JP2022124341A - Topical preparation for skin allergy - Google Patents

Abstract

To provide a composition having the effect of ameliorating allergic dermatitis.SOLUTION: A composition for ameliorating allergic dermatitis contains plasmalogen extracted from the tissue of shellfish, ascidians or birds.SELECTED DRAWING: Figure 1

Description

TECHNICAL FIELD The present invention relates to a composition for improving allergic dermatitis containing plasmalogen.

Allergic dermatitis means eczematous dermatitis caused by allergic inflammation, and representative diseases include contact dermatitis, which is delayed allergic inflammation, and atopic dermatitis, which is immediate and delayed allergic inflammation. Are known. Both dermatitis are associated with locally produced IgE and mast cells in response to allergen invasion. In particular, IgE binds to mast cells and releases histamine, an itch transmitter, by degranulation. Furthermore, mast cells degranulated at the site of inflammation recover, store granules again, and degranulate again. Such degranulation of mast cells not only induces immediate itching but also causes a vicious cycle of itching and scratching (Non-Patent Document 1).

The mainstay of treatment for allergic dermatitis is long-term coping therapy by securing a living environment including avoidance of antigens and pharmacotherapy with anti-inflammatory drugs. Therefore, it is desired to improve QOL by preventing the onset and exacerbation of patients (Non-Patent Document 2), and to take measures to enable patients to receive continuous medical care and to be able to self-manage. ing. In such a situation, therapeutic intervention with anti-IgE antibodies that inhibit the binding of IgE and mast cells has been established for some patients who are not responsive to existing drugs.

Plasmalogen, on the other hand, is a kind of ether-type phospholipid and has a vinyl ether bond. In humans, it is known to be abundant in nerves, cardiovascular systems, immune systems, and the like. As a function of plasmalogen, it has been known to have an anti-central nervous system inflammatory effect (Patent Document 1). Furthermore, it has been reported that plasmalogen exerts an excellent effect on improvement of fatty liver (Patent Document 2), and that plasmalogen orally administered improves memory function of mild Alzheimer's disease patients (Patent Document 2). Non-Patent Document 3).

In addition, "the 1-alkyl ether type phospholipid content in the ether type phospholipid is high and the plasmalogen content is extremely low, that is, it is a natural product with a high 1-alkyl ether type phospholipid content ratio in the ether type phospholipid. Furthermore, the living body itself can be directly used as an extraction source without separating the tissue, and in addition to being abundant in resources and easy to obtain, it also contains astaxanthin, so it has good storage stability. It is possible to obtain a good dermatitis improving/preventing agent (Patent Document 3, paragraph 0012)”. By "it is possible to reduce the IgE concentration in the blood and improve the symptoms of dermatitis (Patent Document 3, paragraph 0037)", "dermatitis, especially atopic dermatitis, is effective A pharmaceutical composition that can effectively prevent or treat dermatitis, and a food and drink composition for preventing or improving dermatitis (Patent Document 3, paragraph 0001)”.

Table 2012/03947 Patent application 2020-063818 Patent 6755530

Igakukai Shimbun No. 2498 2002 Ministry of Health, Labor and Welfare 2019 Immunoallergic Disease 10-Year Strategy (http://www.mhlw.go.jp/newinfo/kobetu/kenkou/ryumachi/index.html)) Fujino T et al, Efficacy and Blood Plasmalogen Changes by Poral Administration of Plasmalogen in Patients with Mild Alzheimer's Disease and Mild Cognitive Impairment: A Multicenter, Randomized, Double-blind, Placebo-controlled Trial” EBiomedicine, [17] (2017) 199- 205

As described above, various pharmacological effects of plasmalogen have been reported. Transdermal administration for dermatitis has not been investigated.

An object of the present invention is to provide a composition for improving allergic dermatitis as a new use of plasmalogen.

The present inventors have found that plasmalogen permeates from the epidermis to the dermis of the skin through the stratum corneum of the skin and reduces the number of mast cells present in the dermis. reached.

That is, the present invention is as follows.
[1] A composition for improving allergic dermatitis, characterized by containing a plasmalogen.
[2] A composition for improving allergic dermatitis, containing plasmalogen in an amount of 0.1% by mass to 10% by mass based on the total composition.
[3] The composition for improving allergic dermatitis according to [1] or [2] above, wherein the plasmalogen is a plasmalogen extracted from animal tissue.
[4] The composition for improving allergic dermatitis according to [3] above, wherein the animal tissue is an animal tissue selected from shellfish, sea squirts and birds.
[5] The composition for improving allergic dermatitis according to [3] or [4], wherein the animal tissue is scallop tissue.

The composition of the present invention has an excellent ameliorating effect on allergic dermatitis as an external preparation for skin.

FIG. 1 shows a chart of a plasmalogen separation method by HPLC and a purified plasmalogen for use in a composition for improving allergic dermatitis. (Example 1) FIG. 2 shows the number of mast cells in the dermis of mice to which a plasmalogen-containing skin coating and a control coating were applied to the skin. (Example 2) FIG. 3 shows the number of mast cells in the dermis of NC/Nga mice fed a control diet and a diet containing plasmalogen. (Comparative example 1) 4 shows the effect of plasmalogen-containing ointment on atopic dermatitis in Example 3. FIG. In FIG. 4, A) shows typical results of the skin of a mouse to which white petrolatum was applied, and B) shows a typical result of the skin of a mouse to which plasmalogen-containing ointment was applied. (Example 3) FIG. 5 shows the effect of the plasmalogen-containing ointment on the redness and bleeding of atopic dermatitis performed in Example 3. (Example 3) 6 shows the effect of plasmalogen-containing ointment on crust formation and dryness in Example 3. FIG. (Example 3)

The composition for improving allergic dermatitis of the present invention is characterized by containing a plasmalogen.
Allergic dermatitis in the present invention means eczema dermatitis developed by allergic inflammation caused by allergen invasion, and includes contact dermatitis and atopic dermatitis. Allergic dermatitis improvement is the concept of relieving eczematous dermatitis. The composition for improving allergic dermatitis of the present invention can prevent the occurrence of itching caused by penetration of allergens into the skin. That is, it can also be used as a composition for preventing eczema dermatitis.

Plasmalogens are a subclass of glycerophospholipids characterized by having a vinyl ether bond at position 1 (sn-1) of the glycerol backbone, and are present at high concentrations in the cell membranes of cells that constitute many mammalian tissues.

The plasmalogen used in the present invention is not particularly limited as long as it is generally classified as a plasmalogen. Examples include choline-type plasmalogen, ethanolamine-type plasmalogen, inositol-type plasmalogen, and serine-type plasmalogen. can be mentioned. Among these, choline-type plasmalogen and ethanolamine-type plasmalogen are preferred, and ethanolamine-type plasmalogen is particularly preferred.

The plasmalogen of the present invention can be extracted from animal tissue. The animal tissue is not particularly limited as long as it contains plasmalogen, and includes aquatic animals such as shellfish, sea squirts, sea cucumbers, salmon, saury, and bonito, and birds. Among these, shellfish, sea squirts, and birds are preferred, and shellfish are particularly preferred. As the site to be used, an edible site (edible site) is preferable. These animal tissues may be cut pieces, but it is preferable to use pulverized pieces because plasmalogen can be extracted more efficiently.

Examples of shellfish include edible bivalves and snails such as scallops, mussels and abalones, with scallops being particularly preferred. Scallops are edible bivalves belonging to the mussel family, and include, for example, those belonging to the genus Mizuhopecten and the genus Pecten. Specifically, scallops collected in Japan (scientific name: Mizuhopectenyessoensis), European scallops collected in Europe (scientific name: Pectenmaximus (Linnaeus)), and the like can be mentioned. Examples of edible parts include scallops, cords (mantle, genital membrane, suspensory membrane) and the like.

The sea squirt is an edible chordate belonging to the Ascidaceae family, and includes those belonging to the genus Ascidia and the genus Ascidia. Concretely, it can be exemplified by Japanese wormwood (scientific name: Halocynthia roretzi), red wormwood (scientific name: Halocynthia aurantium), and the like. The edible part can include the body part (fascial body).

Birds are not particularly limited as long as they are edible birds, and examples thereof include chickens, silky fowls, and ducks. Breast meat rich in plasmalogen is preferable as the edible part.

Extraction of plasmalogen can be performed using water, an organic solvent, or a water-containing organic solvent, and it is preferable to use enzymatic treatment in combination. For example, an ethanol extraction method and a hexane extraction method can be mentioned, and the hexane extraction method is preferred.

The hexane extraction method is not particularly limited as long as it is a method of extraction using hexane. can be done.

The ethanol extraction method is not particularly limited as long as it is a method of extraction using ethanol (including hydrous ethanol). The methods described in JP-A-2012-039472, JP-A-2010-065167, JP-A-2010-063406 and the like can be mentioned.

The composition of the present invention can be used as, for example, pharmaceuticals (including quasi-drugs) and cosmetic ingredients such as external skin preparations whose indication of efficacy has been approved by a predetermined organization.

The composition of the present invention can be used as an external skin preparation or injection. The external preparation for skin is not particularly limited as long as it is used by applying it to the skin. skin external preparations such as agents.

The content of the plasmalogen in the composition of the present invention may be appropriately contained within a range in which the effect is exhibited. Depending on its form, for example, plasmalogen is preferably 0.1% by mass or more, particularly preferably 1.0% by mass or more, based on the total dry mass of the composition of the present invention. Even if the total composition is 10% by mass or more, the effect is not impaired, but the practicality is poor in terms of cost and the like.

The composition of the present invention can be produced by a known method with the addition of other components as long as the effects of the present invention are not impaired. Other ingredients other than the ingredients of the present invention can include, for example, other drugs, vitamins, minerals, proteins, peptides, amino acids, animal oils, and vegetable oils. In addition, water, lower alcohols, film-forming agents, moisturizing agents, ultraviolet absorbers, sterilizers, preservatives, pH adjusters, etc., which are applied to skin coating agents, can be contained.

In order to develop the application of plasmalogen, we applied a plasmalogen-containing skin preparation to the skin of mice. The skin of the application part was collected, mast cells in the skin section were stained with toluidine blue solution, and changes in the number of mast cells in the dermis were compared. As a result, the number of mast cells in the dermis was significantly reduced by applying the plasmalogen-containing skin preparation. This suggests that for allergic dermatitis including atopic dermatitis caused by the release of histamine by degranulation of mast cells, it can be used as an antihistamine drug that inhibits the binding of conventional histamine to H1 receptors and suppresses itching. Plasmalogen is expected to eliminate mast cells from lesions and interrupt the vicious cycle of itching and scratching. In addition, it is expected to be effective against pruritus induced by tryptase, a mast cell enzyme that cannot be suppressed by antihistamines. Therefore, I decided to examine it in detail.

The present invention will be described in more detail below with reference to examples.
The invention is not limited to the following examples.

[Preparation of plasmalogen for improving allergic dermatitis]
A plasmalogen for improving allergic dermatitis was prepared by extracting using hexane from each of scallops and chicken breast meat, for example, using Retable 2016/181491. The main steps for extracting plasmalogen from scallops are shown below.
1. Cochrase P (manufactured by Mitsubishi Chemical Foods Co., Ltd.) and phospholipase A1 (PLA1) (manufactured by Mitsubishi Chemical Foods Co., Ltd.) were added to scallops and digested for 1 hour.
2. Next, hexane/isopropanol was added and stirred, and after allowing to stand still, the supernatant was filtered by suction.
3. An aqueous sodium sulfate solution was added to the filtrate and mixed well.
4. The upper layer was dried on a rotary evaporator.
5. To this, 40 mL of acetone cooled to 4°C was added and mixed.
6. Separation was performed using a centrifuge at 3000 rpm, 10 min, and 4°C.
7. Discard the supernatant and collect the precipitate.
8. Dry overnight in a desiccator.
According to Japanese Patent No. 5430566, separation was performed by HPLC using the mobile phase solvent shown in FIG. 1A, and only the ethanolamine-type plasmalogen was fractionated as shown in FIG. 1C. In addition, FIG. 1B shows the lipid composition in the fractions extracted in steps 1 to 8 above.

Plasmalogen extracted from chicken breast meat was prepared through 7 steps except 5 out of the 8 steps described in [Preparation of Plasmalogen for Ameliorating Allergic Dermatitis]. The resulting extract fraction had a composition containing two types of plasmalogen, ethanolamine type and choline type, and cholesterol, as described in Retable 2016/181491.

[Effect of plasmalogen-containing skin application agent]
The plasmalogen-containing skin application was prepared by dissolving 51 mg of plasmalogen extracted from scallops and 5 mg of fraction extracted from chicken breast meat (plasmalogen content 9.1% by mass) in 3.64 ml of ethanol, followed by distilling 1.56 ml. Water was added to prepare 5.2 ml of plasmalogen solution. A control coating was prepared similarly to the plasmalogen-containing skin coating without dissolving the plasmalogen.

7-week-old male C3H mice were purchased from KBT Oriental Co., Ltd. At 8-week-old, the hair on the dorsal skin was shaved with an electric animal clipper and then removed with a depilatory cream. The subjects were grouped into a group for the application agent (n=5) and a group for the control application agent to which only 70% ethanol was applied (n=5). Application was started 48 hours after shaving. 20 μl/cm ² of the hair-removed area was applied, and the application was continued for 4 weeks at a frequency of 5 times/week. Feed and drinking water were allowed ad libitum during rearing.

After the test, the mice in each group were euthanized under deep anesthesia, and the skin of the applied area was collected. Immediately after chemical fixation with a paraform solution according to a conventional method, paraffin sections were prepared. A total of 20 discontinuous sections were randomly extracted from 5 mice in each group, and mast cells in the skin sections were stained with a toluidine blue solution (pH 4.1).

FIG. 2 shows the results of converting the number of mast cells in the skin on the back of the mouse of the control coating agent subject group and the plasmalogen-containing skin coating agent subject group into 1 mm×1 mm of skin. As a result, the number of mast cells in the skin of the plasmalogen-containing skin application agent subject group compared with the control application agent subject group showed a statistically significant (p<0.05, Student's t-test) reduction change. rice field. Therefore, it was clearly shown for the first time that ethanol-dissolved plasmalogen penetrates the epidermis and reaches the underlying dermis by application to the skin surface, thereby reducing the number of mast cells in the dermis.

Comparative example 1

[Effect of oral ingestion of plasmalogen on dermal mast cell distribution]
A plasmalogen for oral ingestion was prepared from scallops by the steps up to 8 described in the above [Preparation of plasmalogen for improving allergic dermatitis].

Six-week-old male Nc/Nga mice were purchased from KBT Oriental Co., Ltd., divided into two groups, and only one group was given a diet containing 0.1% weight plasmalogen and orally ingested plasmalogen for 12 weeks. bred. Next, a skin section was prepared from a site of atopic dermatitis on the back by the method described in Example 2, and the distribution of mast cells in the dermis was examined. Feed and drinking water were allowed ad libitum during rearing.

As shown in Figure 3, no difference could be found in the number of mast cells in the dermis between mice that received plasmalogen orally (4) and mice that did not (3).

[Effect of plasmalogen-containing ointment on atopic dermatitis]
Therefore, the effect of plasmalogen-containing ointment on atopic dermatitis was verified in an atopic dermatitis model mouse experimental system.

A plasmalogen for ointment was prepared from scallops by the steps up to 8 described in the above [Preparation of plasmalogen for improving allergic dermatitis].
The plasmalogen-containing ointment was prepared by dissolving the extracted plasmalogen in ethanol and heating and mixing with white petrolatum as a base material at 40°C or less. Ethanol was removed by blowing nitrogen gas, and an ointment containing 5% plasmalogen was prepared. The ointment was stored at 4°C.

The effect of plasmalogen-containing ointment on atopic dermatitis was verified using an atopic dermatitis mouse model experiment using a component derived from Dematophagoides farina (Df), a genus of house dust mites, which is an allergen for atopic dermatitis. Feed and drinking water were given freely during the rearing.

6-week-old male Nc/Nga mice were purchased from KBT Oriental Co., Ltd. At 7-week-old, the body hair on the dorsal skin was shaved with an electric animal clipper and then removed with a hair removal cream. The mice were grouped into 3 groups (8 individuals each) shown in 1. The Dermatophagoides mite-derived component was applied twice a week for 3 weeks. At the first time, 100 mg of a component derived from Dermatophagoides leopard mite (trade name: Biosta AD, manufactured by Biosta Co., Ltd.) was applied after hair removal. After the second time, 150 μl of 4% sodium dodecyl sulfate was uniformly applied to the hair-removed part, and after natural drying, 100 mg of the Dermatophagoides mite-derived component was applied.

Figures 4A and 4B are representative results of the skin condition of mice in groups B and C during the course of the experiment. The dorsal skin of mice in groups B and C, to which the Dermatophagoides mite-derived component was applied for 3 weeks, showed localized redness and exfoliation of the keratin.

Next, all mice in groups A, B and C, to which the Dermatophagoides mite-derived component was not applied, were visually observed for redness, bleeding, crust formation, and dryness, which are characteristic symptoms of atopic dermatitis. Determined. The conditions shown in Table 2 were used as indicators for redness/bleeding, and the conditions shown in Table 3 were used as indicators for scab formation/dryness. The condition of each mouse was scored on a 4-point scale, with 0 being no symptoms and 3 being severe.

FIG. 5 is a graph showing the average scored based on Table 2 for the degree of redness/bleeding at the site of atopic dermatitis in 8 mice in each group. Symptoms of redness and bleeding on the dorsal skin of mice in Groups B and C upon completion of application of the Dermatophagoides mite-derived component were more severe than in Group A, which was not applied, and each was statistically significant (††p<0 .01 and †p<0.05) changes.

FIG. 6 shows the average scored based on Table 3 for the degree of crust formation/dryness at the site of atopic dermatitis in 8 mice in each group. The symptoms of crust formation and dryness in Groups B and C at the completion of application of Dermatophagoides mite-derived ingredients were more severe than Group A, which was not applied, and both groups were statistically significant (††p<0 .01). From these results, it was found that an atopic dermatitis mouse model using the Dermatophagoides mite-derived component was established, and it became possible to verify the recovery of symptoms by plasmalogen ointment.

To verify the effect of plasmalogen-containing ointment on atopic dermatitis, mice in group C that developed atopic dermatitis were given 2 earpicks of plasmalogen-containing ointment at a frequency of 5 times/week for 4 times. applied for a week. White petrolatum was applied at the same frequency to group B mice that developed atopic dermatitis. If hair was growing at the time of each application, the hair was clipped with a hair clipper or the like, taking care not to irritate the skin. Furthermore, the skin symptoms were photographed with a digital camera every week.

FIG. 4 shows typical symptoms of the skin of mice in group B to which white petrolatum was applied and group C to which plasmalogen-containing ointment was applied, 3 and 4 weeks after the start of application. As shown in FIG. 4A, redness and exfoliation of keratin were continuously observed in the dorsal skin of the mice in group B to which white petrolatum was applied. However, as shown in FIG. 4B, the symptoms of redness and exfoliation observed in the dorsal skin of mice in group C were alleviated after application of the plasmalogen-containing ointment.

Then, all mice in groups A, B, and C were visually observed, and the degree of redness/bleeding and crust formation/dryness of the dorsal skin was checked every week from 0 to 3 based on Tables 2 and 3 above. It was scored in four stages.

The results are shown in FIG. The degree of redness and bleeding in the dorsal skin of the plasmalogen-containing ointment-applied group in group C compared to group B, which was the white petrolatum-applied group, was statistically significant at 1 and 2 weeks after plasmalogen-containing ointment application. A statistically significant (*p<0.05) improvement change was observed. In addition, as shown in FIG. 6, a statistically significant (*p<0.05) recovery was observed in the C group compared with the B group in terms of the degree of crust formation and dryness.

These results suggest that percutaneous absorption of plasmalogen reduces the number of mast cells in the dermis and blocks the vicious cycle of allergic dermatitis exacerbated by itching and scratching caused by the release of histamine from mast cells. I understand.

Claims (5)

A composition for improving allergic dermatitis, comprising a plasmalogen. A composition for improving allergic dermatitis, comprising 0.1% by mass to 10% by mass of the total composition of a plasmalogen. 2. The composition for improving allergic dermatitis according to claim 1, wherein said plasmalogen is a plasmalogen extracted from animal tissue. 3. The composition for improving allergic dermatitis according to claim 2, wherein said animal tissue is animal tissue selected from shellfish, sea squirts and birds. 4. The composition for improving allergic dermatitis according to claim 2 or 3, wherein the animal tissue is scallop tissue.

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