Ceramide production promoter, filaggrin production promoter, involucrin production promoter

Abstract

To provide a ceramide production promoter, a filaggrin production promoter, and an involucrin production promoter.SOLUTION: The present invention discloses a ceramide production promoter, a filaggrin production promoter, and an involucrin production promoter, containing kale extract, where the kale extract may be ethanol extract.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to a ceramide production promoter, a filaggrin production promoter, and an involucrin production promoter.

Background Art

[0002] A ceramide production promoter containing sphingoids (Patent Document 1) and a ceramide production promoter containing Gennosyouko extract (Patent Document 2) are known. A filaggrin production promoter containing a GPR142 agonist (Patent Document 3), a filaggrin production promoter containing Schisandra chinensis, Ezo-gokogi, and Murraya paniculata (Patent Document 4), and a filaggrin production promoter containing a calcium salt (Patent Document 5) are known. An involucrin production promoter containing dipotassium glycyrrhizinate (Patent Document 6) and a filaggrin production promoter containing Schisandra chinensis, Ezo-gokogi, and Murraya paniculata (Patent Document 4) are known.

Prior Art Documents

Patent Documents

[0003]

Patent Document 1

Patent Document 2

Patent Document 3

Patent Document 4

Patent Document 5

Patent Document 6

Summary of the Invention

Problems to be Solved by the Invention

[0004] To provide a ceramide production promoter, a filaggrin production promoter, and an involucrin production promoter.

Means for Solving the Problems

[0005] The main configuration of the present invention is as follows. 1. A ceramide production promoter, a filaggrin production promoter, and an involucrin production promoter containing kale extract. 2. The production promoter according to 1, wherein the kale extract is an ethanol extract.

Effects of the Invention

[0006] The ceramide production promoter, filaggrin production promoter, and involucrin production promoter of the present invention can promote the production of ceramide, filaggrin, and involucrin in cells. By applying the ceramide production promoter, filaggrin production promoter, and involucrin production promoter of the present invention, preventive and therapeutic effects on atopic dermatitis, psoriasis, ichthyosis, xeroderma, and allergic contact dermatitis can be expected. By applying the ceramide production promoter of the present invention, an inhibitory effect on the growth and metastasis of squamous cell carcinoma can be expected.

Brief Description of the Drawings

[0007]

Figure 1

Figure 2

Figure 3

Modes for Carrying Out the Invention

[0008] Ceramide is an amide derivative that accounts for approximately 50% of the intercellular lipids present in human skin (stratum corneum). Examples of ceramides include N-acylsphingosine (ceramide), N-acyl-dihydrosphingosine (dihydroceramide), N-acylphytosphingosine (phytoceramide), etc. The chemical name of the above-mentioned sphingosine is (2S,3R,4E)-2-amino-4-octadecene-1,3-diol, the chemical name of dihydrosphingosine is (2S,3R)-2-aminooctadecane-1,3-diol, and the chemical name of phytosphingosine is (2S,3S,4R)-2-amino-1,3,4-octadecanetriol. Ceramide is known to control cell growth, differentiation, apoptosis, etc. as a signaling substance. From these facts, substances that promote the production of ceramide can be expected to have effects such as suppressing cell growth, inducing differentiation, and inducing apoptosis, and can be expected to have a therapeutic effect on diseases caused by these abnormalities. Specifically, by promoting the production of ceramide, it can be expected to have an effect of suppressing the growth and metastasis of squamous cell carcinoma, a therapeutic and preventive effect on psoriasis, and a therapeutic effect on atopic dermatitis (Japanese Patent Laid-Open No. 2012-158540).

[0009] Filaggrin is a kind of basic protein produced in epidermal granular cells. Filaggrin is biosynthesized as profilaggrin, which is a precursor. Profilaggrin is a huge protein with a molecular weight of about 400 kDa having a structure in which 10 to 20 filaggrins are linked, and it undergoes phosphorylation to form keratohyalin granules in granular cells. When granular cells migrate to keratinocytes, phosphorylated profilaggrin undergoes dephosphorylation and limited hydrolysis and is decomposed into filaggrin. Filaggrin promotes the fiber formation reaction of keratin fibers proceeding in keratinocytes. By promoting the production of filaggrin, skin diseases associated with reduced filaggrin production can be treated. Specifically, by applying a filaggrin production promoter, a therapeutic effect on atopic dermatitis, ichthyosis vulgaris, metal (nickel) allergic contact dermatitis, and seborrheic dermatitis (senile xerosis) can be expected (WO2014 / 17319).

[0010] Involucrin is the main protein in the stratum corneum and is produced from the upper layer of the spinous layer and below. Involucrin is a soluble protein with a molecular weight of approximately 68,000, but it becomes insoluble by binding to proteins and forms a cornified envelope. By applying an involucrin production promoter, it becomes possible to prevent and treat diseases caused by involucrin deficiency. Specifically, by applying an involucrin production promoter, preventive and therapeutic effects on dry skin diseases such as ichthyosis, psoriasis, atopic dermatitis, and xerosis can be expected (Japanese Patent Laid-Open No. 2009-114146).

[0011] The present invention provides a ceramide production promoter, a filaggrin production promoter, and an involucrin production promoter containing kale extract. Kale is a cruciferous vegetable native to southern Europe and is said to be the original species of cabbage, and its scientific name is Brassica Oleracea L. var. acephala DC. There are various types of kale, such as curly kale, marrow kale, bush kale, tree kale, collard, and Chinese kale, and any of them can be used for the purpose of the present invention. As the part of kale used in the present invention, in addition to those usually used for food such as leaves, the stems, flowers, roots, etc. of kale can also be used, and there is no particular limitation, nor is the cultivation method or cultivation area particularly limited. Kale leaves are used for the production of green juice, but by using the kale residue after squeezing the green juice and containing its extract, the ceramide production promoter, filaggrin production promoter, and involucrin production promoter of the present invention can also be obtained. The kale residue referred to in the present invention means the residue after squeezing green juice from the leaves or stems of kale. As a method for preparing kale residue after squeezing green juice, for example, a method of heating kale leaves with hot water at 60 to 100°C for 5 seconds to 3 minutes, and then squeezing the green juice with a juicer to obtain the kale residue after squeezing can be mentioned. Examples of the extraction solvent for the kale extract of the present invention include hydrophilic solvents such as ethanol and 1,3 - butanediol, and they can also be used as the extraction solvent as a mixture with water. Further, water itself or a hydrophobic solvent such as hexane can also be used as the extraction solvent.

[0012] As a method for preparing the kale extract, for example, ethanol or 1,3 - butanediol is added to kale residue, kale leaves, flowers, stems, or dried products of kale residue, kale leaves, flowers, and stems, heated at 40°C to 70°C for 30 minutes to 5 hours, further immersed and extracted at room temperature for 1 day to 1 month, and filtered to obtain a solution of the kale extract, and then the solvent is evaporated to obtain the kale extract.

[0013] The ceramide production promoter, filaggrin production promoter, and involucrin production promoter of the present invention can be used by being formulated into external skin preparations or oral preparations by conventional methods.

[0014] The content of the kale extract in the ceramide production promoter, filaggrin production promoter, and involucrin production promoter of the present invention is preferably 0.00001% or more and 0.1% or less, and particularly preferably 0.0001% or more and 0.01% or less.

Examples

[0015] <Preparation of kale residue> After heating 100 kg of kale leaves with hot water at 80°C for 30 seconds, the green juice was squeezed with a juicer, and 15 kg of kale residue after squeezing the green juice was obtained.

[0016] <Preparation of kale extract> The kale residue was freeze - dried, 800 g of ethanol was added to 80 g of the obtained dried kale residue, and it was heated at 60°C for 2 hours. After cooling, it was immersed and extracted at room temperature for 2 weeks and filtered to obtain 574.96 g of an ethanol extract. The concentration of the ethanol extract in the ethanol extract was 0.74%, and 4.25 g of the kale extract was contained in 574.96 g of the ethanol extract. In the following quantitative analysis test of gene expression, the above - mentioned ethanol extract was used as the test sample.

[0017] <Quantitative analysis of gene expression> Normal human epidermal keratinocytes (Kurabo) were seeded in a 6-well plate and cultured in HuMedia-KG2 custom GC annex (Kurabo) medium until 100% confluent. Then, test samples were added at 0.05% or 0.5% (concentration of kale extract 0.00037% or 0.0037%), and total RNA was recovered after 24 hours. For RNA recovery, QIAshredder (Qiagen) and Rneasy Mini Kit (Qiagen) were used to extract total RNA. The concentration of the extracted RNA was measured, and based on that value, cDNA synthesis was performed using PrimeScript (Takara). Then, quantitative analysis of gene expression by real-time PCR was performed using SYBR Premix EX taq (Takara) based on the obtained cDNA. The target genes were ceramide synthase (CERS1), filaggrin (FLG), involucrin (IVL), and primers for each were prepared using RPS18 as a housekeeping gene. Also, for the above test, high Ca2+ (1.0 mM) differentiation induction conditions were set. As a control, a sample with the solvent of the test sample added at the same concentration as the final concentration contained in the test sample was used. The results are shown in Tables 1 - 3 and Figures 1 - 3.

[0018] <Gene expression of ceramide synthase (CERS1)>

[0019]

Table 1

[0020] <Gene expression of filaggrin (FLG)>

[0021]

Table 2

[0022] <Gene expression of involucrin (IVL)>

[0023]

Table 3

Claims

【Claim 1】 An involucrin production promoter containing an ethanol extract of kale (however, excluding the use as an agent for improving atopic dermatitis, ichthyosis, psoriasis, xeroderma, or a preventive or therapeutic agent for dry skin diseases).

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