JP2003267880A - Composition for antiallergy and antiinflammation - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain a composition for antiallergy and antiinflammation, which has excellent stability and cost, is safe, has excellent effect on allergy and inflammation of allergic rhinitis, pollinosis, etc., and is useful for antiallergy and antiinflammation. <P>SOLUTION: The interleukin 4 production inhibitor comprises Brassica oleracea acephala and/or Brassica oleracea acephala extract. The composition for antiallergy and the composition for antiinflammation comprise the interleukin 4 production inhibitor and have excellent effect on allergies such as allergic rhinitis, pollinosis, etc. These compositions are orally administered or used as skin care preparations. <P>COPYRIGHT: (C)2003,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an interleukin 4 (IL-4) production inhibitor, an anti-allergic and anti-inflammatory composition containing the same, an oral composition thereof, and a skin external composition.

[0002]

2. Description of the Related Art In recent years, various allergies such as hay fever, food allergies and bronchial asthma have become a problem. Generally, hormone agents having a high pharmacological effect are used, but there is a problem that side effects such as adrenocortical dysfunction and gastrointestinal ulcer are large. Also non-steroidal histamine,
Although leukotriene release inhibitors and IgE antibody production inhibitors are also used, their effects are not sufficient.

IL-4 is a glycoprotein produced by antigen-stimulated T lymphocytes. It is known to act on B lymphocytes to enhance the production of antibodies such as IgE, and to promote the infiltration of inflammatory cells into inflammatory sites. In recent years, it has been considered that if the production of IL-4 associated with this allergic disease can be suppressed, allergic disease and inflammation can be treated in a point different from conventional therapeutic methods and preventive methods. .

IL-4 production inhibitors include sulfonium derivatives (Japan. J. Pharmacol., 61, 27-30.
, 1993), and examples of plants having an IL-4 production inhibitory action include extracts such as nettle, thyme, and yarrow (Japanese Patent Laid-Open No. 10-279491) and inchinko (Japanese Patent Laid-Open No. 10-298090). However, these effects are not sufficient. Therefore, there is a need for an IL-4 production inhibitor that is excellent in stability and safety and has no problem in price.

So far, broccoli, cauliflower, nazuna, tincture, cabbage, apples, etc. have IgE.
Increase suppression effect has been reported (JP 2000-169382).
However, the effect of suppressing IL-4 production in vegetables is not known.

[0006]

DISCLOSURE OF THE INVENTION The present invention provides an IL-4 production inhibitor which is excellent in stability, safety and price, an antiallergic composition containing the same, an antiinflammatory composition or oral use thereof. The present invention is to provide a composition and a composition for external use on the skin.

[0007]

Means for Solving the Problems As a result of intensive studies to solve the above problems, the present inventors have found that the production of IL-4, which is considered to be one of the causes of allergic diseases, in kale and its extract. The inventors have found that it has an inhibitory action and completed the present invention.

That is, the present invention is as follows: An interleukin 4 (IL-4) production inhibitor characterized by containing kale and / or kale extract, and an antiallergic composition comprising the interleukin 4 production inhibitor described in 2.1. 3. An anti-inflammatory composition, which comprises the interleukin 4 production inhibitor according to 3.1. 4. Oral interleukin 4 production inhibitor according to 1 or the antiallergic composition according to 2 or the antiinflammatory composition according to 3, and 5. The present invention relates to the interleukin-4 production inhibitor according to 1, the antiallergic composition according to 2, or the antiinflammatory composition according to 3, which is for external use on the skin.

[0009]

BEST MODE FOR CARRYING OUT THE INVENTION The kale (Bras used in the present invention
sicca Oleracea L.var. acephala DC.) includes kitchen kale, malo kale, bush kale, tree kale, collard and green algae. Kale is a plant of the family Cruciferae originally originating in southern Europe, and is said to be the original cabbage seed. The leaves may be those normally used for food, and the cultivation method and cultivation place are not particularly limited.

As kale and its extract, a dried product obtained by drying the kale itself, a crushed product, a squeezed juice, a crude extract with water or an organic solvent such as alcohol, ether or acetone, and a crude extract are distributed, All of the extract fractions obtained by stepwise purification by various chromatography such as column chromatography can be used. These may be used alone or in combination of two or more.

For example, 3 L of 99.5% ethanol extract was added to 1 kg of dried matter such as leaves, stems, flowers and roots of kale,
The extract obtained by soaking overnight at room temperature may be used as it is as an IL-4 production inhibitor, an antiallergic composition, or an antiinflammatory composition, and the extract is combined with various chromatographies. Then, the purified product may be used.

The kale or its extract has an IL-4 production inhibitory action. IL-4 has the action of enhancing the production of antibodies such as IgE and IgG4, and also has the action of promoting the infiltration of inflammatory cells into the inflammatory site. Furthermore, IL-4 also has a function of differentiating immature T lymphocytes into mature T lymphocytes, and mature T lymphocytes produce more IL-4 than immature T lymphocytes. Moreover, IL-4 is also involved in skin troubles, especially in skin inflammation. Therefore, the IL-4 production inhibitor is effective in the prevention and treatment of allergic diseases such as atopic dermatitis. Further, it can be used for suppressing wrinkles, sagging, itching and the like, which are skin troubles associated with IL-4.

Kale of the present invention, I containing the extract thereof
The L-4 production inhibitor, the antiallergic composition, and the antiinflammatory composition can be produced as an oral food or medicine. It can also be manufactured as a cosmetic or medicine for external use on the skin.

As a method of applying as a medicine, either oral administration or parenteral administration can be adopted. Upon administration, the active ingredient can be mixed with a solid or liquid non-toxic pharmaceutical carrier suitable for administration methods such as oral administration, rectal administration and injection, and then administered in the form of a conventional pharmaceutical preparation.
Examples of such preparations include solid preparations such as tablets, granules, powders and capsules, liquid preparations such as solutions, suspensions and emulsions, and freeze-dried preparations. It can be prepared by conventional means. Examples of the non-toxic carrier for pharmaceutical use include glucose, lactose, sucrose, starch, mannitol, dextrin, fatty acid glyceride, polyethylene glycol, hydroxyethyl starch, ethylene glycol, polyoxyethylene sorbitan fatty acid ester, amino acid, gelatin, Albumin, water, physiological saline and the like can be mentioned. Further, if necessary, conventional additives such as a stabilizer, a wetting agent, an emulsifier, a binder and an isotonicity agent can be appropriately added.

As a food, a kale dried product, an extract or the like as it is, or by adding various nutritional components or by being contained in foods and drinks, is useful as a food or food material for health useful for treating and preventing allergic symptoms. Can be used as For example, starch, lactose, maltose, vegetable oil and fat powder, cocoa butter powder,
After adding a suitable auxiliary agent such as stearic acid, it may be formed into a form suitable for oral use, for example, granules, granules, tablets, capsules, pastes, etc., by using a conventional means and then provided for edible use. Also, various foods, for example, processed meat products such as ham and sausage, seafood processed foods such as kamaboko and chikuwa, bread, confectionery, butter, milk powder, fermented dairy products, water, fruit juice, milk. It may be used by adding it to a beverage such as a soft drink. The blending amount of the present invention is appropriately set depending on the type and condition of the edible composition.

The effective application amount of the kale of the present invention can be appropriately determined according to the age, weight, symptom, application route, application schedule, formulation form of the ingestor, and, for example, in the case of oral administration, as a dry weight. , Usually 0.0
001 to 0.5 g / kg, more preferably 0.001
It is about 0.2 g / kg. It may be administered in several divided doses a day.

Further, the kale of the present invention, an IL-4 production inhibitor containing the extract thereof, an antiallergic composition, and an antiinflammatory composition are produced in the form of cosmetic materials or cosmetics containing them. can do. For example, a dried product or an extract of kale can be added to wheat germ oil or olive oil and used as a cosmetic material.

Further, dried kale or extract can be directly used as a cosmetic ingredient to produce a cosmetic. Cosmetics include oils and fats such as vegetable oils, higher fatty acids, higher alcohols, silicones, anionic surfactants, cationic surfactants, amphoteric surfactants, nonionic surfactants, preservatives, sugars, sequestering agents. , A polymer such as a water-soluble polymer, a thickener, a powder component, an ultraviolet absorber, an ultraviolet blocking agent, a moisturizer such as hyaluronic acid, a fragrance, a pH adjusting agent, a desiccant, etc. can be contained. .
Vitamins, skin stimulants, blood circulation promoters, indigenous bacteria control agents, active oxygen scavengers, anti-inflammatory agents, anti-cancer agents, whitening agents,
Other medicinal ingredients such as bactericides and physiologically active ingredients can also be contained.

As the cosmetics, skin cosmetics such as lotion, emulsion, cream, pack, etc., makeup base lotion, makeup cream, emulsion or cream-like or ointment type foundation, lipstick, eye color,
It can be used as makeup cosmetics such as blush color, body creams such as hand creams, leg creams and body lotions, bath salts, oral cosmetics and hair cosmetics. As the cosmetic containing the composition containing the IL-4 production inhibitory effect obtained by the method of the present invention, from the viewpoint of function, for example, emulsions, cosmetics, face creams, hand creams, lotions, essences and the like are preferable.

Such cosmetics can be manufactured by a conventional method. The amount of the dried product or extract of the plant of the present invention in the cosmetic is not particularly limited, but as an example, it is about 0.01 to 20% by weight of the total weight of the cosmetic in terms of dry weight. Appropriate.

Since the plant of the present invention is a natural product, its toxicity is low and no harmful side effects have been reported. Also,
Since kale is a vegetable that can be obtained by cultivation, kale, the composition of the present invention having a kale extract as an active ingredient,
It is safe and excellent in terms of price.

[Production Example] Kale leaves were dried at 90 ° C. to inactivate the enzyme which is a bitter and astringent ingredient. While heating and refluxing 4 kg of the pulverized product in an electrically heated water bath,
Extraction was performed using 20 L of ethanol (Wako Pure Chemical Industries, Ltd.) to obtain 80 g of kale extract.

[Test Example] The test for the ability to suppress IL-4 production was carried out using En
Based on the method of do et al. (Int archAllergy Immunol 19
93; 101: 425-430). The amount of IL-4 produced was measured using peripheral blood mononuclear cells (PBMC) of asthma patients and healthy subjects.
PBMC were separated from venous blood by density gradient centrifugation.
The obtained PBMC was treated with 10% FCS and 20 μg / ml PHA (phytohemag
glutinin) in RPMI1640 medium
Plated in a 96-well plate (2 x 10 ⁵ cells / 200
μl). At that time, the kale extract obtained in Production Example was added. Dexamethasone was used as a positive control. The culture was carried out at 37 ° C. in the presence of 5% CO ₂ for 5 days. Then, the culture supernatant was centrifuged at 4 ° C. and 400 × g, and the supernatant fraction was subjected to quantification by the ELISA method.
Plate wells with 10 μg / ml mouse anti-human IL-4
Monoclonal antibody was added and incubated overnight at 4 ° C. Then, after blocking with 1% BSA for 30 minutes at room temperature, the dexamethasone fraction and the kale extract fraction were added to the wells and incubated overnight at room temperature. Then, using a rabbit anti-human IL-4 polyclonal antibody,
Incubated at room temperature for 4 hours. Furthermore, after incubation with a goat anti-rabbit IgG antibody for 4 hours, tetramethylbenzidine was added and the absorbance was measured at 450 nm.

The results are shown in Table 1. The inhibitory rate of IL-4 production in the kale extract [100− (IL-4 production amount when the kale extract was added / IL-4 production amount of the control of the culture solution only) × 100] was calculated, and from this, IC was calculated.
_{A 50} value (concentration that inhibits IL-4 production by 50%) was determined. The IC ₅₀ value of dexamethasone, which is a positive control in this experimental system, was 1.46 μg / ml.

[0025]

[Table 1]

The results in Table 1 show that the kale extract has an excellent IL-4 production inhibitory action.

(Formulation Example 1) [Production of tablet] Using the ethanolic extract of kale obtained in the production example, a tablet having the following composition was produced according to a conventional method. (Composition) (Composition: wt%) Kale extract 24 Lactose 63 Corn starch 12 Guar gum 1

A double-blind test is carried out using this tablet to examine the preventive and palliative effects of hay fever. For cedar pollinosis patients, tablets are administered for 10 weeks, divided into a test product intake group and a placebo intake group. The intake is 5 in each group
Granules / day, lactose was added as a component of the placebo, and the entire tablet was 100%. During the test period, the subject is asked to fill out an allergy diary, and the effects are confirmed by evaluating the symptoms based on the "Allergic rhinitis severity classification plan" of the Japanese Society of Allergology.

(Formulation Example 2) [Production of juice] Using the ethanolic extract of kale obtained in the production example, a juice having the following composition was produced according to a conventional method. (Composition) (Composition: wt%) Frozen concentrated Unshu mandarin orange juice 5.0 5.0 Fructose glucose liquid sugar 11.0 Citric acid 0.2 L-Ascorbic acid 0.02 Perfume 0.2 Pigment 0.1 Kale extract 0. 2 Water 83.28

A double-blind test is carried out using this juice as a test product to examine the preventive and palliative effects of hay fever. For cedar pollinosis patients, we divided into test product intake group and placebo intake group,
Administration is performed for 10 weeks. Juice intake is 3 for each group
The amount of water was increased to 100 ml of juice as a placebo component. During the test period, the subjects are asked to complete an allergy diary, and their symptoms are evaluated based on the "Allergic rhinitis severity classification plan" of the Japanese Society of Allergology to confirm the symptom improving and alleviating effects of hay fever.

(Formulation Example 3) [Production of face cream] Using the ethanolic extract of kale obtained in the production example, a face cream having the following composition was produced in accordance with a conventional method. (Composition) (Composition: wt%) Isopropyl isostearate 8.0 Jojoba oil 6.0 Cetanol 8.0 Stearyl alcohol 2.0 Polyoxyethylene lauryl ether 1.5 Propylene glycol 6.0 Sorbitol 1.0 Paraben 0.0 4 Kale extract 0.5 Vitamin E 0.5 Fragrance 0.1 Purified water 66.0

A double-blind test is conducted using this face cream as a test product to examine the effect of improving atopic dermatitis. Atopic dermatitis patients are divided into a test product intake group and a placebo intake group, and a 10-week test is conducted. The amount of cream used is about 5g / day for each group, and the amount of purified water is increased to 100% for the whole cream as a placebo ingredient.
And Eruptions are scored on a scale of 5 with "most severe 4, severe 3, moderate 2, mild 1, none 0", and the degree of itching is evaluated on a scale of 10. By analyzing these evaluations, the atopic dermatitis improving effect of the face cream is confirmed.

[0033]

EFFECTS OF THE INVENTION The present invention can provide an interleukin production inhibitor having an action of suppressing the production of interleukin-4, which is one of inflammatory cytokines, and can provide allergic rhinitis, allergy such as hay fever, and the like. It is possible to provide a composition which has an excellent effect on inflammation and is useful for anti-allergy and anti-inflammatory purposes.

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61P 17/00 A61P 17/00 29/00 29/00 37/08 37/08 43/00 111 43/00 111 F term (reference) 4C083 AA111 AA112 AA122 AC072 AC132 AC182 AC342 AC482 AD662 BB41 BB51 CC02 CC03 CC04 CC05 CC07 CC11 CC12 CC13 CC14 CC25 CC31 CC41 DD14 DD15 DD16 DD17 DD21 DD22 DD23 DD31 DD41 EE07 EE10 EE12 AC032 BA09 BA10 CA05 CA06 CA07 CA14 MA17 MA22 MA23 MA35 MA37 MA41 MA43 MA44 MA52 MA60 MA63 MA66 NA06 NA14 ZA34 ZA59 ZA89 ZB13 ZC41

Claims (5)

[Claims] 1. An agent for suppressing interleukin 4 production, which comprises kale and / or kale extract. 2. An antiallergic composition comprising the agent for suppressing interleukin production according to claim 1. 3. An anti-inflammatory composition comprising the agent for suppressing interleukin production according to claim 1. 4. The agent for suppressing interleukin 4 production according to claim 1, which is for oral use, the antiallergic composition according to claim 2, or the antiinflammatory composition according to claim 3. 5. The agent for suppressing interleukin 4 production according to claim 1, which is for external use on the skin, the antiallergic composition according to claim 2, or the antiinflammatory composition according to claim 3.