Atopic dermatitis improving agent, interleukin production inhibitor, and interleukin gene expression inhibitor

Abstract

To provide an atopic dermatitis ameliorating agent or the like capable of alleviating atopic dermatitis.SOLUTION: Provided is an atopic dermatitis ameliorating agent or the like comprising a glyceryl ascorbic acid acylated derivative having a specific molecular structure.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to an agent for improving atopic dermatitis, an interleukin production inhibitor, and an interleukin gene expression inhibitor.

Background Art

[0002] Atopic skin diseases are chronic diseases in which eczema accompanied by itching repeatedly worsens and improves, and are considered to develop due to the complex interaction of various causes. Atopic skin diseases are thought to worsen, for example, by the production of lipid mediators that induce allergic reactions, but it is not thought that this is the only cause of the worsening of atopic skin diseases.

[0003] Conventionally, various agents for improving atopic dermatitis for improving atopic skin diseases as described above are known.

[0004] As this type of agent for improving atopic dermatitis, for example, those containing an ointment containing docosahexaenoic acid (DHA) and / or eicosapentaenoic acid (EPA) and tacrolimus ointment are known (Patent Document 1).

[0005] The agent for improving atopic dermatitis described in Patent Document 1 can suppress the production of leukotriene B4, which is one of the lipid mediators that induce allergic reactions, and thus can improve the symptoms of atopic skin diseases.

Prior Art Documents

Patent Documents

[0006]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0007] However, formulations that can improve atopic skin diseases have not yet been sufficiently investigated.

[0008] Therefore, the object of the present invention is to provide an atopic dermatitis improving agent, a topical skin preparation, and a cosmetic product that can improve atopic skin diseases. [Means for solving the problem]

[0009] The atopic dermatitis improving agent according to the present invention is characterized by containing a glyceryl ascorbate acylated derivative represented by the following formula (I). [ka]

[0010] The interleukin production inhibitor according to the present invention comprises a glyceryl ascorbate acylated derivative represented by the above formula (I), and is intended to suppress the production of interleukin 33 and interleukin 1.

[0011] The interleukin gene expression inhibitor according to the present invention comprises a glyceryl ascorbate acylated derivative represented by the above formula (I), and is intended to suppress the expression of interleukin 33 and interleukin 1 genes. [Effects of the Invention]

[0012] The atopic dermatitis improving agent, interleukin production inhibitor, and interleukin gene expression inhibitor of the present invention can improve atopic skin diseases. [Brief explanation of the drawing]

[0013] [Figure 1] This graph shows the results of an in vitro study evaluating the effects of interleukin-33 (IL-33) on gene expression. [Figure 2] This graph shows the results of an in vitro study evaluating the effect of interleukin-1α (IL-1α) on gene expression. [Figure 3] A graph representing the test results (in vitro test) evaluating the effect on the gene expression of interleukin 1β (IL-1β). [Figure 4] A graph representing the test results (in vitro test) evaluating the effect on the gene expression of interleukin 33 (IL-33). [Figure 5] A graph representing the test results (in vitro test) evaluating the effect on the gene expression of interleukin 1α (IL-1α). [Figure 6] A graph representing the test results (in vitro test) evaluating the effect on the gene expression of interleukin 1β (IL-1β).

Mode for Carrying Out the Invention

[0014] One embodiment of the atopic dermatitis improver, interleukin production inhibitor, and interleukin gene expression inhibitor (hereinafter, also simply referred to as the preparation) according to the present invention will be described below.

[0015] The preparation of this embodiment contains a glyceryl ascorbic acid acylated derivative represented by the following formula (I).

[0016]

Chemical Formula

[0017] Since the preparation of this embodiment contains a glyceryl ascorbic acid acylated derivative represented by the above formula (I), it can exert an inhibitory effect on the production of specific inflammatory cytokines related to atopic skin diseases, and thus can improve atopic skin diseases.

[0018] The compound represented by the above formula (I) may be referred to as 2-O-glyceryl-6-hexadecanoyl ascorbic acid. Compounds similar to the compound represented by formula (I) above, such as 2-O-glyceryl-6-ethanolascorbic acid, 2-O-glyceryl-6-butanoylascorbic acid, 2-O-glyceryl-6-octanoylascorbic acid (2-O-glyceryl-6-(2-ethylhexanoyl)ascorbic acid), 2-O-glyceryl-6-tetradecanoylascorbic acid, 2-O-glyceryl-6-octadecanoylascorbic acid, or 2-O-glyceryl-6-docosanoylascorbic acid, cannot necessarily adequately improve atopic skin diseases.

[0019] The formulation of this embodiment contains a glyceryl ascorbate acylated derivative represented by formula (I), and can therefore suppress the production of cytokines in response to inflammatory stimuli in epidermal cells. For example, it can suppress the production of specific cytokines such as interleukin-33 (IL-33) and interleukin-1 (IL-1α or IL-1β). This can improve atopic skin diseases.

[0020] Interleukin-33 (IL-33) is a cytokine involved in the pathogenesis of allergic diseases, and in skin allergic diseases, it is a cytokine that particularly exacerbates the pathogenesis of atopic dermatitis. Specifically, interleukin-33 (IL-33) is a type of cytokine produced by epidermal cells, and it is overexpressed in the epidermal cells of patients with atopic dermatitis. More specifically, interleukin-33 (IL-33) activates Th2 cells (helper T cells) and type 2 innate lymphoid cells (ILC2), and induces Th2 cytokines such as interleukin-4 (IL-4) and interleukin-13 (IL-13), thereby causing type 2 inflammatory responses, mainly in atopic dermatitis. Interleukin-1 (IL-1) is an inflammatory cytokine involved in various inflammations, and is also involved in the exacerbation of atopic dermatitis. For example, interleukin-1 (IL-1) is produced by epidermal cells in response to various stimuli, causing inflammation. The IL-1 receptor in mature helper T cells is present only on Th2 cells. IL-1 can cause Th2 cells to proliferate, potentially leading to the development or exacerbation of atopic dermatitis.

[0021] The formulation of this embodiment can suppress the expression of inflammatory cytokine genes produced by epidermal cells as described above, and therefore is also an inhibitor of the gene expression of IL-33, IL-1α, and IL-1β.

[0022] The compound represented by formula (I) above can be synthesized, for example, by the following synthesis method described in Japanese Patent Application Publication No. 2011-079772. (Intermediate: Synthesis of 2-O-glyceryl ascorbate) Under an argon atmosphere, L-ascorbic acid and sodium bicarbonate are added to water and stirred at room temperature for 30 minutes, then glycidol is added. The mixture is heated to 60°C and stirred for 5 hours. Methanol is added, the mixture is filtered, and the filtrate is concentrated under reduced pressure. The resulting residue is subjected to silica gel column chromatography. Elution is performed with chloroform / methanol / water = 6 / 4 / 1, and the mixture is concentrated under reduced pressure to obtain 2-O-glyceryl ascorbic acid. (Synthesis of 2-O-glyceryl-6-O-hexadecanoyl ascorbic acid) Under an argon atmosphere, pyridine is added to the synthesized 2-O-glyceryl ascorbic acid, followed by n-hexadecanoic anhydride, and the mixture is stirred at 60°C for 3 hours. Then, water is added and extraction is performed with ethyl acetate. The extract is concentrated under reduced pressure, and 134 mg of the resulting residue is subjected to silica gel column chromatography. The residue is eluted and purified with a chloroform / methanol / water mixture (7 / 3 / 0.3), and concentrated under reduced pressure to obtain 2-O-glyceryl-6-O-hexadecanoyl ascorbic acid.

[0023] In the formulation of this embodiment, the total concentration of the compound represented by formula (I) may be, for example, 0.001% by mass or more and 5% by mass or less, and more preferably 0.01% by mass or more and 2.0% by mass or less. The above-mentioned concentration has the advantage of being able to further improve atopic skin diseases.

[0024] The above-mentioned formulations typically contain water and may further contain, in addition to the above-mentioned ingredients, thickeners, surfactants, preservatives, etc.

[0025] The properties of the formulation of this embodiment are not particularly limited, but for example, it may be liquid. The formulation of this embodiment may also be solid.

[0026] The formulation of this embodiment can be manufactured by a general method. For example, the above formulation can be manufactured by mixing and stirring each component to be blended. A general-purpose stirring device can be used. If necessary, stirring may be done while heating.

[0027] The above-mentioned preparations are preferably topical skin preparations (transdermal administration preparations) or cosmetics. Such topical skin preparations or cosmetics are usually applied to the skin. The above-mentioned topical skin preparations or cosmetics may be applied to, for example, the skin of the face, neck, limbs, scalp, hair, and mucous membranes such as the nostrils, lips, ears, genitals, and anus. Furthermore, the above-mentioned topical skin preparations or cosmetics may be used as bath additives or as skin patches.

[0028] The formulation of this embodiment is not necessarily bound by the classification of cosmetics, quasi-drugs, pharmaceuticals, etc. under the Pharmaceuticals and Medical Devices Act.

[0029] The atopic dermatitis improving agent, interleukin production inhibitor, and interleukin gene expression inhibitor of the present invention are as illustrated above, but the present invention is not limited to the embodiments described above. Furthermore, the present invention can employ various forms commonly used in topical skin compositions and oral administration compositions, as long as they do not impair the effects of the present invention.

[0030] The matters disclosed herein include the following: (1) An atopic dermatitis improving agent containing a glyceryl ascorbate acylated derivative represented by the following formula (I). [ka] (2) An interleukin production inhibitor comprising a glyceryl ascorbate acylated derivative represented by the above formula (I). (3) An interleukin gene expression inhibitor comprising a glyceryl ascorbate acylated derivative represented by the above formula (I). [Examples]

[0031] The present invention will now be described in more detail with reference to examples, but the present invention is not limited to these examples.

[0032] (Test Example 1) Formulations for each test example (for example, those usable as topical skin preparations) were prepared by dissolving 2-O-glyceryl-6-hexadecanoylascorbic acid (a compound represented by formula (I)) in a solvent such as water. The concentration was set to 10 μM, 20 μM, etc., each time.

[0033] (Test Example 2) The formulation was prepared in the same manner as in Test Example 1, except that 2-O-glyceryl-6-dodecanoylascorbic acid was used instead of the compound represented by formula (I). (Test Example 3) The formulation was prepared in the same manner as in Test Example 1, except that 2-O-glyceryl-6-tetradecanoylascorbic acid was used instead of the compound represented by formula (I). (Test example 4) The formulation was prepared in the same manner as in Test Example 1, except that 2-O-glyceryl-6-octadecanoylascorbic acid was used instead of the compound represented by formula (I). (Test Example 5) The formulation was prepared in the same manner as in Test Example 1, except that ascorbic acid was used instead of the compound represented by formula (I). (Test Example 6) The formulation was prepared in the same manner as in Test Example 1, except that ascorbyl palmitate was used instead of the compound represented by formula (I).

[0034] <Evaluation of cytokine production suppression (in vitro test)> Details of the in vitro test are as follows: Normal human epidermal keratinocytes (NHEK Kurabo Corporation) were placed in a 12-well plate in a 6 × 10⁶ arrangement. 4 Cells were seeded and cultured for 72 hours at 37°C under 5% CO2 conditions. After removing the culture medium, it was replaced with a medium containing the following inflammatory stimulants (final concentration 25 ng / mL) and each compound used in the above test example, and cultured for a further 3 hours at 37°C under 5% CO2 conditions. Subsequently, total RNA was purified from the cells according to the instructions for the product name "PureLink RNA Mini kit" (invitrogen), and the purified RNA was reverse transcribed into cDNA using the test kit "ReverTra Ace™ qPCR RT Master Mix (TOYOBO)". The mRNA expression level of IL-33 in the above cDNA was quantitatively evaluated by real-time PCR. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as an internal standard correction. In each test example, the compounds used were dissolved in dimethyl sulfoxide (DMSO) to prepare stocks, and then the compounds used in each test example were dissolved in culture medium to a concentration of 10-20 μM. At that time, the DMSO concentration was diluted to a level that would not cause cytotoxicity (1% or less). ·Inflammatory stimulant (Phorbol12-myristate13-acetate) PMA • Negative control (without NC PMA addition) • Positive control (steroidal anti-inflammatory drugs) (Dexamethasone)DEX concentration 10μM Similarly, evaluations were also conducted for interleukin-1α (IL-1α) and interleukin-1β (IL-1β).

[0035] Figures 1 to 3 show graphs illustrating the results of in vitro tests evaluating the suppression of cytokine production for Test Examples 1 to 4. Figures 4 to 6 show graphs illustrating the results of in vitro tests evaluating the suppression of cytokine production for test examples 1, 5, and 6. As can be seen from Figures 1 to 6, formulations containing 2-O-glyceryl-6-hexadecanoyl ascorbic acid suppressed the expression of cytokine genes strongly associated with the exacerbation of atopic skin disease. Therefore, the atopic dermatitis improving agent of this embodiment (interleukin production inhibitor and interleukin gene expression inhibitor) can improve atopic skin diseases. [Industrial applicability]

[0036] The atopic dermatitis improving agent of the present invention can be used, for example, in topical skin preparations or cosmetics. The interleukin production inhibitor and the interleukin gene expression inhibitor of the present invention are applied to the skin and used, for example, to prevent and alleviate dry skin caused by atopic skin diseases, or to prevent and alleviate inflammatory skin roughness caused by atopic skin diseases. The atopic dermatitis improving agent of the present invention can be applied directly to the stratum corneum and is suitably used in pharmaceuticals, quasi-drugs, or cosmetics.

Claims

1. An atopic dermatitis treatment agent containing a glyceryl ascorbate acylated derivative represented by the following formula (I) as an active ingredient. 【Chemistry 1】

2. An interleukin production inhibitor comprising a glyceryl ascorbate acylated derivative represented by the following formula (I) as an active ingredient, for suppressing the production of interleukin 33 and interleukin 1. 【Chemistry 2】

3. An interleukin gene expression inhibitor comprising a glyceryl ascorbate acylated derivative represented by the following formula (I) as an active ingredient, for suppressing the expression of interleukin 33 and interleukin 1 genes. 【Transformation 3】

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