JP2001064192A - Migration inhibitor for langerhans cell and antigen presentation inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an inhibitor capable of inhibiting the migration of Langerhans cells or inhibiting antigen presentation to T-lymphocytes, and thereby especially useful for the prophylaxis or therapy of allergy associated with type IV allergy by making the inhibitor include a matrix metalloproteinase-9 inhibitor. SOLUTION: This inhibitor contains a matrix metalloproteinase-9 inhibitor. The inhibitor is preferably at least one kind selected from the extracts of mugwort, chamomile, red clover, wild pansy, balsam, perilla, aloe, licorice and the like, and the extract can be obtained, for example, by using the whole or part of a raw or dried plant as a raw material, and extracting the raw material by using water, a water-soluble organic solvent such as methanol, etc., according to a conventional method.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to an agent for inhibiting the migration of Langerhans cells and an agent for inhibiting antigen presentation.

[0002]

2. Description of the Related Art Recently, matrix metalloproteinases (MMPs; eg, collagenase, gelatinase)
Is known to be deeply involved in the formation of blood vessels, metastasis and invasion of cancer, and its inhibitors are being applied as excellent anticancer agents that suppress cancer cell metastasis and inhibit angiogenesis of cancer tissues . It is also known that various MMPs are involved in diseases associated with tissue destruction such as rheumatism and periodontal disease, and matrix metalloproteinase inhibitors are
Application to the treatment of these diseases is also expected.

[0003]

An object of the present invention is to provide a new use of a matrix proteinase-9 inhibitor.

[0004]

Means for Solving the Problems First, the inventor of the present invention determined that Langerhans cells, which are antigen-presenting cells present in skin and mucous membrane tissues, were used to reduce allergic diseases such as allergic contact dermatitis and atopic dermatitis. Mechanism was involved in the occurrence of the disease.

[0005] Langerhans cells are differentiated from bone marrow stem cells, and after their precursor cells reach the skin and mucous membrane tissues of the whole body along the blood flow, they settle on the epidermis of the skin and mucous membrane tissues. Then, antigenic substances, for example, metals such as nickel, certain low-molecular compounds (so-called "haptens"), proteins and peptides such as mite antigens,
When invading through the skin or mucous membranes, the Langerhans cells capture these antigens, then migrate to the regional lymph nodes and present them to T lymphocytes within the lymph nodes. As a result, the T lymphocytes recognize the antigen carried from the locality by the Langerhans cells, memorize the antigen, proliferate monoclonally,
Eventually, it spreads throughout the body along the flow of blood and lymph (establishment of sensitization). The mechanism of this sensitization is type I and IV
Common to allergic types.

[0006] The mechanism by which an allergic reaction is induced differs depending on the type of allergy. In the case of type IV allergy, when the antigen re-enters after sensitization,
Again, Langerhans cells capture the antigen and circulate in the T lymphocytes (T lymphocytes that store antigen information specific to that antigen).
To the lymphocytes) again (inducing allergic reactions). As a result, T lymphocytes are activated, release various mediators of inflammation, and cause local infiltration of other inflammatory cells such as macrophages, resulting in inflammation involving many inflammatory mediators and cells. Is caused to cause skin allergic diseases such as allergic contact dermatitis and atopic dermatitis.

As described above, Langerhans cells contact T lymphocytes in the following two stages of the development of allergic inflammation and present antigen information to T lymphocytes. (1) Capture the invading antigen, carry it to the lymph node, and stimulate T lymphocytes specific to that antigen (= establish sensitization, in case of type I and type IV allergy); (2) From the second time When the antigen invades, the T lymphocyte is stimulated again by presenting the antigen to circulating T lymphocytes (= allergic reaction, only in case of type IV allergy).

[0008] At this time, it is extremely important that the T lymphocyte itself, even if it encounters (or comes into contact with) an antigen stored in the T lymphocyte, if the antigen is present alone, the antigen Cannot be recognized, and the antigen can be recognized only through an antigen presenting cell such as a Langerhans cell.

Specifically, antigen presenting cells such as Langerhans cells express a class II histocompatibility antigen such as HLA-DR on the cell surface, and the cells directly (or alternatively) Macromolecules such as proteins are bound on this class II histocompatibility antigen (after digestion inside the cell), exposing the antigen to the cell surface. T lymphocytes then bind T cell receptors, also expressed on their cell surface, to histocompatible antigens on the cell surface of antigen presenting cells,
Recognize antigen information presented (exposed) on histocompatible antigens.

[0010] Langerhans cells, which are antigen presenting cells, usually express a class II histocompatibility antigen on their cell surface, and have a dendritic shape with elongated long projections. Interestingly, contact of the Langerhans cells with the antigen significantly increases the expression of histocompatible antigens on the cell surface and extends the dendrites longer. That is,
It makes contact with more T lymphocytes in a wider area and changes itself so that antigen information can be more efficiently transmitted to T lymphocytes (antigen presentation).

[0011] In the course of studying the function of Langerhans cells, the present inventors have found that the cells produce MMP-9. Surprisingly, it has been revealed that MMP-9 is deeply involved in various functions of Langerhans cells, in particular, essential functions as antigen presenting cells.

First, if a mouse is treated with a substance that inhibits the activity of MMP-9 before applying a substance that causes allergy to the mouse (before sensitization), Langerhans cells migrate to the lymph nodes. Have been found to be able to prevent

That is, as described above, Langerhans cells capture antigens that have entered through the skin or mucous membranes, transport them to regional lymph nodes, and stimulate T lymphocytes specific to the antigens to cause allergy. As an essential first step, substances that inhibit the activity of MMP-9 inhibit Langerhans cells from migrating (migrating) to lymph nodes,
It has been found that the chance of contact between Langerhans cells and T lymphocytes is reduced, and as a result, the establishment of allergy (establishment of sensitization) is inhibited.

Second, MMP-9 is applied to already sensitized mice before application of the substance that causes allergy again.
When mice are treated with a substance that inhibits the activity of, the phenomena that occur when Langerhans cells come into contact with antigens (the phenomenon that increases the expression level of histocompatibility antigens on the cell surface, Any of the stretching phenomena)
However, it was found that it was suppressed remarkably.

That is, when an allergic reaction is induced, the Langerhans cells come into contact with the locally infiltrated T lymphocytes again,
As a result, activation of T lymphocytes triggers inflammation in type IV allergy.
Inhibitor 9 significantly suppresses the increase in the expression level of histocompatibility antigen on the surface of Langerhans cells, which is a direct contact with T lymphocytes, and also suppresses the elongation of dendrites of Langerhans cells. As a result, the present inventors have found that the frequency of contact between T lymphocytes and Langerhans cells (frequency of presentation or delivery of antigen) is reduced, thereby inhibiting the onset of allergy.

The present inventor has argued that the MMP-9 inhibitor inhibits the migration of Langerhans cells, thereby preventing the development of allergy, and furthermore, the MMP-9 inhibitor suppresses antigen presentation to T lymphocytes. Thus, the present invention was found to be useful for the prevention or treatment of allergy involving T lymphocytes, particularly allergy involving type IV allergy, and completed the present invention.

That is, the present invention provides the inventions described in the following items.

Item 1 An agent for inhibiting migration of Langerhans cells, comprising a matrix metalloproteinase-9 inhibitor.

Item 2 The matrix metalloproteinase-9 inhibitor is selected from the group consisting of mugwort, chamomile, red clover,
Wild pansies, Balsam pear, Perilla, Aloe, Licorice, Gardenia, Kumazasa, Clara, Mulberry, Birch, Horsetail, Achillea millefolium, Japanese Artemisia, Carrot, Hamamelis, Rose, Loofah, Mint, Momo, Reishi, Cedar, Rosemary, Apricot, Echina , Burdock,
Hibamata, salvia, hops, oysters, calendula,
Vetch, eucalyptus, melissa, cornflower, thyme,
Item 3. The migration inhibitor according to item 1, wherein the agent is at least one selected from the group consisting of turmeric, rooibos, ogon, peonies, ginger, touki, pleasure, button, chickweed and marigold extracts.

Item 3 A pharmaceutical composition for preventing allergies comprising the antimigration agent according to Item 1 or 2 as an active ingredient.

Item 4. A cosmetic composition for preventing allergies comprising the migration inhibitor according to Item 1 or 2 as an active ingredient.

Item 5. An allergy-preventive food containing the migration inhibitor according to Item 1 or 2 as an active ingredient.

Item 6. An inhibitor of Langerhans cell antigen presentation comprising a matrix metalloproteinase-9 inhibitor.

Item 7. A matrix metalloproteinase-9 inhibitor is selected from the group consisting of mugwort, chamomile, red clover,
Wild pansies, Balsam pear, Perilla, Aloe, Licorice, Gardenia, Kumazasa, Clara, Mulberry, Birch, Horsetail, Achillea millefolium, Japanese Artemisia, Carrot, Hamamelis, Rose, Loofah, Mint, Momo, Reishi, Cedar, Rosemary, Apricot, Echina , Burdock,
Hibamata, salvia, hops, oysters, calendula,
Vetch, eucalyptus, melissa, cornflower, thyme,
Item 7. The antigen presentation inhibitor according to Item 6, wherein the agent is at least one selected from the group consisting of extracts of turmeric, rooibos, ogon, peonies, ginger, touki, pleasure, button, chickweed and marigold.

Item 8 A pharmaceutical composition for preventing or treating allergy, comprising the antigen presentation inhibitor according to Item 6 or 7 as an active ingredient.

Item 9. A cosmetic composition having the effect of improving skin roughness comprising the antigen presentation inhibitor according to Item 6 or 7 as an active ingredient.

Item 10: An allergy-preventive food containing the antigen presentation inhibitor according to item 6 or 7 as an active ingredient.

[0028]

BEST MODE FOR CARRYING OUT THE INVENTION Matrix metalloproteinase-9 (hereinafter sometimes referred to as "MMP-9")
Is a type of matrix metalloproteinase that is an extracellular matrix-degrading enzyme, also called gelatinase B, and is an enzyme that uses gelatin, type III, type IV or type V collagen, elastin, or the like as a substrate.

An inhibitor of matrix metalloproteinase-9 (hereinafter sometimes referred to as an “MMP-9 inhibitor”) has the effect of inhibiting the activity of matrix metalloproteinase-9 and / or inhibiting the activity of matrix metalloproteinase-9. It has an action of suppressing production, and in the present invention, compounds and extracts which have been known to have such an action can be used. MMP-9 inhibitors in the present invention include M
It is only necessary to have an action of inhibiting the activity of MP-9 and / or an action of suppressing the production thereof, and it is possible to use a matrix metalloproteinase other than MMP-9 (for example, matrix metalloproteinase-1, -2, -3, etc.). They may also have an inhibitory effect.

Examples of the MMP-9 inhibitor used in the present invention include synthetic peptides having an MMP-9 inhibitory action such as batimastat; antibiotics having an MMP-9 inhibitory action such as tetracycline, minocycline and doxycycline; inhibitors of protein kinase scan -C such as -C; chelating agents such as EDTA, 1,10-phenanthroline and derivatives thereof; cytokine such as interferon γ other, alpha ₂ microglobulin, ursolic acid, acetyl Cysteine and the like.

Furthermore, MMP-9 inhibitors include mugwort, chamomile, red clover, wild pansy,
Balsam pear, perilla, aloe, licorice, gardenia, kumazasa, clara, mulberry, birch, horsetail, horseshoe millefolium, scorpionflower, carrot, hammerellis, rose, loofah, mint, peach, reishi, cedar, rosemary, apricot, chinensis, burdock Hibamata, salvia,
Extracts such as hops, oysters, calendula officinalis, eucalyptus, eucalyptus, melissa, cornflower, thyme, turmeric, rooibos, gougon, peonies, gingerbread, touki, squirrel, button, chickweed, marigold and the like are exemplified.

The MMP-9 inhibitors used in the present invention include, among these extracts, mugwort, red clover, wild pansy, licorice, horsetail, horse chestnut, achillea millefolium, valerium, reishi, cedar, rosemary, burdock, chickweed and marie Gold extract is preferred; mugwort, red clover, wild pansy,
Extracts of licorice, horsetail, sagebrush, cedar, chickweed and marigold are more preferred; extracts of sagebrush, wild pansies, sagebrush and marigolds are even more preferred; extracts of sagebrush and wild pansies are most preferred.

The above-mentioned plant extract can be prepared, for example, by using a raw or dried product of the whole or part of a plant (eg, flowers, fruits, leaves, branches, roots, etc.) as a raw material and using an appropriate extraction solvent in a conventional manner. It can be obtained by extracting according to It is preferable to use a crushed or crushed dried plant material as a raw material. As the extraction solvent, for example, water, a water-soluble organic solvent, or a mixed solvent thereof can be used. Examples of the aqueous organic solvent include lower alcohols such as methanol, ethanol, and isopropanol. The use of an aqueous organic solvent is preferred from the viewpoint of preventing the decay of the extract and the efficiency of extraction. The ratio in the case of using a mixed solvent of water and an organic solvent as the extraction solvent can be appropriately set according to the type of the raw material and the like. For example, the organic solvent is 50% (v / v) or less.
Preferably, it can be 30% (v / v) or less. The extraction is performed by extracting the above-mentioned extraction solvent, for example, from 1 to 10 of the raw material.
Using about 0 times the volume, preferably about 2 to 5 times the volume, the mixture can be heated as needed, and can be repeatedly extracted two to three times. These extracts may be used as the extract itself, or may be used as a concentrate or a dry powder. These extracts may be used by performing a treatment using activated carbon or the like for the purpose of decolorization or deodorization, if necessary. These plant extracts are easily available as commercial products.

The MMP-9 inhibitors can be used alone or in combination of two or more. Further, the MMP-9 inhibitor is a matrix metalloproteinase inhibitor other than matrix metalloproteinase-9 (for example, MM
P-1 inhibitor, MMP-2 inhibitor, MMP-3 inhibitor, etc.).

The MMP-9 inhibitor can be taken as a medicine or food, or can be used as a cosmetic.

(1) Pharmaceutical Composition The MMP-9 inhibitor can be used as a general pharmaceutical preparation form by using it together with a suitable pharmaceutically acceptable carrier. As the form of the pharmaceutical preparation, various forms can be selected according to the purpose of treatment or prevention, for example, solid preparations such as extracts, tablets, pills, granules, capsules, and troches; Liquid preparations such as liquid preparations, emulsions, injections (liquid preparations and suspensions), syrups, aerosols, eye drops, infusions, decoctions, etc .; ointments, plasters, lotions, cataplasms, etc. Such external preparations are exemplified. These administration methods are not particularly limited, and can be performed according to various forms. For example, powders, granules, tablets, capsules, solutions and the like can be administered orally, and injections can be administered intravenously. External preparations can be applied to the skin, powdered preparations such as powder,
It can also be used by applying it in advance to the inner surface of a rubber glove or the like and wearing it.

[0037] The above-mentioned preparations are mixed with additives (eg, binders, disintegrants, surfactants, absorption promoters, adsorbents, lubricants, fillers, etc.) which are usually used in pharmaceuticals, if necessary. Can do things. When used as an external preparation, components such as an ultraviolet absorber and an ultraviolet scattering agent can be added. Further, a drug other than the MMP-9 inhibitor, for example, a known antiallergic or antiinflammatory agent may be added.

The content of the MMP-9 inhibitor in the above preparation can be appropriately set according to the type of the MMP-9 inhibitor, the form of the preparation, and the like. For example, 0.001 to 99.99
It can be about 9% by weight. Alternatively, when an extract is used as the MMP-9 inhibitor, a drug comprising only the extract may be used. For the dosage of the above formulation,
MP-9 inhibitor type, formulation form, patient symptoms, age,
It is appropriately selected according to gender and other conditions. For example,
When the MMP-9 inhibitor is an extract, the amount of the MMP-9 inhibitor is 0.04% of the dry weight of the extract per adult day.
About 001 to 300 mg, preferably 0.001 to 10
The dose is preferably about 0 mg, and it can be administered once a day or divided into 2 to 4 times. MMP
The dose when the -9 inhibitor is other than an extract can be appropriately set by referring to the above dose according to its inhibitory ability.

Since the above-mentioned preparation contains an MMP-9 inhibitor, it can be used as a migration inhibitor of Langerhans cells. When such a cell migration inhibitor is used as an active ingredient, the occurrence of allergy is suppressed by inhibiting the migration of Langerhans cells, thereby, for example, type IV allergic diseases such as allergic contact dermatitis and atopic dermatitis Alternatively, it can be used for prevention of allergy such as type I allergic disease such as hay fever and urticaria.
Therefore, the present invention includes a pharmaceutical composition for preventing allergies, which comprises, as an active ingredient, a migration inhibitor of Langerhans cells containing an MMP-9 inhibitor. The pharmaceutical composition may be the Langerhans cell migration inhibitor itself formulated as described above, or may further contain other components in addition to the migration inhibitor depending on the purpose of administration. Is also good.

Further, since the above-mentioned preparation contains an MMP-9 inhibitor, it can be used as an antigen presentation inhibitor of Langerhans cells. When such an antigen presentation inhibitor is used as an active ingredient, it exerts an action of suppressing the presentation of antigens of Langerhans cells to T lymphocytes, thereby producing an allergic disease such as allergic contact dermatitis, atopic dermatitis and the like. It can be used for the prevention or treatment of type allergic diseases, and can alleviate or eliminate the symptoms of a subject who has developed such allergic diseases, and can prevent the deterioration of the symptoms. Alternatively, it is possible to prevent a subject who has not developed an allergic disease from developing it. Therefore, the present invention includes a pharmaceutical composition for preventing or treating allergy, which comprises an inhibitor of Langerhans cell antigen presentation containing an MMP-9 inhibitor as an active ingredient.
The pharmaceutical composition may be an antigen presentation inhibitor itself of Langerhans cells formulated as described above,
Depending on the purpose of administration, an antigen presentation inhibitor may further contain other components.

(2) Cosmetic Composition The MMP-9 inhibitor can be prepared and used in the form of commonly used cosmetics. As the form of cosmetics, various forms can be selected according to the purpose, and specifically, soap,
Facial cleansers, lotions, emulsions, foundations, lipsticks, lip balms, cleansing creams, massage creams, packs, hand creams, hand powders, body shampoos, body lotions, body creams, bath cosmetics and the like can be mentioned. The cosmetics can be prepared according to a conventional method corresponding to each form. The cosmetic composition of the present invention can be used by applying it to the skin by the usual use in each form. In addition, powdery cosmetics such as hand powder can be used by applying it in advance to the inner surface of a rubber glove or the like and wearing it.

The cosmetic composition of the present invention may optionally contain components commonly used in cosmetics, such as humectants, fragrances, ultraviolet absorbers, and ultraviolet scattering agents, as long as the effects of the present invention are not impaired. Can do things.

The content of the MMP-9 inhibitor in the cosmetic composition of the present invention can be appropriately set according to the type of the MMP-9 inhibitor, the form of the cosmetic, and the like.
It can be about 99.999% by weight. The above cosmetics can be applied to the skin once a day or in two to four times an appropriate amount depending on the type of MMP-9 inhibitor, the form of the cosmetic, the degree of rough skin, and the like.

Since the composition prepared as described above contains an MMP-9 inhibitor, it can be used as a migration inhibitor of Langerhans cells. That is, when such a cell migration inhibitor is used as an active ingredient, by inhibiting the migration of Langerhans cells to suppress the occurrence of allergy, for example, type IV allergic diseases such as allergic contact dermatitis and atopic dermatitis Alternatively, it can be preferably used to prevent allergic diseases such as type I allergic diseases such as urticaria. Also,
It can also be used to prevent rough skin, especially rough skin caused by these allergic diseases. Therefore, the present invention includes a cosmetic composition for preventing allergies comprising, as an active ingredient, a migration inhibitor of Langerhans cells containing an MMP-9 inhibitor. The cosmetic composition may be the migration inhibitor itself prepared as a cosmetic as described above, or may be a composition in which other components are further added to the migration inhibitor.

Further, the composition prepared in cosmetics as described above contains an MMP-9 inhibitor, and thus can be used as an antigen presentation inhibitor of Langerhans cells. That is, if such an antigen presentation inhibitor is used as an active ingredient,
To prevent allergic diseases, especially allergic diseases (allergic contact dermatitis, atopic dermatitis, etc.) involving allergies, especially type IV allergy, by exerting the action of suppressing the presentation of antigens of Langerhans cells to T lymphocytes It can be preferably used, and can also be used for improving skin roughness, especially skin roughness due to these allergies.
Therefore, the present invention includes a cosmetic composition having an effect of improving skin roughness, which comprises an inhibitor of Langerhans cell antigen presentation containing an MMP-9 inhibitor as an active ingredient. The cosmetic composition may be the antigen presentation inhibitor itself prepared as a cosmetic as described above, or may be a mixture of the antigen presentation inhibitor and other components.

(3) Food MMP-9 inhibitors can be prepared and used in the form of commonly used foods. Examples of the form of the food include liquid beverages such as juice, soft drink, tea; powdered beverages such as powdered juice and powdered soup; chocolate, candy, chewing gum, ice cream, jelly,
Cookies, biscuits, corn flakes, tablets,
Examples thereof include solid or semi-solid foods such as chewable tablets, gummy, wafers, and rice crackers.

The foods described above are blended with additives (eg, sweeteners, coloring agents, antioxidants, vitamins, flavors, etc.) usually contained in the foods, as long as the effects of the present invention are not impaired. It can be prepared according to a conventional method according to each form.

The content of the MMP-9 inhibitor was as follows:
It can be appropriately set according to the type of the inhibitor, the form of the food, and the like, and can be, for example, about 0.001 to 99.999% by weight. Alternatively, when an extract is used as the MMP-9 inhibitor, the food may be composed of only the extract. The intake amount is appropriately selected according to conditions such as the type of the MMP-9 inhibitor, the form of the food, the age and gender of the ingestor. For example, when the MMP-9 inhibitor is an extract, the amount of the MMP-9 inhibitor is about 0.0001 to 300 mg, preferably about 0.001 to 100 mg per adult day, as a dry weight of the extract. It can be taken once or twice or four times a day. The intake amount when the MMP-9 inhibitor is other than the extract can be appropriately set by referring to the above intake amount according to its inhibitory ability.

Since the above food contains an MMP-9 inhibitor, it can be used as a migration inhibitor of Langerhans cells. That is, if such a cell migration inhibitor is an active ingredient, by inhibiting the migration of Langerhans cells, by suppressing the occurrence of allergy, for example,
A subject who can be used for the prevention of type IV allergic diseases such as allergic contact dermatitis and atopic dermatitis, or type I allergic diseases such as hay fever and urticaria, and who has developed such allergic diseases In addition to suppressing the further progression of the symptoms, it is possible to prevent subjects who have not developed the symptoms from developing the allergic symptoms as described above. Therefore, the present invention includes an allergy-preventive food containing a Langerhans cell migration inhibitor containing an MMP-9 inhibitor as an active ingredient. The food may be the migration inhibitor itself prepared as a food as described above, or may be a mixture of the migration inhibitor and other components according to the purpose of ingestion.

Further, since the above food contains an MMP-9 inhibitor, it can be used as an antigen presentation inhibitor of Langerhans cells. That is, if such an antigen presentation inhibitor is used as an active ingredient, it exerts an action of suppressing the antigen presentation of Langerhans cells to T lymphocytes,
Allergic diseases, for example, allergic contact dermatitis, can be used for the prevention of type IV allergic diseases such as atopic dermatitis, not only can suppress the further progression of the symptoms of subjects who have developed such allergic diseases In addition, it is possible to prevent a subject who has not developed symptoms from developing allergic symptoms as described above. Therefore, the present invention includes an allergy-preventive food containing an inhibitor of Langerhans cell antigen presentation containing an MMP-9 inhibitor as an active ingredient. The food may be the antigen presentation inhibitor itself prepared as a food as described above, or may be a mixture of the antigen presentation inhibitor and other components according to the purpose of ingestion.

The inhibitor of Langerhans cell migration or antigen presentation, or the allergy-preventive food of the present invention is
It can be used as a functional food having these effects, such as a health food, a food for specified health use, a dietary supplement, and a food for the sick.

[0052]

The present invention will be described in more detail with reference to Examples, Production Examples, Pharmacological Test Examples and Formulation Examples. Unless otherwise specified, "%" indicates "% by weight".

Example 1 Langerha with MMP-9 inhibitor
Intradermally migration inhibition Balb / c mice to Nsu lymph node cells, the anti-M to inhibit MMP-9 activity
An MP-9 antibody (10 μg) or a control antibody (10 μg) having no MMP-9 inhibitory activity was injected, respectively. After 30 minutes, the sensitizing substance rhodamine B (2
%) 25 ul was applied to the pinna of the mouse to induce migration of Langerhans cells from the skin to the lymph nodes.

The number of Langerhans cells per mm ² of epidermis and the number of Langerhans cells per lymph node before application of rhodamine B (before sensitization) and 18 hours after application of rhodamine B were measured under a microscope. Table 1 shows the results.
The values in the table are shown as mean ± standard deviation.

[0055]

[Table 1]

The Langerhans cells in the epidermis were isolated before sensitization.
In the case of no treatment, the number was 1247 per mm ² , and the number was 926 18 hours after the application of rhodamine B.
6% decreased. On the other hand, before applying Rhodamine B, anti-MMP
When the -9 antibody was injected, the number of Langerhans cells in the epidermis was 1170, which was reduced by only about 6%. Therefore, it was revealed that administration of an anti-MMP-9 antibody (MMP-9 inhibitor) reduces the number of Langerhans cells in the epidermis, that is, suppresses the migration of Langerhans cells. On the other hand, when the control antibody was injected before the sensitization, the number of cells was 910, and a decrease in the number of cells (migration of cells) was observed, which was comparable to that in the case of no treatment.

Similarly, the number of Langerhans cells in the lymph node was 2640 per lymph node before sensitization, but was 7557 by rhodamine B application in the case of no treatment.
Increased to pieces. When the cells were treated with the anti-MMP-9 antibody before the sensitization, the number of Langerhans cells after the sensitization was 3156, and the number of the cells increased only slightly. On the other hand, when the control antibody was injected before the sensitization, the Langerhans cells after the sensitization increased to the same extent as in the case of no treatment.

As described above, it was revealed that the migration of Langerhans cells was suppressed by the MMP-9 inhibitor.

Example 2 Langerha with MMP-9 inhibitor
Inhibition of antigen cell presentation on the abdominal cells of the abdominal skin of Balb / c mice.
The mice were sensitized by applying 0.1 ml of 5% oxazolone. Five days later, an anti-MMP that inhibits MMP-9 activity
-9 antibody (10 μg) or a control antibody having no MMP-9 inhibitory activity (10 μg) was injected into the auricle, and 3
After 0 minute, 25 μl of 0.25% oxazolone was applied to the pinna to induce allergic contact dermatitis. 24 of provocation
After a lapse of time, the morphology of Langerhans cells in the epidermis was observed, and the difference between the group to which the control antibody was administered and the group to which the anti-MMP-9 antibody was administered was compared and examined. As a result, in the group to which the control antibody was administered, activation of Langerhans cells, that is, marked extension of dendrites and class
II Expression of histocompatibility antigen was enhanced, but MMP
In the group to which the anti-MMP-9 antibody that inhibits -9 activity was administered, such activation of Langerhans cells was not observed. Therefore, it was revealed that the activation of Langerhans cells was suppressed by the MMP-9 inhibitor.

The thickness of the pinna 24 hours after the induction was measured. As a result, the thickness of the pinna in the group to which the control antibody was administered was 0.3370 ± 0.038 mm, which was remarkable as compared with the thickness before the induction. Swelling was noted. In contrast, MMP
The thickness of the auricle in the group to which the -9 antibody was administered was 0.250 ± 0.2.
006 mm, almost no change before the induction, and no swelling was observed. Therefore, it was confirmed that the MMP-9 inhibitor suppressed the activation of Langerhans cells, as a result, the antigen presenting ability was suppressed, and allergic contact dermatitis was prevented.

Production Example 1 About 1 liter of water was added to 1 kg of dried mugwort (Artemisia vulgaris L.), and the mixture was heated and refluxed for 2 hours to perform extraction. Thereafter, extraction was carried out by heating and refluxing for 2 hours in the same manner using the same amount of water. Then, the extracts were filtered and combined, and the solvent was distilled off under reduced pressure, concentrated to about 1 liter, and reduced in a desiccator. Dry overnight under brown powdery substance 19g
I got

Preparation Example 2 400 g of dried red clover petals were immersed in 5 liters of a 30% (V / V) aqueous ethanol solution, and the mixture was heated and refluxed for 2 hours to perform extraction. Then the same amount of ethanol /
After similarly extracting with heating and refluxing for 2 hours using a water mixture, the extracts were filtered and combined, and then evaporated to dryness under reduced pressure to obtain 17 g of a brown powdery substance.

Example of Pharmacological Test <Measurement of anti-MMP-9 activity> Balb / c mice or untreated Balb / c mice or skin to which a 1% picryl chloride-acetone solution was applied 4 hours before was subjected to 0.25% The cells were treated with trypsin at 37 ° C. for 90 minutes, and the obtained epidermal cells (containing 2 to 4% of Langerhans cells) were suspended in RPMI-1640 medium containing 10% fetal bovine serum, respectively.
The mixture was dispensed into the wells of the culture plate at 1 × 10 ⁵ .
Each test substance shown in Table 2 below (all in water extract) was added to the medium of the epidermal cell suspension prepared from the picryl chloride-treated mouse at a predetermined concentration and cultured for 24 hours. After completion of the culture, the activity of matrix metalloproteinase-9 (MMP-9) in the culture supernatant was confirmed by gelatin enzymography, and the activity was quantified using a densitometer.

The MMP-9 activity in the culture supernatant of the epidermal cells obtained from untreated mice (without addition of the test substance) was determined by (A); The MMP-9 activity in the culture supernatant was defined as (B); and the MMP-9 activity in the culture supernatant of epidermal cells prepared from a mouse treated with picryl chloride and added with a test substance was defined as (C). The inhibition rate was calculated by the following formula.

[0065]

(Equation 1)

The results are shown in Table 2 below.

[0067]

[Table 2]

[0068]

[Table 3]

Each test substance described in Table 2 had an MMP-9 inhibitory action.

A pharmaceutical composition, a cosmetic composition, and a food composition were prepared according to the following formulation.

Formulation Example 1: Ointment Ingredients Compounding amount (%) Artemisia extract 2.5 Propylene glycol # 400 15.0 Macrogol ointment 82.5 The artemisia extract obtained in Production Example 1 was replaced with propylene glycol # 4
After uniformly dispersing the mixture in 00, Macrogol ointment was added and mixed to obtain an ointment.

Formulation Example 2: Lotion Component Content (%) Red Clover Extract 1.0 Wild Pansy Extract 1.0 Glycerin 6.0 Ethanol 9.0 Polyoxyethylene Hardened Castor Oil 0.8 Methyl Paraben 0.05 Citric Acid 0.05 Sodium citrate 0.07 Fragrance 0.1 Purified water Remainder Glycerin, citric acid, sodium citrate,
An aqueous solution was prepared by dissolving a red clover water extract and a wild pansy 30% ethanol extract. Polyoxyethylene hydrogenated castor oil (60.EO), methyl paraben and fragrance were separately dissolved in ethanol, added to the above aqueous solution, solubilized, and filtered to obtain a lotion.

Formulation Example 3: Cream Ingredients Amount (%) Ingredient (A) Polyglycerin fatty acid ester 4.0 Cetanol 2.0 Stearic acid 1.0 Isopropyl myristate 5.0 Olive oil 2.0 Squalane 9.0 Self-emulsification Glyceryl monostearate 3.0 Paraben 0.3 Component (B) Marigold extract 0.5 Kawamurai extract 0.5 Glycerin 5.0 Trimethylglycine 1.0 Purified water 60.0 Component (C) Potassium hydroxide aqueous solution 3 0.0 Purified Water The remainder The component (A) was dissolved by heating to 80 ° C. The component (B) was dissolved by heating and kept at 80 ° C., and the component (A) was added thereto with stirring and mixed well. Further, the component (C) was added, the mixture was cooled while stirring, and a flavor (0.2%) was added and mixed to obtain a cream.

Formulation Example 4: Granules Component Content (%) Mugwort Extract 65 Lactose 13 Corn Corn 12 Carboxymethylcellulose Calcium 5.5 Methylcellulose 4.5 According to the above formula, mugwort extract, lactose obtained in Production Example 1,
Corn starch and carboxymethylcellulose calcium were sufficiently mixed, and the mixture was granulated according to a conventional method using a methylcellulose aqueous solution.

Formulation Example 5: Capsule Ingredients Ingredients (%) Wild Pansy Extract 28 Crystalline Cellulose 45 Corn Starch 22 Talc 3 Magnesium Stearate 2 According to the above ingredients, each ingredient is mixed to form a uniform mixture and then orally administered. This was filled into a gelatin capsule for use to obtain the desired capsule.

Formulation Example 6: Chewable tablets Compounding amount (%) Mugwort extract 50 Xylitol 30 Aspartame 9.4 Magnesium stearate 10.5 Fragrance 0.1 After mixing each component according to the above-mentioned composition so as to form a uniform mixture. And pressed to obtain the desired chewable tablet.

Examples 3 and 4 According to the above pharmacological test examples, MM
Effect on allergic contact dermatitis (Example 3) and atopic dermatitis (Example 4) by using an antigen presentation inhibitor of Langerhans cells containing an extract in which the inhibitory effect of P-9 activity was confirmed. It was confirmed.

The ointment of Formulation Example 1 or the cream of Formulation Example 3 was applied to each of 10 patients with allergic contact dermatitis and atopic dermatitis for 1 week, and the condition of erythema, kayumi and eczema was observed. Later, a comprehensive judgment was made based on changes in each symptom. The results are shown in Tables 3 and 4 below.

[0079]

[Table 4]

[0080]

[Table 5]

According to the comprehensive judgment results shown in Tables 3 and 4, the ointment of Formulation Example 1 and the cream of Formulation Example 3 each exhibited an effect of suppressing the antigen presentation of Langerhans cells, thereby causing allergic reactions. It was confirmed that excellent effects were exhibited on both contact dermatitis and atopic dermatitis.

[0082]

According to the present invention, an MMP-9 inhibitor comprises
It was clarified that Langerhans cells had the effect of suppressing migration to lymph nodes and antigen presentation to T cells. Therefore, the inhibitory effect of Langerhans cells on migration to lymph nodes causes allergic diseases (eg, type IV allergic diseases such as allergic contact dermatitis and atopic dermatitis, and type I allergic diseases such as hay fever and hives). Therefore, medicines and foods containing the migration inhibitor of the present invention as an active ingredient can be used for allergic diseases (for example, type IV allergic diseases such as allergic contact dermatitis and atopic dermatitis). Or type I allergic diseases such as hay fever and urticaria). Alternatively, the preventive or therapeutic effect of allergy involving T lymphocytes (eg, type IV allergic disease such as allergic contact dermatitis and atopic dermatitis) is exerted by the action of Langerhans cells to suppress antigen presentation to T cells. Therefore, the medicine and food containing the antigen presentation inhibitor of the present invention as an active ingredient prevent or treat allergic diseases (for example, type IV allergic diseases such as allergic contact dermatitis and atopic dermatitis). Can be used for

Similarly, according to the cosmetic comprising the Langerhans cell migration inhibitor of the present invention as an active ingredient, allergies (eg, type I allergic diseases such as urticaria, allergic contact dermatitis, atopic dermatitis, etc.) Of type IV allergic disease), and the effect of preventing rough skin, especially the rough skin due to these allergies. Furthermore, according to cosmetics containing an antigen presentation inhibitor of Langerhans cells as an active ingredient, prevention of allergy (for example, type IV allergic disease such as allergic contact dermatitis and atopic dermatitis),
The effect of improving skin roughness, especially skin roughness due to these allergies is obtained.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61P 43/00 111 A61P 43/00 111 F term (Reference) 4B018 LB01 LB08 LE02 MD48 MD61 MD66 ME07 MF01 4C083 AA111 AA112 AA122 AB032 AB442 AC022 AC072 AC102 AC122 AC132 AC242 AC302 AC352 AC422 AC432 AC482 AC582 AD042 AD092 AD112 AD212 AD242 AD262 AD272 AD412 BB51 CC03 CC04 CC05 DD14 DD15 DD16 DD22 DD23 DD27 DD31 EE12 EE13 4C084 AA02 DC32 MA12 Z152 Z672 ZB262 ZC202 4C088 AA13 AA18 AB03 AB12 AB14 AB19 AB24 AB25 AB26 AB29 AB34 AB38 AB40 AB41 AB51 AB52 AB57 AB58 AB59 AB60 AB76 AB81 AB86 AB99 AC01 AC03 AC04 AC05 AC06 AC11 AC13 BA08 BA09 BA10 MA52 MA63 NA14 ZA36 ZA67 ZBZ ZBZ

Claims (10)

[Claims] 1. Matrix metalloproteinase-9
An inhibitor of migration of Langerhans cells containing an inhibitor. 2. Matrix metalloproteinase-9
Inhibitors include mugwort, chamomile, red clover, wild pansy, balsam, perilla, aloe, licorice,
Gardenia, Kumazasa, Clara, Mulberry, Birch, Japanese horsetail, Achillea millefolium, Japanese Artemisia, Carrot,
Hamamelis, Rose, Loofah, Mint, Peach, Reishi, Cedar, Rosemary, Apricot, Echinashi, Burdock, Hibamata, Salvia, Hop, Oyster, Tokinsenka, Astragalus, Eucalyptus, Melissa, Cornflower, Thyme, Turmeric, Rooibos, Orgon, Peonies, ginger,
The migration inhibitor according to claim 1, wherein the migration inhibitor is at least one member selected from the group consisting of extracts of crested squirrels, brooms, buttons, chickweeds, and marigolds. 3. A pharmaceutical composition for preventing allergies comprising the migration inhibitor according to claim 1 as an active ingredient. 4. A cosmetic composition for preventing allergies comprising the migration inhibitor according to claim 1 or 2 as an active ingredient. 5. An allergy preventive food comprising the migration inhibitor according to claim 1 or 2 as an active ingredient. 6. An agent for suppressing antigen presentation of Langerhans cells, comprising a matrix metalloproteinase-9 inhibitor. 7. Matrix metalloproteinase-9
Inhibitors include mugwort, chamomile, red clover, wild pansy, balsam, perilla, aloe, licorice,
Gardenia, Kumazasa, Clara, Mulberry, Birch, Japanese horsetail, Achillea millefolium, Kawamurai, Carrot,
Hamamelis, Rose, Loofah, Mint, Peach, Reishi, Japanese cedar, Rosemary, Apricot, Echinashi, Burdock, Hibamata, Salvia, Hop, Oyster, Tokinsenka, Astragalus, Eucalyptus, Melissa, Cornflower, Thyme, Turmeric, Rooibos, Orgon, Peonies, ginger,
The antigen presentation inhibitor according to claim 6, wherein the agent is at least one member selected from the group consisting of extracts of touki, broom, button, chickweed and marigold. 8. A pharmaceutical composition for preventing or treating allergy, comprising the antigen presentation inhibitor according to claim 6 as an active ingredient. 9. A cosmetic composition having the effect of improving skin roughness, comprising the antigen presentation inhibitor according to claim 6 as an active ingredient. 10. An allergy preventive food comprising the antigen presentation inhibitor according to claim 6 or 7 as an active ingredient.