactions are classified into four types from type I to type IV depending on the mechanism. Among them, allergic diseases such as allergic rhinitis, bronchial asthma and urticaria usually occur due to type I allergic reaction. The type I allergic reaction is also called immediate allergy, and mast cells in tissues and Fc on the surface of blood basophils.
Allergen-specific IgE antibody binds to ε receptor,
Then, when the allergen binds to the IgE antibody, chemical mediators such as histamine and leukotriene are excessively released from mast cells and basophils, and various allergic reactions are excited. Therefore, for the treatment of type I allergic diseases, antihistamines and antiallergic agents having an action of suppressing the release of chemical mediators from mast cells are used. However, such antihistamines and antiallergic agents often have side effects, and safety is a problem when used continuously for a long period of time.

[0004] Diseases such as allergic contact dermatitis are usually caused by a type IV allergic reaction also called delayed allergy. Steroids are used to treat type IV allergic disease. Steroids suppress cytokine production and are particularly effective in treating eczema. However, when used in large quantities or for a long period of time, there is a high possibility of causing serious side effects, and safety is a problem.

On the other hand, it has been attempted to improve allergic diseases by consuming a large amount of general foods showing an antiallergic effect instead of administration of these antiallergic drugs, for which safety is a problem. ing. As general foods having an antiallergic effect, perilla extract, bean tea extract, and the like are known, and the market of allergy reducing foods to which these are added is expanding. However, these are not effective in fundamentally improving allergy. In addition, it is known that lactic acid bacteria cells have an inhibitory effect on IgE production and an antiallergic effect in vitro (Japanese Patent Application Laid-Open No. 9-1997).
No. 2959) has not yet been put to practical use as an antiallergic agent.

[0006] Therefore, there is a long-awaited demand for foods which are safe, have no side effects, can be eaten every day, and have an effective antiallergic effect.

[0007]

An object of the present invention is to provide an antiallergic agent which is derived from natural products, has no side effects, is highly safe, and can be used in foods and drinks such as pharmaceuticals and foods for specified health use and health foods. Is to do.

[0008]

According to the present invention, there is provided an antiallergic agent characterized by containing sour milk whey.

Further, according to the present invention, there is provided the above antiallergic agent which is effective for immediate allergy or delayed allergy.

Further, according to the present invention, there is provided the above-mentioned antiallergic agent administered orally and / or transdermally.

[0011]

BEST MODE FOR CARRYING OUT THE INVENTION The antiallergic agent of the present invention contains sour milk whey.

As the sour milk, fermented milk obtained by fermenting a raw material for fermentation using a fermenting bacterium containing various lactic acid bacteria can be used. As the lactic acid bacterium, specifically, for example, Lactobacillus helveticus can be used. Also, fermentation can be performed by mixing and culturing this with other lactic acid bacteria species or co-cultivating with yeast.

As the Lactobacillus helveticus, specifically, for example, Lactobacillus helveticus JCM1003, JCM1006, JCM106
2, ATCC 15009, ATCC 12046, ATC
C8001, ATCC10797, ATCC12046
And the like. Particularly, the JCM1003 strain is preferable.

The fermentation raw materials include animal milk and vegetable milk. Examples of the animal milk include whole-fat milk, skim milk, whey, powdered milk, and reduced milk, and examples of the vegetable milk include those derived from soybeans. As the fermentation raw material, yeast extract, vitamins, amino acids, salts, saccharides (oligosaccharides such as glucose, sucrose or raffinose, stachyose, etc.), stabilizers and the like are optionally added as necessary as secondary components. What was done can be used.

The fermentation of the fermentation raw material can be performed, for example, as follows. The fermentation raw material is sterilized by heating, and after cooling, a starter prepared by preculturing the fermentation bacterium such as Lactobacillus helveticus in advance is added. The amount of the starter to be added was 10 ⁵
The number can be from 10 ⁷ / ml to 10 ⁷ / ml. The fermentation temperature can be 25 to 50 ° C, preferably 30 to 45 ° C, and the culture time can be 3 to 48 hours, preferably 8 to 24 hours. Lactic acid bacteria count is more than 10 ⁸ / ml, pH
The fermentation can be stopped at the point of time of 4.0 or less, and the fermented milk can be obtained.

The obtained fermented milk is sterilized by a method such as heating the lactic acid bacteria alive or at a temperature of 80 ° C. or the like, and then subjected to a centrifugal separation operation or the like to obtain an acid-coagulated casein protein. The sour milk whey from which is removed is obtained in a separated state, and can be used as a component of the antiallergic agent of the present invention. Alternatively, the obtained fermented milk can be used as it is as a material containing acid milk whey without separation as a component of the antiallergic agent of the present invention. Further, the acid milk whey or the material containing the acid milk whey is concentrated by decompression concentration or the like, or dried by a method such as freeze-drying or spray drying to obtain a slurry or powdered processed product, and the antiallergic substance of the present invention is obtained. It can also be used as a component of an agent.

The antiallergic agent of the present invention can be composed of only the above-mentioned sour milk whey, the above-mentioned fermented milk, the above-mentioned processed product, or a mixture thereof. You can also. Specifically, for example, one or more extenders can be included. In addition, food ingredients such as sugars, proteins, lipids, vitamins, minerals, pigments, and flavors and / or food additives can be included. The mixing ratio of the acid milk whey in the antiallergic agent of the present invention can be appropriately selected, and is usually 5 to 100.
A weight percent range is appropriate.

The form of the antiallergic agent of the present invention may be any of fermented milk products such as yogurt, cheese and lactic acid bacteria beverages, processed foods and drinks containing fermented milk, capsules, granules, tablets and the like. it can. In addition, as a dosage form such as an ethanol solution, it can be administered transdermally by application to the skin surface or the like, and it can also be administered in combination with these administration methods.

The dose of the antiallergic agent of the present invention can be appropriately adjusted depending on the age and symptoms of the patient. In the case of oral administration, for example,
The effect of the present invention can be obtained by adjusting the amount to 0.1 g / kg / day or more, preferably 10 g / kg / day or more.

[0020]

The antiallergic agent of the present invention is derived from natural foods, has no side effects, is highly safe, has high nutritional value, has a good taste, and has inflammation and allergic rhinitis derived from allergic reactions. It can improve, suppress or prevent symptoms of allergic diseases such as atopic dermatitis and the like, and is useful as an antiallergic agent which can be used in pharmaceuticals and foods and drinks such as foods for specified health use and health foods.

[0021]

The present invention will be described in more detail with reference to the following Examples, but it should not be construed that the invention is limited thereto.

The antiallergic effect of acid milk whey was examined.

[0023]

Example 1 An ICR mouse was used as an experimental animal, and active abdominal anaphylaxis (ovalbumin, OA) was used as an antigen.
AA. ) A test was performed to evaluate the immediate allergic suppression effect of acid milk whey.

(Preparation of samples etc.) Preparation method of test feed: Lactobacillus helveticus JCM1003 strain and Saccharomyces cerevisiae A in 1 kg of skim milk (solid content 9% by weight) sterilized at 90 ° C.
40 g of a symbiotic starter fermented solution pre-cultured with TCC2565 was inoculated and cultured at 37 ° C. for 24 hours to obtain an acid milk. 7000 g of this sour milk was centrifuged for 10 minutes to remove solids, and 902 g of the supernatant (whey) obtained was freeze-dried to prepare 60.5 g of sour milk whey powder. Using the whey powder, a 10% by weight sour milk whey meal having the composition shown in Table 1 was prepared.

As a feed to be administered to the control group, a control diet containing no sour milk whey as shown in Table 1 was prepared.

[0026]

[Table 1]

Preparation of stock solution of allergen: Ovalbumin (Chicken egg; grade V, SIGMA CHE)
MICAL CO. (USA)) in sterile saline at 2.0 mg /
It was dissolved at a concentration of ml and filtered through a sterilized 0.45 μm filter (manufactured by Toyo Roshi Kaisha, Ltd.) to obtain an allergen stock solution.・ Preparation of sensitive allergen emulsion: The above allergen stock solution and Freund's incomplete adjuvant (DIFCO LABORAT)
ORIES (USA)) in a volume ratio of 1: 1 and emulsified to obtain a sensitive allergen emulsion. Induction allergen solution: The above allergen stock solution was diluted 20 times with sterile physiological saline to obtain an allergen solution for induction. Evans blue solution: Evans blue (manufactured by Wako Pure Chemical Industries, Ltd.) was dissolved in sterile physiological saline to a final concentration of 1% by weight or 0.5% by weight, respectively, and filtered with filter paper. The filtrate was further filtered with a 0.2 μm sterile filter (manufactured by Toyo Roshi Kaisha, Ltd.).

(Evaluation of immediate allergic suppression effect) Male I
5 CR-aged mice (n = 5, 2 groups; SLC Japan) were used. During the pre-breeding period, the solid feed (CE-
2; made by Nippon Clea Co., Ltd.) and water.
After three days of preliminary breeding, a group of mice was intraperitoneally in the lower abdomen,
50 μl of the sensitive allergen emulsion per animal
1 injection and ovalbumin sensitization was performed. After sensitization,
The sour milk whey diet and water were allowed ad libitum. As a control, another group of mice were treated similarly except that they received a control diet instead of the acid milk whey diet. Hereinafter, the group receiving the sour milk whey diet is referred to as a sample diet group, and the group receiving the control diet is referred to as a control diet group.

After sensitization to ovalbumin, each of the test feeds was fed freely for 12 days, and then the mice of each group were injected with 100 μl of the 1% by weight Evans blue solution into the tail vein.
1 injection. Subsequently, ether anesthesia was performed, and the skin of the abdomen was immediately peeled off to expose the abdominal wall. Six minutes after the administration of the Evans blue solution, the allergen solution for induction 5 was placed in the abdominal wall.
0 μl was injected. Exactly 7 minutes after the intraperitoneal injection, the mice were sacrificed by cervical dislocation and the abdominal wall containing the pigment extravasation site was cut off. The excised abdominal wall was attached to a transparent petri dish, the major axis and minor axis of the pigment extravasation site were measured, and the area was calculated. The average value and standard error of the experimental results of each group were determined, and St
A significant difference test between groups was performed by the t-test of the dent. The results are shown in FIG. As is clear from FIG. 1, the extravasation of the pigment was significantly suppressed in the sample diet group as compared with the control diet group. That is, production of a specific IgE antibody against ovalbumin used as an allergen was suppressed.

[0030]

Example 2 Passive cutaneous anaphylaxis using ovalbumin as an antigen (hereinafter referred to as P)
Called CA. ) A test was conducted to evaluate the immediate allergic suppression effect of acid milk whey.

In the same manner as in Example 1, two groups of mice (n = 5 mice) were sensitized with ovalbumin and allowed to freely take an acid milk whey diet or a control diet and water. After the end of the 12-day free intake period, blood was collected from the mice using a glass capillary tube (manufactured by Terumo Corporation) by a method of collecting blood from the veins of the eyes, and mouse serum was obtained by a known method.

Nine aged female Sprague-Dowley (SD) rats, each group of 10 rats (Japan SLC Co., Ltd.) were anesthetized, and the back hair was carefully shaved with an electric clipper. 0.1 ml of the above-mentioned mouse serum diluted at various dilution rates was injected intradermally at the shaved site of the rat. After standing for 24 hours, ovalbumin (Chicken egg; grade V, SIGMA CHEMICAL
CO. (USA)) in sterile physiological saline 10.0 mg / ml
1 ml of a mixed solution of 0.3 ml of the PCA allergen solution and 2.7 ml of the 0.5% by weight Evans blue solution, which was dissolved at a concentration of 0.35 μm and filtered through a 0.45 μm sterilized filter (manufactured by Toyo Roshi Kaisha, Ltd.) Rats were injected via the tail vein and the rats were sacrificed 30 minutes after injection. The back skin of the rat was peeled off, the major axis and minor axis of the blue stained circle on the inner surface of the skin were measured, and the average diameter and area were calculated. The average diameter of 5 mm or more was defined as positive, and the positive and negative at each dilution ratio of the sample diet group and the control diet group were determined. Table 2 shows the results. The mean value and standard error of each group were determined for the area of the blue stained circle in the experiment with a dilution ratio of 1/4, and a significant difference test between groups was performed by Student's t-test. The results are shown in FIG.

[0033]

[Table 2]

As is apparent from FIG. 2, the extravasation of the pigment was significantly suppressed in the sample diet group as compared with the control diet group. That is, production of a specific IgE antibody against ovalbumin used as an allergen was suppressed.

[0035]

Example 3 Inhibitory Effect of Acid Milk Whey on Type IV Allergy Female ICR 6-week-old Mice (n = 5, 2 groups; Japan SLC
Co., Ltd.). These mice were given solid feed (C
E-2) and water were ingested freely and preliminarily reared for 3 days.
After the preliminary breeding period, 150 μl of an ethanol solution containing 3% by weight of oxazolone was applied to the abdomen, and eating of the test feed was started at the same time. As the test feed, the same sample food or control food as in Example 1 was used for each group, and the test feed and water were freely taken. Seven days after the sensitization, an ethanol solution containing 1% by weight of oxazolone was applied to the auricle of the mouse to cause auricle-contact dermatitis. The thickness of the pinna 24, 48 and 72 hours after application is measured using a dial thickness gauge (R1-A, Ozaki MF
G, Tokyo, Japan), and the difference from the auricle thickness measured before induction of dermatitis was determined. The average value and standard error of each measurement result of the sample diet group and the control diet group were determined, and a significant difference test between groups was performed by Student's t-test. The results are shown in FIG.

As apparent from FIG. 3, ear edema due to oxazolone was significantly suppressed in the sample diet group as compared with the control diet group. That is, the inhibitory effect on type IV allergy was confirmed.

[0037]

Example 4 Inhibitory Effect of Type IV Allergy by Applying Acid Milk Whey For delayed allergy, an ethanol solution such as acid milk whey was applied to the auricle in advance before allergy induction, and the antiallergic effect of the application was examined.

Two groups of mice of the same species and same age as those used in Example 3 were used. These mice were given chow (CE)
-2) and water were given freely, and the animals were preliminarily reared for 3 days. After the pre-breeding period, 150 μl of an ethanol solution containing 3% by weight of oxazolone was applied to the abdomen to sensitize. 7 from sensitization
A day later, a 25% by volume aqueous ethanol solution containing 5% by weight of the sour milk whey powder obtained in Example 1 was applied as a control application group by applying 10 μl of a 25% by volume aqueous ethanol solution to the front and back of each ear of a group of mice. The sample was similarly applied to another group of mice to obtain a sample application group. Ten minutes after the application, 10 μl of an ethanol aqueous solution containing 1% by weight of oxazolone was applied to the front and back of the pinna of the mouse to cause pinna contact dermatitis. The thickness of the pinna 24, 48 and 72 hours after this application is measured with a dial thickness gauge
(R1-A, Ozaki MFG, Tokyo, Japan), and the difference from the pinna thickness measured before dermatitis induction was determined. The average value and standard error of each measurement result of the sample application group and the control application group were determined, and the Student's t-
A significant difference test between groups was performed by a test. FIG. 4 shows the results.

As is clear from FIG. 4, ear edema due to oxazolone was significantly suppressed in the sample application group as compared with the control application group. That is, the inhibitory effect on type IV allergy was confirmed.

[Brief description of the drawings]

FIG. 1 is a graph showing the difference in production of specific antigen in an AAA reaction test using ovalbumin in a sample diet group mouse and a control diet group mouse that ingested an acid milk whey diet in Example 1. is there.

FIG. 2 is a graph showing a difference in specific antigen production in a PCA reaction test using ovalbumin between a sample diet group mouse and a control diet group mouse that ingested an acid milk whey diet in Example 2. is there.

FIG. 3 is a graph showing the difference in the degree of auricle-contact dermatitis caused by oxazolone in a sample diet group mouse and a control diet group mouse ingesting an acid milk whey diet in Example 3.

FIG. 4 is a graph showing the difference in the degree of auricle-contact dermatitis caused by oxazolone in the mouse to which the sample was applied and the mouse to which the control was applied in Example 4.

Continued on the front page F term (reference) 4B001 AC06 AC31 AC32 BC14 EC05 4B018 LB07 LE01 LE02 LE03 LE04 MD81 MD86 MD91 ME07 MF13 4C087 AA01 AA02 BB39 CA07 MA01 MA52 MA55 NA14 ZB13

Claims (3)

[Claims] 1. An antiallergic agent comprising sour milk whey. 2. The antiallergic agent according to claim 1, which is effective for immediate allergy or delayed allergy. 3. The antiallergic agent according to claim 1, which is administered orally and / or transdermally.