WO2005092357A1

WO2005092357A1 - Antiallergic agent and antiinflammatory agent

Info

Publication number: WO2005092357A1
Application number: PCT/JP2005/005181
Authority: WO
Inventors: Hiroko Isoda; Shinichi Yokota; Yukuo Abe
Original assignee: Kaneka Corporation
Priority date: 2004-03-29
Filing date: 2005-03-23
Publication date: 2005-10-06
Also published as: JPWO2005092357A1

Abstract

An antiallergic agent, an antiinflammatory agent, a basophil degranulation inhibitor, a mast cell degranulation inhibitor or a β-hexosaminidase release inhibitor containing a plant belonging to the genus Lycium or a polar solvent extract thereof. By using the plant belonging to the genus Lycium which is a natural material having been taken by humans for a long time, an antiallergic agent and an antiinflammatory agent with a high safety can be provided by a convenient method. Moreover, these antiallergic agent and antiinflammatory agent can be added to foods, drinks, cosmetics, bathing preparations, drugs and so on.

Description

Specification

Antiallergic and anti-inflammatory agents

Technical field

The present invention relates to an anti-allergy agent and an anti-inflammatory agent that can be used as foods and drinks for humans or animals, pharmaceuticals, and cosmetics.

Background art

[0002] Living organisms are inherently provided with an immune function for eliminating extraneous substances from the outside and maintaining homeostasis. However, this immune function sometimes acts to damage the body, and a type of disorder reaction caused by this excessive immune function is particularly called allergy. Allergies are activated basophils, mast cells, T-lymphocytes, B-lymphocytes, etc. due to an over-reaction of immunocompetent cells with a causative substance derived from microorganisms, plants, animals, drugs, chemicals, foods, etc. Tissues in the body are damaged by physiologically active substances released from the body. The allergy responds quickly to the onset of the allergic reaction, depending on the time it takes for the reaction to occur. It is divided into progressive delayed allergy.

[0003] Basophils present in blood and mast cells present around blood vessels and connective tissues have granules containing histamine and the like, and degranulation of these cells causes / 3-hexosaminidase and the like together with histamine. It has been known to release enzymes and induce immediate allergic reactions such as asthma and atopic eczema.

[0004] Note that an allergic disease is not the same as an atopic disease. Allergy is involved in some atopic diseases, but microbial infections and genetic factors play a complex role in the onset of atopic diseases, as typified by atopic dermatitis .

[0005] In addition, substances released by degranulation of basophils and mast cells are well known as substances causing inflammation.

[0006] Thus, it is considered that mast cells and basophils can reduce allergy by inhibiting degranulation of these cells, which are strongly involved in immediate allergy. Therefore, an effective basophil cell degranulation inhibitor or mast cell degranulation inhibitor It is desirable to develop agents to provide useful antiallergic agents.

[0007] Anti-allergic agents include agents that suppress degranulation from mast cells and release of chemical mediators in type I allergic reactions that are immediate allergies, and agents that inhibit the binding of histamine to HI receptors. Drugs that inhibit the lipid-type chemical mediator thromboxane A2 and drugs that suppress leukotriene have been developed, and have side effects such as central nervous system depression such as drowsiness and sedation, and cystitis-like symptoms such as hematuria. No drug is satisfactory enough. In particular, the safety of antiallergic drugs is particularly important because they are often used not only for treatment but also for prophylactic purposes over a long period of time.

[0008] Lycium plants are plants of the Solanaceae family, the leaves of which are called licorice leaves, the fruits are licorice leaves, and the root bark is skeletal bark. It is said to have the effect of preventing presbyopia.

[0009] A wolfberry plant contains β-sitosterol, linolenic acid, zeaxanthin, betaine and fuesaliene, in addition to carotenes and vitamins. As a pharmacological effect, it has been reported that betaine has an effect on lipid metabolism and anti-fatty liver, and an anti-fatty liver action of a water extract on the liver is known.

[0010] Regarding the antiallergic effect of wolfberry, a 5-lipoxygenase activity inhibitor containing an ingredient that is easily soluble in a low-polarity organic solvent as an active ingredient and an antiallergic food containing the same have been reported. The components have obvious anti-allergic effects, and they have been identified (Patent Document 1).

[0011] Furthermore, the present invention refers to the effect on the atopic dermatitis, which is reported to have an effect on the atopic disease of a wolfberry plant, and its basophil cell degranulation inhibitory activity or mast cell degranulation. No antiallergic effect due to inhibitory activity was found (Patent Document 2). In addition, there has been no report that basophil cell degranulation inhibitory activity, mast cell degranulation inhibitory activity, and / 3-hexosaminidase release inhibitory activity are caused by a wolfberry plant or a polar solvent thereof.

Patent Document 1: JP 2001-233778

Patent Document 2: JP 2004-67526

Disclosure of the invention Problems the invention is trying to solve

[0012] An object of the present invention is to provide a new antiallergic agent and an antiinflammatory agent which can be easily produced with high safety and can be blended in cosmetics and foods and drinks.

Means for solving the problem

[0013] The present inventors have conducted intensive studies to solve the above-mentioned problems, and as a result, an extract obtained by extracting a Lycium plant with a polar solvent has a β-hexosaminidase inhibitory activity, The present inventors have found that they have a basophil cell degranulation inhibitory activity and a mast cell degranulation inhibitory activity, and have an antiallergic action and an anti-inflammatory action, thereby completing the present invention.

[0014] That is, the present invention relates to an antiallergic agent comprising a Lycium plant or an extract thereof as an active ingredient.

[0015] The present invention also relates to a basophil cell degranulation inhibitor containing a Lycium plant or an extract thereof.

[0016] The present invention also relates to a mast cell degranulation inhibitor comprising a Lycium plant or an extract thereof.

[0017] The present invention also relates to a j3-hexosaminidase release inhibitor, which comprises an extract of a Lycium plant.

[0018] The antiallergic agent and the antiinflammatory agent of the present invention may be either a preventive agent or an ameliorating agent, or may be a combination of both.

The invention's effect

[0019] The wolfberry extract described in the present specification is useful as an antiallergic agent and an antiinflammatory agent. Since the wolfberry extract of the present invention is highly safe and can be prepared by a simple method, it is highly applicable to pharmaceuticals, foods and drinks, cosmetics, bath preparations, etc., and its application range is extremely wide. Expected.

Brief Description of Drawings

FIG. 1 is a graph showing a dilution ratio of a wolfberry hot water extract and an inhibition rate of β-hexosaminidase release of human basophils. BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, embodiments of the present invention will be described in detail.

[0022] As described above, the genus Lycium is a plant belonging to the Solanaceae family, and is a species of Lycium chinense (also referred to as Lycium chmensis: ¾ may be self-prone), Lycium barb arum Lycium turcomanicum, Lycium potaninii, Lycium dasystemum, etc. Exists. The wolfberry plant used for the antiallergic agent, antibasophil cell degranulation inhibitor, mast cell degranulation inhibitor or β-hexosaminidase release inhibitor of the present invention is not particularly limited as long as it belongs to the genus Lycium. chmense3; 7 lysium Lycium barbarum power is preferred, Lycmm chinense power is nuna mash.

[0023] The method for producing a Lycium plant used in the present invention is not particularly limited. The wolfberry plant used in the present invention may be not only a naturally grown plant but also an artificially cultivated plant. In addition, there are no particular restrictions on the timing of harvesting, growing years, cultivation method, and cultivation period of the wolfberry plant used in the present invention.

[0024] The wolfberry plant used in the present invention may be used by selecting only one kind from among various kinds of production methods, or may be used by combining a plurality of kinds.

[0025] An extract of a wolfberry plant is obtained by extracting various parts of a plant body (whole plant, flower, leaf, etc.) as they are or after pulverizing them and extracting them with an extraction solvent. The portion used for extraction is not particularly limited, but is more preferably a fruit (Guyuko), which is favored by leaves (Gyugia leaves), fruits (Guyako), and root bark (skeletal bark).

[0026] As the extraction solvent, a polar solvent can be used. Examples of the polar solvent include polar organic solvents such as water and alcohols (eg, lower alcohols such as ethanol and methanol, and polyhydric alcohols such as propylene glycol). Any combination of more than one species can be used. Preferably, it is water alone or a mixed solvent of water and a polar organic solvent.

[0027] In the case of a mixed solvent of water and a polar organic solvent, the mixing ratio of water and the polar organic solvent is not particularly limited, but it is preferable that the main component is water, that is, the volume ratio of water is 50% or more. As the polar organic solvent to be mixed with water, methanol or ethanol is more preferable, since lower alcohols are preferable.

[0028] In the case of extracting mainly with water, the components to be extracted are different because the polarity as the solvent is different as compared with the case of extracting mainly with an organic solvent. The amount of extraction solvent used 0.5 weight to 50 weight is preferable to the genus plant, and 2 weight to 10 weight is more preferable.

[0029] The extraction method is not particularly limited. Usually, extraction may be performed at normal temperature and normal pressure using an extraction solvent. In this case, the extraction time is not particularly limited. 14 days is preferred. After extraction, it may be concentrated, dried, or made into a solution, paste, gel, or powder with oils and fats. In some cases, extraction can be performed under a condition of 30 to 120 ° C., or extraction can be performed under supercritical conditions using an autoclave, carbon dioxide, or the like. The extraction time may be appropriately set. If necessary, it can be further purified by an optional operation using an activated carbon column or an ion exchange resin.

[0030] In one embodiment of the present invention, an extract obtained by extracting a wolfberry plant with hot water can be used. The amount of hot water used for the extraction is preferably an amount that can sufficiently dissolve the active ingredient contained in the wolfberry plant.

[0031] In the present invention, extraction of a wolfberry plant is carried out by washing the collected wolfberry plant to remove dirt and organisms attached to the surface, and then cutting, ball milling, ultrasonic treatment, a homogenizer and the like. It can be extracted after crushing. Prior to crushing, air drying or freeze drying may be performed. The crushed wolfberry plant is preferably extracted repeatedly several times with an extraction solvent such as hot water while stirring. This extract can be used as it is as a basophil cell degranulation inhibitor, mast cell degranulation inhibitor, -hexosaminidase release inhibitor, antiallergic agent or antiinflammatory agent.

[0032] The extract of the genus wolfberry of the present invention can be adjusted to an appropriate concentration by diluting or concentrating as necessary. For example, the extract of the present invention can be diluted with water or alcohol by adding it to be used as a water-soluble substance, or can be mixed with fats and oils to form a cream. Furthermore, the extract is spray-dried to obtain a powdered basophil cell degranulation inhibitor, mast cell degranulation inhibitor, / 3-hexosaminidase release inhibitor, anti-allergic agent or anti-inflammatory agent You can also. As described above, the concentration and shape of the extract of the present invention are not particularly limited, and can be appropriately determined according to the use.

[0033] The extract of the present invention may contain components other than an extract of a wolfberry plant. Since the extract of the present invention uses an extract of a wolfberry plant used for food as an active ingredient, the extract is highly safe for a living body. For this reason, the extract of the present invention is used for pharmaceuticals, functional foods, Products, quasi-drugs, cosmetics, bath preparations and a wide range of other products.

Since the wolfberry plant or the extract of the wolfberry plant of the present invention has β xosaminidase release inhibitory activity, it has an effect of inhibiting degranulation from basophils or mast cells. It can be used as a basophil cell degranulation inhibitor or a mast cell degranulation inhibitor, and is also useful as an antiallergic or antiinflammatory agent. That is, / 3 xosaminidase is one of the substances released from basophils or obesity cells upon degranulation. Therefore, anti-allergic and anti-inflammatory effects can be evaluated by measuring the / 3 xosaminidase released by degranulation from cells using the / 3 xosaminidase release assay.

In the present invention, basophil cell degranulation inhibitory activity or mast cell degranulation inhibitory activity is measured by the activity of -hexosaminidase released by degranulation. β-Hexosaminidase activity was released by sensitizing cells with a mouse anti-DNP-IgE antibody using basophil cells such as KU812 shown in Example 3 below, and then using the antigen DNP-BSA. The i3 xosaminidase activity is measured. In addition, the enriched cells may be used for measuring i3_hexosaminidase activity. If wolfberry genus having high basophil cell degranulation inhibitory activity or mast cell degranulation inhibitory activity is used in combination, stronger antiallergic and anti-inflammatory effects can be expected.

[0036] The anti-allergic agent and anti-inflammatory agent of the present invention contain an active ingredient of a wolfberry plant or an extract of a wolfberry plant, and may be an extract solution or an extract itself, or a carrier or auxiliary agent known in the art. It may be a composition to which an agent, a food or drink material, another pharmaceutically acceptable formulation material, or the like is added. The amount of the extract is not particularly specified, but the concentration of the extract of the present invention in these products can be appropriately selected within a range in which a desired effect is obtained. When administered to the human body, it can be administered in one to several doses, for example, at a dose of 0.2 to 100 mg / Kg body weight per day. Depending on the purpose of administration and the route of administration, the dosage form of the drug may be tablets, capsules, injections, drops, powders, suppositories, granules, ointments, suspensions, emulsions, syrups , Creams and the like.

[0037] Further, in this composition, binders, excipients, lubricants, disintegrants, Additives such as agents, stabilizers, emulsifiers and buffers can be included. Preferred examples of the binder include starch, trehalose, dextrin, gum arabic powder, and the like. Preferable examples of the lubricant include stearic acid, talc, sucrose fatty acid ester, polyethylene glycol and the like. Preferred examples of the disintegrant include starch, carboxymethyl cellulose, corn starch and the like. Preferable examples of the stabilizer include fats and oils, propylene glycol and the like. Preferable examples of the emulsifier include an anionic surfactant, a nonionic surfactant, and polybutyl alcohol. Preferred examples of the buffer include buffers such as phosphate, carbonate, and citrate.

[0038] Further, the antiallergic agent or antiinflammatory agent of the present invention can be contained in foods and drinks, cosmetics and bath preparations, and the content thereof is not particularly limited, but is usually 0.0001% by weight or more, preferably 0.01% by weight or more. 100% by weight is good. When the extract of the present invention is contained in cosmetics and used as an antiallergic agent or an anti-inflammatory agent, the content can be adjusted to 0.01 to 10% by weight.

[0039] When the antiallergic agent or antiinflammatory agent of the present invention is included in a food, the food can be made a functional food for the purpose of preventing and / or improving allergy and inflammation. The type of food to be targeted is not particularly limited as long as the extract does not inhibit the anti-allergic and anti-inflammatory effects. For example, beverages such as juice, soft drinks, tea, etc., processed foods such as bread and rice cake, sweets such as candy, instant foods such as cup ramen, fats and oils such as butter and salad oil, dressings, mayonnaise, sauces, and soy sauce It can be included in a wide range of foods and drinks, such as seasonings such as yamirin, sprinkles, and miso. Of course, instead of the extract, it is possible to grind the wolfberry plant as it is, and ingest it directly into foods and the like.

Example

Hereinafter, the present invention will be described more specifically with reference to Production Examples, Examples, and Formulation Examples. However, the scope of the present invention is not limited to these examples.

(Example 1)

After cutting and crushing the dry weight lg of Kukushiko (Lycium chinense), Milli-Q 10 ml of water was added, and the mixture was autoclaved at 105 ° C for 15 minutes. The mixture was centrifuged at 3000 rpm at 4 ° C for 5 minutes, and the supernatant was filtered through a 0.22 / m filter to obtain a hot water extract Sampnore. The extract was further diluted 100 times, 1000 times, and 10000 times with a cell culture medium before use.

(Example 2)

After cutting and crushing lkg of dry weight of Kukushiko (from Lycium chinense), 10 L of Milli-Q water was added, and 105. C, Autoclave for 15 minutes. Four. The mixture was centrifuged at 3000 rpm for 5 minutes, and the supernatant was filtered through a 0.22 zm filter to obtain an extract. The obtained liquid was concentrated to dryness to obtain about 87 g of a solid.

(Example 3) / 3-Hexosaminidase-releasing Atsushi

Human immature basophils (KU812) cells were seeded on a medium prepared with 10% FBS, MEM 4.7 g / 500 ml, 200 mM L-gunorethamine 5 ml / 500 ml, 10% NaHCO (pH 7.0.7.4). Incubate at 37 ° C under 5% CO. Next, the cultured KU812 was peeled from the flask, and adjusted to a cell concentration of 5 × 10 ⁵ cells / ml using the same medium as above, and finally 0.3 ml of lmg / ml mouse anti-DNP_IgE antibody was added. / ml. After suspending well, the cells were seeded at 96μΙ / well in a 96-well plate, and incubated at 37 ° C under 5% CO in a carbon dioxide gas incubator to sensitize the cells. The next day, the Itoda spores were washed twice with 200 / il PBS, and then-hexosaminidase release buffer (116.9 mM NaCl, 5.4 mM KC1, 0.8 mM MgSO-7H〇, 5.6 mM gnorecose, 25 mM HEPES

, 2. OmM CaCl, 1. Omg / ml BSA dissolved in 500 ml of water, p with 10% NaHCO

Adjust to H7.7 and filter sterilize) 60 / l with diluted sample of wolfberry extract (diluted 1/100, 1/1000, 1/10000) or control (PBS) prepared in Example 1 with 5 Incubate for 10 minutes in a carbon dioxide gas incubator at 37 ° C under 5% CO and 1% cal. Next, 4 z gZml DNP_BSA a 5 ₁ l month卩tut, 5% C_〇 under 37. After incubation in a carbon dioxide incubator with C for 1 hour, add 205.5 μl of the substrate solution (85.5 mg of p_nitrophenyto N-acetylb-D_glucosamide to 50 ml of 50 mM citrate buffer (pH 4.5; Add 80 μL of the solution (added to pH) with acid) and incubate at 37 ° C for 30 minutes. Then, the reaction stop solution (100 mM carbonate buffer; Na C03 1.06 g)

/ 100ml, NaHC〇3 0.84g / l00ml) After heating 200 μl, read the microplate reader Measure the absorbance (A) at 450 nm at first. At this time, the inhibition rate of -hexosaminidase release is calculated by the following equation.

[0044] Inhibition rate (%) = {1- (S-B) / (C-b)} X100

S: A when sample is added to cells

B: A when sample was added in the absence of cells

C: A of control (when PBS was added to cells)

b: A when control was added in the absence of cells

The results are shown in Figure 1.

[0045] The wolfberry extract of the present invention significantly inhibited the release of / 3 xosaminidase from basophils, which is an indicator of antiallergic action.

(Example 4) Production of wolfberry extract-containing tablet

45 parts by weight of the wolfberry extract obtained in Example 2

Lactose 35 parts by weight

15 parts by weight of crystalline cellulose

5 parts by weight of sucrose fatty acid ester

The above composition was mixed and tableted to produce a dietary tablet containing a wolfberry extract.

(Example 5) Production of soft capsule containing wolfberry extract

40 parts by weight of the wolfberry extract obtained in Example 2

Olive oil 55 parts by weight

Glycerin fatty acid ester 5 parts by weight

The above composition was mixed, pulverized, and filled into a gelatin capsule to produce a dietary power capsule containing a wolfberry extract.

(Example 6) Production of soft drink containing wolfberry extract

1 part by weight of the wolfberry extract obtained in Example 2

Liquid sugar 30 parts by weight

0.1 parts by weight of citric acid

0.5 parts by weight of fragrance

Water 150 weight 水 Mixing with the above composition to produce soft drink containing wolfberry extract

(Example 7) Preparation of wolfberry extract-containing cracker

Wolfberry extract obtained in Example 2

120 parts by weight of flour

Salt

Baking powder

/ '~

water

_C where a conventional manner to prepare a wolfberry extract-containing crackers above composition

(Example 8) Preparation of dressing containing wolfberry extract

10 parts by weight of the wolfberry extract obtained in Example 2

Olive oil

Vinegar

Salt

pepper

Lemon water

A dressing containing wolfberry extract was prepared by the usual method with the above composition

(Example 9) Preparation of chewing gum containing wolfberry extract

0.5 parts by weight of the wolfberry extract obtained in Example 2

Gum base

sugar

0.5 parts by weight of fragrance

Water

A wolfberry extract-containing chewing gum having the above composition was prepared by a conventional method.

(Example 10) Preparation of tea containing wolfberry extract

Dissolve an appropriate amount of wolfberry extract obtained in Example 2 in hot water

Claims

The scope of the claims

[I] An antiallergic agent comprising a Lycium plant or a polar solvent extract thereof as an active ingredient.

[2] A basophil cell degranulation inhibitor containing a Lycium plant or a polar solvent extract thereof.

[3] An inhibitor of mast cell degranulation containing a Lycium plant or a polar solvent extract thereof

[4] A β-hexosaminidase release inhibitor comprising a Lycium plant or a polar solvent extract thereof.

[5] An anti-allergic agent comprising one or more of the degranulation inhibitor according to claim 24 or the hexosaminidase release inhibitor.

[6] The anti-allergic agent according to claim 1 or 5, which is Lycium chinense, a Lycium plant power.

[7] The method according to any one of claims 15 to 6, wherein the Lycium extract is an extract obtained by extracting wolfberry with water or a mixed solvent of water and a polar organic solvent. The antiallergic agent as described.

[8] A food or drink characterized by comprising the antiallergic agent according to any one of claims 15 6 and 7.

[9] A cosmetic product comprising the antiallergic agent according to [1] or [15] or [7].

[10] A bath preparation comprising the antiallergic agent according to [1] or [16] or [7].

[II] A pharmaceutical product characterized by comprising the antiallergic agent according to claim 1 or any one of claims 1 to 6.

[12] A method for preventing or treating allergy using the anti-allergic agent according to [1] or [15] or [7].

[13] An anti-inflammatory agent comprising at least one of the degranulation inhibitor or the xosaminidase release inhibitor according to claim 2-4.

14. The anti-inflammatory agent according to claim 13, which is Lycium chinense, a Lycium plant power.

15. The method according to claim 13, wherein the Lycium extract is an extract obtained by extracting wolfberry with water or a mixed solvent of water and a polar organic solvent. Anti-inflammatory drugs.

[16] Food and drink characterized by comprising the anti-inflammatory agent according to [4] in any one of [13] to [15].

P

Po

[17] A cosmetic comprising the anti-inflammatory agent according to [4] of claim 13-15.

P

PPo

[18] A bath preparation comprising the anti-inflammatory agent according to [4] of [13] to [15].

[19] A medicament characterized by comprising the anti-inflammatory agent according to claim 4 in any one of claims 13 to 15.

P

PPo

[20] A method for preventing or treating inflammation using the anti-inflammatory agent according to any one of claims 13 to 15.